ALBERT

Notes: 1 We have investigated the actions of the α1D-adrenoceptor selective antagonist BMY 7378 in comparison with yohimbine at α1- and α2-adrenoceptors. 2 In rat aorta (α1D-adrenoceptor), BMY 7378 (pA2 of 8.67) was about 100 times more potent than yohimbine (pA2 of 6.62) at antagonizing the contractile response to noradrenaline. 3 In human saphenous vein (α2C-adrenoceptor), BMY 7378 (pA2 of 6.48) was approximately 10 times less potent than yohimbine (pA2 of 7.56) at antagonizing the contractile response to noradrenaline. 4 In prostatic portions of rat vas deferens, BMY 7378 (10 μm) did not significantly affect the concentration-dependent inhibition of single pulse nerve stimulation-evoked contractions by xylazine (an action at prejunctional α2D-adrenoceptors). 5 In ligand-binding studies, BMY 7378 showed 10-fold selectivity for α2C-adrenoceptors (pKi of 6.54) over other α2-adrenoceptors. 6 It is concluded that BMY 7378, in addition to α1D-adrenoceptor selectivity in terms of α1-adrenoceptors, shows selectivity for α2C-adrenoceptors in terms of α2-adrenoceptors.

Notes: 1 To investigate anti-inflammatory activity of organic germanium, we measured the effect of germanium-concentrated yeast on arachidonic acid release, prostaglandin E2 (PGE2) production, histamine release, and intracellular H2O2 or hydroperoxide generation in RBL 2H3 cells, and carrageenan-induced paw oedema in rats. 2 Germanium-concentrated yeast dose-dependently inhibited carrageenan-induced paw oedema, suggesting that germanium-concentrated yeast has anti-inflammatory activity in acute inflammation. 3 Germanium-concentrated yeast significantly inhibited melittin-induced arachidonic acid release and PGE2 production in RBL 2H3 cells. 4 Germanium-concentrated yeast did not affect melittin-induced histamine release and silica-induced intracellular H2O2 or hydroperoxide generation in RBL 2H3 cells. 5 These results suggest that anti-inflammatory activity of germanium-concentrated yeast appears partly to be related to the inhibition of arachidonic acid release and PGE2 production in RBL 2H3 cells.

Notes: 1 The pressor action of noradrenaline and its blockade by selective α1-adrenoceptor antagonists in the pithed mouse were evaluated. 2 Chloroethylclonidine (α1B/D-adrenoceptor alkylating agent) or BMY 7378 (α1D-adrenoceptor antagonist), both at 1 mg kg−1, did not block the increase in blood pressure induced by noradrenaline. 3 5-Methylurapidil (α1A-adrenoceptor antagonist), at 0.1 mg kg−1, displaced the dose–response curve approximately six-fold to the right. 4 The results support the idea that the pithed mouse vasculature express α1A-adrenoceptors and suggest that it is a good model to study the roles of α1-adrenoceptors in gene knockout or overexpression.

Notes: 1 The aim of this paper was to determine the different signalling cascades involved in contraction of the rat urinary bladder detrusor muscle mediated via muscarinic acetylcholine receptors (muscarinic AChR). Contractile responses, phosphoinositides (IPs) accumulation, nitric oxide synthase (NOS) activity and cyclic GMP (cGMP) production were measured to determine the reactions associated with the effect of cholinergic agonist carbachol. The specific muscarinic AChR subtype antagonists and different inhibitors of the enzymatic pathways involved in muscarinic receptor-dependent activation of NOS and cGMP were tested. 2 Carbachol stimulation of M3 and M4 muscarinic AChR increased contractility, IPs accumulation, NOS activity and cGMP production. All of these effects were selectively blunted by 4-DAMP and tropicamide, M3 and M4 antagonists respectively. 3 The inhibitors of phospholipase C (PLC), calcium/calmodulin (CaM), neuronal NOS (nNOS) and soluble guanylate cyclase, but not of protein kinase C and endothelial NOS (eNOS), inhibited the carbachol action on detrusor contractility. These inhibitors also attenuated the muscarinic receptor-dependent increase in cGMP and activation of NOS. 4 In addition, sodium nitroprusside and 8-bromo-cGMP, induced negative relaxant effect. 5 The results obtained suggest that carbachol activation of M3 and M4 muscarinic AChRs, exerts a contractile effect on rat detrusor that is accompanied by an increased production of cGMP and nNOS activity. The mechanism appears to occur secondarily to stimulation of IPs turnover via PLC activation. This in turn, triggers cascade reactions involving CaM, leading to activation of nNOS and soluble guanylate cyclase. They, in turn, exert a modulator inhibitory cGMP-mediated mechanism limiting the effect of muscarinic AChR stimulation of the bladder.

Notes: 1 The objective was to investigate a possible contribution of a nerve-derived hyperpolarizing factor to the differences between non-adrenergic non-cholinergic (NANC) nerve-mediated relaxations in different states of active tone in the rat gastric fundus. 2 NANC relaxations induced by electrical field stimulation (ES: 0.1, 0.5 and 1 Hz; 25 V; 1 ms; 10 s) in 40% contracted strips (S40) were greater when compared with those in 80% contracted strips (S80). 3 ES-induced relaxations were effectively attenuated by Nω-nitro-l-arginine (l-NNA; 100 μm) in S40 and S80. Percentage reduction of the responses obtained in the presence of l-NNA in S40 group was less than that of S80. 4 In S40 group, nifedipine (0.5–1 μm) and verapamil (0.5–1 μm) inhibited the responses to 0.1 and 0.5 Hz. Nifedipine (1 μm) and verapamil (0.5 μm) caused no change in the responses to ES in S80. 5 In S40, when l-NNA (100 μm) and nifedipine or verapamil, either in 1 μm concentration, were administered together, the inhibition on the electrical relaxations were more than that of each drug alone. 6 In conclusion, NANC nerve-mediated relaxations are increased when studied in an active state of 40%, and a factor, sensitive to nifedipine seems to be responsible for this distinction.

Notes: 1 Experiments were carried out to characterize the possible adrenergic properties of the 5-HT1A antagonists WAY 100635 and MM-77 using the mouse isolated vasa deferentia preparation. 2 When vasa deferentia were preincubated for 10 min in the presence of MM-77 (10−8–10−6 m) or WAY100635 (10−8–7 × 10−7 m), a concentration-dependent inhibition of the contractile response to submaximal electrical field stimulation (10 Hz, 50 V, 50 ms) was observed with pIC50 values of 7.05 ± 0.01 and 6.85 ± 0.1 respectively. 3 MM-77 (10−8–10−6 m) antagonized the contractile responses of the vasa deferentia to phenylephrine (PE) (10−6–10−3 m) in a concentration-dependent manner. Schild plots of these data were linear and yielded a mean ρA2 value of 6.81 ± 0.084. The mean slope was 1.42 ± 0.22. 4 WAY100635 (10−8–10−6 m) antagonized the contractile responses of the vasa deferentia to PE (10−6–10−3 m) in a concentration-dependent manner. Schild plots of these data were linear and yielded a mean ρA2 value of 7.05 ± 0.08. The mean slope was 0.97 ± 0.1. 5 The results suggest that while WAY100635 acts as a competitive antagonist at α1-adrenoceptors, MM-77 displays non-competitive antagonist characteristics at this receptor subtype. 6 These results may have important implications for the use of these compounds as 5-HT1A receptor antagonists in in vivo studies.

Notes: 1 The aim of the present study was to examine the modulator influence of muscarinic M2 receptors on responses of rat urinary bladder detrusor muscle evoked by endogenous stimuli, i.e. by stimulation of the bladder innervation. 2 Responses were evoked by electrical field stimulation (EFS; 2–20 Hz, 0.8 ms, 60 V) of isolated strip preparations mounted in organ baths. The tension of the muscle strips was recorded digitally. EFS was performed by applying stimulation with either a short duration (5 s) or a longer duration (to reach peak response; approximately 20 s). 3 Effects of muscarinic receptor antagonists (muscarinic M1/M3 receptor selective: 4-diphenylacetoxy-N-methylpiperidine methobromide (4-DAMP); muscarinic M2 receptor selective: methoctramine), a β-adrenergic antagonist (propranolol) and an adenosine receptor antagonist (8-p-sulfophenyltheophylline) were assessed on contractile activity and on poststimulatory relaxations. 4 Low concentrations of methoctramine (10−8 m) reduced or tended to reduce the EFS-induced contraction, e.g. at 2 Hz by 12% while methoctramine at 10−7 m had no significant effect. In addition, in the presence of 4-DAMP (10−9 m), which tended to inhibit contractions at all frequencies (2–20 Hz; −17 to −25%), methoctramine at 10−8 and 10−7 m induced a further reduction of the contractile responses (−5 to −10%; 2–20 Hz). 5 The β-adrenergic receptor antagonist propranolol (10−6 m) and the adenosine receptor antagonist 8-p-sulfophenyltheophylline (10−6 m) both increased contractile responses by 9–21% (2–10 Hz, long duration; P 〈 0.05–0.001) as a consequence of antagonizing relaxatory stimuli. Neither antagonist affected the contractile responses to EFS with the short duration stimulation. Poststimulatory relaxations were reduced by 30–60% (P 〈 0.05) by propranolol and by 40–60% (P 〈 0.001) by 8-p-sulfophenyltheophylline, but for 8-p-sulfophenyltheophylline only after stimulation with the short duration. 6 In the presence of methoctramine (10−7 m), the 8-p-sulfophenyltheophylline-induced increases of the contractile response to long duration EFS were significantly enhanced at 10 Hz (+12 ± 4%; P 〈 0.05), whereas no such enhancement of the propranolol inhibitory effect occurred in the presence of methoctramine. However, poststimulatory β-adrenoceptor-evoked relaxations after short duration EFS were increased by about 35% in the presence of methoctramine, but not those after long duration. 7 Thus, muscarinic M2 receptor activation inhibits adenosine receptor- and β-adrenoceptor-evoked relaxations of the rat detrusor muscle. The inhibition occurs via a transient postjunctional mechanism that mainly affects responses with a short latency.

