ALBERT

Description: by Jialiang Guo, Qi Zhang, Jia Li, Yansong Liu, Zhiyong Hou, Wei Chen, Lin Jin, Ye Tian, Linlin Ju, Bo Liu, Tianhua Dong, Fei Zhang, Yingze Zhang Non-union is a major clinical problem in the healing of fractures, especially in patients with osteoporosis. The systemic administration of drugs is time consuming and large doses are demanding and act slowly, whereas local release acts rapidly, increases the quality and quantity of the bone tissue. We hypothesize that local delivery demonstrates better therapeutic effects on an osteoporotic fracture. The aim of this paper is to investigate the effect of the local application of ibandronate loaded with a collagen sponge on regulating bone formation and remodeling in an osteoporotic rat model of fracture healing. We found that the local delivery of ibandronate exhibited excellent effects on improving the bone microarchitecture and suppressed effects on bone remodeling. At 4 weeks, more callus formation and improvement of mechanical character and microstructure were observed in a local delivery via μCT, mechanical test, histological research and serum analysis. The suppression of bone remodeling was compared with a systemic treatment at 12 weeks, and the structural mechanical properties and microarchitecture were also improved with local delivery. This research identifies an earlier, safer and integrated approach for local delivery of ibandronate with collagen and provides a better strategy for the treatment of osteoporotic fracture in rats.

Description: by Maia P. Smith, Marie Standl, Joachim Heinrich, Holger Schulz Background Because of unreliable self-report, accelerometry is increasingly used to objectively monitor physical activity (PA). However, results of accelerometric studies vary depending on the chosen cutpoints between activity intensities. Population-specific activity patterns likely affect the size of these differences. To establish their size and stability we apply three sets of cutpoints, including two calibrated to a single reference, to our accelerometric data and compare PA estimates. Methods 1402 German adolescents from the GINIplus and LISAplus cohorts wore triaxial accelerometers (Actigraph GT3x) for one week (mean 6.23 days, 14.7 hours per day) at the hip. After validation of wear, we applied three sets of cutpoints for youth, including the most common standard (Freedson, 2005) and two calibrated to a single reference, (Romanzini uni- and triaxial, from Romanzini, 2014) to these data, estimating daily sedentary, light, moderate, vigorous and moderate-to-vigorous PA (MPA, VPA, MVPA). Stability of differences was assessed by comparing Romanzini’s two sets of cutpoints. Results Relative agreement between cutpoints was closer for activity of lower intensities (largest difference for sedentary behaviour 9%) but increased for higher intensities (largest difference for light activity 40%, MPA 102%, VPA 88%; all p

Description: by Bertha J. Vandegrift, Chang You, Rosalba Satta, Mark S. Brodie, Amy W. Lasek Gender differences in psychiatric disorders such as addiction may be modulated by the steroid hormone estrogen. For instance, 17β-estradiol (E2), the predominant form of circulating estrogen in pre-menopausal females, increases ethanol consumption, suggesting that E2 may affect the rewarding properties of ethanol and thus the development of alcohol use disorder in females. The ventral tegmental area (VTA) is critically involved in the rewarding and reinforcing effects of ethanol. In order to determine the role of E2 in VTA physiology, gonadally intact female mice were sacrificed during diestrus II (high E2) or estrus (low E2) for electrophysiology recordings. We measured the excitation by ethanol and inhibition by dopamine (DA) of VTA DA neurons and found that both excitation by ethanol and inhibition by dopamine were greater in diestrus II compared with estrus. Treatment of VTA slices from mice in diestrus II with an estrogen receptor antagonist (ICI 182,780) reduced ethanol-stimulated neuronal firing, but had no effect on ethanol-stimulated firing of neurons in slices from mice in estrus. Surprisingly, ICI 182,780 did not affect the inhibition by DA, indicating different mechanisms of action of estrogen receptors in altering ethanol and DA responses. We also examined the responses of VTA DA neurons to ethanol and DA in ovariectomized mice treated with E2 and found that E2 treatment enhanced the responses to ethanol and DA in a manner similar to what we observed in mice in diestrus II. Our data indicate that E2 modulates VTA neuron physiology, which may contribute to both the enhanced reinforcing and rewarding effects of alcohol and the development of other psychiatric disorders in females that involve alterations in DA neurotransmission.

Description: by Chelsea M. Rochman, J. Mark Parnis, Mark A. Browne, Sebastian Serrato, Eric J. Reiner, Matthew Robson, Thomas Young, Miriam L. Diamond, Swee J. Teh We examined whether environmentally relevant concentrations of different types of microplastics, with or without PCBs, directly affect freshwater prey and indirectly affect their predators. Asian clams ( Corbicula fluminea ) were exposed to environmentally relevant concentrations of polyethylene terephthalate (PET), polyethylene, polyvinylchloride (PVC) or polystyrene with and without polychlorinated biphenyls (PCBs) for 28 days. Their predators, white sturgeon ( Acipenser transmontanus ), were exposed to clams from each treatment for 28 days. In both species, we examined bioaccumulation of PCBs and effects (i.e., immunohistochemistry, histology, behavior, condition, mortality) across several levels of biological organization. PCBs were not detected in prey or predator, and thus differences in bioaccumulation of PCBs among polymers and biomagnification in predators could not be measured. One of the main objectives of this study was to test the hypothesis that bioaccumulation of PCBs would differ among polymer types. Because we could not answer this question experimentally, a bioaccumulation model was run and predicted that concentrations of PCBs in clams exposed to polyethylene and polystyrene would be greater than PET and PVC. Observed effects, although subtle, seemed to be due to microplastics rather than PCBs alone. For example, histopathology showed tubular dilation in clams exposed to microplastics with PCBs, with only mild effects in clams exposed to PCBs alone.

Description: by Deisy Barrios, Raquel Morillo, Ina Guerassimova, Esther Barbero, Héctor Escobar-Morreale, Alexander T. Cohen, Cecilia Becattini, Victor Tapson, Roger Yusen, David Jimenez Background We sought to examine sex-related differences in the characteristics and outcome in patients presenting with acute symptomatic pulmonary embolism (PE). Methods We conducted a retrospective cohort study of 2,096 patients diagnosed with acute PE. The characteristics were recorded at presentation. Treatment was at the discretion of patients’ physicians. The primary study outcome, all-cause mortality, and the secondary outcomes of PE-specific mortality, recurrent venous thromboembolism, and major bleeding were assessed during the first month of follow-up after PE diagnosis. Results Overall, the women were older than the men and had significantly higher rates of immobilization. They had significantly lower rates of chronic obstructive pulmonary disease and cancer. Women had a higher prevalence of syncope and elevated brain natriuretic peptide levels. Thirty-day all-cause mortality was similar between women and men (7.1% versus 6.2%; P = 0.38). Male gender was not independently significantly associated with PE-related death (adjusted odds ratio [OR] 1.02; 95% CI, 0.50 to 2.07; P = 0.96). Restricting the analyses to haemodynamically stable patients (n = 2,021), female gender was an independent predictor of all-cause (adjusted OR 1.56; 95% CI, 1.07 to 2.28; P = 0.02) and PE-specific mortality (adjusted OR 1.85; 95% CI, 1.02 to 3.33; P = 0.04). Compared with men, women were 2.05 times more likely to experience a major bleed. Conclusions Women and men with PE had different clinical characteristics, presentation, and outcomes. Women receiving anticoagulation have a significantly higher risk of major bleeding, suggesting the need for careful monitoring of anticoagulant intensity in women.

Description: by Friederike Hammersen, Philip Lewin, Judith Gebauer, Ilonka Kreitschmann-Andermahr, Georg Brabant, Alexander Katalinic, Annika Waldmann This study investigated sleep quality and health-related quality of life (HRQOL) among long-term survivors of Hodgkin (HL) and non-Hodgkin lymphoma (NHL). The aim was to explore the impact of personal and health-related factors on sleep quality as well as associations between sleep quality and HRQOL. For the postal survey, participants with a minimum age of 18 years initially treated between 1998 and 2008 were recruited via the population-based cancer registry in Schleswig-Holstein, Northern Germany. Questionnaires included amongst others the Pittsburg Sleep Quality Index (PSQI) and the 36-Item Short Form Health Survey (SF-36v1). Descriptive and comparative statistics were performed. Additionally, a regression analysis was conducted to identify predictors of sleep quality. In total, we recruited 515 participants (398 NHL, 117 HL) with a mean age of 63.1 years. Approximately half of the survivors were classified as good sleepers. HRQOL scores differed between good and poor sleepers with lower scores in poor sleepers. In a prediction model, self-reported depression, exhaustion, higher age, inability to work, endocrinological disorders and female gender classified as predictors of sleep quality. This study highlights the impact of sleep quality on HRQOL in long-term survivors of NHL and HL. Thus, sleep quality should be routinely assessed during follow-up of cancer survivors with special attention to patients with potential risk factors.

Description: by Nuria García-Marchena, Daniel Silva-Peña, Ana Isabel Martín-Velasco, María Ángeles Villanúa, Pedro Araos, María Pedraz, Rosa Maza-Quiroga, Pablo Romero-Sanchiz, Gabriel Rubio, Estela Castilla-Ortega, Juan Suárez, Fernando Rodríguez de Fonseca, Antonia Serrano, Francisco Javier Pavón The identification of growth factors as potential biomarkers in alcohol addiction may help to understand underlying mechanisms associated with the pathogenesis of alcohol use disorders (AUDs). Previous studies have linked growth factors to neural plasticity in neurocognitive impairment and mental disorders. In order to further clarify the impact of chronic alcohol consumption on circulating growth factors, a cross-sectional study was performed in abstinent AUD patients (alcohol group, N = 91) and healthy control subjects (control group, N = 55) to examine plasma concentrations of brain-derived neurotrophic factor (BDNF), insulin-like growth factor-1 (IGF-1) and IGF-1 binding protein-3 (IGFBP-3). The association of these plasma peptides with relevant AUD-related variables and psychiatric comorbidity was explored. The alcohol group was diagnosed with severe AUD and showed an average of 13 years of problematic use and 10 months of abstinence at the moment of participating in the study. Regarding common medical conditions associated with AUD, we observed an elevated incidence of alcohol-induced liver and pancreas diseases (18.7%) and psychiatric comorbidity (76.9%). Thus, AUD patients displayed a high prevalence of dual diagnosis (39.3%) [mainly depression (19.9%)] and comorbid substance use disorders (40.7%). Plasma BDNF and IGF-1 concentrations were significantly lower in the alcohol group than in the control group (p

Description: by Stephen P. Cohen, Hongxia Liu, Cristiana T. Argueso, Andy Pereira, Casiana Vera Cruz, Valerie Verdier, Jan E. Leach Plant disease is a major challenge to agriculture worldwide, and it is exacerbated by abiotic environmental factors. During some plant-pathogen interactions, heat stress allows pathogens to overcome host resistance, a phenomenon which could severely impact crop productivity considering the global warming trends associated with climate change. Despite the importance of this phenomenon, little is known about the underlying molecular mechanisms. To better understand host plant responses during simultaneous heat and pathogen stress, we conducted a transcriptomics experiment for rice plants (cultivar IRBB61) containing Xa7 , a bacterial blight disease resistance ( R ) gene, that were infected with Xanthomonas oryzae , the bacterial blight pathogen of rice, during high temperature stress. Xa7- mediated resistance is unusual relative to resistance mediated by other R genes in that it functions better at high temperatures. Using RNA-Seq technology, we identified 8,499 differentially expressed genes as temperature responsive in rice cultivar IRBB61 experiencing susceptible and resistant interactions across three time points. Notably, genes in the plant hormone abscisic acid biosynthesis and response pathways were up-regulated by high temperature in both mock-treated plants and plants experiencing a susceptible interaction and were suppressed by high temperature in plants exhibiting Xa7- mediated resistance. Genes responsive to salicylic acid, an important plant hormone for disease resistance, were down-regulated by high temperature during both the susceptible and resistant interactions, suggesting that enhanced Xa7- mediated resistance at high temperature is not dependent on salicylic acid signaling. A DNA sequence motif similar to known abscisic acid-responsive cis- regulatory elements was identified in the promoter region upstream of genes up-regulated in susceptible but down-regulated in resistant interactions. The results of our study suggest that the plant hormone abscisic acid is an important node for cross-talk between plant transcriptional response pathways to high temperature stress and pathogen attack. Genes in this pathway represent an important focus for future study to determine how plants evolved to deal with simultaneous abiotic and biotic stresses.

Description: by Yang-yang Liu, Han-yan Sun, Wei Li, Yun-lei Xia, Yuan-yu Deng, Ai-xiang Zhang, Huai-gu Chen In China, Fusarium head blight is caused mainly by the Fusarium graminearum species complex (FGSC), which produces trichothecene toxins. The FGSC is divided into three chemotypes: 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), and nivalenol (NIV). In order to predict the geographical changes in the distribution of these chemotype populations in major winter wheat-producing areas in China, the biological characteristics of twenty randomly selected isolates from each of the three chemotypes were studied. No significant difference was exhibited in the growth rate of 3-ADON, 15-ADON, and NIV isolates at 15°C. At 20°C and 25°C, the growth rate of 15-ADON isolates was the highest. At 30°C, the growth rate of NIV and 3-ADON isolates was significantly higher than that of 15-ADON isolates. The 15-ADON isolates produced the highest quantities of perithecia and two to three days earlier than the other two populations at each temperature, and released more ascospores at 18°C. The aggressiveness test on wheat seedlings and ears indicated there was no significant difference between the 3-ADON and 15-ADON isolates. However, the aggressiveness of NIV isolates was significantly lower than that of the 3-ADON and 15-ADON isolates. The DON content in grains from heads inoculated with the 3-ADON isolates was higher than the content of 15-ADON and NIV isolates. The results showed that 15-ADON population had the advantage in perithecia formation and ascospore release, and the 3-ADON population produced more DON in wheat grains. We suggested that distribution of these three chemotype populations may be related to these biological characteristics.

Description: by Marek Zak, Malgorzata Biskup, Pawel Macek, Halina Krol, Szymon Krupnik, Anna Opuchlik Objective Breast cancer treatment, including radical surgery, is also pursued as late as the 7th - 8th decade of women’s lives. Standard physical rehabilitation procedures offered to those women are predominantly focused on attenuating specific functional deficits of the upper limb and trunk. Seldom do they entail any regimens specifically aimed at recovering overall functionality, and reducing exposure to falls-risk.The study aimed to assess potential interrelationships between the self-reported falls, individual functional capabilities and appreciably reducing exposure to falls-risk in a group of post-menopausal, post-surgical breast cancer survivors. Methods The study recruited 102 women (aged 65–79; mean age 70.2), post-surgical breast cancer survivors. The subjects were stratified by age into three groups: Group 1 (65–69 years); Group 2 (70–74 years), and Group 3 (75–79 years). Individual functional capabilities were assessed with Eight-foot up & go test (8UG), chair stand test (CST), and 2-minute step test (2ST). Tinetti POMA test was applied to assess gait and balance disorders. Self-reported falls in the past year were ascertained through a questionnaire. Results Assessment of individual aerobic endurance (2ST) also demonstrated a clear deficit in the mean scores category in all respective age sub-groups, as compared against the reference values. The deficits ranged from 4.86 to 15.90 steps less than the normative values; the oldest subjects demonstrating the largest deficit. The aerobic endurance tests results significantly impacted the ultimate assessment of an individual falls-risk in the oldest group. The analysis of the number of falls sustained within the recent year indicated that 43.67% of the subjects fell victim of such incidents. Conclusion An individual exposure to falls-risk was found to be appreciably more dependent upon individual aerobic endurance rather than overall strength of the lower part of the body in the breast cancer survivors over 75.

Description: by Jae Ha Gwak, So Young Sohn Background Recently, the need for rapid wound-healing has significantly increased because of the increasing number of patients who are diagnosed with diabetes and obesity. These conditions have contributed to a surge in the number of patients with chronic wounds worldwide. Furthermore, many cost-effective wound-healing technologies have been developed in order to keep up with the increased demand. In this paper, we performed a quantitative study of the trends associated with wound-healing technologies using patent data. Methodology We analyzed the trends considering four different groups of patent applicants: firms, universities, research institutes, and individuals using a structural topic model. In addition, we analyzed the knowledge flow between patent applicants using citation analysis, and confirmed the role of applicants in the knowledge-flow network using k-means clustering. As a result, the primary wound-healing technology patents applied for by the four groups varied considerably, and we classified the roles of patent applicants were found in the knowledge-flow network. Conclusions Our results showed the organizations that are leading each area of wound-healing technology. Furthermore, from the results, we identified specific institutions that are efficient for spreading knowledge related to wound-healing technology based on the patents. This information can contribute to the planning of investment strategies and technology policies related to wound-healing.

Description: by Xiyang Liu, Jiewei Jiang, Kai Zhang, Erping Long, Jiangtao Cui, Mingmin Zhu, Yingying An, Jia Zhang, Zhenzhen Liu, Zhuoling Lin, Xiaoyan Li, Jingjing Chen, Qianzhong Cao, Jing Li, Xiaohang Wu, Dongni Wang, Haotian Lin Slit-lamp images play an essential role for diagnosis of pediatric cataracts. We present a computer vision-based framework for the automatic localization and diagnosis of slit-lamp images by identifying the lens region of interest (ROI) and employing a deep learning convolutional neural network (CNN). First, three grading degrees for slit-lamp images are proposed in conjunction with three leading ophthalmologists. The lens ROI is located in an automated manner in the original image using two successive applications of Candy detection and the Hough transform, which are cropped, resized to a fixed size and used to form pediatric cataract datasets. These datasets are fed into the CNN to extract high-level features and implement automatic classification and grading. To demonstrate the performance and effectiveness of the deep features extracted in the CNN, we investigate the features combined with support vector machine (SVM) and softmax classifier and compare these with the traditional representative methods. The qualitative and quantitative experimental results demonstrate that our proposed method offers exceptional mean accuracy, sensitivity and specificity: classification (97.07%, 97.28%, and 96.83%) and a three-degree grading area (89.02%, 86.63%, and 90.75%), density (92.68%, 91.05%, and 93.94%) and location (89.28%, 82.70%, and 93.08%). Finally, we developed and deployed a potential automatic diagnostic software for ophthalmologists and patients in clinical applications to implement the validated model.

Description: by The PLOS ONE Staff

Description: by David L. Dickinson, Sean P. A. Drummond, Todd McElroy Chronic sleep restriction (SR) increases sleepiness, negatively impacts mood, and impairs a variety of cognitive performance measures. The vast majority of work establishing these effects are tightly controlled in-lab experimental studies. Examining commonly-experienced levels of SR in naturalistic settings is more difficult and generally involves observational methods, rather than active manipulations of sleep. The same is true for analyzing behavioral and cognitive outcomes at circadian unfavorable times. The current study tested the ability of an at-home protocol to manipulate sleep schedules (i.e., impose SR), as well as create a mismatch between a subject’s circadian preference and time of testing. Viability of the protocol was assessed via completion, compliance with the SR, and success at manipulating sleepiness and mood. An online survey was completed by 3630 individuals to assess initial eligibility, 256 agreed via email response to participate in the 3-week study, 221 showed for the initial in-person session, and 184 completed the protocol (175 with complete data). The protocol consisted of 1 week at-home SR (5-6 hours in bed/night), 1 week wash-out, and 1 week well-rested (WR: 8-9 hours in bed/night). Sleep was monitored with actigraphy, diary, and call-ins. Risk management strategies were implemented for subject safety. At the end of each experimental week, subjects reported sleepiness and mood ratings. Protocol completion was 83%, with lower depression scores, higher anxiety scores, and morning session assignment predicting completion. Compliance with the sleep schedule was also very good. Subjects spent approximately 2 hours less time in bed/night and obtained an average of 1.5 hours less nightly sleep during SR, relative to WR, with 82% of subjects obtaining at least 60 minutes less average nightly sleep. Sleepiness and mood were impacted as expected by SR. These findings show the viability of studying experimental chronic sleep restriction outside the laboratory, assuming appropriate safety precautions are taken, thus allowing investigators to significantly increase ecological validity over strictly controlled in-lab studies.

Description: by Orly Dorot, Hermann Steller, Daniel Segal, Mia Horowitz

Description: by Melissa J. Armstrong, C. Daniel Mullins, Gary S. Gronseth, Anna R. Gagliardi Background Patient and consumer engagement in clinical practice guideline development is internationally advocated, but limited research explores mechanisms for successful engagement. Objective To investigate the perspectives of potential patient/consumer guideline representatives on topics pertaining to engagement including guideline development group composition and barriers to and facilitators of engagement. Setting and participants Participants were guideline-naïve volunteers for programs designed to link community members to academic research with diverse ages, gender, race, and degrees of experience interacting with health care professionals. Methods Three focus groups and one key informant interview were conducted and analyzed using a qualitative descriptive approach. Results Participants recommended small, diverse guideline development groups engaging multiple patient/consumer stakeholders with no prior relationships with each other or professional panel members. No consensus was achieved on the ideal balance of patient/consumer and professional stakeholders. Pre-meeting reading/training and an identified contact person were described as keys to successful early engagement; skilled facilitators, understandable speech and language, and established mechanisms for soliciting patient opinions were suggested to enhance engagement at meetings. Conclusions Most suggestions for effective patient/consumer engagement in guidelines require forethought and planning but little additional expense, making these strategies easily accessible to guideline developers desiring to achieve more meaningful patient and consumer engagement.

Description: by Jie Wang, Suying Chang, Liyun Zhao, Wentao Yu, Jian Zhang, Qingqing Man, Li He, Yifan Duan, Hui Wang, Robert Scherpbier, Shi-an Yin Background Poor growth and micronutrient deficiency mainly attack older infants and young children. Some countries have adopted clinically effective measures to combat malnutrition, but the compliance and improvement in efficacy of intervention vehicles in national programs require evaluation. Methods Baseline and follow-up cross-sectional surveys were conducted before and after a nutrition intervention program in 3 national poverty counties in China. Soybean-based complementary food supplements called Yingyangbao (YYB) in Chinese and training materials on child feeding were distributed to households with children aged 6–23 months for 18 months. Representative children were selected by probability proportional to size sampling methods to assess compliance of YYB and the intervention efficacy. A questionnaire was designed to collect data on basic characteristics of children, breastfeeding, 24-hour dietary intake, and consumption and appetite of YYB. Anthropometrics and hemoglobin were measured in the field, and anemia prevalence was evaluated. Venous blood was drawn from children aged 12–35 months to evaluate micronutrient status. Logistic regression was used to identify the risk factors for children’s anemia. Results Of the children involved in the follow-up survey (n = 693), the P 50 ( P 25, P 75) intake of YYB was 6.7 (3.5, 7.0) sachets weekly, and 54.7% of the children liked the taste of YYB. Compared with the baseline situation (n = 823), the proportion of children fed a diverse diet and foods rich in iron or vitamin A increased ( P 〈 0.01) in the follow-up study. The prevalence of stunting and underweight decreased ( P 〈 0.05), the prevalence of anemia decreased from 28.0% to 19.9% ( P 〈 0.01), and the prevalence of vitamin B 12 deficiency decreased from 26.8% to 15.4% ( P 〈 0.01). For children aged 12–23 months, those who liked YYB and consumed 6 or more sachets of YYB weekly were at lower risk for anemia (OR = 0.34, 95% CI 0.13–0.90, P 〈 0.05), but the risk of stunting was associated with a non-diverse diet (OR = 1.48, 95% CI 1.06–2.07, P 〈 0.05). Conclusion The quality of diet and nutritional status of children aged 6–23 months are significantly improved by the intervention of YYB and nutrition education, and good compliance to YYB contributes to a low risk for anemia. Trial Registration Chinese Clinical Trial Registry ChiCTR-OOC-16008846

Description: by Saeam Shin, Sung Ran Cho, Sinyoung Kim, Jong Rak Choi, Kyung-A Lee Optimal timing of apheresis initiation is important for maximizing the hematopoietic stem cell (HSC) yield. This study aimed to identify useful parameters from automatic hematology analyzers for predicting the peripheral blood CD34+ cell count after mobilization. We prospectively enrolled 53 healthy donors and 72 patients, and evaluated 43 cell morphology parameters from Unicel DxH800 (Beckman Coulter, USA) and Advia 2120i (Siemens Healthcare Diagnostics, USA). The correlation of each parameter with the CD34+ cell count in pre-apheresis blood samples was analyzed. The delta neutrophil index (DNI) from Advia 2120i, standard deviation of volume of neutrophils and monocytes (SD-V-NE and SD-V-MO), standard deviation of conductivity of neutrophils and monocytes (SD-C-NE and SD-C-MO), mean conductivity of neutrophils and monocytes (MN-C-NE and MN-C-MO), and standard deviation of axial light loss of neutrophils and monocytes (SD-AL2-NE and SD-AL2-MO) from DxH800 showed significant correlations with the CD34+ cell count. SD-V-NE, SD-C-NE, and SD-C-MO showed good or fair area under the curve values for the prediction of the CD34+ cell count. SD-V-NE, SD-C-NE, and SD-C-MO from DxH800 will provide rapid, useful information for the initiation of apheresis after mobilization.

Description: by Shankar Raj Devkota, Eunju Kwon, Sung Chul Ha, Hyeun Wook Chang, Dong Young Kim Bacillus subtilis SigW is localized to the cell membrane and is inactivated by the tight interaction with anti-sigma RsiW under normal growth conditions. Whereas SigW is discharged from RsiW binding and thus initiates the transcription of its regulon under diverse stress conditions such as antibiotics and alkaline shock. The release and activation of SigW in response to extracytoplasmic signals is induced by the regulated intramembrane proteolysis of RsiW. As a ZAS (Zinc-containing anti-sigma) family protein, RsiW has a CHCC zinc binding motif, which implies that its anti-sigma activity may be regulated by the state of zinc coordination in addition to the proteolytic cleavage of RsiW. To understand the regulation mode of SigW activity by RsiW, we determined the crystal structures of SigW in complex with the cytoplasmic domain of RsiW, and compared the conformation of the CHCC motif in the reduced/zinc binding and the oxidized states. The structures revealed that RsiW inhibits the promoter binding of SigW by interacting with the surface groove of SigW. The interaction between SigW and RsiW is not disrupted by the oxidation of the CHCC motif in RsiW, suggesting that SigW activity might not be regulated by the zinc coordination states of the CHCC motif.

Description: by Aikichi Iwamoto, Rikizo Taira, Yoshiyuki Yokomaku, Tomohiko Koibuchi, Mahbubur Rahman, Yoko Izumi, Kenji Tadokoro Japan has been known as a low HIV-prevalence country with a concentrated epidemic among high-risk groups. However, it has not been determined whether Japan meets the 90-90-90 goals set by the Joint United Nations Programme on HIV/AIDS (UNAIDS)/World Health Organization (WHO). Moreover, to date, the HIV care cascade has not been examined. We estimated the total number of diagnosed people living with HIV/AIDS (PLWHA) (n = 22,840) based on legal reports to the Ministry of Health, Labour and Welfare by subtracting the number of foreigners who left Japan (n = 2,273) and deaths (n = 2,321) from the cumulative diagnosis report (n = 27,434). The number of total undiagnosed PLWHA was estimated by age and sex specific HIV-positive rates observed among first-time blood donors between 2011–2015 in Japan. Our estimates show that 14.4% (n = 3,830) of all PLWHA (n = 26,670) were undiagnosed in Japan at the end of 2015. The number of patients retained in care (n = 20,615: 77.3% of PLWHA), the percentage of those on antiretroviral therapy (n = 18,921: 70.9% of PLWHA) and those with suppressed viral loads (

Description: by David Rodrigues, Diniz Lopes, Madoka Kumashiro When individuals are highly committed to their romantic relationship, they are more likely to engage in pro-relationship maintenance mechanisms. The present research expanded on the notion that commitment redirects self-oriented goals to consider broader relational goals and examined whether commitment interacts with a promotion and prevention focus to activate derogation of attractive alternatives. Three studies used cross-sectional and experimental approaches. Study 1 showed that romantically involved individuals predominantly focused on promotion, but not prevention, reported less initial attraction to an attractive target than single individuals, especially when highly committed to their relationship. Study 2 showed that romantically involved individuals induced in a promotion focus, compared to those in prevention focus, reported less initial attraction, but only when more committed to their relationship. Regardless of regulatory focus manipulation, more committed individuals were also less likely to perceive quality among alternative scenarios and to be attentive to alternative others in general. Finally, Study 3 showed that romantically involved individuals induced in promotion focus and primed with high commitment reported less initial attraction, than those primed with low commitment, or than those induced in prevention focus. Once again, for these latter no differences occurred according to commitment prime. Together, the findings suggest that highly committed promotion focused individuals consider broader relationship goals and activate relationship maintenance behaviors such as derogation of attractive alternatives to promote their relationship.

Description: by Kyosuke Uno, Toh Miyazaki, Kengo Sodeyama, Yoshiaki Miyamoto, Atsumi Nitta Shati/Nat8L significantly increased in the nucleus accumbens (NAc) of mice after repeated methamphetamine (METH) treatment. We reported that Shati/Nat8L overexpression in mouse NAc attenuated METH-induced hyperlocomotion, locomotor sensitization, and conditioned place preference. We recently found that Shati/Nat8L overexpression in NAc regulates the dopaminergic neuronal system via the activation of group II mGluRs by elevated N -acetylaspartylglutamate following N- acetylaspartate increase due to the overexpression. These findings suggest that Shati/Nat8L suppresses METH-induced responses. However, the mechanism by which METH increases the Shati/Nat8L mRNA expression in NAc is unclear. To investigate the regulatory mechanism of Shati/Nat8L mRNA expression, we performed a mouse Shati/Nat8L luciferase assay using PC12 cells. Next, we investigated the response of METH to Shati/Nat8L expression and CREB activity using mouse brain slices of NAc, METH administration to mice, and western blotting for CREB activity of specific dopamine receptor signals in vivo and ex vivo. We found that METH activates CREB binding to the Shati/Nat8L promoter to induce the Shati/Nat8L mRNA expression. Furthermore, the dopamine D1 receptor antagonist SCH23390, but not the dopamine D2 receptor antagonist sulpiride, inhibited the upregulation of Shati/Nat8L and CREB activities in the mouse NAc slices. Thus, the administration of the dopamine D1 receptor agonist SKF38393 increased the Shati/Nat8L mRNA expression in mouse NAc. These results showed that the Shati/Nat8L mRNA was increased by METH-induced CREB pathway via dopamine D1 receptor signaling in mouse NAc. These findings may contribute to development of a clinical tool for METH addiction.

Description: by Na Zhang, Richard Fielding, Inda Soong, Karen K. K. Chan, Conrad Lee, Alice Ng, Wing Kin Sze, Janice Tsang, Victor Lee, Wendy Wing Tak Lam Objective The eight-item Brief Illness Perception Questionnaire (B-IPQ) supposedly evaluates cognitive and emotional representations of illness. This study examined the validity and reliability of a traditional Chinese version of the B-IPQ in Hong Kong Chinese breast cancer survivors. Methods 358 Chinese breast cancer survivors who had recently ended their primary treatment completed this B-IPQ Chinese version. Confirmatory factor analysis (CFA) tested the factor structure. The internal consistency, construct, predictive and convergent validities of the scale were assessed. Results CFA revealed that the original three-factor (cognitive-emotional representations and illness comprehensibility) structure of the B-IPQ poorly fitted our sample. After deleting one item measuring illness coherence, seven-item gave an optimal two-factor (cognitive-emotional representations) structure for the B-IPQ (B-IPQ-7). Cronbach’s alpha for the two subscales were 0.653 and 0.821, and for the overall seven-item scale of B-IPQ was 0.783. Correlations of illness perception and physical symptom distress, anxiety, depression and known-group comparison between different treatment status suggested acceptable construct validity. The association between baseline illness perception and psychological distress at 3-month follow up supported predictive validity. Conclusions B-IPQ-7 appears to be a moderately valid measure of illness perception in cancer population, potentially useful for assessing illness representations in Chinese women with breast cancer.