Notes: 1 To clarify the effects of doxapram on the baroreflex, we recorded carotid sinus nerve (CSN) activity in isolated and perfused carotid artery bifurcations of rabbits. 2 The CSN activity due to chemoreceptor stimulation was blocked by resection of the nerve branches from the carotid body. After the resection, the CSN activity was correlated to increase of carotid sinus (CS) pressure. 3 Administration of doxapram reduced the CSN activity originating from baroreceptors. The effect of doxapram on baroreceptors was dose dependent and reversible. 4 It is unlikely that doxapram altered CS wall mechanics because CS pressure did not change in the presence of the drug. 5 We conclude that doxapram acts on the cardiovascular system in part by inhibiting the negative feedback loop that originates in CS baroreceptors.

Notes: 1 Cardiac glycosides have been used for centuries as therapeutic agents for the treatment of heart diaseases. In patients with heart failure, digoxin and the other glycosides exert their positive inotropic effect by inhibiting Na+–K+-ATPase, thereby increasing intracellular sodium, which, in turn, inhibits the Na+/Ca2+ exchanger and increases intracellular calcium levels. As the therapeutic index of digitalis is narrow, arrhythmias are common problems in clinical practice. The mechanisms and mediators of these arrhythmias, however, are not completely understood. 2 The involvement of the sympathetic and parasympathetic nervous system in digitalis cardiac toxicity is reviewed. 3 Receptors, channels, exchange systems or other cellular components involved in digitalis-induced cardiotoxicity are also reviewed. 4 Possible mediators of digitalis-induced cardiac toxicity are discussed. 5 Management of digitalis toxicity in patients is summarized. 6 The determination of the possible mediators of digitalis-induced cardiac toxicity will enhance our knowledge and lead to the development of new therapeutic strategies to treat these lethal arrhythmias.

Notes: 1 Microinjection of peptide YY (PYY) (0.23–2.3 nmol) into the posterior hypothalamic nucleus (PHN) of conscious rats evokes a dose-dependent pressor response and a bradycardia. 2 The increase in mean arterial pressure evoked by 2.3 nmol of PYY was not blocked by intravenous pretreatment with: (i) the nicotinic ganglionic receptor antagonist pentolinium (PENT, 10 mg kg−1) alone, or in combination with the muscarinic receptor antagonist methylatropine (MeATR, 1 mg kg−1); (ii) the α1-adrenoceptor antagonist prazosin (PRAZ, 0.2 mg kg−1); (iii) the V1-vasopressin receptor antagonist [d(CH2)5Tyr(Me)]AVP (AVPX, 20 μg kg−1); (iv) the combination of AVPX, PENT and MeATR; (v) the combination of PRAZ, AVPX, PENT, MeATR, and the α2-adrenoceptor antagonist yohimbine (0.3 mg kg−1); or (vi) the angiotensin II type 1 receptor antagonist ZD 7155 (1 mg kg−1). 3 Adrenal demedullation inhibited the PYY-evoked responses of drug-naïve rats, and rats pretreated with the combination of PENT, MeATR and AVPX. 4 Transection of the splanchnic nerve innervating the adrenal medullae attenuated the bradycardia, as did ZD 7155, but not the PYY-evoked pressor response. 5 Systemic pretreatment of rats with the neuropeptide Y1 receptor antagonist BIBP 3226 (1 mg kg−1) blocked the PYY-evoked cardiovascular changes, but not those evoked by microinjection of carbachol (5.5 nmol) into the PHN. 6 These results suggest that the cardiovascular changes evoked from the PHN by PYY requires the presence of the adrenal medullae, which are stimulated by: (i) a hormone to release an NPY-like substance that evokes the pressor response, and (ii) the splanchnic nerve to evoke the release of a substance that results in the bradycardia.

Notes: 1 We examined whether extremely low frequency electromagnetic fields (ELF-EMF) affect the basal level of cardiovascular parameters and influence of drugs acting on the sympathetic nervous system. 2 Male rats were exposed to sham control and EMF (60 Hz, 20 G) for 1 (MF-1) or 5 days (MF-5). We evaluated the alterations of blood pressure (BP), pulse pressure (PP), heart rate (HR), and the PR interval, QRS interval and QT interval on the electrocardiogram and dysrhythmic ratio in basal level and dysrhythmia induced by β-adrenoceptor agonists. 3 In terms of the basal levels, there were no statistically significant differences among control, MF-1 and MF-5 in PR interval, QRS interval, mean BP, HR and PP. However, the QT interval, representing ventricular repolarization, was significantly reduced by MF-1 (P 〈 0.05). 4 (−)-Dobutamine (β1-adrenoceptor-selective agonist)-induced tachycardia was significantly suppressed by ELF-EMF exposure in MF-1 for the increase in HR (ΔHR), the decrease in QRS interval (ΔQRS) and the decrease in QT (ΔQT) interval. Adrenaline (nonselective β-receptor agonist)-induced dysrhythmia was also significantly suppressed by ELF-EMF in MF-1 for the number of missing beats, the dysrhythmic ratio, and the increase in BP and PP. 5 These results indicated that 1-day exposure to ELF-EMF (60 Hz, 20 G) could suppress the increase in HR by affecting ventricular repolarization and may have a down-regulatory effect on responses of the cardiovascular system induced by sympathetic agonists.

Notes: 1 Cyclic AMP formation has consistently been reported to be desensitized in various tissues including heart of animal models of end-stage renal failure (ESRF). In contrast, reports on desensitization of cAMP formation in ESRF patients remain contradictory. Whether this discrepancy results from a difference between human ESRF and its animal models or from the use of circulating blood cells in the human and various solid tissues in the animal studies, remains unclear. Therefore, we performed three studies with heart and platelets of ESRF patients undergoing haemodialysis or continuous ambulatory peritoneal dialysis and age- and gender-matched controls with normal renal function (n = 11–13 each). 2 In platelets from haemodialysis patients adenylyl cyclase activity in response to receptor-dependent and -independent agonists was reduced by ≈30%, and this could be explained by an alteration at the level of adenylyl cyclase itself. However, no such desensitization was seen in platelets from peritoneal dialysis patients. 3 In hearts from ESRF patients undergoing haemodialysis, β-adrenoceptor density and subtype distribution, cAMP formation in response to the β-adrenoceptor agonist isoprenaline or various receptor-independent stimuli, were very similar to those in control patients but activity of G-protein-coupled receptor kinase was increased by ≈20%. 4 We conclude that conflicting reports on the desensitization of cAMP formation between ESRF patients and ESRF animal models are not explained by the use of solid tissues in animal studies vs. circulating blood cells in patient studies. Rather desensitization of cAMP formation seems to be a less consistent feature of human ESRF than of its animal models.

Notes: 1 Our aim was to study the role of nitric oxide (NO) and arachidonic acid pathways in the α1-adrenoceptor-mediated vasoconstriction in mesenteric resistance arteries from 3–4 and 22 to 23-month-old Sprague-Dawley rats. 2 The expression of NO synthase (NOS), cyclooxygenase (COX) isoforms, soluble guanylate cyclase, superoxide dismutase and the NAD(P)H oxidase subunits p22phox and p47phox were determined. 3 The NG-nitro-l-arginine methyl ester, a non-selective NOS inhibitor, shifted to the left but indomethacin and NS 398, non-selective and selective COX-2 inhibitors, shifted to the right the concentration-response curve for the vasoconstriction by phenylephrine in both age groups. 4 Ageing up-regulated endothelial NOS and p22phox expression but did not modify COX, soluble guanylate cyclase, superoxide dismutase and p47phox expression. 5 These data suggest that the observed enhancement of eNOS protein expression could constitute a compensatory mechanism to counter-regulate a chronic loss of NO possibly through increased superoxide anion production from NAD(P)H oxidase induced by age.

Notes: 1 The effect of WAY 405 ((R)-N-(2-methyl-(4-indolyl-1-piperazinyl)ethyl)-N-(2-pyridinyl) cyclohexane carboxamide), a putative 5-HT1A receptor antagonist, on cardiovascular function was studied. 2 In anaesthetized rats, the i.v. injection of WAY 405 did not significantly modify basal heart rate nor blood pressure at doses of 1, 3, 10 and 30 μg kg−1; while the antagonist dose dependently antagonized the 5-HT1A receptor agonist, 8-OH-DPAT (8-hydroxy-2-(di-n-propylamino)tetralin)-induced hypotension and bradycardia. 3 WAY 405 antagonized noradrenaline-induced contraction in isolated arteries, with pKB values of 6.6 ± 0.1, 6.5 ± 0.1 and 6.5 ± 0.1, for rat tail artery (α1A-adrenoceptors), rabbit aorta (α1B-adrenoceptors), and rat aorta (α1D-adrenoceptors) respectively. 4 The results show that in the control of blood pressure the new compound, WAY 405, behaves as a silent 5-HT1A receptor antagonist in the anaesthetized rat, also having low affinity for vascular α1-adrenoceptors.