Description: by Jing Meng, Zuoshan Li, Lin Shen Individuals with autism-spectrum disorder (ASD) exhibit impairments in response to others’ pain. Evidence suggests that features of autism are not restricted to individuals with ASD, and that autistic traits vary throughout the general population. To investigate the association between autistic traits and the responses to others’ pain in typically developing adults, we employed the Autism-Spectrum Quotient (AQ) to quantify autistic traits in a group of 1670 healthy adults and explored whether 60 participants (30 males and 30 females) with 10% highest AQ scores (High-AQ) would exhibit difficulties in the responses to others’ pain relative to 60 participants (30 males and 30 females) with 10% lowest AQ scores (Low-AQ). This study included a Visual Task and an Auditory Task to test behavioral differences between High-AQ and Low-AQ groups’ responses to others’ pain in both modalities. For the Visual Task, participants were instructed to respond to pictures depicting others’ pain. They were instructed to judge the stimuli type (painful or not), judge others’ pain intensity, and indicate the unpleasantness they personally felt. For the Auditory Task, experimental procedures were identical to the Visual Task except that painful voices were added. Results showed the High-AQ group was less accurate than the Low-AQ group in judging others’ pain. Moreover, relative to Low-AQ males, High-AQ males had significantly longer reaction times in judging others’ pain in the Auditory Task. However, High-AQ and Low-AQ females showed similar reaction times in both tasks. These findings demonstrated identification of others’ pain by healthy adults is related to the extent of autistic traits, gender, and modality.

Description: by Fei Chen, Yuan Zhang, Ashley B. Daugherty, Zunyi Yang, Ryan Shaw, Mengxing Dong, Stefan Lutz, Steven A. Benner One research goal for unnatural base pair (UBP) is to replicate, transcribe and translate them in vivo . Accordingly, the corresponding unnatural nucleoside triphosphates must be available at sufficient concentrations within the cell. To achieve this goal, the unnatural nucleoside analogues must be phosphorylated to the corresponding nucleoside triphosphates by a cascade of three kinases. The first step is the monophosphorylation of unnatural deoxynucleoside catalyzed by deoxynucleoside kinases (dNK), which is generally considered the rate limiting step because of the high specificity of dNKs. Here, we applied a Drosophila melanogaster deoxyribonucleoside kinase (DmdNK) to the phosphorylation of an UBP (a pyrimidine analogue (6-amino-5-nitro-3-(1’-b-d-2’-deoxyribofuranosyl)-2(1H)-pyridone, Z) and its complementary purine analogue (2-amino-8-(1’-b-d-2’-deoxyribofuranosyl)-imidazo[1,2-a]-1,3,5-triazin-4(8H)-one, P). The results showed that DmdNK could efficiently phosphorylate only the dP nucleoside. To improve the catalytic efficiency, a DmdNK-Q81E mutant was created based on rational design and structural analyses. This mutant could efficiently phosphorylate both dZ and dP nucleoside. Structural modeling indicated that the increased efficiency of dZ phosphorylation by the DmdNK-Q81E mutant might be related to the three additional hydrogen bonds formed between E81 and the dZ base. Overall, this study provides a groundwork for the biological phosphorylation and synthesis of unnatural base pair in vivo .

Description: by Karina Ribeiro Silva, Isis Côrtes, Sally Liechocki, João Regis Ivar Carneiro, Antônio Augusto Peixoto Souza, Radovan Borojevic, Clarissa Menezes Maya-Monteiro, Leandra Santos Baptista Background/Objectives The pathological condition of obesity is accompanied by a dysfunctional adipose tissue. We postulate that subcutaneous, preperitoneal and visceral obese abdominal white adipose tissue depots could have stromal vascular fractions (SVF) with distinct composition and adipose stem cells (ASC) that would differentially account for the pathogenesis of obesity. Methods In order to evaluate the distribution of SVF subpopulations, samples of subcutaneous, preperitoneal and visceral adipose tissues from morbidly obese women (n = 12, BMI: 46.2±5.1 kg/m 2 ) were collected during bariatric surgery, enzymatically digested and analyzed by flow cytometry (n = 12). ASC from all depots were evaluated for morphology, surface expression, ability to accumulate lipid after induction and cytokine secretion (n = 3). Results A high content of preadipocytes was found in the SVF of subcutaneous depot ( p = 0.0178). ASC from the three depots had similar fibroblastoid morphology with a homogeneous expression of CD34, CD146, CD105, CD73 and CD90. ASC from the visceral depot secreted the highest levels of IL-6, MCP-1 and G-CSF ( p = 0.0278). Interestingly, preperitoneal ASC under lipid accumulation stimulus showed the lowest levels of all the secreted cytokines, except for adiponectin that was enhanced ( p = 0.0278). Conclusions ASC from preperitoneal adipose tissue revealed the less pro-inflammatory properties, although it is an internal adipose depot. Conversely, ASC from visceral adipose tissue are the most pro-inflammatory. Therefore, ASC from subcutaneous, visceral and preperitoneal adipose depots could differentially contribute to the chronic inflammatory scenario of obesity.

Description: by Daijiro Hori, Brittany Dunkerly-Eyring, Yohei Nomura, Debjit Biswas, Jochen Steppan, Jorge Henao-Mejia, Hideo Adachi, Lakshmi Santhanam, Dan E. Berkowitz, Charles Steenbergen, Richard A. Flavell, Samarjit Das Background Endothelial dysfunction and arterial stiffening play major roles in cardiovascular diseases. The critical role for the miR-181 family in vascular inflammation has been documented. Here we tested whether the miR-181 family can influence the pathogenesis of hypertension and vascular stiffening. Methods and results qPCR data showed a significant decrease in miR-181b expression in the aorta of the older mice. Eight miR-181a1/b1 -/- mice and wild types (C57BL6J:WT) were followed weekly for pulse wave velocity (PWV) and blood pressure measurements. After 20 weeks, the mice were tested for endothelial function and aortic modulus. There was a progressive increase in PWV and higher systolic blood pressure in miR-181a1/b1 -/- mice compared with WTs. At 21 weeks, aortic modulus was significantly greater in the miR-181a1/b1 -/- group, and serum TGF-β was found to be elevated at this time. A luciferase reporter assay confirmed miR-181b targets TGF-βi (TGF-β induced) in the aortic VSMCs. In contrast, wire myography revealed unaltered endothelial function along with higher nitric oxide production in the miR-181a1/b1 -/- group. Cultured VECs and VSMCs from the mouse aorta showed more secreted TGF-β in VSMCs of the miR-181a1/b1 -/- group; whereas, no change was observed from VECs. Circulating levels of angiotensin II were similar in both groups. Treatment with losartan (0.6 g/L) prevented the increase in PWV, blood pressure, and vascular stiffness in miR-181a1/b1 -/- mice. Immunohistochemistry and western blot for p-SMAD2/3 validated the inhibitory effect of losartan on TGF-β signaling in miR-181a1/b1 -/- mice. Conclusions Decreased miR-181b with aging plays a critical role in ECM remodeling by removing the brake on the TGF-β, pSMAD2/3 pathway.

Description: by Marc Duez, Mathieu Giraud, Ryan Herbert, Tatiana Rocher, Mikaël Salson, Florian Thonier

Description: by Xiao Gao, Xiao Deng, Xin Wen, Ying She, Petra Corianne Vinke, Hong Chen

Description: by Igor Govorov, Signe Löfgren, Roza Chaireti, Margareta Holmström, Katarina Bremme, Miriam Mints

Description: by Andreas Meier, Kai Higashigaito, Katharina Martini, Moritz Wurnig, Burkhardt Seifert, Dagmar I. Keller, Thomas Frauenfelder, Hatem Alkadhi

Description: by Enrique Maguilla, Marcial Escudero

Description: by The PLOS ONE Journal Staff

Description: by Christopher Thompson, Sahar Rahim, Jeremiah Arnold, Abigail Hielscher The extracellular matrix (ECM) is abnormal in breast tumors and has been reported to contribute to breast tumor progression. One factor, which may drive ongoing matrix synthesis in breast tumors, is the loss of stromal caveolin-1 (cav-1), a scaffolding protein of caveolae, which has been linked to breast tumor aggressiveness. To determine whether loss of cav-1 results in the abnormal expression of matrix proteins, mammary glands from cav- 1-/- and cav- 1 +/+ mice were investigated for differences in expression of several ECM proteins. In addition, the presence of myofibroblasts, changes in the vessel density, and differences in duct number and size were assessed in the mammary glands of both animal models. Using immunohistochemistry, expression of fibronectin, tenascin-C, collagens and αSMA were significantly increased in the mammary glands of cav-1-/- mice. Second harmonic generation revealed more organized collagen fibers in cav-1 -/- glands and supported immunohistochemical analyses of increased collagen abundance in the glands of cav-1 -/- mice. Analysis of the ductal structure demonstrated a significant increase in the number of proliferating ducts in addition to significant increases in the duct circumference and area in cav-1 -/- glands compared to cav- 1 +/+ glands. Differences in microvessel density weren’t apparent between the animal models. In summary, we found that the loss of cav-1 resulted in increased ECM and α-SMA protein expression in murine mammary glands. Furthermore, we found that an abnormal ductal architecture accompanied the loss of cav-1. These data support a role for cav-1 in maintaining mammary gland structure.

Description: by Wei Dong, Amber Farooqui, Alberto J. Leon, David J. Kelvin Influenza viruses cause mild to severe respiratory infections in humans. Due to efficient means of transmission, the viruses infect human population on a large scale. Apart from vaccines, antiviral drugs are used to control infection; neuraminidase inhibitors are thought to be the first choice of treatment, particularly for severe cases. Rapidly evolving and emerging influenza viruses with increased frequency of viral resistance to these drugs stress the need to explore novel antiviral compounds. In this study, we investigated antiviral activity of ginseng extract and ginsenosides, the ginseng-derived triterpene and saponin compounds, against 2009 pandemic H1N1 virus in vitro and in vivo. Our data showed that treatment of mice with ginsenosides protected the animals from lethal 2009 pandemic H1N1 infection and lowered viral titers in animal lungs. Mechanistic studies revealed that ginsenosides interact with viral hemagglutinin protein and prevent the attachment of virus with α 2–3’ sialic acid receptors present on host cell surfaces. The interference in the viral attachment process subsequently minimizes viral entry into the cells and decreases the severity of the viral infection. We also describe that sugar moieties present in ginsenosides are indispensible for their attachment with viral HA protein. On the basis of our observations, we can say that ginsenosides are promising candidates for the development of antiviral drugs for influenza viruses.

Description: by Vladyslav Oles, Alexander Panchenko, Andrei Smertenko Rise of atmospheric CO 2 is one of the main causes of global warming. Catastrophic climate change can be avoided by reducing emissions and increasing sequestration of CO 2 . Trees are known to sequester CO 2 during photosynthesis, and then store it as wood biomass. Thus, breeding of trees with higher wood yield would mitigate global warming as well as augment production of renewable construction materials, energy, and industrial feedstock. Wood is made of cellulose-rich xylem cells produced through proliferation of a specialized stem cell niche called cambium. Importance of cambium in xylem cells production makes it an ideal target for the tree breeding programs; however our knowledge about control of cambium proliferation remains limited. The morphology and regulation of cambium are different from those of stem cell niches that control axial growth. For this reason, translating the knowledge about axial growth to radial growth has limited use. Furthermore, genetic approaches cannot be easily applied because overlaying tissues conceal cambium from direct observation and complicate identification of mutants. To overcome the paucity of experimental tools in cambium biology, we constructed a Boolean network CARENET (CAmbium REgulation gene NETwork) for modelling cambium activity, which includes the key transcription factors WOX4 and HD-ZIP III as well as their potential regulators. Our simulations predict that: (1) auxin, cytokinin, gibberellin, and brassinosteroids act cooperatively in promoting transcription of WOX4 and HD-ZIP III ; (2) auxin and cytokinin pathways negatively regulate each other; (3) hormonal pathways act redundantly in sustaining cambium activity; (4) individual cambium cells can have diverse molecular identities. CARENET can be extended to include components of other signalling pathways and be integrated with models of xylem and phloem differentiation. Such extended models would facilitate breeding trees with higher wood yield.

Description: by Àngela Domínguez, Núria Soldevila, Diana Toledo, Núria Torner, Luis Force, María José Pérez, Vicente Martín, Lourdes Rodríguez-Rojas, Jenaro Astray, Mikel Egurrola, Francisco Sanz, Jesús Castilla, Working Group of the Project PI12/02079 Pneumococcal pneumonia is a serious cause of morbidity and mortality in the elderly, but investigation of the etiological agent of community-acquired pneumonia (CAP) is not possible in most hospitalized patients. The aim of this study was to estimate the effect of pneumococcal polysaccharide vaccination (PPSV23) in preventing CAP hospitalization and reducing the risk of intensive care unit admission (ICU) and fatal outcomes in hospitalized people aged ≥65 years. We made a multicenter case-control study in 20 Spanish hospitals during 2013–2014 and 2014–2015. We selected patients aged ≥65 years hospitalized with a diagnosis of pneumonia and controls matched by sex, age and date of hospitalization. Multivariate analysis was performed using conditional logistic regression to estimate vaccine effectiveness and unconditional logistic regression to evaluate the reduction in the risk of severe and fatal outcomes. 1895 cases and 1895 controls were included; 13.7% of cases and 14.4% of controls had received PPSV23 in the last five years. The effectiveness of PPSV23 in preventing CAP hospitalization was 15.2% (95% CI -3.1–30.3). The benefit of PPSV23 in avoiding ICU admission or death was 28.1% (95% CI -14.3–56.9) in all patients, 30.9% (95% CI -32.2–67.4) in immunocompetent patients and 26.9% (95% CI -38.6–64.8) in immunocompromised patients. In conclusion, PPSV23 showed a modest trend to avoidance of hospitalizations due to CAP and to the prevention of death or ICU admission in elderly patients hospitalized with a diagnosis of CAP.

Description: by Mirsha Quinto-Sánchez, Celia Cintas, Caio Cesar Silva de Cerqueira, Virginia Ramallo, Victor Acuña-Alonzo, Kaustubh Adhikari, Lucía Castillo, Jorge Gomez-Valdés, Paola Everardo, Francisco De Avila, Tábita Hünemeier, Claudia Jaramillo, Williams Arias, Macarena Fuentes, Carla Gallo, Giovani Poletti, Lavinia Schuler-Faccini, Maria Cátira Bortolini, Samuel Canizales-Quinteros, Francisco Rothhammer, Gabriel Bedoya, Javier Rosique, Andrés Ruiz-Linares, Rolando González-José

Description: by Hiroshi Kishikawa, Kayoko Kimura, Asako Ito, Kyoko Arahata, Sakiko Takarabe, Shogo Kaida, Takanori Kanai, Soichiro Miura, Jiro Nishida

Description: by Kenichi Suda, Leslie Rozeboom, Hui Yu, Kim Ellison, Christopher J. Rivard, Tetsuya Mitsudomi, Fred R. Hirsch Small cell lung cancer (SCLC) is a highly aggressive malignancy with few therapeutic advances in the treatment in recent decades. Based on a recent study that identified the spliceosome as a therapeutic vulnerability in MYC-driven breast cancers, we evaluated the efficacy of a spliceosome inhibitor in SCLC cell lines and analyzed the correlation with MYC status. Among 23 SCLC cell lines examined, eight showed high MYC protein expression (〉 80% positive cells) by immunohistochemistry (IHC), while 10 cell lines demonstrated no staining for MYC. The remaining five cell lines showed weak staining ( 〈 40% positive cells). All four cell lines that were previously demonstrated to have MYC gene amplification were positive for MYC by IHC. Four cell lines with high MYC expression and four with low MYC expression were used in further analysis. A spliceosome inhibitor, pladienolide B, showed high efficacy (IC 50 〈 12nM) in all eight cell lines tested, irrespective of the MYC IHC or MYC gene amplification status. We observed that the four cell lines with higher sensitivity to the spliceosome inhibitor were established from patients with prior chemotherapy. Therefore we chronically treated H1048 cells, that were established from a treatment-naïve patient, with cisplatin for 4 weeks, and found that H1048-cisplatin treated cells became more sensitive to pladienolide B. In conclusion, our in vitro results indicate that spliceosome inhibitors would be promising molecular target drugs in SCLC irrespective of the MYC status, especially in the second-line settings after an effective front-line chemotherapy.

Description: by Philipp A. Reuken, Dorian Torres, Michael Baier, Bettina Löffler, Christoph Lübbert, Norman Lippmann, Andreas Stallmach, Tony Bruns

Description: by Francesca Necchi, Allan Saul, Simona Rondini Serum Bactericidal Activity (SBA) assay is the method of choice to evaluate the complement-mediated functional activity of both infection- and vaccine-induced antibodies. To perform a typical SBA assay, serial dilutions of sera are incubated with target bacterial strains and complement. The conventional SBA assay is based on plating on agar the SBA reaction mix and counting the surviving bacterial colony forming units (CFU) at each serum dilution. Even with automated colony counting, it is labor-intensive, time-consuming and not amenable for large-scale studies. Here, we have developed a luminescence-based SBA (L-SBA) method able to detect surviving bacteria by measuring their ATP. At the end of the SBA reaction, a single commercially available reagent is added to each well of the SBA plate, and the resulting luminescence signal is measured in a microplate reader. The signal obtained is proportional to the ATP present, which is directly proportional to the number of viable bacteria. Bactericidal activity is subsequently calculated. We demonstrated the applicability of L-SBA with multiple bacterial serovars, from 5 species: Citrobacter freundii , Salmonella enterica serovars Typhimurium and Enteritidis, Shigella flexneri serovars 2a and 3a, Shigella sonnei and Neisseria meningitidis . Serum bactericidal titers obtained by the luminescence readout method strongly correlate with the data obtained by the conventional agar plate-based assay, and the new assay is highly reproducible. L-SBA considerably shortens assay time, facilitates data acquisition and analysis and reduces the operator dependency, avoiding the plating and counting of CFUs. Our results demonstrate that L-SBA is a useful high-throughput bactericidal assay.

Description: by Kai Zhang, Hua Zheng, Falin Chen, Ruida Li, Miao Yang, Zhiyun Ouyang, Jun Lan, Xuewu Xiang Nitrogen (N) fertilization is necessary to sustain productivity in eucalypt plantations, but it can increase the risk of greenhouse gas emissions. However, the response of soil greenhouse gas emissions to N fertilization might be influenced by soil characteristics, which is of great significance for accurately assessing greenhouse gas budgets and scientific fertilization in plantations. We conducted a two-year N fertilization experiment (control [CK], low N [LN], middle N [MN] and high N [HN] fertilization) in two eucalypt plantations with different soil characteristics (higher and lower soil organic carbon sites [HSOC and LSOC]) in Guangxi, China, and assessed soil–atmosphere greenhouse gas exchanges. The annual mean fluxes of soil CO 2 , CH 4 , and N 2 O were separately 153–266 mg m -2 h -1 , -55 –-40 μg m -2 h -1 , and 11–95 μg m -2 h -1 , with CO 2 and N 2 O emissions showing significant seasonal variations. N fertilization significantly increased soil CO 2 and N 2 O emissions and decreased CH 4 uptake at both sites. There were significant interactions of N fertilization and SOC level on soil CO 2 and N 2 O emissions. At the LSOC site, the annual mean flux of soil CO 2 emission was only significantly higher than the CK treatment in the HN treatment, but, at the HSOC site, the annual mean flux of soil CO 2 emission was significantly higher for both the LN (or MN) and HN treatments in comparison to the CK treatment. Under the CK and LN treatments, the annual mean flux of N 2 O emission was not significantly different between HSOC and LSOC sites, but under the HN treatment, it was significantly higher in the HSOC site than in the LSOC site. Correlation analysis showed that changes in soil CO 2 and N 2 O emissions were significantly related to soil dissolved organic carbon, ammonia, nitrate and pH. Our results suggested significant interactions of N fertilization and soil characteristics existed in soil–atmosphere greenhouse gas exchanges, which should be considered in assessing greenhouse gas budgets and scientific fertilization strategies in eucalypt plantations.

Description: by Kelvin Kamfwa, Dongyan Zhao, James D. Kelly, Karen A. Cichy Common bean ( Phaseolus vulgaris L.) fixes atmospheric nitrogen (N 2 ) through symbiotic nitrogen fixation (SNF) at levels lower than other grain legume crops. An understanding of the genes and molecular mechanisms underlying SNF will enable more effective strategies for the genetic improvement of SNF traits in common bean. In this study, transcriptome profiling was used to identify genes and molecular mechanisms underlying SNF differences between two common bean recombinant inbred lines that differed in their N-fixing abilities. Differential gene expression and functional enrichment analyses were performed on leaves, nodules and roots of the two lines when grown under N-fixing and non-fixing conditions. Receptor kinases, transmembrane transporters, and transcription factors were among the differentially expressed genes identified under N-fixing conditions, but not under non-fixing conditions. Genes up-regulated in the stronger nitrogen fixer, SA36, included those involved in molecular functions such as purine nucleoside binding, oxidoreductase and transmembrane receptor activities in nodules, and transport activity in roots. Transcription factors identified in this study are candidates for future work aimed at understanding the functional role of these genes in SNF. Information generated in this study will support the development of gene-based markers to accelerate genetic improvement of SNF in common bean.

Description: by Emily Saintas, Liam Abrahams, Gulshan T. Ahmad, Anu-Oluwa M. Ajakaiye, Abdulaziz S. H. A. M. AlHumaidi, Candice Ashmore-Harris, Iain Clark, Usha K. Dura, Carine N. Fixmer, Chinedu Ike-Morris, Mireia Mato Prado, Danielle Mccullough, Shishir Mishra, Katia M. U. Schöler, Husne Timur, Maxwell D. C. Williamson, Markella Alatsatianos, Basma Bahsoun, Edith Blackburn, Catherine E. Hogwood, Pamela E. Lithgow, Michelle Rowe, Lyto Yiangou, Florian Rothweiler, Jindrich Cinatl jr., Richard Zehner, Anthony J. Baines, Michelle D. Garrett, Campbell W. Gourlay, Darren K. Griffin, William J. Gullick, Emma Hargreaves, Mark J. Howard, Daniel R. Lloyd, Jeremy S. Rossman, C. Mark Smales, Anastasios D. Tsaousis, Tobias von der Haar, Mark N. Wass, Martin Michaelis The formation of acquired drug resistance is a major reason for the failure of anti-cancer therapies after initial response. Here, we introduce a novel model of acquired oxaliplatin resistance, a sub-line of the non-MYCN-amplified neuroblastoma cell line SK-N-AS that was adapted to growth in the presence of 4000 ng/mL oxaliplatin (SK-N-AS r OXALI 4000 ). SK-N-AS r OXALI 4000 cells displayed enhanced chromosomal aberrations compared to SK-N-AS, as indicated by 24-chromosome fluorescence in situ hybridisation. Moreover, SK-N-AS r OXALI 4000 cells were resistant not only to oxaliplatin but also to the two other commonly used anti-cancer platinum agents cisplatin and carboplatin. SK-N-AS r OXALI 4000 cells exhibited a stable resistance phenotype that was not affected by culturing the cells for 10 weeks in the absence of oxaliplatin. Interestingly, SK-N-AS r OXALI 4000 cells showed no cross resistance to gemcitabine and increased sensitivity to doxorubicin and UVC radiation, alternative treatments that like platinum drugs target DNA integrity. Notably, UVC-induced DNA damage is thought to be predominantly repaired by nucleotide excision repair and nucleotide excision repair has been described as the main oxaliplatin-induced DNA damage repair system. SK-N-AS r OXALI 4000 cells were also more sensitive to lysis by influenza A virus, a candidate for oncolytic therapy, than SK-N-AS cells. In conclusion, we introduce a novel oxaliplatin resistance model. The oxaliplatin resistance mechanisms in SK-N-AS r OXALI 4000 cells appear to be complex and not to directly depend on enhanced DNA repair capacity. Models of oxaliplatin resistance are of particular relevance since research on platinum drugs has so far predominantly focused on cisplatin and carboplatin.

Description: by Joaquin Paillamanque, Ana Sanchez-Tusie, Emerson M. Carmona, Claudia L. Treviño, Carolina Sandoval, Francisco Nualart, Nelson Osses, Juan G. Reyes Arachidonic acid (AA), a compound secreted by Sertoli cells (SC) in a FSH-dependent manner, is able to induce the release of Ca 2+ from internal stores in round spermatids and pachytene spermatocytes. In this study, the possible site(s) of action of AA in round spermatids, the signalling pathways associated and the intracellular Ca 2+ stores targeted by AA-induced signalling were pharmacologically characterized by measuring intracellular Ca 2+ using fluorescent Ca 2+ probes. Our results suggest that AA acts by interacting with a fatty acid G protein coupled receptor, initiating a G protein signalling cascade that may involve PLA2 and ERK activation, which in turn opens intracellular ryanodine-sensitive channels as well as NAADP-sensitive channels in acidic intracellular Ca 2+ stores. The results presented here also suggest that AMPK and PKA modulate this AA-induced Ca 2+ release from intracellular Ca 2+ stores in round spermatids. We propose that unsaturated free fatty acid lipid signalling in the seminiferous tubule is a novel regulatory component of rat spermatogenesis.

Description: by Nai-si Huang, Chen-lian Quan, Miao Mo, Jia-jian Chen, Ben-long Yang, Xiao-yan Huang, Jiong Wu Objectives The current study aims to summarize breast anthropomorphic measurement features in Chinese patients with breast diseases and to investigate their potential correlations with demographic factors. Materials and methods Fifteen breast anthropomorphic parameters of 605 Chinese female patients were collected prospectively. Breast ptosis status was scaled by two methods and breast volume was calculated according to a modified formula of BREAST-V. Results Among 1210 breasts, the average breast volume was 340.0±109.1 ml (91.8–919.2 ml). The distance from the nipple to the inframammary fold was 7.5±1.6 cm in the standing position. The width of the breast base was 14.3±1.4 cm (8.5–23.5 cm). The incidence of breast ptosis was 22.8% (274/1204), of which 37 (23.5%) and 79 (31.7%) women had severe ptosis assessed by different criteria. Increased height (OR[odds ratio] = 1.500, P

Description: by Lingmei Sun, Kai Liao, Chengcheng Hang, Dayong Wang Objective To investigate the effects of honokiol on induction of reactive oxygen species (ROS), antioxidant defense systems, mitochondrial dysfunction, and apoptosis in Candida albicans . Methods To measure ROS accumulation, 2′,7′-dichlorofluorescein diacetate fluorescence was used. Lipid peroxidation was assessed using both fluorescence staining and a thiobarbituric acid reactive substances (TBARS) assay. Protein oxidation was determined using dinitrophenylhydrazine derivatization. Antioxidant enzymatic activities were measured using commercially available detection kits. Superoxide dismutase (SOD) genes expression was measured using real time RT-PCR. To assess its antifungal abilities and effectiveness on ROS accumulation, honokiol and the SOD inhibitor N,N′-diethyldithiocarbamate (DDC) were used simultaneously. Mitochondrial dysfunction was assessed by measuring the mitochondrial membrane potential (mtΔψ). Honokiol-induced apoptosis was assessed using an Annexin V-FITC apoptosis detection kit. Results ROS, lipid peroxidation, and protein oxidation occurred in a dose-dependent manner in C . albicans after honokiol treatment. Honokiol caused an increase in antioxidant enzymatic activity. In addition, honokiol treatment induced SOD genes expression in C . albicans cells. Moreover, addition of DDC resulted in increased endogenous ROS levels and potentiated the antifungal activity of honokiol. Mitochondrial dysfunction was confirmed by measured changes to mtΔψ. The level of apoptosis increased in a dose-dependent manner after honokiol treatment. Conclusions Collectively, these results indicate that honokiol acts as a pro-oxidant in C . albicans . Furthermore, the SOD inhibitor DDC can be used to potentiate the activity of honokiol against C . albicans .

Description: by Benjamin Udoka Nwosu, Louise Maranda, Ninfa Candela Importance Irritable bowel syndrome (IBS) is associated with significant morbidity in children and adolescents, and the therapeutic efficacy of available treatment options is limited. The role of vitamin D supplementation in pediatric IBS is unclear as the vitamin D status of pediatric patients with IBS is unknown. Equally, the relationship of vitamin D status with psychosomatic symptoms in children and adolescents is unclear. Aim To characterize the vitamin D status of pediatric patients with IBS using a case-control study design. Hypothesis Serum 25-hydroxyvitamin D [25(OH)D] concentration will be similar between patients with IBS and controls. Subjects and methods A retrospective case-controlled study of 116 controls (age 14.6 ± 4.3 y), female (n = 67; 58%) and 55 subjects with IBS (age 16.5 ± 3.1y), female (n = 44; 80%). Overweight was defined as BMI of ≥85 th but

Description: by Yun-Peng Huang, Cheng-Fei Du, Cheng-Kung Cheng, Zheng-Cheng Zhong, Xuan-Wei Chen, Gui Wu, Zhe-Chen Li, Jin-Duo Ye, Jian-Hua Lin, Li Zhen Wang

Description: by Jennifer Dykxhoorn, Simon Hatcher, Marie-Hélène Roy-Gagnon, Ian Colman Background Understanding suicidality has proven challenging given the complex aetiology in early childhood. Being able to accurately predict groups at increased risk of developing suicidal thoughts may aid in the development of targeted prevention programs that mitigate increased vulnerability. Further, the predictors of suicidal thoughts may be shared with other outcomes in adolescence. Previous research has linked many factors to suicidality, so the objective of this study was to consider how these factors may act together to increase risk of suicidal thoughts and other non-mental health outcomes. Methods Two longitudinal datasets were used in this analysis: the National Longitudinal Survey of Children and Youth (NLSCY) and the Avon Longitudinal Survey of Parents and Children (ALSPAC). A Classification and Regression Tree model comprised of 75 factors describing early childhood was constructed to identify subgroups of adolescents at high risk of suicidal thoughts in the NLSCY and was validated in ALSPAC. These subgroups were investigated to see if they also had elevated rates of antisocial behaviour, substance misuse, poor physical health, poor mental health, risky health behaviours, and/or poor academic performance. Results The sensitivity was calculated to be 22·7%, specificity was 89·2%, positive predictive value 17·8%, and negative predictive value 91·8% and had similar accuracy in the validation dataset. The models were better at predicting other adverse outcomes compared to suicidal thoughts. Conclusion There are groups of risk factors present in early life that can predict higher risk of suicidality in adolescence. Notably, these factors were also predictive of a range of adverse outcomes in adolescence.

Description: by Hui-Ru Chen, Tirawat Rairat, Shih-Hurng Loh, Yu-Chieh Wu, Thomas W. Vickroy, Chi-Chung Chou The present study was carried out to demonstrate novel use of pharmacokinetic approaches to characterize drug behaviors/movements in the vegetables with implications to food safety. The absorption, distribution, metabolism and most importantly, the elimination of tetracycline (TC) and sulfamethoxazole (SMX) in edible plants Brassica rapa chinensis and Ipomoea aquatica grown hydroponically were demonstrated and studied using non-compartmental pharmacokinetic analysis. The results revealed drug-dependent and vegetable-dependent pharmacokinetic differences and indicated that ephemeral vegetables could have high capacity accumulating antibiotics (up to 160 μg g -1 for TC and 38 μg g -1 for SMX) within hours. TC concentration in the root (C max ) could reach 11 times higher than that in the cultivation fluid and 3–28 times higher than the petioles/stems. Based on the volume of distribution (Vss), SMX was 3–6 times more extensively distributed than TC. Both antibiotics showed evident, albeit slow elimination phase with elimination half-lives ranging from 22 to 88 hours. For the first time drug elimination through the roots of a plant was demonstrated, and by viewing the root as a central compartment and continuous infusion without a loading dose as drug administration mode, it is possible to pharmacokinetically monitor the movement of antibiotics and their fate in the vegetables with more detailed information not previously available. Phyto-pharmacokinetic could be a new area worth developing new models for the assessment of veterinary drugs in edible plants.

Description: by Hannah L. Albritton, Pamela A. Kozlowski, Rebecca A. Lillis, Chris L. McGowin, Julia D. Siren, Stephanie N. Taylor, Joyce A. Ibana, Lyndsey R. Buckner, Li Shen, Alison J. Quayle Chlamydia trachomatis (CT) is the leading sexually transmitted bacterial infection. The continued global burden of CT infection strongly predicates the need for a vaccine to supplement current chlamydial control programs. The correlates of protection against CT are currently unknown, but they must be carefully defined to guide vaccine design. The localized nature of chlamydial infection in columnar epithelial cells of the genital tract necessitates investigation of immunity at the site of infection. The purpose of this study was to develop a sensitive whole bacterial enzyme-linked immunosorbent assay (ELISA) to quantify and compare CT-specific IgG and IgA in sera and genital secretions from CT-infected women. To achieve this, elementary bodies (EBs) from two of the most common genital serovars (D and E) were attached to poly-L-lysine-coated microtiter plates with glutaraldehyde. EB attachment and integrity were verified by the presence of outer membrane antigens and the absence of bacterial cytoplasmic antigens. EB-specific IgG and IgA standards were developed by pooling sera with high titers of CT-specific antibodies from infected women. Serum, endocervical and vaginal secretions, and endocervical cytobrush specimens from CT-infected women were used to quantify CT-specific IgG and IgA which were then normalized to total IgG and IgA, respectively. Analyses of paired serum and genital samples revealed significantly higher proportions of EB-specific antibodies in genital secretions compared to sera. Cervical and vaginal secretions and cytobrush specimens had similar proportions of EB-specific antibodies, suggesting any one of these genital sampling techniques could be used to quantify CT-specific antibodies when appropriate normalization methodologies are implemented. Overall, these results illustrate the need to investigate genital tract CT antibody responses, and our assay provides a useful quantitative tool to assess natural immunity in defined clinical groups and CT vaccine trials.