Notes: 1 Chronic cold exposure of rats (7 days in a cold room at 4 °C) attenuated the sympathetic nerve stimulation (NS)-induced overflow of noradrenaline (NE) (measured by high-performance liquid chromatography, coupled to electrochemical detection) appearing in the perfusate/superfusate of the perfused mesenteric arterial bed as well as the increase in the perfusion pressure. 2 The same type of cold exposure resulted in an increase in tyrosine hydroxylase (TH) gene expression measured in the superior cervical ganglion and NE content measured in the mesenteric artery obtained from cold-exposed rats. 3 Addition of sodium nitroprusside, a nitric oxide (NO) donor, to the buffer perfusing the mesenteric arterial bed obtained from rats maintained at room temperature also resulted in an attenuation of the NS-induced overflow of NE and increase in perfusion pressure. 4  N c-nitro-l-arginine methyl ester (l-NAME), an NO synthase inhibitor, placed in the drinking water prevented the attenuation of the pre- and post-junctional responses to NS of the mesenteric arterial bed obtained from cold-exposed rats. 5  l-NAME treatment also increased the cold-induced elevation of blood pressure seen in whole animals. 6 The present results are consistent with the idea that cold exposure leads to a concomitant increase in sympathetic nerve activity and production of NO. We hypothesize that the increase in production and release of NO results in a decrease in the biologically active form of NE despite increased synthesis and release of the catecholamine. 7 It is concluded that the above-mentioned interactions serve as a protective mechanism offsetting the increased release and action of NE from sympathetic nerves and thus preventing the development of hypertension.

Notes: 1 G-protein-coupled receptor signalling, including transactivation of receptor tyrosine kinases (RTKs), has been implicated in vascular pathology. However, the role of specific RTKs in the development of diabetes-induced cardiovascular complications is not known. 2 We investigated the ability of a chronic administration of genistein, a broad-spectrum inhibitor of tyrosine kinases (TKs), AG1478, a specific inhibitor of epidermal growth factor receptor (EGFR) TK activity, and AG825, a specific inhibitor of Erb2, to modulate the altered vasoreactivity of isolated carotid artery ring segments to common vasoconstrictors and vasodilators in streptozotocin (STZ)-induced diabetes. 3 In diabetic carotid artery, the vasoconstrictor responses induced by noradrenaline (NE), endothelin-1 (ET-1), and angiotensin II (Ang II), were significantly increased whereas vasodilator responses to carbachol and histamine were significantly reduced. Inhibition of TKs, EGFR or Erb2 pathway did not affect the body weight or agonist-induced vasoconstrictor and vasodilator responses in the non-diabetic control animals. However, inhibition of TKs by genistein, EGFR TK by AG1478 or Erb2 by AG825 treatment produced a significant normalization of the altered agonist-induced vasoconstrictor responses without affecting blood glucose levels. Treatment with diadzein, an inactive analogue of genistein, did not affect the vasoconstrictor and vasodilator responses in the diabetic animals. 4 Treatment with genistein, AG1478 or AG825 resulted in a significant improvement in diabetes-induced impairment in endothelium-dependent relaxation to carbachol and histamine. 5 These data suggest that activation of TK-mediated pathways, including EGFR TK signalling and Erb2 pathway, are involved in the development of diabetic vascular dysfunction in the carotid artery.

Notes: 1  The aim of the current study was to investigate the existence of P2Y4 purinergic receptors in the HT-29 human colon cancer cell line. 2  We utilized Western blots and immunocytochemistry for the analysis. 3  Western blotting demonstrated two bands that could not be found after the antibody had been preabsorbed with the control peptide, suggesting that both bands are related to the P2Y4 purinergic receptor. 4  Immunocytochemistry showed immunoreactivity for the P2Y4 purinergic receptor localized in the cytoplasm of the HT-29 cells. 5  This is the first demonstration of the protein expression of P2Y4 purinergic receptors in a human colon cancer cell line.

Notes: 1 The purpose of this study was to examine the effect of inhibition of the formation of cytochrome P450 metabolites of arachidonic acid with 1-aminobenzotriazole (ABT) on the development of hypertension and end-organ damage in spontaneously hypertensive rats (SHR) chronically treated with nitric oxide synthesis inhibitor l-NAME (SHR-l-NAME). 2 Administration of l-NAME in drinking water (80 mg l−1) to SHR for 3 weeks significantly elevated mean arterial blood pressure (MABP) (223 ± 4 mmHg) as compared to SHR controls drinking regular water (165 ± 3 mmHg). The administration of ABT (50 mg kg−1 i.p. alt diem) for 6 days significantly attenuated elevation of blood pressure in SHR-l-NAME (204 ± 4 mmHg). 3 l-NAME-induced increase in urine volume and protein was significantly lower in ABT-treated animals. 4 The impaired vascular responsiveness to noradrenaline and isoprenaline in the perfused mesenteric vascular bed of SHR-l-NAME-treated animals was significantly improved by ABT treatment. 5 Morphological studies of the kidneys and hearts showed that treatment with ABT minimized the extensive arterial fibrinoid necrosis, arterial thrombosis, significant narrowing of arterial lumen with marked arterial hyperplastic arterial changes that were observed in vehicle treated SHR-l-NAME. 6 In isolated perfused hearts, recovery of left ventricular function from 40 min of global ischaemia was significantly better in ABT-treated SHR-l-NAME. 7 These results suggest that in hypertensive individuals with endothelial dysfunction and chronic NO deficiency, inhibitors of 20-HETE synthesis may be able to attenuate development of high blood pressure and end-organ damage.

Notes: 1 The airways of patients with asthma are hyperresponsive to adenosine. The phenomenon can be mimicked in the actively sensitized Brown Norway rat by exposure to allergen or lipopolysaccharide (LPS). We wondered whether combined treatment with allergen and endotoxin would result in additive effects or synergism with respect to increasing the sensitivity of the airways of the Brown Norway rat to adenosine. 2 Animals actively sensitized to ovalbumin and challenged intratracheally with allergen or endotoxin manifested increased bronchoconstrictor responses to adenosine. A combination of ovalbumin and endotoxin also increased the response to adenosine but the effects were at best additive. 3 Changes in the response to adenosine were selective as responses to 5-hydroxytryptamine were unaltered following ovalbumin or LPS either alone or in combination. 4 Thus, endotoxin and allergen acting together could play a role in up-regulating the response of the human asthmatic airway to adenosine. However, our data suggest that the interaction would be additive rather than synergistic.

Notes: 1 We have characterized the α1-adrenoceptor subtypes present in isolated aorta of the α1D-adrenoceptor knockout (KO) mice, by chloroethylclonidine (CEC)-induced alkylation and their protection by selective α1-adrenoceptor antagonists. 2 The α1D-adrenoceptor is involved in the contractile response to noradrenaline in wild type (WT) mouse aorta. 3 In WT mice 5-methylurapidil (5-MU, an α1A-adrenoceptor antagonist) or BMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl] ethyl]-8-azaspiro[4.5] decane-7,9 dione, a selective α1D-adrenoceptor antagonist), protected the receptors from CEC-induced (α1B/D-adrenoceptor) alkylation, the combination of both antagonists resulted in complete protection, while AH11110A (1-[biphenyl-2-yloxy]-4-imino-4-piperidin-1-yl-butan-2-ol, an α1B-adrenoceptor antagonist) did not protect. 4 In aorta of KO mice there was a 19-fold rightward shift in noradrenaline effective concentration (EC50) compared with WT; while 5-MU alone or in combination with AH11110A protected α1-adrenoceptors to the same extent. 5 The data indicate that α1A-adrenoceptors mediate contraction and suggest their role in maintaining homeostasis in the α1D-adrenoceptors KO mice.

Notes: 1 This study evaluated the inhibitory action of apigenin-7-O-β-d-glucuronopyranoside (AGC), apigenin, and omeprazole on reflux oesophagitis and gastritis in rats. AGC was isolated from Clerodendron trichotomum leaves. 2 Oesophagitis and gastritis were induced by surgical procedure and the administration of indomethacin, respectively. The intraduodenal (i.d.) administration of AGC decreased the volume of gastric juice and increased the gastric pH compared with apigenin and omeprazole. The acid output was more inhibited by AGC in a dose-dependent manner than by apigenin and omeprazole. Compared with apigenin and omeprazole, AGC significantly decreased the size of gastric lesions, which were induced by exposure of the gastric mucosa to indomethacin. 3 Malondialdehyde (MDA) content, which is the end product of lipid peroxidation, was increased significantly after the induction of reflux oesophagitis. The MDA content was decreased by AGC (i.d. 3 mg kg−1), but not by either apigenin or omeprazole. This suggests that AGC has an antioxidative effect. In the oesophagitis group, the mucosal levels of glutathione (GSH) were significantly lower than that in the normal group. However, the GSH levels were preserved after administering the AGC, suggesting that AGC possesses scavenging activity. 4 In summary, AGC is more potent than apigenin and omeprazole at inhibiting reflux oesophagitis and gastritis and may therefore be a promising drug for their treatment.