Description: by Jeffrey Funk Although high degrees of reliability have been found for many types of forecasts purportedly due to the existence of accountability, public forecasts of technology are rarely assessed and continue to have a poor reputation. This paper’s analysis of forecasts made by MIT’s Technology Review provides a rare assessment and thus a means to encourage accountability. It first shows that few of the predicted “breakthrough technologies” currently have large markets. Only four have sales greater than $10 billion while eight technologies not predicted by Technology Review have sales greater than $10 billion including three with greater than $100 billion and one other with greater than $50 billion. Second, possible reasons for these poor forecasts are then discussed including an over emphasis on the science-based process of technology change, sometimes called the linear model of innovation. Third, this paper describes a different model of technology change, one that is widely used by private companies and that explains the emergence of those technologies that have greater than $10 billion in sales. Fourth, technology change and forecasts are discussed in terms of cognitive biases and mental models.

Description: by Weiling Xu, Suzy A. A. Comhair, Allison J. Janocha, Abigail Lara, Lori A. Mavrakis, Carole D. Bennett, Satish C. Kalhan, Serpil C. Erzurum Aims Arginine metabolism via inducible nitric oxide synthase (iNOS) and arginase 2 (ARG2) is higher in asthmatics than in healthy individuals. We hypothesized that a sub-phenotype of asthma might be defined by the magnitude of arginine metabolism categorized on the basis of high and low fraction of exhaled nitric oxide (F E NO). Methods To test this hypothesis, asthmatics ( n = 52) were compared to healthy controls ( n = 51) for levels of F E NO, serum arginase activity, and airway epithelial expression of iNOS and ARG2 proteins, in relation to clinical parameters of asthma inflammation and airway reactivity. In parallel, bronchial epithelial cells were evaluated for metabolic effects of iNOS and ARG2 expression in vitro . Results Asthmatics with high F E NO (≥ 35 ppb; 44% of asthmatics) had higher expression of iNOS ( P = 0.04) and ARG2 ( P = 0.05) in the airway, indicating F E NO is a marker of the high arginine metabolic endotype. High F E NO asthmatics had the lowest FEV 1 % ( P 〈 0.001), FEV 1 /FVC ( P = 0.0002) and PC 20 ( P 〈 0.001) as compared to low F E NO asthmatics or healthy controls. Low F E NO asthmatics had near normal iNOS and ARG2 expression (both P 〉 0.05), and significantly higher PC 20 ( P 〈 0.001) as compared to high F E NO asthmatics. In vitro studies to evaluate metabolic effects showed that iNOS overexpression and iNOS+ARG2 co-expression in a human bronchial epithelial cell line led to greater reliance on glycolysis with higher rate of pyruvate going to lactate. Conclusions The high F E NO phenotype represents a large portion of the asthma population, and is typified by greater arginine metabolism and more severe and reactive asthma.

Description: by Christian Thaler, Tanja Schneider, Jan Sedlacik, Daniel Kutzner, Jan-Patrick Stellmann, Christoph Heesen, Jens Fiehler, Susanne Siemonsen Purpose In multiple sclerosis (MS) the sensitivity for detection of contrast enhancing lesions (CEL) in T1-weighted scans is essential for diagnostics and therapy decisions. The purpose of our study was to evaluate the sensitivity of T1w MPRAGE scans in comparison to T1w dark blood technique (T1-DB) for CEL in MS. Materials and methods 3T MR imaging was performed in 37 MS patients, including T2-weighted imaging, T1w MPRAGE before and after gadolinium injection (unenhanced-T1 and T1-CE) and T1-DB imaging. After gadolinium application, the T1-DB scan was performed prior to T1-CE. From unenhanced-T1 and T1-CE scans, subtraction images (T1-SUB) were calculated. The number of CEL was determined separately on T1-CE and T1-DB by two raters independently. Lesions only detected on T1-DB scans then were verified on T1-SUB. Only lesions detected by both raters were included in further analysis. Results In 16 patients, at least one CEL was detected by both rater, either on T1-CE or T1-DB. All lesions that were detected on T1-CE were also detected on T1-DB images. The total number of contrast enhancing lesions detected on T1-DB images (n = 54) by both raters was significantly higher than the corresponding number of lesions identified on T1-CE (n = 27) (p = 0.01); all of these lesions could be verified on SUB images. In 21 patients, no CEL was detected in any of the sequences. Conclusions The application of T1-DB technique increases the sensitivity for CEL in MS, especially for those lesions that show only subtle increase in intensity after Gadolinium application but remain hypo- or iso-intense to surrounding tissue.

Description: by Gunjita Singh, Olivier Da Ines, Maria Eugenia Gallego, Charles I. White The ploidy of eukaryote gametes must be halved to avoid doubling of numbers of chromosomes with each generation and this is carried out by meiosis, a specialized cell division in which a single chromosomal replication phase is followed by two successive nuclear divisions. With some exceptions, programmed recombination ensures the proper pairing and distribution of homologous pairs of chromosomes in meiosis and recombination defects thus lead to sterility. Two highly related recombinases are required to catalyse the key strand-invasion step of meiotic recombination and it is the meiosis-specific DMC1 which is generally believed to catalyse the essential non-sister chromatid crossing-over, with RAD51 catalysing sister-chromatid and non-cross-over events. Recent work in yeast and plants has however shown that in the absence of RAD51 strand-exchange activity, DMC1 is able to repair all meiotic DNA breaks and surprisingly, that this does not appear to affect numbers of meiotic cross-overs. In this work we confirm and extend this conclusion. Given that more than 95% of meiotic homologous recombination in Arabidopsis does not result in inter-homologue crossovers, Arabidopsis is a particularly sensitive model for testing the relative importance of the two proteins—even minor effects on the non-crossover event population should produce detectable effects on crossing-over. Although the presence of RAD51 protein provides essential support for the action of DMC1, our results show no significant effect of the absence of RAD51 strand-exchange activity on meiotic crossing-over rates or patterns in different chromosomal regions or across the whole genome of Arabidopsis, strongly supporting the argument that DMC1 catalyses repair of all meiotic DNA breaks, not only non-sister cross-overs.

Description: by Wenyang Hou, Swati Gupta, Marie-Claude Beauchamp, Libin Yuan, Loydie A. Jerome-Majewska The transmembrane emp24 domain/p24 (TMED) family are essential components of the vesicular transport machinery. Members of the TMED family serve as cargo receptors implicated in selection and packaging of endoplasmic reticulum (ER) luminal proteins into coatomer (COP) II coated vesicles for anterograde transport to the Golgi. Deletion or mutations of Tmed genes in yeast and Drosophila results in ER-stress and activation of the unfolded protein response (UPR). The UPR leads to expression of genes and proteins important for expanding the folding capacity of the ER, degrading misfolded proteins, and reducing the load of new proteins entering the ER. The UPR is activated in non-alcoholic fatty liver disease (NAFLD) in human and mouse and may contribute to the development and the progression of NAFLD. Tmed2 , the sole member of the vertebrate Tmed β subfamily, exhibits tissue and temporal specific patterns of expression in embryos and developing placenta but is ubiquitously expressed in all adult organs. We previously identified a single point mutation, the 99J mutation, in the signal sequence of Tmed2 in an N-ethyl-N-nitrosourea (ENU) mutagenesis screen. Histological and molecular analysis of livers from heterozygous mice carrying the 99J mutation, Tmed2 99J/+ , revealed a requirement for TMED2 in liver health. We show that Tmed2 99J/+ mice had decreased levels of TMED2 and TMED10, dilated endoplasmic reticulum membrane, and increased phosphorylation of eIF2α, indicating ER-stress and activation of the UPR. Increased expression of Srebp1a and 2 at the newborn stage and increased incidence of NAFLD were also found in Tmed2 99J/+ mice. Our data establishes Tmed2 99J/+ mice as a novel mouse model for NAFLD and supports a role for TMED2 in liver health.

Description: by Emre Bektik, Adrienne Dennis, Prateek Prasanna, Anant Madabhushi, Ji-Dong Fu The direct reprogramming of cardiac fibroblasts into induced cardiomyocyte (CM)-like cells (iCMs) holds great promise in restoring heart function. We previously found that human fibroblasts could be reprogrammed toward CM-like cells by 7 reprogramming factors; however, iCM reprogramming in human fibroblasts is both more difficult and more time-intensive than that in mouse cells. In this study, we investigated if additional reprogramming factors could quantitatively and/or qualitatively improve 7-factor-mediated human iCM reprogramming by single-cell quantitative PCR. We first validated 46 pairs of TaqMan ® primers/probes that had sufficient efficiency and sensitivity to detect the significant difference of gene expression between individual H9 human embryonic stem cell (ESC)-differentiated CMs (H9CMs) and human fibroblasts. The expression profile of these 46 genes revealed an improved reprogramming in 12-week iCMs compared to 4-week iCMs reprogrammed by 7 factors, indicating a prolonged stochastic phase during human iCM reprogramming. Although none of additional one reprogramming factor yielded a greater number of iCMs, our single-cell qPCR revealed that additional HAND2 or microRNA-1 could facilitate the silencing of fibroblast genes and yield a better degree of reprogramming in more reprogrammed iCMs. Noticeably, the more HAND2 expressed, the higher-level were cardiac genes activated in 7Fs+HAND2-reprogrammed iCMs. In conclusion, HAND2 and microRNA-1 could help 7 factors to facilitate the early progress of iCM-reprogramming from human fibroblasts. Our study provides valuable information to further optimize a method of direct iCM-reprogramming in human cells.

Description: by Susan Horton, Hellen Gelband, Dean Jamison, Carol Levin, Rachel Nugent, David Watkins Background Cost-effectiveness rankings of health interventions are useful inputs for national healthcare planning and budgeting. Previous comprehensive rankings for low- and middle- income countries were undertaken in 2005 and 2006, accompanying the development of strategies for the Millennium Development Goals. We update the rankings using studies published since 2000, as strategies are being considered for the Sustainable Development Goals. Methods Expert systematic searches of the literature were undertaken for a broad range of health interventions. Cost-effectiveness results using Disability Adjusted Life-Years (DALYs) as the health outcome were standardized to 2012 US dollars. Results 149 individual studies of 93 interventions qualified for inclusion. Interventions for Reproductive, Maternal, Newborn and Child Health accounted for 37% of interventions, and major infectious diseases (AIDS, TB, malaria and neglected tropical diseases) for 24%, consistent with the priorities of the Millennium Development Goals. More than half of the interventions considered cost less than $200 per DALY and hence can be considered for inclusion in Universal Health Care packages even in low-income countries. Discussion Important changes have occurred in rankings since 2006. Priorities have changed as a result of new technologies, new methods for changing behavior, and significant price changes for some vaccines and drugs. Achieving the Sustainable Development Goals will require LMICs to study a broader range of health interventions, particularly in adult health. Some interventions are no longer studied, in some cases because they have become usual care, in other cases because they are no longer relevant. Updating cost-effectiveness rankings on a regular basis is potentially a valuable exercise.

Description: by Gianni Bussolati, Laura Annaratone, Enrico Berrino, Umberto Miglio, Mara Panero, Marco Cupo, Patrizia Gugliotta, Tiziana Venesio, Anna Sapino, Caterina Marchiò Tissue fixation in phosphate buffered formalin (PBF) remains the standard procedure in histopathology, since it results in an optimal structural, antigenic and molecular preservation that justifies the pivotal role presently played by diagnoses on PBF-fixed tissues in precision medicine. However, toxicity of formaldehyde causes an environmental concern and may demand substitution of this reagent. Having observed that the reported drawbacks of commercially available glyoxal substitutes of PBF (Prefer, Glyo-fix, Histo-Fix, Histo-CHOICE, and Safe-Fix II) are likely related to their acidity, we have devised a neutral fixative, obtained by removing acids from the dialdehyde glyoxal with an ion-exchange resin. The resulting glyoxal acid-free (GAF) fixative has been tested in a cohort of 30 specimens including colon (N = 25) and stomach (N = 5) cancers. Our results show that GAF fixation produces a tissue and cellular preservation similar to that produced by PBF. Comparable immuno-histochemical and molecular (DNA and RNA) analytical data were obtained. We observed a significant enrichment of longer DNA fragment size in GAF-fixed compared to PBF-fixed samples. Adoption of GAF as a non-toxic histological fixative of choice would require a process of validation, but the present data suggest that it represents a reliable candidate.

Description: by Takayuki Onojima, Keiichi Kitajo, Hiroaki Mizuhara Neural oscillation is attracting attention as an underlying mechanism for speech recognition. Speech intelligibility is enhanced by the synchronization of speech rhythms and slow neural oscillation, which is typically observed as human scalp electroencephalography (EEG). In addition to the effect of neural oscillation, it has been proposed that speech recognition is enhanced by the identification of a speaker’s motor signals, which are used for speech production. To verify the relationship between the effect of neural oscillation and motor cortical activity, we measured scalp EEG, and simultaneous EEG and functional magnetic resonance imaging (fMRI) during a speech recognition task in which participants were required to recognize spoken words embedded in noise sound. We proposed an index to quantitatively evaluate the EEG phase effect on behavioral performance. The results showed that the delta and theta EEG phase before speech inputs modulated the participant’s response time when conducting speech recognition tasks. The simultaneous EEG-fMRI experiment showed that slow EEG activity was correlated with motor cortical activity. These results suggested that the effect of the slow oscillatory phase was associated with the activity of the motor cortex during speech recognition.

Description: by Tatiane Macedo-Silva, Rosana Beatriz Duque Araujo, Kamila Anna Meissner, Wesley Luzetti Fotoran, Márcia Melo Medeiros, Mauro Ferreira de Azevedo, Gerhard Wunderlich The genome of the malaria parasite Plasmodium falciparum contains the surf gene family which encodes large transmembrane proteins of unknown function. While some surf alleles appear to be expressed in sexual stages, others occur in asexual blood stage forms and may be associated to virulence-associated processes and undergo transcriptional switching. We accessed the transcription of surf genes along multiple invasions by real time PCR. Based on the observation of persistent expression of gene surf 4.1, we created a parasite line which expresses a conditionally destabilized SURFIN4.1 protein. Upon destabilization of the protein, no interference of parasite growth or morphological changes were detected. However, we observed a strong increase in the transcript quantities of surf 4.1 and sometimes of other surf genes in knocked-down parasites. While this effect was reversible when SURFIN4.1 was stabilized again after a few days of destabilization, longer destabilization periods resulted in a transcriptional switch away from surf 4.1. When we tested if a longer transcript half-life was responsible for increased transcript detection in SURFIN4.1 knocked-down parasites, no alteration was found compared to control parasite lines. This suggests a specific feedback of the expressed SURFIN protein to its transcript pointing to a novel type of regulation, inedited in Plasmodium .

Description: by Marcos Oliveira, Carmelo Bastos-Filho, Ronaldo Menezes Crime is a major threat to society’s well-being but lacks a statistical characterization that could lead to uncovering some of its underlying mechanisms. Evidence of nonlinear scaling of urban indicators in cities, such as wages and serious crime, has motivated the understanding of cities as complex systems—a perspective that offers insights into resources limits and sustainability, but that usually neglects details of the indicators themselves. Notably, since the nineteenth century, criminal activities have been known to occur unevenly within a city; crime concentrates in such way that most of the offenses take place in few regions of the city. Though confirmed by different studies, this concentration lacks broad analyses on its characteristics, which hinders not only the comprehension of crime dynamics but also the proposal of sounding counter-measures. Here, we developed a framework to characterize crime concentration which divides cities into regions with the same population size. We used disaggregated criminal data from 25 locations in the U.S. and the U.K., spanning from 2 to 15 years of longitudinal data. Our results confirmed that crime concentrates regardless of city and revealed that the level of concentration does not scale with city size. We found that the distribution of crime in a city can be approximated by a power-law distribution with exponent α that depends on the type of crime. In particular, our results showed that thefts tend to concentrate more than robberies, and robberies more than burglaries. Though criminal activities present regularities of concentration, we found that criminal ranks have the tendency to change continuously over time—features that support the perspective of crime as a complex system and demand analyses and evolving urban policies covering the city as a whole.

Description: by Julie Rivière, Yann Le Strat, Pascal Hendrikx, Barbara Dufour Bovine tuberculosis (bTB) is a common disease in cattle and wildlife, with health, zoonotic and economic implications. Infected wild animals, and particularly reservoirs, could hinder eradication of bTB from cattle populations, which could have an important impact on international cattle trade. Therefore, surveillance of bTB in wildlife is of particular importance to better understand the epidemiological role of wild species and to adapt the control measures. In France, a bTB surveillance system for free-ranging wildlife, the Sylvatub system, has been implemented since 2011. It relies on three surveillance components (SSCs) (passive surveillance on hunted animals (EC-SSC), passive surveillance on dead or dying animals (SAGIR-SSC) and active surveillance (PSURV-SSC)). The effectiveness of the Sylvatub system was previously assessed, through the estimation of its sensitivity ( i . e . the probability of detecting at least one case of bTB infection by each SSC, specie and risk-level area). However, to globally assess the performance of a surveillance system, the measure of its sensitivity is not sufficient, as other factors such as economic or socio-economic factors could influence the effectiveness. We report here an estimation of the costs of the surveillance activities of the Sylvatub system, and of the cost-effectiveness of each surveillance component, by specie and risk-level, based on scenario tree modelling with the same tree structure as used for the sensitivity evaluation. The cost-effectiveness of the Sylvatub surveillance is better in higher-risk departments, due in particular to the higher probability of detecting the infection (sensitivity). Moreover, EC-SSC, which has the highest unit cost, is more efficient than the surveillance enhanced by the SAGIR-SSC, due to its better sensitivity. The calculation of the cost-effectiveness ratio shows that PSURV-SSC remains the most cost-effective surveillance component of the Sylvatub system, despite its high cost in terms of coordination, sample collection and laboratory analysis.

Description: by Yuna Noh, Yoonjin Yoon Introduction Aging has long been regarded as one of the most critical factors affecting crash injury outcomes. In South Korea, where the elderly population is projected to reach 35.9% by 2050, the implications of an increasing number of elderly vehicle users on road safety are evident. In this research, the confounding effect of occupant age in a vehicle in terms of seat position and seatbelt use was investigated. In addition, elderly occupants were divided into a younger-old group aged between 65 and 74 years and an older-old group aged 75 years and older in an effort to assess whether the conventional elderly age standard of 65 years should be reconsidered. Methods A multinomial logit framework was adopted to predict two-level injury severity using collision data between 2008 and 2015. Predictor variables included gender, age group, seat position, seatbelt, road type, road slope, road surface, road line, and type of vehicle. Five models, a base model with no interactions and four interaction models which were combinations of age group, seatbelt use and seat position, were devised and evaluated. Results With no interacting term, age was the most prominent predictor. Elderly occupants were most likely to suffer from severe injury without a seatbelt in all seat positions, and the use of a seatbelt reduced this likelihood the most in the elderly group as well. Front passenger seats had the highest risk to elderly occupants, while the driver seat was statistically insignificant. When the elderly group was divided into the younger-old group and the older-old group, the older-olds were found to be much more vulnerable compared to the younger-olds. In particular, older drivers were five times more likely to suffer a severe injury without a seatbelt. Conclusions The degree of injury severity of elderly occupants was reduced the most with the use of a seatbelt, demonstrating the importance of using seat restraints. The sharp increase in the risk of injury of the older-old group suggests that the age standard of 65 years as the elderly group with regard to traffic safety may require reconsideration due to the growing number of elderly vehicle users on the road. Our results provide practical evidence with which to formulate new safety policies, including mandatory seatbelt use, driving age limits and insurance pricing.

Description: by Sheila Castro-Sánchez, María Álvarez-Satta, Mohamed A. Tohamy, Sergi Beltran, Sophia Derdak, Diana Valverde Ciliopathies are a group of rare disorders characterized by a high genetic and phenotypic variability, which complicates their molecular diagnosis. Hence the need to use the latest powerful approaches to faster identify the genetic defect in these patients. We applied whole exome sequencing to six consanguineous families clinically diagnosed with ciliopathy-like disease, and for which mutations in predominant Bardet-Biedl syndrome ( BBS ) genes had previously been excluded. Our strategy, based on first applying several filters to ciliary variants and using many of the bioinformatics tools available, allowed us to identify causal mutations in BBS2 , ALMS1 and CRB1 genes in four families, thus confirming the molecular diagnosis of ciliopathy. In the remaining two families, after first rejecting the presence of pathogenic variants in common cilia-related genes, we adopted a new filtering strategy combined with prioritisation tools to rank the final candidate genes for each case. Thus, we propose CORO2B , LMO7 and ZNF17 as novel candidate ciliary genes, but further functional studies will be needed to confirm their role. Our data show the usefulness of this strategy to diagnose patients with unclear phenotypes, and therefore the success of applying such technologies to achieve a rapid and reliable molecular diagnosis, improving genetic counselling for these patients. In addition, the described pipeline also highlights the common pitfalls associated to the large volume of data we have to face and the difficulty of assigning a functional role to these changes, hence the importance of designing the most appropriate strategy according to each case.

Description: by Daniel Belstrøm, Maria Lynn Sembler-Møller, Maria Anastasia Grande, Nikolai Kirkby, Sean Liam Cotton, Bruce J. Paster, Palle Holmstrup Objectives The purpose of this study was to compare microbial profiles of saliva, pooled and site-specific subgingival samples in patients with periodontitis. We tested the hypotheses that saliva can be an alternative to pooled subgingival samples, when screening for presence of periopathogens. Design Site specific subgingival plaque samples (n = 54), pooled subgingival plaque samples (n = 18) and stimulated saliva samples (n = 18) were collected from 18 patients with generalized chronic periodontitis. Subgingival and salivary microbiotas were characterized by means of HOMI NGS (Human Oral Microbe Identification using Next Generation Sequencing) and microbial community profiles were compared using Spearman rank correlation coefficient. Results Pronounced intraindividual differences were recorded in site-specific microbial profiles, and site-specific information was in general not reflected by pooled subgingival samples. Presence of Porphyromonas gingivalis , Treponema denticola , Prevotella intermedia , Filifactor alocis , Tannerella forsythia and Parvimona micra in site-specific subgingival samples were detected in saliva with an AUC of 0.79 (sensitivity: 0.61, specificity: 0.94), compared to an AUC of 0.76 (sensitivity: 0.56, specificity: 0.94) in pooled subgingival samples. Conclusions Site-specific presence of periodontal pathogens was detected with comparable accuracy in stimulated saliva samples and pooled subgingival plaque samples. Consequently, saliva may be a reasonable surrogate for pooled subgingival samples when screening for presence of periopathogens. Future large-scale studies are needed to confirm findings from this study.

Description: by Yuanzheng Zhang, Shuo Fang, Jiezhi Dai, Lei Zhu, Hao Fan, Weiya Tang, Yongjie Fan, Haiying Dai, Peipei Zhang, Ying Wang, Xin Xing, Chao Yang To explore the repairing effect of combination of adipose stem cells (ASCs) and composite scaffolds on CWR, the electrospun Poly 1, 8-octanediol-co-citric acid (POC)-poly-L-lactide acid (PLA) composite scaffolds were prepared, followed by in vitro and in vivo biocompatibility evaluation of the scaffolds. Afterwards, ASCs were seeded on POC-PLA to construct the POC-PLA-ASCs scaffolds, and the POC-PLA, POC-PLA-ASCs, and traditional materials expanded polytetrafluoroethylene (ePTFE) were adopt for CWR in New Zealand white (NZW) rabbit models. As results, the POC-PLA-ASCs patches possessed good biocompatibility as the high proliferation ability of cells surrounding the patches. Rabbits in POC-PLA-ASCs groups showed better pulmonary function, less pleural adhesion, higher degradation rate and more neovascularization when compared with that in other two groups. The results of western blot indicated that POC-PLA-ASCs patches accelerated the expression of VEGF and Collagen I in rabbit models. From the above, our present study demonstrated that POC-PLA material was applied for CWR successfully, and ASCs seeded on the sheets could improve the pleural adhesions and promote the reparation of chest wall defects.

Description: by Margret Paar, Christine Rossmann, Christoph Nusshold, Thomas Wagner, Axel Schlagenhauf, Bettina Leschnik, Karl Oettl, Martin Koestenberger, Gerhard Cvirn, Seth Hallström Albumin is the most abundant plasma protein. Critical illness is often associated with altered, predominately decreased, serum albumin levels. This hypoalbuminaemia is usually corrected by administration of exogenous albumin. This study aimed to track the concentration-dependent influence of albumin on blood coagulation in vitro. Whole blood (WB) samples from 25 volunteers were prepared to contain low (19.3 ± 7.7 g/L), physiological (45.2 ± 7.8 g/L), and high (67.5 ± 18.1 g/L) levels of albumin. Haemostatic profiling was performed using a platelet function analyzer (PFA) 200, impedance aggregometry, a Cone and Platelet analyzer (CPA), calibrated automated thrombogram, and thrombelastometry (TEM). Platelet aggregation-associated ATP release was assessed via HPLC analysis. In the low albumin group, when compared to the physiological albumin group, we found: i) shortened PFA 200-derived closure times indicating increased primary haemostasis; ii) increased impedance aggregometry-derived amplitudes, slopes, ATP release, as well as CPA-derived average size indicating improved platelet aggregation; iii) increased TEM-derived maximum clot firmness and alpha angles indicating enhanced clot formation. TEM measurements indicated impaired clot formation in the high albumin group compared with the physiological albumin group. Thus, albumin exerted significant anticoagulant action. Therefore, low albumin levels, often present in cancer or critically ill patients, might contribute to the frequently occurring venous thromboembolism.

Description: by Melody Wiseheart, Annalise A. D’Souza, Jacey Chae Spacing effects during retention of verbal information are easily obtained, and the effect size is large. Relatively little evidence exists on whether motor skill retention benefits from distributed practice, with even less evidence on complex motor skills. We taught a 17-note musical sequence on a piano to individuals without prior formal training. There were five lags between learning episodes: 0-, 1-, 5-, 10-, and 15-min. After a 5-min retention interval, participants’ performance was measured using three criteria: accuracy of note playing, consistency in pressure applied to the keys, and consistency in timing. No spacing effect was found, suggesting that the effect may not always be demonstrable for complex motor skills or non-verbal abilities (timing and motor skills). Additionally, we taught short phrases from five songs, using the same set of lags and retention interval, and did not find any spacing effect for accuracy of song reproduction. Our findings indicate that although the spacing effect is one of the most robust phenomena in the memory literature (as demonstrated by verbal learning studies), the effect may vary when considered in the novel realm of complex motor skills such as piano performance.

Description: by Lipei Zhao, Liang Zhao, Buchang Zhang, Jason M. Robotham, Kenneth H. Roux, Hengli Tang Allergy to peanuts has become a common and severe problem, especially in westernized countries. In this study, we evaluated the target and epitope specificity of the capture and detection mouse monoclonal antibodies (mAbs) used in a commercial peanut allergen detection platform. We first identified the target of these antibodies as Ara h 3 and then used an overlapping peptide array of Ara h 3 to determine the antibody-binding epitopes. Further amino acids critical for the binding via alanine substitutions at individual amino acid residues within the epitope were mapped. Finally, inhibition ELISA and inhibition immunoblotting using a recombinant Ara h 3 protein were performed to confirm these results. Surprisingly, the capture and detection mAbs showed identical binding characteristics and were presumed to represent two isolates of the same clone, a notion supported by both isoelectric focusing electrophoresis and Liquid chromatography–mass spectrometry experiments. The simultaneous binding of a pair of identical mAbs to an individual allergen such as Ara h3 is attributed to the multivalency of the analyte and has implications for developing diagnostic assays for additional multimeric allergens.

Description: by Kritika Sadh, Priyanka Rai, Roop Mallik Lipid droplets (LDs) are cellular stores of neutral fat that facilitate lipid and protein trafficking in response to metabolic cues. Unlike other vesicles, the phospholipid membrane on the LD is a monolayer. Interestingly, this monolayer membrane has free cholesterol, and may therefore contain lipid microdomains that serve as a platform for assembling proteins involved in signal transduction, cell polarity, pathogen entry etc. In support of this, cell culture studies have detected microdomain-associated “raftophilic” proteins on LDs. However, the physiological significance of this observation has been unclear. Here we show that two proteins (Flotillin-1 and SNAP23) that bind to membrane microdomains associate differently with LDs purified from rat liver depending on the feeding/fasting state of the animal. Flotillin-1 increases on LDs in the fed state, possibly because LDs interact with the endoplasmic reticulum (ER), facilitating supply of flotillin-1 from ER to LDs. Interestingly, this increase in flotillin-1 is correlated with an increase in free cholesterol on the LDs in fed state. In opposite behaviour to flotillin-1, SNAP23 increases on LDs in the fasted state and this appears to mediate LD-mitochondria interactions. Such LD-mitochondria interactions may provide fatty acids to mitochondria for promoting beta-oxidation in hepatocytes in response to fasting. Our work brings out physiologically relevant aspects of lipid droplet biology that are different from, and may not be entirely possible to replicate and study in cell culture.

Description: by Linqing Zhong, Hongmei Song, Wei Wang, Ji Li, Mingsheng Ma Objective The aim of the current study was to determine the contributions of several common mutations in the Mediterranean fever (MEFV) gene, namely, E148Q, M680I, M694V and V726A, to ankylosing spondylitis (AS) susceptibility. Methods Two investigators independently searched the literature regarding the association of MEFV with AS in the PubMed, EMBASE, Web of Science, and Scopus databases. They independently selected eligible articles and then extracted data from the included studies. The associations between MEFV mutations and AS risk were assessed with odds ratios (ORs) and 95% confidence intervals (95% CI). Further analyses were conducted with STATA 12.0 software (Stata Corp.; College Station, Texas, USA). Results Four mutations (E148Q, M680I, M694V and V726A) were genotyped in 869 AS cases and 879 controls from the 8 eligible studies. Of the four mutations, M694V (pooled OR: 3.330, 95% CI: 2.129–5.208) was found to be associated with AS through overall analysis. However, the other mutations demonstrated no relation with AS (pooled ORs: 1.295, 1.258, 1.778; 95% CI: 0.886–1.891, 0.688–2.298 and 0.938–3.371). No significant publication bias was discovered in the meta-analysis. Conclusions The present study indicates that the MEFV M694V mutation may contribute to the pathogenesis of AS. The associations between the other mutations and AS need to be validated with more relevant and well-designed studies.

Description: by Xiaocui Fang, Hanyi Xie, Hongyang Duan, Ping Li, Maryam Yousaf, Haiyan Xu, Yanlian Yang, Chen Wang Cancer is the leading cause of death worldwide, and metastasis is the main attribute to cancer death. CXCR4 and its natural ligand CXCL12 have been known to play a critical role in tumorigenesis, angiogenesis and metastasis. Therefore, designing a new CXCR4 antagonist to prevent tumor metastasis will be of great significance. Herein, a novel chemically synthesized peptide (E5) that has an ability to target CXCR4/CXCL12 axis was loaded in micelle glycol-phosphatidylethanolamine (PEG-PE) block copolymer to form micelle-encapsulated E5 (M-E5). We demonstrated that M-E5 exhibited higher affinity for CXCR4-overexpressing MCF-7 and HepG2 tumor cells as compared to free E5, and efficiently inhibited the tumor cells migration. Mechanistic studies implied that PEG-PE micelle can encapsulate E5 and improve E5 targeting efficiency for CXCR4 by accumulating E5 on the tumor cell membrane. Furthermore, through encapsulation of chemotherapeutic drug doxorubicin (Dox) in PEG-PE micelle, we proved that PEG-PE micelle could serve as a co-carrier for both E5 and Dox (M-E5-Dox). M-E5 enhanced the efficiency of Dox by down-regulating the phosphorylation level of Akt, Erk and p38/MAPK proteins. In conclusion, PEG-PE micelle demonstrated a promising delivery system for E5, and M-E5 is expected to be a potential therapeutic agent that will help to improve the clinical benefits in current therapies used for solid tumors.