Notes: 1 The fish somatostatin receptor 3 (fsst3) is one of the few somatostatin (SRIF) receptors cloned from a non-mammalian species so far. Here we extended our earlier characterization of this receptor by investigating the guanine nucleotide sensitivity of agonist radioligand binding at the fsst3 receptor recombinantly expressed in CCL39 (Chinese hamster lung fibroblast) cells. Further, we measured somatostatin (SRIF) and cortistatin (CST) analogues stimulated GTPγS binding, inhibition of forskolin-stimulated adenylate cyclase (FSAC) and stimulation of phospholipase C (PLC) activities. The present transductional data were then compared with previous radioligand binding and/or second messenger features determined for fsst3 and/or human SRIF receptors (hsst2, hsst3 and hsst5). 2 The GTP analogue guanylylimidodiphosphate (GppNHp) inhibited binding of [125I]CGP 23996 and [125I][Tyr3octreotide by 72 and 83% suggesting preferential labelling of G-protein-coupled fsst3 receptors. By contrast, [125I]LTT-SRIF28 and [125I][Tyr10]CST14 binding was rather GppNHp insensitive (42 and 35% inhibition) suggesting labelling of both coupled and non-coupled receptor states. These results might explain the apparent higher receptor densities determined in saturation experiments with [125I]LTT-SRIF28 and [125I][Tyr10]CST14 (4470 and 4030 fmol mg−1) compared with [125I]CGP 23996 and [125I][Tyr3]octreotide (3420 and 1520 fmol mg−1). 3 SRIF14 (10 μm)-stimulated specific [35S]GTPγS binding by three-fold; SRIF28 and octreotide displayed full agonism, whereas most other ligands displayed 60–80% intrinsic activity compared with SRIF14. SRIF14 and SRIF28 inhibited forskolin-stimulated AC (FSAC) activity by 60%; all tested ligands except BIM 23056 inhibited FSAC with comparable high intrinsic activities. SRIF14 stimulated PLC activity five- to six-fold, as determined by measuring total [3H] IPx accumulation; it was rather insensitive to pertussis toxin (PTX, 100 ng ml−1, 21% inhibition), which suggests the Gq-family proteins couple to PLC activity. SRIF14, SRIF28 and [Tyr10]CST14 showed full agonism at PLC, whereas all other ligands behaved as partial agonists (20–70% intrinsic activity). BIM 23056, which showed weak partial or no agonism, antagonized SRIF14-induced total [3H]-IPx production (pKB = 6.83), but failed to block competitively agonist-stimulated [35S]GTPγS binding or agonist-induced inhibition of FSAC activity. 4 Comparison of the pharmacological profiles of fsst3 receptors established in GTPγS binding, FSAC inhibition and PLC stimulation resulted in low correlations (r = 0.410–0.594). Both rank orders of potency and rank orders of relative efficacy varied in the three second messenger experiments. Significant, although variable correlations were obtained comparing GTPγS binding and inhibition of FSAC activity with previously reported affinity profiles of [125I]LTT-SRIF28, [125I][Tyr10]CST14, [125I]CGP 23996, [125I][Tyr3]octreotide (r = 0.75–0.83; 0.68–0.89). By contrast, the PLC stimulation and radioligand-binding profiles did not correlate. 5 Comparison of the functional data (GTPγS binding, FSAC inhibition, PLC stimulation) of fsst3 receptors with those of human sst2, sst3, sst5 receptors expressed in CCL39 cells resulted in highest correlation with the hsst5 receptor (r = 0.94, 0.97, 0.49) 〉 hsst2 (0.80, 0.50, n.d.) 〉 hsst3 (0.25, 0.19, 0.17). 6 In summary, fsst3 receptors expressed in CCL39 cells are involved in signalling cascades similar to those reported for mammalian SRIF receptors, suggesting SRIF receptors to be highly conserved in evolution. Binding and functional data showed highest similarity of fsst3 receptors with the human sst5 receptor subtype. Different affinities, receptor densities and GppNHp-sensitivities determined with the four radioligands (agonists) are assumed to results from ligand-specific states of the fsst3-ligand complex. The differences in the rank orders of potency and relative efficacy in the various signalling cascades may be explained by agonist-induced receptor trafficking.

Notes: The field of lymphatic research has been recently invigorated by the identification of genes and mechanisms that control various aspects of lymphatic development. We are beginning to understand how, starting from a subgroup of embryonic venous endothelial cells, the whole lymphatic system forms in a stepwise manner. The generation of genetically engineered mice with defects in different steps of the lymphangiogenic program has provided models that are increasing our understanding of the lymphatic system in health and disease. This knowledge, in turn, should lead to the development of better diagnostic methods and treatments of lymphatic disorders and tumor metastasis.

Notes: In this review I describe the several stages of my research career, all of which were driven by a desire to understand the basic mechanisms responsible for the complex and beautiful organization of the eukaryotic cell. I was originally trained as an electron microscopist in Argentina, and my first major contribution was the introduction of glutaraldehyde as a fixative that preserved the fine structure of cells, which opened the way for cytochemical studies at the EM level. My subsequent work on membrane-bound ribosomes illuminated the process of cotranslational translocation of polypeptides across the ER membrane and led to the formulation, with Gunter Blobel, of the signal hypothesis. My later studies with many talented colleagues contributed to an understanding of ER structure and function and aspects of the mechanisms that generate and maintain the polarity of epithelial cells. For this work my laboratory introduced the now widely adopted Madin-Darby canine kidney (MDCK) cell line, and demonstrated the polarized budding of envelope viruses from those cells, providing a powerful new system that further advanced the field of protein traffic.

Notes: Chromatin can be differentiated by the deposition of variant histones at centromeres, active genes, and silent loci. Variant histones are assembled into nucleosomes in a replication-independent manner, in contrast to assembly of bulk chromatin that is coupled to replication. Recent in vitro studies have provided the first glimpses of protein machines dedicated to building and replacing alternative nucleosomes. They deposit variant H2A and H3 histones and are targeted to particular functional sites in the genome. Differences between variant and canonical histones can have profound consequences, either for delivery of the histones to sites of assembly or for their function after incorporation into chromatin. Recent studies have also revealed connections between assembly of variant nucleosomes, chromatin remodeling, and histone post-translational modification. Taken together, these findings indicate that chromosome architecture can be highly dynamic at the most fundamental level, with epigenetic consequences.

Notes: Plants shape their organs with a precision demanded by optimal function; organ shaping requires control over cell wall expansion anisotropy. Focusing on multicellular organs, I survey the occurrence of expansion anisotropy and discuss its causes and proposed controls. Expansion anisotropy of a unit area of cell wall is characterized by the direction and degree of anisotropy. The direction of maximal expansion rate is usually regulated by the direction of net alignment among cellulose microfibrils, which overcomes the prevailing stress anisotropy. In some stems, the directionality of expansion of epidermal cells is controlled by that of the inner tissue. The degree of anisotropy can vary widely as a function of position and of treatment. The degree of anisotropy is probably controlled by factors in addition to the direction of microfibril alignment. I hypothesize that rates of expansion in maximal and minimal directions are regulated by distinct molecular mechanisms that regulate interactions between matrix and microfibrils.

Notes: Over the past decades, intravital microscopy (IVM), the imaging of cells in living organisms, has become a valuable tool for studying the molecular determinants of lymphocyte trafficking. Recent advances in microscopy now make it possible to image cell migration and cell-cell interactions in vivo deep within intact tissues. Here, we summarize the principal techniques that are currently used in IVM, discuss options and tools for fluorescence-based visualization of lymphocytes in microvessels and tissues, and describe IVM models used to explore lymphoid and non-lymphoid organs. The latter will be introduced according to the physiologic itinerary of developing and differentiating T and B lymphocytes as they traffic through the body, beginning with their development in bone marrow and thymus and continuing with their migration to secondary lymphoid organs and peripheral tissues.

Notes: Centrosomes, spindle pole bodies, and related structures in other organisms are a morphologically diverse group of organelles that share a common ability to nucleate and organize microtubules and are thus referred to as microtubule organizing centers or MTOCs. Features associated with MTOCs include organization of mitotic spindles, formation of primary cilia, progression through cytokinesis, and self-duplication once per cell cycle. Centrosomes bind more than 100 regulatory proteins, whose identities suggest roles in a multitude of cellular functions. In fact, recent work has shown that MTOCs are required for several regulatory functions including cell cycle transitions, cellular responses to stress, and organization of signal transduction pathways. These new liaisons between MTOCs and cellular regulation are the focus of this review. Elucidation of these and other previously unappreciated centrosome functions promises to yield exciting scientific discovery for some time to come.

Notes: The combined use of the new technologies of multiphoton-based intravital imaging, the chemotaxis-mediated collection of invasive cells, and high sensitivity expression profiling has allowed the correlation of the behavior of invasive tumor cells in vivo with their gene expression patterns. New insights have resulted including a gene expression signature for invasive cells and the tumor microenvironment invasion model. This model proposes that tumor invasion and metastasis can be studied as a problem resembling normal morphogenesis. We discuss how these new insights may lead to a better understanding of the molecular basis of the invasive behavior of tumor cells in vivo, which may result in new strategies for the diagnosis and treatment of metastasis.

Notes: Secretory and transmembrane proteins enter the secretory pathway through the protein-conducting Sec61 channel in the membrane of the endoplasmic reticulum. In the endoplasmic reticulum, proteins fold, are frequently covalently modified, and oligomerize before they are packaged into transport vesicles that shuttle them to the Golgi complex. Proteins that misfold in the endoplasmic reticulum are selectively transported back across the endoplasmic reticulum membrane to the cytosol for degradation by proteasomes. Depending on the topology of the defect in the protein, cytosolic or lumenal chaperones are involved in its targeting to degradation. The export channel for misfolded proteins is likely also formed by Sec61p. Export may be powered by AAA-ATPases of the proteasome 19S regulatory particle or Cdc48p/p97. Exported proteins are frequently ubiquitylated prior to degradation and are escorted to the proteasome by polyubiquitin-binding proteins.

Notes: Phagocytosis, the process by which cells engulf large particles, requires a substantial contribution of membranes. Recent studies have revealed that intracellular compartments, including endocytic organelles and the endoplasmic reticulum (ER), can engage in fusion events with the plasma membrane at the sites of nascent phagosomes. The finding that ER proteins are delivered to phagosomes, where degraded peptides are loaded onto major histocompatibility complex (MHC) class II molecules, has significantly enhanced our understanding of the immune functions associated with these organelles. Although it is well known that pathogens are killed in phagosomes, the contribution of ER proteins to phagosomes has provided a novel pathway for the loading of exogenous peptides onto MHC class I molecules, a process known as cross-presentation. Thus, phagocytosis has evolved from a nutritional function in unicellular organisms to play key roles in both innate and adaptive immunity in vertebrates.

Notes: The sensory and motor components of nervous systems are connected topographically and contain neural maps of the external world. The paradigm for such maps is the precisely ordered wiring of the output cells of the eye to their synaptic targets in the tectum of the midbrain. The retinotectal map is organized in development through the graded activity of Eph receptor tyrosine kinases and their ephrin ligands. These signaling proteins are arrayed in complementary expression gradients along the orthogonal axes of the retina and tectum, and provide both input and recipient cells with Cartesian coordinates that specify their location. Molecular genetic studies in the mouse indicate that these coordinates are interpreted in the context of neuronal competition for termination sites in the tectum. They further suggest that order in the retinotectal map is determined by ratiometric rather than absolute difference comparisons in Eph signaling along the temporal-nasal and dorsal-ventral axes of the eye.

Notes: The cytoskeleton plays important roles in plant cell shape determination by influencing the patterns in which cell wall materials are deposited. Cortical microtubules are thought to orient the direction of cell expansion primarily via their influence on the deposition of cellulose into the wall, although the precise nature of the microtubule-cellulose relationship remains unclear. In both tip-growing and diffusely growing cell types, F-actin promotes growth and also contributes to the spatial regulation of growth. F-actin has been proposed to play a variety of roles in the regulation of secretion in expanding cells, but its functions in cell growth control are not well understood. Recent work highlighted in this review on the morphogenesis of selected cell types has yielded substantial new insights into mechanisms governing the dynamics and organization of cytoskeletal filaments in expanding plant cells and how microtubules and F-actin interact to direct patterns of cell growth. Nevertheless, many important questions remain to be answered.

Notes: The establishment of the Angiosperm root apical meristem is dependent on the specification of a stem cell niche and the subsequent development of the quiescent center at the presumptive root pole. Distribution of auxin and the establishment of auxin maxima are early formative steps in niche specification that depend on the expression and distribution of auxin carriers. Auxin specifies stem cell niche formation by directly and indirectly affecting gene activities. Part of the indirect regulation by auxin may involve changes in redox, favoring local, oxidized microenvironments. Formation of a QC is required for root meristem development and elaboration. Many signals likely pass between the QC and the adjacent root meristem tissues. Disappearance of the QC is associated with roots becoming determinate. Given the many auxin feedback loops, we hypothesize that roots evolved as part of an auxin homeostasis mechanism.

Notes: Polarization is a feature common to many cell types. Epithelial cells, for example, exhibit a characteristic apical-basolateral polarity that is critical for their function. In addition to this ubiquitous form of polarity, whole fields of cells are often polarized in a plane perpendicular to the apical-basal axis. This form of polarity, referred to as planar cell polarity (PCP), exists in all adult Drosophila cuticular tissues, as well as in numerous vertebrate tissues, including the mammalian skin and inner ear epithelia. Recent advances in the study of PCP establishment are beginning to unravel the molecular mechanisms underlying this cellular process. This review discusses new developments in the molecular understanding of PCP in Drosophila and vertebrates and integrates the current data in a model to illustrate how interactions between PCP factors might function to generate planar polarity.

Notes: In contrast to other tissues, the nervous system is enriched in the polyunsaturated fatty acids (PUFAs): arachidonic acid (AA, 20:4 n-6) and docosahexaenoic acid (DHA, 22:6 n-3). Despite their abundance in the nervous system, AA and DHA cannot be synthesized de novo by mammals; they, or their precursors, must be ingested from dietary sources and transported to the brain. During late gestation and the early postnatal period, neurodevelopment is exceptionally rapid, and substantial amounts of PUFAs, especially DHA, are critical to ensure neurite outgrowth as well as proper brain and retina development. Here, we review the various functions of DHA in the nervous system, the proteins involved in its internalization and metabolism into phospholipids, and its relationship to several neurological disorders, including Alzheimer's disease and depression.

Notes: Bacteria communicate with one another using chemical signal molecules. As in higher organisms, the information supplied by these molecules is critical for synchronizing the activities of large groups of cells. In bacteria, chemical communication involves producing, releasing, detecting, and responding to small hormone-like molecules termed autoinducers . This process, termed quorum sensing, allows bacteria to monitor the environment for other bacteria and to alter behavior on a population-wide scale in response to changes in the number and/or species present in a community. Most quorum-sensing-controlled processes are unproductive when undertaken by an individual bacterium acting alone but become beneficial when carried out simultaneously by a large number of cells. Thus, quorum sensing confuses the distinction between prokaryotes and eukaryotes because it enables bacteria to act as multicellular organisms. This review focuses on the architectures of bacterial chemical communication networks; how chemical information is integrated, processed, and transduced to control gene expression; how intra- and interspecies cell-cell communication is accomplished; and the intriguing possibility of prokaryote-eukaryote cross-communication.

Notes: Brassinosteroids (BRs), the polyhydroxylated steroid hormones of plants, regulate the growth and differentiation of plants throughout their life cycle. Over the past several years, genetic and biochemical approaches have yielded great progress in understanding BR signaling. Unlike their animal counterparts, BRs are perceived at the plasma membrane by direct binding to the extracellular domain of the BRI1 receptor S/T kinase. BR perception initiates a signaling cascade, acting through a GSK3 kinase, BIN2, and the BSU1 phosphatase, which in turn modulates the phosphorylation state and stability of the nuclear transcription factors BES1 and BZR1. Microarray technology has been used extensively to provide a global view of BR genomic effects, as well as a specific set of target genes for BES1 and BZR1. These gene products thus provide a framework for how BRs regulate the growth of plants.

Notes: ʼ̛?‚ heterodimeric integrins mediate dynamic adhesive cell-cell and cell-extracellular matrix (ECM) interactions in metazoa that are critical in growth and development, hemostasis, and host defense. A central feature of these receptors is their capacity to change rapidly and reversibly their adhesive functions by modulating their ligand-binding affinity. This is normally achieved through interactions of the short cytoplasmic integrin tails with intracellular proteins, which trigger restructuring of the ligand-binding site through long-range conformational changes in the ectodomain. Ligand binding in turn elicits conformational changes that are transmitted back to the cell to regulate diverse responses. The publication of the integrin ʼ̛V?‚3 crystal structure has provided the context for interpreting decades-old biochemical studies. Newer NMR, crystallographic, and EM data, reviewed here, are providing a better picture of the dynamic integrin structure and the allosteric changes that guide its diverse functions.

Notes: This review surveys what is known about the structure and function of the subnuclear domains called Cajal bodies (CBs). The major focus is on CBs in mammalian cells but we provide an overview of homologous CB structures in other organisms. We discuss the protein and RNA components of CBs, including factors recently found to associate in a cell cycle-dependent fashion or under specific metabolic or stress conditions. We also consider the dynamic properties of both CBs and their molecular components, based largely on recent data obtained thanks to the advent of improved in vivo detection and imaging methods. We discuss how these data contribute to an understanding of CB functions and highlight major questions that remain to be answered. Finally, we consider the interesting links that have emerged between CBs and alterations in nuclear structure apparent in a range of human pathologies, including cancer and inherited neurodegenerative diseases. We speculate on the relationship between CB function and molecular disease.

Notes: Phosphoinositide phosphates (PIPs) correspond to phosphorylated derivatives of phosphatidylinositol (PI). Despite their relatively low abundance in the plasma membrane, PIPs play a crucial role as precursors of second messengers and are themselves important signaling and targeting molecules. Indeed, modulation of levels of PIPs affects, for example, cortical actin organization, membrane dynamics, and cell migration. The focus of this review is on selected interesting targets of PIPs. Those proteins that bind PIPs and are involved in regulation of actin assembly, actin membrane linkage, and actin contractility are discussed, as well as those that are involved in signaling, such as small GTPases, protein kinases, and phosphatases, or in regulation of membrane dynamics.

Notes: In eukaryotes, the entwined pathways of RNA transport and local translational regulation are key determinants in the spatio-temporal articulation of gene expression. One of the main advantages of this mechanism over transcriptional control in the nucleus lies in the fact that it endows local sites with independent decision-making authority, a consideration that is of particular relevance in cells with complex cellular architecture such as neurons. Localized RNAs typically contain codes, expressed within cis-acting elements, that specify subcellular targeting. Such codes are recognized by trans-acting factors, adaptors that mediate translocation along cytoskeletal elements by molecular motors. Most transported mRNAs are assumed translationally dormant while en route. In some cell types, especially in neurons, it is considered crucial that translation remains repressed after arrival at the destination site (e.g., a postsynaptic microdomain) until an appropriate activation signal is received. Several candidate mechanisms have been suggested to participate in the local implementation of translational repression and activation, and such mechanisms may target translation at the level of initiation and/or elongation. Recent data indicate that untranslated RNAs may play important roles in the local control of translation.

Notes: Five human teratoma cell lines have been characterized for the presence of a certain number of marker antigens whose presence or absence has been shown to be characteristic of mouse embryonal carcinoma (EC) cells. Four out of the five lines have been shown to respond to at least some of the criteria associated with murine EC cells even though only limited in vitro differentiation could be demonstrated. The significance of certain unusual marker antigen combinations present on the cell line Tera I and its clones and so far unobserved for the murine model is discussed. The observation in Tera I populations of cells carrying simultaneously both the F9 and β2-microglobulin or HLA antigens, suggest that the human cell lines may represent a novel material for the study of mammalian differentiation.