Description: by Wenhui Zeng, Riyuan Wang, Tianyang Zhang, Chunying Gong, Weidong Zuo, Rongpeng Liu, Yao Ou, Hanfu Xu The transcriptional coactivator Yorkie(Yki), is a critical downstream effector of the Hippo(Hpo) signaling pathway that controls organ size through the regulation of cell proliferation and apoptosis. During the past ten years the biological function of Yki has been studied extensively in Drosophila and a few other insects, however, little is known about it in the silkworm, Bombyx mori , a major research model of lepidopteran insect. Here, we describe the isolation, characterization and expression of the B . mori Yki ortholog, BmYki. The coding sequence of the BmYki was 1314 bp in length, encoding a protein of 437 amino acids containing two conserved WW domains. BmYki transcripts were ubiquitous but not abundant in all detected tissues and developmental stages. Comparatively, it was expressed at pretty high level in silk glands and at the stage of fifth-instar day-3 larvae. Overexpression of BmYki in cultured B . mori embryonic cells significantly promoted transcription of genes associated with cell proliferation and apoptosis, indicating that BmYki functions in the regulation of organ growth-related biological processes. Interestingly, transcription of silk protein-coding genes and transcription factors regulating the synthesis of silk proteins was downregulated remarkably, suggesting that BmYki was involved in the regulation of silk protein synthesis. This study provides new insights into the role of BmYki in Hpo pathway regulation in silkworm.

Description: by Hiroki Kawamoto, Aiko Hirata, Shigeyuki Kawano When Microbotryum lychnidis-dioicae infects a male Silene latifolia , M . lychnidis-dioicae smut spores develop in the pollen sac instead of pollen. In contrast, when M . lychnidis-dioicae infects a female S . latifolia , the female flowers become male-like, promoting stamen formation. However, it is unclear when and how M . lychnidis-dioicae invades the anther. It is important to investigate not only whether hyphae exist when the apical meristem tissue differentiates into flowers and anthers, but also whether hyphae exist when stamen filaments form. We used Grocott’s methenamine silver stain and lectin stain, which stain chitin in the fungal cell wall, to search for M . lychnidis-dioicae in flower tissues. A few M . lychnidis-dioicae hyphae were observed intercellularly in the center of the connective of vascular bundles at the early anther developmental stage. Subsequently, large numbers of deeply stained M . lychnidis-dioicae hyphae were observed intercellularly in the cells surrounding the pollen sac, as well as in the center of the pollen sac. Hyphae stained with lectin were observed intercellularly in all of the stamen filaments at flower development stages. Hyphae were observed in the peduncle connecting the flower and stem. It is thought that M . lychnidis-dioicae invaded the anther via the stamen filament over a long period. Additionally, in total, 163 sections of connective were obtained, and the cell structure of each anther was colored and subjected to three-dimensional reconstruction. The M . lychnidis-dioicae hyphae observed in the connective were mainly old hyphae with large vacuoles or dead hyphae (S1 Fig). These hyphae branched out, towards the pollen sac, while growing between the cells. We also observed that the host cells that collapsed near the hyphae had thick cell walls and teliospores. Cell wall collapse and cell degeneration were observed only around hyphae with thick cell walls.

Description: by Sanghoon Lee, Xia Jiang Background The dysregulation of microRNAs (miRNAs) alters expression level of pro-oncogenic or tumor suppressive mRNAs in breast cancer, and in the long run, causes multiple biological abnormalities. Identification of such interactions of miRNA-mRNA requires integrative analysis of miRNA-mRNA expression profile data. However, current approaches have limitations to consider the regulatory relationship between miRNAs and mRNAs and to implicate the relationship with phenotypic abnormality and cancer pathogenesis. Methodology/Findings We modeled causal relationships between genomic expression and clinical data using a Bayesian Network (BN), with the goal of discovering miRNA-mRNA interactions that are associated with cancer pathogenesis. The Multiple Beam Search (MBS) algorithm learned interactions from data and discovered that hsa-miR-21, hsa-miR-10b, hsa-miR-448, and hsa-miR-96 interact with oncogenes, such as, CCND2, ESR1, MET, NOTCH1, TGFBR2 and TGFB1 that promote tumor metastasis, invasion, and cell proliferation. We also calculated Bayesian network posterior probability (BNPP) for the models discovered by the MBS algorithm to validate true models with high likelihood. Conclusion/Significance The MBS algorithm successfully learned miRNA and mRNA expression profile data using a BN, and identified miRNA-mRNA interactions that probabilistically affect breast cancer pathogenesis. The MBS algorithm is a potentially useful tool for identifying interacting gene pairs implicated by the deregulation of expression.

Description: by Ivy Y. Choi, Olga N. Karpus, Jason D. Turner, Debbie Hardie, Jennifer L. Marshall, Maria J. H. de Hair, Karen I. Maijer, Paul P. Tak, Karim Raza, Jörg Hamann, Christopher D. Buckley, Danielle M. Gerlag, Andrew Filer Introduction Previous studies have shown increased expression of stromal markers in synovial tissue (ST) of patients with established rheumatoid arthritis (RA). Here, ST expression of stromal markers in early arthritis in relationship to diagnosis and prognostic outcome was studied. Methods ST from 56 patients included in two different early arthritis cohorts and 7 non-inflammatory controls was analysed using immunofluorescence to detect stromal markers CD55, CD248, fibroblast activation protein (FAP) and podoplanin. Diagnostic classification (gout, psoriatic arthritis, unclassified arthritis (UA), parvovirus associated arthritis, reactive arthritis and RA), disease outcome (resolving vs persistent) and clinical variables were determined at baseline and after follow-up, and related to the expression of stromal markers. Results We observed expression of all stromal markers in ST of early arthritis patients, independent of diagnosis or prognostic outcome. Synovial expression of FAP was significantly higher in patients developing early RA compared to other diagnostic groups and non-inflammatory controls. In RA FAP protein was expressed in both lining and sublining layers. Podoplanin expression was higher in all early inflammatory arthritis patients than controls, but did not differentiate diagnostic outcomes. Stromal marker expression was not associated with prognostic outcomes of disease persistence or resolution. There was no association with clinical or sonographic variables. Conclusions Stromal cell markers CD55, CD248, FAP and podoplanin are expressed in ST in the earliest stage of arthritis. Baseline expression of FAP is higher in early synovitis patients who fulfil classification criteria for RA over time. These results suggest that significant fibroblast activation occurs in RA in the early window of disease.

Description: by Dorien F. Bangma, Anselm B. M. Fuermaier, Lara Tucha, Oliver Tucha, Janneke Koerts Objectives Financial decision-making (FDM) is crucial for independent living. Due to cognitive decline that accompanies normal aging, older adults might have difficulties in some aspects of FDM. However, an improved knowledge, personal experience and affective decision-making, which are also related to normal aging, may lead to a stable or even improved age-related performance in some other aspects of FDM. Therefore, the present explorative study examines the effects of normal aging on multiple aspects of FDM. Methods One-hundred and eighty participants (range 18–87 years) were assessed with eight FDM tests and several standard neuropsychological tests. Age effects were evaluated using hierarchical multiple regression analyses. The validity of the prediction models was examined by internal validation (i.e. bootstrap resampling procedure) as well as external validation on another, independent, sample of participants (n = 124). Multiple regression and correlation analyses were applied to investigate the mediation effect of standard measures of cognition on the observed effects of age on FDM. Results On a relatively basic level of FDM (e.g., paying bills or using FDM styles) no significant effects of aging were found. However more complex FDM, such as making decisions in accordance with specific rules, becomes more difficult with advancing age. Furthermore, an older age was found to be related to a decreased sensitivity for impulsive buying. These results were confirmed by the internal and external validation analyses. Mediation effects of numeracy and planning were found to explain parts of the association between one aspect of FDM (i.e. Competence in decision rules) and age; however, these cognitive domains were not able to completely explain the relation between age and FDM. Conclusion Normal aging has a negative influence on a complex aspect of FDM, however, other aspects appear to be unaffected by normal aging or improve.

Description: by Ying-Chou Chen, Wei-Che Lin Background Osteoporotic vertebral fractures adversely impact quality of life and also increase the risk of infection and mortality. Alendronate treatment increases bone mass and reduces the risk of fractures in patients with osteoporosis by suppressing bone resorption. We investigated the relationship between alendronate treatment and infection-related death in patients with osteoporotic vertebral fractures. Methods We retrospectively reviewed patients with osteoporosis and vertebral fractures from January 2001 to December 2007. The use of alendronate, glucocorticoid and medical factors including smoking, alcohol consumption, diabetes, hypertension, stroke, liver disease, heart disease, and pulmonary disease were analyzed. Cox regression was used to analyze the factors associated with life-threatening infections. Results A total of 210 patients (161 females and 49 males) were included with a mean age of 74.06±7.43 years. Among them, 87 had life-threatening infections and 123 did not. In Cox regression analysis, the patients who used alendronate had a significantly lower risk of life-threatening infections ( p = 0.006, HR = 0.845, 95% CI 0.750–0.954), while glucocorticoid users had higher risk of death ( p = 0.010, HR = 2.037, 95% CI 1.187–3.498). Conclusions Osteoporosis was associated with a high rate of life-threatening infections, and the use of alendronate had a lower rate of infection-related death. Therefore, we suggest that alendronate be used after vertebral fractures in these patients.

Description: by Robert Czajkowski, Anna Smolarska, Zofia Ozymko Dickeya solani is one of the most important pectinolytic phytopathogens responsible for high losses in potato, especially in seed potato production in Europe. Lytic bacteriophages can affect the structure of the host population and may influence spread, survival and virulence of the pathogen and in consequence, infection of the plant. In this study, we aimed to acquire information on the viability of the broad host lytic bacteriophage ΦD5 on potato, as well as to apprehend the specific effect of this bacteriophage on its host D . solani type-strain in different settings, as a preliminary step to target co-adaptation of phages and host bacteria in plant environment. Viability of the ΦD5 phage in tuber extract, on tuber surface, in potting compost, in rainwater and on the leaf surface, as well as the effect of copper sulfate, were examined under laboratory conditions. Also, the interaction of ΦD5 with the target host D . solani in vitro and in compost-grown potato plants was evaluated. ΦD5 remained infectious in potato tuber extract and rain water for up to 72 h but was inactivated in solutions containing 50 mM of copper. The phage population was stable for up to 28 days on potato tuber surface and in potting compost. In both, tissue culture and compost-grown potato plants, ΦD5 reduced infection by D . solani by more than 50%. The implications of these findings are discussed.

Description: by Maha Sellami, Wissem Dhahbi, Lawrence D. Hayes, Johnny Padulo, Fatma Rhibi, Hanen Djemail, Anis Chaouachi The aim of this investigation was to compare serum growth hormone (GH), insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-binding protein-3 (IGFBP-3) in response to a combined sprint and resistance training (CSRT) program in young and middle-aged men.Thirty-eight healthy, moderately trained men participated in this study. Young and middle-aged men were randomly assigned to, a young training group (YT = 10, 21.4±1.2yrs) ora young control group (YC = 9, 21.6±1.8 yrs), a middle-aged training group (MAT = 10, 40.4±2.1 yrs) or a middle-aged control group (MAC = 9, 40.5±1.8 yrs). Participants performed the Wingate Anaerobic Test (WAnT) before and after a 13-week CSRT program (three sessions per week). Blood samples were collected at rest, after warm-up, immediately post-WAnT, and 10 min post-WAnT. CSRT induced increases in GH at rest and in response to the WAnT in YT and MAT (P

Description: by Ronald A. Bos, Dick H. Plettenburg The addition of a cosmetic glove to an upper limb prosthesis has a distinct effect on the cosmetic value, but its viscoelastic behaviour adds a substantial amount of stiffness and hysteresis to the system. As a result, the overall usability of the prosthesis is degraded. A novel negative stiffness element is designed to compensate for the cosmetic glove's stiffness. A combination of linear helical springs and the concept of rolling link mechanisms has resulted in a Rolling Stiffness Compensation Mechanism (RSCM). Results show that the RSCM is capable of exerting a progressive negative stiffness characteristic and can be built small enough to fit inside a 33 mm diameter wrist. Using the RSCM, an otherwise voluntary opening toddler-sized prosthesis is converted into a voluntary closing device, reducing maximum operation forces down to 40 N with a combined efficiency of 52%. Further adjustments to the design are possible to further improve the efficiency of the mechanism. Moreover, changes in geometric relations of the mechanism offers possibilities for a wide range of prostheses and other applications.

Description: by Kohei Ikezoe, Tomohiro Handa, Kiminobu Tanizawa, Toyofumi F. Chen-Yoshikawa, Takeshi Kubo, Akihiro Aoyama, Hideki Motoyama, Kyoko Hijiya, Shinsaku Tokuda, Yoshinari Nakatsuka, Yuko Yamamoto, Ayako Oshima, Shin-ichi Harashima, Sonoko Nagai, Toyohiro Hirai, Hiroshi Date, Kazuo Chin Objective Young patients with advanced interstitial lung disease (ILD) are potential candidates for cadaveric lung transplantation. This study aimed to examine clinical features, outcomes, and prognostic factors in Japanese ILD patients awaiting lung transplantation. Methods We investigated the clinical features and outcomes of 77 consecutive candidates with ILD who were referred to Kyoto University Hospital and subsequently actively listed for lung transplant in the Japan Organ Transplant Network between 2010 and 2014. Results Of the 77 candidates, 33 had idiopathic pulmonary fibrosis (IPF) and 15 had unclassifiable ILD. During the observational period, 23 patients (30%) received lung transplantations and 49 patients (64%) died before transplantation. Of the 33 patients with IPF, 13 (39%) had a family history of ILD and 13 (39%) had an “inconsistent with usual interstitial pneumonia pattern” on high-resolution computed tomography (HRCT). The median survival time from registration was 16.7 months, and mortality was similar among patients with IPF, unclassifiable ILD, and other ILDs. Using a multivariate stepwise Cox proportional hazards model, 6-min walking distance was shown to be an independent prognostic factor in candidates with ILD (per 10 m, hazard ratio (HR): 0.97; 95% confidence interval (CI): 0.95–0.99, p

Description: by Haider Mussawy, Lennart Viezens, Gerrit Hauenherm, Malte Schroeder, Christian Schaefer Organ-specific microcirculation plays a central role in tumor growth, tumor cell homing, tissue engineering, and wound healing. Mouse models are widely used to study these processes; however, these mouse strains often possess unique microhemodynamic parameters, making it difficult to directly compare experiments. The full functional characterization of bone and striated muscle microcirculatory parameters in non-obese diabetic-severe combined immunodeficiency/y-chain; NOD-Prkds IL2rg (NSG) mice has not yet been reported. Here, we established either a dorsal skinfold chamber or femur window in NSG mice (n = 23), allowing direct analysis of microcirculatory parameters in vivo by intravital fluorescence microscopy at 7, 14, 21, and 28 days after chamber preparation. Organ-specific differences were observed. Bone had a significantly lower vessel density but a higher vessel diameter than striated muscle. Bone also showed higher effective vascular permeability than striated muscle. The centerline velocity values were similar in the femur window and dorsal skinfold chamber, with a higher volumetric blood flow in bone. Interestingly, bone and striated muscle showed similar tissue perfusion rates. Knowledge of physiological microhemodynamic values of bone and striated muscle in NSG mice makes it possible to analyze pathophysiological processes at these anatomic sites, such as tumor growth, tumor metastasis, and tumor microcirculation, as well as the response to therapeutic agents.

Description: by Romana J. R. Yanez, Renate Lamprecht, Milaid Granadillo, Brandon Weber, Isis Torrens, Edward P. Rybicki, Inga I. Hitzeroth High-risk human papillomaviruses (hr-HPVs) cause cervical cancer, the fourth most common cancer in women worldwide. A HPV-16 candidate therapeutic vaccine, LALF 32-51 -E7, was developed by fusing a modified E7 protein to a bacterial cell-penetrating peptide (LALF): this elicited both tumour protection and regression in pre-clinical immunization studies. In the current study, we investigated the potential for producing LALF 32-51 -E7 in a plant expression system by evaluating the effect of subcellular localization and usage of different expression vectors and gene silencing suppressors. The highest expression levels of LALF 32-51 -E7 were obtained by using a self-replicating plant expression vector and chloroplast targeting, which increased its accumulation by 27-fold compared to cytoplasmic localization. The production and extraction of LALF 32-51 -E7 was scaled-up and purification optimized by affinity chromatography. If further developed, this platform could potentially allow for the production of a more affordable therapeutic vaccine for HPV-16. This would be extremely relevant in the context of developing countries, where cervical cancer and other HPV-related malignancies are most prevalent, and where the population have limited or no access to preventative vaccines due to their typical high costs.