Notes: Principles of Gene Manipulation. Studies in Microbiology

Notes: A healthy 22-year-old woman was noted to have erythrocytes of the Pk phenotype: a strong Pk antigen, no detectable P antigen and anti-P antibody in her serum. Her erythrocytes contained four to six times as much Pk glycolipid (globotriaosylceramide or CTH) and approximately half as much P glycolipid (globotertraosylceramide or globoside) as normal red cells. The structures of CTH and globoside were characterized by analysis of permethylated sugars and complement fixation in addition to chromaographic mobility and sugar composition. Inasmuch as the erythrocytes of two Pk individuals that were analysed previously (Marcus et al., 1976) contained no detectable globoside, these abnormalities appear o represent a new phenotype in the P blood group system.

Notes: We studied 201 unrelated French Basque individuals for HLA and Bf polymorphisms. The haplotypes of eighty-seven of them were deduced from family studies. The results show the frequency of the Bf F1 allele (0.1393) which is the highest one currently reported. They confirm the high frequencies of HLA-Aw19.2 and B18 previously reported in that population and show that a whole haplotype with strong linkage disequilibria, namely Aw19.2, Cw5, B18, Bf F1, DRw3 is frequent. On the other hand, the gene frequency of Bf S is decreased (0.5497) as compared with the other European Caucasoïd populations, while a slight increase in the Bf F gene frequency (0.2960) appears. These results point out that it is of importance to consider the genetic background in choosing the population where linkage disequilibria are to be studied.

Notes: The genetic control of hybrid resistance to BALB/c fibrosarcoma Meth-A was investigated. A Meth-A tumour grew slower in (BALB/c X C57BL/6)F1 and reciprocal hybrid mice than in syngeneic BALB/c mice and was also found to grow slower in females than in males. Significant F1 resistance was demonstrated after both subcutaneous and intraperitoneal injection of tumour cells. However, (BALB/c X DBA/2)F1 mice did not show any significant resistance to Meth-A. In H-2 linkage studies of [BALB/c X (BALB/c X C57BL/6)] backcross mice, no statistically significant differences in the resistance of H-2 heterozygotes and homozygotes to Meth-A were observed. These results indicated that F1 hybrid resistance to Meth-A was controlled by non-H-2-linked resistance factor(s). No linkage was observed between resistance to Meth-A and coat colour c- and b-loci.

Notes: CBA/N mice have an X-linked B cell defect which prevents them from responding to non-mitogenic thymic independent (TI-II) antigens such as dinitrophenylated (DNP-AGG) Ficoll. The F1 male progeny of CBA/N female mice express the same defect. Spleen cell suspensions from such defective mice (CBA/N X C3H/HeN F1 males) could not respond to DNP-AGG-Ficoll following in vitro immunization and subsequent transfer into irradiated, syngeneic, F1 male recipients as expected. In contrast, normal CBA/N X C3H/HeN F1 female spleen cells could respond and effect a ‘rescue'; they mounted strong plaque-foriming cell 7 days after in vitro exposure to DNP-AGG-Ficoll and subsequent transfer into irradiated F1 male recipients. Defective F1 male spleen cells could bind significant quantities of DNP-AGG-Ficoll, however, after, in vitro exposure. Extensive washing of these spleen cells could not reverse this binding. Such DNP-AGG-Ficoll-exposed and washed F1 male spleen cells could, after transfer, aid normal untreated F1 female cells in their rescue function. The defective F1 male spleen cells could convey immunogenic quantities of DNP-AGG-Ficoll to the ‘rescuing’ F1 female cells.Mitomycin treatment of F1 male cells did not interfere with their conveyor function. Goat anti-mouse μ serum impeded the passive antigen conveyor function of defective F1 male cells as did prior exposure to high concentrations of free DNP-AGG hapten. Our data support the view that the B cell defect of CBA/N X C3H/HeN F1 male mice does not relate to antigen binding, but rather to an inability to be effectively triggered by certain cell-bound polymeric antigens.

Notes: Results of a population study with all currently available B-cell specific alloantisera indicate that eight antigens controlled by the RhLA-linked DR locus can now be identified. This leaves a gene frequency of about 0.15 for unidentified or ‘blank’ antigens of that locus. Of the nine identifiable la antigens which are not controlled by the DR locus, three or four may form the basis of a second series which is probably also controlled by the RhLA region.

Notes: Induction of tolerance to bovine serum albumin was studied in mice selected for high (H) or low (L) antibody responsiveness and in their F1 hybrids. No high or low zone tolerances were obtained in H mice whereas L mice were susceptible to tolerance induction by the two schedules. H mice were immunized by repeated injections of tolerogenic BSA for low zone tolerance induction but not after the administration of a single high dose of tolerogenic BSA. Resistance to tolerance induction is dominant in F1 hybrids.

Notes: An attempt is made to account for immunoglobulin chain synthesis in terms of genetic events involving IS or controlling elements analogous to those found in bacteria, maize and drosophila. Transposition of variable and constant genes and normal immunoglobulin chain synthesis as well as qualitative and quantitative abnormalities might be explained by such regulatory elements. Intrachromosomal transpositions over short distances would be expressed as apparent hypermutability or redundancy of the variable DNA segment. The constant gene might comprise four sequences coding for the three homology domains and the hinge, separated by intervening sequences. A strong preference for shortrange transposition on the same chromosome and immobilization of the controlling element in the end might account for allelic exclusion.

Notes: Fab2 fragments from antisera raised in rabbits with partially purified cellular and serum HLA antigens were tested for their ability to block the cytolytic activity of operationally specific HLA-A, B alloantisera. One Fab2 fragment preparation blocked the cytolytic activity of all the HLA-A,B alloantisera tested; the remaining nine inhibited the lytic activity of alloantisera to certain HLA-A,B allospecificities, suggesting that these xenoantisera contain antibody to certain HLA-A,B allotype determinants or to closely associated structures. In contrast to previous reports in the literature none of the xenoantisera contained significant amounts of antibodies to human β2-microglobulin.

Notes: NIM-M8 is a monoclonla IgM antibody, specific for the LWab antigen as shown by its reaction with red cells of all donors except those lacking LWa, LWb and LWab. Indirect immunofluorescent staining and cell sorter analyses have shown that LWab is present on a subpopulation of human lymphoctes. Cell fractionation studies indicate that subsets of both B and T cells express LWab and it may, therefore, provide a further marker for heterogeneity in these lymphocyte populations.

Notes: Immunoprecipitation using a monoclonal antibody showed that the Wrb antigen is present on the abnormal (δ-α) hybrid sialoglycoprotein of Sta-positive human erythrocytes but not on the abnormal (δ-α) hybrid sialoglycoprotein of Dantu-positive erythrocytes. These results provide further information regarding the nature and location of the Wrb antigen on the normal erythrocyte sialoglycoprotein α.

Notes: We studied structural and functional characteristics of lymphocytes from adult and fetal baboons (Papio cynocephalus). Flow cytometry with monoclonal antibodies to human lymphocyte antigens and plant lectins was used to define expression of surface antigens on lymphocytes from adult and 140 day fetal baboons (term = 180 days). Major T cell antigenic determinants on adult and fetal baboon lymphocytes were the Tp50, Tp32-45, and p45 glycoproteins detected by monoclonal reagents T11, OKT8, and OKT10 respectively. Baboon T lymphocytes did not react with the OKT3/anti-Leu4 or OKT4/ anti-Leu3a reagents which detect, respectively, Tp19-29 and Tp55, major surface glycoproteins on human T lymphocytes. OKT6, which identifies the human TL antigen equivalent on thymocytes, did not react with baboon thymocytes. These data demonstrate major evolutionary divergence between human and baboon T lymphocytes. By contrast, baboon lymphocytes resembled human peripheral lymphocytes in reactivities with several non-T cell reagents. Lectin binding studies revealed substantially fewer peanut agglutinin-and wheat germ agglutinin-binding cells in suspensions of baboon fetal splenocytes and adult peripheral lymphocytes compared with fetal thymocytes. Thereffore, maturation of baboon T lymphocytes is associated with loss of surface carbohydrate structures that bind these lectins. Adult and fetal baboon lymphocytes resembled human and murine lymphocytes in their capabilities to respond to mitogens and to produce interleukin-2. As in oter species, adult, but not fetal baboon lymphocytes, mediated NK activity against a variety of nucleated target cells. Despite divergence in lymphocyte antigen epression, babbon lymphocyte functional development colsely parallels that seen in humans.

Notes: The Gm, Am and Km immunoglobulin allotypes and ABO blood groups were studied in three groups of Tunisian Berbers.The results showed that the actual Berbers of Tunisia present certain heterogeneity and their ancestors were probably the first inhabitants of North Africa. Indeed, although their Gm-Am haplotypes are mainly Caucasoid, some of them are typically African.The group of Kesra village, the most Caucasoid, shows frequencies of Gm-Am haplotypes very close to those of South European populations, particularly the Spanish, who are probably of the same origin. The gene frequencies of the ABO groups in the three Berber groups were similar to those recorded in European populations with a relatively high frequency of the O genes typical of the Berbers.

Notes: Monoclonal antibody 212.i.4.2 mediated complement-dependent lysis of spleen and lymph node cells carrying the tw1, tw12, tw71, t6, tw73, and tLub1 haplotypes, while cells from mice carrying 11 other t haplotypes were not lysed. The antibody also detected an epitope controlled by genes in the H-2Dd region of non-t mice. A molecule of 46,000 molecular weight was immunoprecipitated by 212.i.4.2 from detergent extracts of 125I-labelled spleen cells of +/tw12 and B10.D2 mice. The H-2dm2 mutation did not alter the expression of the epitope recognized by 212.i.4.2. However, the H-2dm1 mutation decreased the reactivity of lymphoid cells with the antibody in cytotoxicity tests, and 212.i.4.2 immunoprecipitated little or no protein from extracts of B10.D2(R106) spleen cells which carry the H-2dm1 mutation.

Notes: Spleen cells from Balb/c mice given multiple injections of intact human erythrocytes (group O, NN) were fused with NS1 myeloma cells. Culture fluids from the resulting hybrid cells were screened for agglutinating antibody against a panel of erythrocytes. One cell line, 2/23, secreted an IgM antibody which reacted more strongly with NN than with MM cells. Neuraminidase or papain treatment of erythrocytes abolished agglutination whereas trypsin treatment did not. Reactions with U-erythrocytes of different MN phenotypes confirmed the anti-N specificity of monoclonal antibody 2/23. This is the first report of monoclonal anti-N stimulated by the immunization of mice with intact erythrocytes.

Notes: Spleen cells from 30 individual murine irradiation chimeras of the type (P1 x P2)F1→ P1 were compared in a rosetting assay for H-2K and H-2D cell surface antigen expression with normal (P1 x P2)F1 hybrid controls. Eleven out of the 30 chimeras were in the normal range, but the other 19 differed from F1 controls by 4- to 100-fold in endpoint titre for at least one H-2K or H-2D antigen. Every possible class of variation was found, i.e. up or down variation of H-2K or H-2D antigens of P1 or P2 type. This evidence, together with data from T6 chromosome marker experiments which also showed full reconstitution of lethally irradiated P1 recipients by (P1 x P2)F1 donor lymphomyloid stem cells, suggested that incomplete reconstitution was not the cause of H-2 antigenic variation.Low expression of P2 H-2 antigens on spleen cells derived from (P1 x P2)F1→ P1 chimeras was investigated further. Fifteen lethally irradiated (P1 x P2)F1 recipients of bone marrow cells from two such chimeras were all of normal F1 H-2 phenotype when tested 10-12 weeks after reconstitution, thus excluding stable, low P2 H-2-expressing variant F1 stem cells as a cause of the phenomenon. If P1 recipients were hyperimmunized against P2 cells before lethal irradiation and reconstitution with (P1 x P2)F1 stem cells, there were significantly fewer Till-McCulloch colonies in their spleens 10 days after reconstitution than in spleens of unimmunized controls. Also 〉 90% of immunized recients died by 6 weeks after stem cell injection but two survivors both showed very low levels of P2 H-2K and H-2D antigens. These results together with previously published evidence of anti-P2 Tc cell activity and P2 skin graft rejection in (P1 x P2)F1→ P1 chimeras suggested that residual anti-P2 immunological capability in lethally irradiated P1 recipients may be associated with low P2 H-2 expression on their F1-derived spleen cells, although the mechanism does not involve selection of stable, variant F1 stem cells. The mechanism(s) of other classes of variation in H-2 expression in (P1 x P2)F1→ P1 chimeras were not investigated.

Notes: RNA was extracted from the splenocytes of Brucella abortus antigen stimulated mice and of control mice. The proportion of chromatographically separated polyadenylated 11.2S mRNA, was determined. With the technique used, only stimulated mice exhibited significant amounts of this RNA species. The highest level was reached 1 day after the stimulation, and the decay from this level presented an oscillatory form during the 4 weeks following the injection.In two different genetic backgrounds, H-2b mice did not respond to the stimulus, in contrast to H-2a and H-2f mice. H-2b/H-2f heterozygotes behaved roughly as intermediate between H-2b and H-2f mice. This genetic control seems to parallel the genetic control of some Brucella-induced, thymus-dependent events previously described.

Notes: Book Reviews in this ArticleR. WITKOWSKI and O. PROKOP: Genetik erblicher Syndrome und Mgbildungen. Worterbuch fur die Familienberatung.

Notes: Monoclonal anti-Ia inhibition experiments were conducted to confirm and extend genetic mapping data of I-A gene control of immunity to human haemoglobin (Hb). It was found that the Aβ gene is of critical importance in conferring immunity to the α-chain and β-chain subunits of Hb. A possible involvement of I-E region genes in B10.D2 mice to β-chain is discussed. Through the use of an α-chain specific T cell clone data, is obtained indicating that an intact Ia.8+ Aβ chain is necessary for antigen presentation in vitro.

Notes: C4 is composed of two tightly linked genes (C4A and C4B) lying within the major histocompatibility complex of chromosome 6 that can be demonstrated by agarose gel electrophoresis. Seven alleles and five alleles at the C4A and C4B loci, respectively, were detected in 169 black individuals from the southeastern United States. Furthermore, the phenotypic frequencies of C4A6, C4A5, C4A4, C4B4, C4B3, and C4BQO were significantly different between black and white Americans.

Notes: Two B complex genotypes, B1B1 and B19B19, of outbred line S1, were tested for low and high immune response to GAT, from which four recombinants were recovered: B1B1 GAT-hi and lo, and B19B19 GAT-hi and -lo. Also included in the study were birds of B2B2 genotype with an intermediate level of immune response to GAT.A total of 225 birds of these groups were challenged with the Bryan strain of Rous Sarcoma virus subgroup C, RSV (RAV-7), by inoculation into the wing web at five weeks of age. The B1B1 genotype had the lowest percentage of regressors (17.6%), B19B19 had the highest (42.2%), and the B2B2 genotype was intermediate (23.7%). Combining the results of GAT response over the B1B1 and B19B19 genotypes, 14.0% of GAT-lo and 37.8% of GAT-hi regressed their tumours, respectively. The highly significant (P ≤ 0.01) difference between the combined GAT-hi and -lo groups would suggest that the Rs locus controlling tumour regression induced by the subgroup C virus is closely linked to the region controlling immune response to GAT, but the data also provides evidence that the B-F region of the B complex also plays an important role in RSV-induced tumour regression.

Notes: Surface immunoglobulin on spleen cells from NZB and NZB/W mice and congenic mice bearing the nude or X-linked immune defective (Xid) gene was examined by flow microfluorometry with regard to both the frequency of positive cells and density expressed on the cell. These data indicate that although the frequency of unseparated sIg+ B lymphocytes is equivalent among all of these groups of mice, the densities of sIgM and sIgD are different. Spleen cells from these mice were also separated by free-flow electrophoresis and analyzed in a similar manner. This analysis demonstrated the absence of a subpopulation of B lymphocytes with a low electrophoretic mobility and low expression of sIgM. These studies suggest that maturational and/or activation states of the B cells in mice bearing the Xid or nude genes are different from those seen in the parent strains of mice. Such alterations in cell-surface antigens correlate with differences in the natural history of immunopathology of the autoimmune disease in these congenic colonies of New Zealand mice.

Notes: Fifty-five Caucasoid patients with polymalgia rheumatica (PMR) or giant cell arteritis (GCA) were immunoglobulin (Gm) allotyped for this study. Forty-four of these patients had been previously HLA-A,B,C and DR locus allotyped. The incidence of the immunoglobulin allotypic marker Glm(2) was significantly increased in the GCA group (50.00% v. controls 18.75%, P= 〈0.01). There was a similar but insignificant rise of this Gm marker in the PMR group (27.24% v. 18.75%, NS). The increase in Glm(2) in the GCA group was not accompanied by a corresponding rise in the number of people homozygous for Glm(2), i.e., all the increase could be attributed to patients with the Glm(1,2,3): G3m(5,10,21) phenotype.

Notes: The strains B 10.S(7R), B 10.S(23 R) and B 10.S(24R), all thought to be genetically identical, differ in levels of susceptibility to infection with Trichinella spiralis. In a series of nine independent experiments, B 10.S(7R) was shown to be more susceptible than the other two strains. In another series of seven experiments, the strain B 10.A(18R) was shown to be more susceptible to infection with T. spiralis than the strains B 10.S(21R) or B 10.BAR-5, all of which were thought to share common H-2 alleles. These results indicate that a gene mapping between the S and D loci influences susceptibility to infection with T. spiralis. Typing of these strains for Qa and Tl loci rule out the possibility of a double crossover accounting for the differences observed. The new gene is designated Ts-2. Previously published data have also been reinterpreted and another gene Ts-1 is shown to be associated with the Aβ locus. When the d allele is expressed at the Ts-2 locus, strains of mice expressing s, q, f or b alleles at Ts-1 are rendered more susceptible to infection.

Notes: IgG and IgA heavy chain allotypes were determined in the sera of 483 Caucasian Type 1 diabetes patients and 503 Caucasian healthy controls. There was no significant difference between patients and controls neither on the level of Gm phenotype frequencies nor on the level of Gm three-locus and two-locus haplotype frequencies. A selective IgA deficiency was found in 14 patients (2.9%) but in none of the control individuals (P〈10-4).

Notes: Gm allotypes were detected and quantitated by radioimmunoassay (RIA) in paired serum and CSF samples from patients suffering from various neurological diseases. Of 115 patients with neurological disorders (65 MS and 50 others), seven subjects displayed one or two allotypes in their CSF which were absent in serum. The Gm phenotype in the patient's serum allowed us to infer the genotype without the need of familial data. A comparison of the regression curves obtained in RIA from the unexpected allotype in CSF and the counterpart in a normal serum pool argued for an identity of the Gm antigen carried by both inhibitory molecules. The unexpected allotype(s) in CSF can be considered as the product of a latent Gm gene which may be activated by either immune perturbations due to the disease per se or some particular immune regulations in the central nervous system.

Notes: Book reviewed in this article:N. Catsimpoolas; Cell Analysis. Plenum Publishing CorpJ.W.Shay: Techniques In Somatic Cell Genetics

Notes: Immunoprecipitaon studies of the rhesus monkey major histocompatibility system have shown that the RhLA-DR locus codes for class II antigens with molecular features that are homologous to the class II antigens coded for by the human HLA-dR locus, The products of another alloantigenic RhLA-linked locus of the rhesus monkey, called ‘48’, is provisionally characterized as a class I system.