Description: by Paulina Nowicka-Krawczyk, Joanna Żelazna-Wieczorek, Tomasz Koźlecki Aerial algae are an important biological factor causing the biodegradation of building materials and facades. Conservation procedures aimed at the protection of historic and utility materials must be properly designed to avoid an increase of the degradation rate. The aim of the present study was to investigate the effect of silver nanoparticles (AgNP) synthetized with features contributing to the accessibility and toxicity (spherical shape, small size) on the most frequently occurring species of green algae in aerial biofilms and thus, the most common biodegradation factor– Apatococcus lobatus . Changes in the chloroplasts structure and the photosynthetic activity of the cells under AgNP exposure were made using confocal laser microscopy and digital image analysis and the estimation of growth inhibition rate was made using a biomass assay. In the majority of cases, treatment with AgNP caused a time and dose dependant degradation of chloroplasts and decrease in the photosynthetic activity of cells leading to the inhibition of aerial algae growth. However, some cases revealed an adaptive response of the cells. The response was induced by either a too low, or—after a short time—too high concentration of AgNP. Taken together, the data suggest that AgNP may be used as a biocide against aerial algal coatings; however, with a proper caution related to the concentration of the nanoparticles.

Description: by Atsuhiro Yamane, Hiroki Kohno, Tsubomi Ikeda, Kumi Kaneko, Atsushi Ugajin, Toshiyuki Fujita, Takekazu Kunieda, Takeo Kubo

Description: by Ulrike Flierl, Jörn Tongers, Dominik Berliner, Jan-Thorben Sieweke, Florian Zauner, Christoph Wingert, Christian Riehle, Johann Bauersachs, Andreas Schäfer Early use of mechanical circulatory support, e.g. veno-arterial extracorporeal membrane oxygenation (ECMO) or left ventricular unloading by microaxial pump in refractory cardiogenic shock is recommended in current guidelines. Development of acquired von Willebrand Syndrome (AVWS) in patients with left ventricular assist devices (LVADs) and ECMO has been reported. There is an increasing number of patients treated with the Impella ® CP microaxial pump for left ventricular unloading. However, the prevalence of AVWS in these high risk patients is unknown and needs to be determined. We therefore screened 21 patients (68 ± 11years) treated with Impella ® (17 for cardiogenic shock, 4 for protected PCI) for the presence of AVWS by determining von Willebrand factor multimers, VWF collagen binding capacity and VWF antigen. During the time course of Impella ® support, 20/21 patients (95%) developed AVWS (mean duration of support: 135 ± 114 hours, mean time from device implantation to first diagnosis of AVWS: 10.6 ± 10.8 hours). Our data indicate that AVWS is a common phenomenon during left ventricular unloading via microaxial pump support. Thus, AVWS has to be considered as contributing factor for potential bleeding complications in this high risk patient population, especially in the context of dual antiplatelet therapy.

Description: by Bing Mao, Hong Yin, Yan Wang, Tian-Hong Zhao, Rong-Rong Tian, Wei Wang, Jia-Shu Ye Enhanced ultraviolet radiation (UV) and elevated tropospheric ozone (O 3 ) may individually cause reductions in the growth and productivity of important agricultural crops. However, research regarding their combined effects on important agricultural crops is still scarce, especially on changes in secondary metabolites and endogenous hormones, which are important protective substances and signal components that control plant responses to environment stresses. In this study, using an experimental setup of open top chambers, we monitored the responses of seed yield per plant, leaf secondary metabolites and leaf endogenous hormones under the stress of elevated O 3 and enhanced UV radiation individually, as well as their combined stress. The results indicated that elevated O 3 (110 ± 10 nmol mol -1 for 8 hours per day) and enhanced UV radiation (1.73 kJ h -1 m -2 ) significantly decreased seed yield per plant. Concentrations of rutin, queretin and total flavonoids were significantly increased under the elevated O 3 treatment or the enhanced UV radiation treatment or the combination treatment at flowering and podding stages, and concentrations of rutin, queretin and total flavonoids showed significant correlations with seed yield per plant. Concentrations of ABA and IAA decreased under the three treatments. There was a significant positive correlation between the ABA concentration and seed yield and a negative correlation between the IAA concentration and seed yield. We concluded that the combined stress of elevated O 3 and UV radiation significantly decreased seed yield per plant. Yield reduction was associated with changes in the concentrations of flavonoids, ABA and IAA in soybean leaves. The effects of the combined O 3 and UV stress were always greater than those of the individual stresses alone.

Description: by Shun Lu, Shuya Liu, Astrid Wietelmann, Baktybek Kojonazarov, Ann Atzberger, Cong Tang, Ralph Theo Schermuly, Hermann-Josef Gröne, Stefan Offermanns GPR116 (ADGRF5) and ELTD1 (ADGRL4) belong to different subfamilies of the adhesion G-protein-coupled receptor group but are both expressed in endothelial cells. We therefore analyzed their functions in mice lacking these receptors. While loss of GPR116 or ELTD1 alone had no obvious effect on cardiovascular or kidney function, mice lacking both, GPR116 and ELTD1, showed malformations of the aortic arch arteries and the cardiac outflow tract leading to perinatal lethality in about 50% of the mutants. In addition to cardiovascular malformations, surviving mice developed renal thrombotic microangiopathy as well as hemolysis and splenomegaly, and their lifespan was significantly reduced. Loss of GPR116 and ELTD1 specifically in endothelial cells or neural crest-derived cells did not recapitulate any of the phenotypes observed in GPR116-ELTD1 double deficient mice, indicating that loss of GPR116 and ELTD1 expressed by other cells accounts for the observed cardiovascular and renal defects.

Description: by Na Wang, Hong Ma, Zhe Li, Yalei Gao, Xuezhao Cao, Yanhua Jiang, Yongjian Zhou, Sidan Liu Methods Sprague-Dawley adult male rats (12–14 weeks old) were exposed to 14-day CUS and then subjected to partial hepatectomy 24 h after the last stress session. The rats were pretreated with an antagonist of the glucocorticoids (GCs) receptor RU486 (30 mg/kg, i.p.) 1 h prior to stress exposure. The behavioral changes were evaluated with open field test and elevated plus-maze test. The hippocampal cytokines interleukin (IL)-1β and IL-6 were measured on postoperative days 1, 3 and 7. Ionized calcium binding adaptor protein (Iba)-1, microglial M2 phenotype marker Arg1, brain derived neurotrophic factor (BDNF) and CD200 were also examined at each time point. Results CUS exacerbated surgery-induced sickness behavior. Exposure to CUS alone failed to alter the levels of pro-inflammatory cytokines in the brain. However, CUS exaggerated surgery-induced pro-inflammatory cytokines expression (e.g. IL-1β and IL-6) and upregulated the levels of Iba-1 on postoperative days 1 and 3. An additional significant decreased BDNF, CD200 and a lower level of Arg1 were also observed in the stressed rats following surgical procedure. Pretreatment with RU486 blunted the potentiating effects of CUS on surgery-induced sickness behavior and neuroinflammatory responses. Conclusion Chronic unpredictable stress enhanced surgery-induced sickness behavior and neuroinflammatory responses. Stress-induced GCs played a pivotal role in enhancing surgery-induced neuroinflammatory processes by modulation of microglia functions.

Description: by Marie-Thérèse Linossier, Liubov E. Amirova, Mireille Thomas, Myriam Normand, Marie-Pierre Bareille, Guillemette Gauquelin-Koch, Arnaud Beck, Marie-Claude Costes-Salon, Christine Bonneau, Claude Gharib, Marc-Antoine Custaud, Laurence Vico Background Dry immersion (DI), a ground-based model of microgravity previously used in Russia, has been recently implemented in France. The aim of this study was to analyze early events in a short-term DI model in which all conditions are met to investigate who is first challenged from osteo- or adipo-kines and to what extent they are associated to insulin-regulating hormones. Methods Twelve healthy men were submitted to a 3-day DI. Fasting blood was collected during pre-immersion phase for the determination of the baseline data collection (BDC), daily during DI (DI 24h , DI 48H and DI 72h ), then after recovery (R +3h and R +24h ). Markers of bone turnover, phosphocalcic metabolism, adipokines and associated factors were measured. Results Bone resorption as assessed by tartrate-resistant acid phosphatase isoform 5b and N-terminal crosslinked telopeptide of type I collagen levels increased as early as DI 24h . At the same time, total procollagen type I N- and C-terminal propeptides and osteoprotegerin, representing bone formation markers, decreased. Total osteocalcin [OC] was unaffected, but its undercarboxylated form [Glu-OC] increased from DI 24h to R +3h . The early and progressive increase in bone alkaline phosphatase activities suggested an increased mineralization. Dickkopf-1 and sclerostin, as negative regulators of the Wnt-β catenin pathway, were unaltered. No change was observed either in phosphocalcic homeostasis (calcium and phosphate serum levels, 25-hydroxyvitamin D, fibroblast growth factor 23 [FGF23]) or in inflammatory response. Adiponectemia was unchanged, whereas circulating leptin concentrations increased. Neutrophil gelatinase-associated lipocalin [lipocalin-2], a potential regulator of bone homeostasis, was found elevated by 16% at R +3h compared to DI 24h . The secretory form of nicotinamide phosphoribosyl-transferase [visfatin] concentrations almost doubled after one day of DI and remained elevated. Serum insulin-like growth factor 1 levels progressively increased. Fasting insulin concentrations increased during the entire DI, whereas fasting glucose levels tended to be higher only at DI 24h and then returned to BDC values. Changes in bone resorption parameters negatively correlated with changes in bone formation parameters. Percent changes of ultra-sensitive C-reactive protein positively correlated with changes in osteopontin, lipocalin-2 and fasting glucose. Furthermore, a positive correlation was found between changes in FGF23 and Glu-OC, the two main osteoblast-/osteocyte-derived hormones. Conclusion Our results demonstrated that DI induced an unbalanced remodeling activity and the onset of insulin resistance. This metabolic adaptation was concomitant with higher levels of Glu-OC. This finding confirms the role of bone as an endocrine organ in humans. Furthermore, visfatin for which a great responsiveness was observed could represent an early and sensitive marker of unloading in humans.

Description: by Alex Harwig, Zita Kruize, Zhenhuang Yang, Tobias Restle, Ben Berkhout The RNA interference (RNAi) pathway was recently expanded by the discovery of multiple alternative pathways for processing of natural microRNA (miRNA) and man-made short hairpin RNA (shRNA) molecules. One non-canonical pathway bypasses Dicer cleavage and requires instead processing by Argonaute2 (Ago2), which also executes the subsequent silencing step. We named these molecules AgoshRNA, which generate only a single active RNA strand and thus avoid off-target effects that can be induced by the passenger strand of a regular shRNA. Previously, we characterized AgoshRNA processing by deep sequencing and demonstrated that—after Ago2 cleavage—AgoshRNAs acquire a short 3’ tail of 1–3 A-nucleotides and are subsequently trimmed, likely by the poly(A)-specific ribonuclease (PARN). As a result, the mature single-stranded AgoshRNA may dock more stably into Ago2. Here we set out to analyze the activity of different synthetic AgoshRNA processing intermediates. Ago2 was found to bind preferentially to partially single-stranded AgoshRNA in vitro . In contrast, only the double-stranded AgoshRNA precursor associated with Ago2 in cells, correlating with efficient intracellular processing and reporter knockdown activity. These results suggest the presence of a cellular co-factor involved in AgoshRNA loading into Ago2 in vivo . We also demonstrate specific AgoshRNA loading in Ago2, but not Ago1/3/4, thus further reducing unwanted side effects.

Description: by Rodolphe Carpentier, Anne Platel, Helena Maiz-Gregores, Fabrice Nesslany, Didier Betbeder Atmospheric pollution is mainly composed of volatile pollutants and particulate matter that strongly interact. However, their specific roles in the induction of cellular toxicity, in particular the impact of the vectorization of atmospheric pollutants by ultrafine particles, remains to be fully elucidated. For this purpose, non-toxic poly-lactic co-glycolic acid (PLGA) nanoparticles were synthesized and three pollutants (benzo(a)pyrene, naphthalene and di-ethyl-hexyl-phthalate) were adsorbed on the surface of the nanoparticles in order to evaluate the toxicity (cytotoxicity, genotoxicity and ROS induction) of these complexes to a human airway epithelial cell line. The adsorption of the pollutants onto the nanoparticles was confirmed by HPLC analysis. Interestingly, the cytotoxicity assays (MTT, LDH and CellTox Green) clearly demonstrated that the vectorization by nanoparticles decreases the toxicity of the adsorbed pollutants. Genotoxicity was assessed by the micronucleus test and the comet assay and showed no increase in primary DNA damage or in chromosomal aberrations of nanoparticle vectorized pollutants. Neither cytotoxicity nor genotoxicity was correlated with ROS induction. To conclude, our results indicate that the vectorization of pollutants by nanoparticles does not potentiate the toxicity of the pollutants studied and that, on the contrary, adsorption onto nanoparticles could protect cells against pollutants’ toxicity.

Description: by Cheng-Ping Shih, Hung-Che Lin, Chi-Hsiang Chung, Po-Jen Hsiao, Chih-Hung Wang, Jih-Chin Lee, Wu-Chien Chien Tinnitus mostly results from central and peripheral auditory pathology. Chronic kidney disease (CKD) is a major risk factor for cerebrovascular disease. However, no studies have evaluated the association between tinnitus and CKD. The aim of this study is to investigate the risk of tinnitus in patients with CKD. This retrospective cohort study was conducted using Taiwan National Health Insurance Research Database from 2000 to 2010. We established a CKD group (n = 185,430) and a non-CKD comparison group (n = 556,290) to investigate the incidence of tinnitus. Cox proportional hazard regression analysis was used to evaluate the effects of CKD on tinnitus risk. The results showed CKD significantly increased the risk of tinnitus (adjusted hazard ratio, 3.02; 95% CI, 2.655–3.456, P

Description: by Arunrat Tangmunkongvorakul, Patou Masika Musumari, Kriengkrai Srithanaviboonchai, Veruree Manoyos, Teeranee Techasrivichien, S. Pilar Suguimoto, Masako Ono-Kihara, Masahiro Kihara, Suwat Chariyalertsak Background Many young migrant workers move across the border to Chiang Mai, a major city in Northern Thailand, in search of work opportunities. This study describes their sexual behavior, lifestyles, relationships and experiences with youth-friendly sexual and reproductive health (SRH) services. Methods This is the qualitative arm of a mixed methods study using focus group discussions (FGDs) among young MWs aged 15–24 years in urban Chiang Mai. We conducted 6 FGDs with 84 participants (43 males, 41 females) organized in groups of 10–15 people, including 3 groups of males, 2 groups of females, and 1 group of both males and females. Results We found that the lack of parental control, pressure to assimilate into Thai society, access to social media and modern communication technologies, and limited knowledge and access to sexual and reproductive health (SRH) services interplayed to shape lifestyle and sexual behaviors, including low condom use among young migrants. Conclusion The present study helped discern the vulnerability of young migrants to adverse SRH outcomes. This particular group of youth needs urgent intervention to improve their knowledge on SRH and access to a youth-friendly clinic to help them personalize risk of HIV and other adverse SRH outcomes.

Description: by Nobuyuki Okuma, Makiko Saita, Noriyuki Hoshi, Tomoyoshi Soga, Masaru Tomita, Masahiro Sugimoto, Katsuhiko Kimoto Background This study characterized the changes in quality and quantity of saliva, and changes in the salivary metabolomic profile, to understand the effects of masticatory stimulation. Methods Stimulated and unstimulated saliva samples were collected from 55 subjects and salivary hydrophilic metabolites were comprehensively quantified using capillary electrophoresis-time-of-flight mass spectrometry. Results In total, 137 metabolites were identified and quantified. The concentrations of 44 metabolites in stimulated saliva were significantly higher than those in unstimulated saliva. Pathway analysis identified the upregulation of the urea cycle and synthesis and degradation pathways of glycine, serine, cysteine and threonine in stimulated saliva. A principal component analysis revealed that the effect of masticatory stimulation on salivary metabolomic profiles was less dependent on sample population sex, age, and smoking. The concentrations of only 1 metabolite in unstimulated saliva, and of 3 metabolites stimulated saliva, showed significant correlation with salivary secretion volume, indicating that the salivary metabolomic profile and salivary secretion volume were independent factors. Conclusions Masticatory stimulation affected not only salivary secretion volume, but also metabolite concentration patterns. A low correlation between the secretion volume and these patterns supports the conclusion that the salivary metabolomic profile may be a new indicator to characterize masticatory stimulation.