Notes: The relationships between the antigens recognized by four monoclonal anti-human ‘Ia’-like antibodies were investigated using sequential immunoprecipitation and capping techniques. Two of the antibodies were ‘monomorphic’ and have previously been shown to recognize epitopes in which carbohydrate residues are involved, whereas the two ‘polymorphic’ antibodies recognized protein-defined epitopes—one of these epitopes being present on MB+DR- molecules. In the absence of an indisputable anti-DR monoclonal antibody, it was not possible to conclusively verify which ‘Ia’-encoded antigens were detected by the anti-‘Ia’-like monoclonal antibodies. Nevertheless, several firm conclusions could be drawn: (a) so-called ‘monomorphic’ antibodies do not necessarily react with all ‘Ia’ molecules encoded by a single locus—from the results using the two monomorphic antibodies, B5.1 and 3F1.1, described herein, two populations of antigens being B5.1+3F1.1+ and B5.1+3F1.1- were identified; (b) cross-reactivity of a polymorphic determinant expressed on antigenically-separable ‘Ia’ molecules was noted—using the two polymorphic antibodies, 26.1 and F5C9, molecules which were 26.1+F5C9+ and 26.1-F5C9+ were identified; and (c) the data clearly point to the existence of at least two loci coding for ‘Ia’-like antigens (one of which may or may not be the HLA-DR locus). Given that polymorphisms can now include protein- and carbohydrate-defined epitopes, that cross-reactions occur and that the definition of DR itself by monoclonal antibodies is not clear, the complexity of the human ‘Ia’ antgens is apparent.

Notes: We report HLA genotypes in four familial cases of Hodgkin's disease (HD), Nodular Sclerosis (NS) histological subtype, where all patients showed B18 antigen. This finding, although statistically not supported, confirms the possible correlation between HD and B18 antigen which carries a high relative risk in international data.

Notes: Previous studies from this laboratory have resulted in the determination of the antigenic structure of sperm-whale myoglobin (Mb). In the present work, we have investigated the fine specificity requirements for T-cell recognition of one of the Mb antigenic sites (antigenic site 5). The antigenic site (peptide 145-153) and seven progressively longer peptides, increasing in length stepwise by two residues at a time, up to 22 residues in length (peptide 132-153), were synthesized. In addition, four truncated peptides were synthesized with intentional deletions at Tyr- 151 and Ala- 144. The T-cell recognition of these purified synthetic peptides was examined here in detail in three strains of mice (BALB/cByJ, B10.D2/n and SJL/J). Mb-primed mice afforded T-cells which proliferated to smaller peptides (two or four residues longer than the site; i.e. peptides 145-153 and 143-153) and more so to the longer peptides 135-153 and 132-153 and to Mb. No response was obtained to the truncated peptides, thus underscoring the fine specificity T-cells. No response was obtained also to intermediate-sized peptides. The latter result, due to an unfavourable mode of folding, suggested a conformational dependency in T-lymphocyte recognition.

Notes: Serum blood samples from 563 of the total 700 Nganasans, members of the isolate in the northern-most part of Siberia were tested for G1m, (z,a,x,f), G2m (n), G3m (g,b0,b1,b3,b5,s,t), and km (1) allotypic determinants. Additionally, 78 Yenisey Samoyeds (Entsi) who are the Nganasan's western neighbours were studied. Both populations are remarkable for high frequency of ‘Northern Oriental’Gm (za;.;b0b3b5st) which appears to be the most frequent haplotype in the Nganasans (0.486), and is the second frequent in Yenisey Samoyeds (0.276). The Gm (f;b) generalized haplotype which used to be considered as an indicator of Caucasian gene flow occurred in the Nganasans in the very low frequency of 0.008, versus 0.045 revealed in adjoining Yenisey Samoyeds. Both populations also differ in the frequency of Km1 which is two times lower in the Nganasans (0.048), than in Yenisey Samoyeds (0.103). When segregation ratios for the Gm locus were inspected in 67 Nganasan families, no apparent deviations from Mendelian expectations, and no recombinant phenotypes were observed.

Notes: Genetic control of PQ prolongation of the electrocardiogram (ECG) in the mouse, immunized with killed group A streptococci, was studied by using various congenic mice. Mice of H-2a, H-2k and H-2f haplotypes showed high frequencies of PQ prolongation, while haplotypes of H-2b, H-2d and H-2s showed low frequencies of PQ prolongation. Studies using various recombinant mice revealed that at least one immune-associated (Ir) gene mapped in the left side of the I-B subregion. High responsiveness of F1 hybrids of H-2b and H-2d, as well as B1O.A(5R) and B10.A(3R), suggests the existence of a complementing gene. In addition, the differences between C3H and CKB, as well as differences between C3H.SW and CWB, indicate that another Ir gene maps in the immunoglobulin heavy chain (Igh) coding loci. Repeated injections of anti-I-J or anti-I-A antisera also modified this PQ prolongation. These results suggested that both the major histocompatibility complex (MHC) and immunoglobulin (Igh) loci seem to be playing important roles in the pathogenesis of PQ prolongation.

Notes: The growth and reproduction complex contains recessive genes (grc) which influence body weight and gonadal development. Homozygous males are sterile, and they have an arrest of spermatogenesis at the primary spermatocyte stage. Homozygous females are fertile but have a reduced reproductive capacity. The data presented in this paper show that the latter defect is associated with a decrease in the relative number of secondary ovarian follicles and an increase in the number of atretic follicles. This finding indicates that most of the primary follicles do not mature properly. Thus, the genetic defect in gametogenesis controlled by the grc appears to occur at the same stage of development in both females and males.

Notes: Graves’ disease (GD) is a common, autoimmune disease involving the thyroid gland, and it has been previously suggested that pro-inflammatory cytokines are involved in the disease's pathogenesis. The aim of this study was to test whether the interleukin (IL)-6 gene promoter region, or tumour necrosis factor (TNF)-α or IL-8 gene 3′-untranslated region (3′-UTR) polymorphisms could provide useful genetic markers for an individual's susceptibility to GD. A normal control group of 60 healthy people and 95 patients featuring GD were examined. Polymerase chain reaction (PCR)-based restriction analysis was performed for the three gene polymorphisms using endonucleases BsrBI, NcoI and ApaLI, respectively. We found no significant difference between the frequencies of genotype and allelic variants for the IL-6 gene promoter (−572 G/C), the TNF-α gene promoter (−308 A/G) and the IL-8 gene 3′-UTR (2767 A/G) for GD patients and for normal controls. Cytokines are a large group of proteins that may elicit multiple effects upon immunological reactions. It still appears to be very worthwhile to continue to aggressively search for cytokine gene polymorphisms in order to predict the development of such disease.

Notes: Two novel interleukin-24 (IL-24) splice variants were identified in normal human melanocytes by sequencing cloned polymerase chain reaction (PCR) products that are not expressed in metastatic melanoma. These gene products have been generated by differential skipping of exons 3 (IL-24 delE3) and 5 (IL-24 delE5). IL-24 delE3 has limited sequence identity to the IL-24-interacting protein mda-7s, and IL-24 delE5 is homologous to IL-24.

Notes: Human leukocyte antigen (HLA) gene products have been implicated in the pathogenesis of an increasing number of eye diseases, mainly inflammatory in nature. This perspective reviews the current hypotheses for why HLA polymorphisms are associated with specific eye diseases. Statistical problems in studies involving HLA associations are discussed, and possible solutions outlined. The relevance of HLA testing in routine ophthalmic practice, its practical and cost implications is also assessed.

Notes: Nuclear factor κB (NF-κB) designates a group of critical transcription factors involved in a variety of immunologic and/or inflammatory processes. Conceivably, genes involved in the NF-κB pathway make interesting candidate genes for chronic inflammatory disorders, including the inflammatory bowel diseases (IBD), Crohn's disease (CD) and ulcerative colitis (UC). In two mouse models of colitis, strong linkage has been observed with a locus on chromosome 3 that harbours the Nfkb1 gene. In addition, a polymorphism in the promoter region of the human NFKB1 gene was found to be associated with susceptibility to UC. In this study, we searched to confirm this previously found association in IBD in a different population. Allele and genotype frequencies of the −94 ins/delATTG polymorphism were determined in 266 unrelated Dutch Caucasian IBD patients (127 UC, 139 CD), and 155 matched healthy controls. The allele frequency of the deletion was significantly higher in UC patients (P = 0.019), but not in CD patients, compared to healthy controls, and the UC patients homozygous for the −94 ATTG deletion had a younger age of onset. Our findings confirm the previously found association between this polymorphism and susceptibility to UC in an independent study population and adds further evidence for the role of this gene in disease susceptibility.

Notes: The polymorphic T-cell receptor Vβ (TRBV) genes encode much of the variable region of the T-cell receptor β chain. Analysis of allele frequencies of three closely linked polymorphic TRBV genes, TRBV7-3, TRBV9 and TRBV6-4, was undertaken in several populations. The frequencies of these alleles are not significantly different in populations of Caucasians, African Americans and Western Africans. However, Chinese population is extremely homogenous at all three loci. The current study identifies the existence of haplotypic relationships between alleles of these genes in the Caucasian population. The ORF allele TRBV7-3*A3 is found exclusively on chromosomes bearing TRBV9*A2 and TRBV6-4*A2 in this cohort. In contrast, TRBV7-3*A1 and the null allele TRBV7-3*A2 are associated only with TRBV9*A1 and TRBV6-4*A1. This pattern of linkage disequilibrium (LD) is altered in the African American and Western African populations. In these cohorts, there is a marked reduction in LD between alleles of TRBV7-3 and TRBV9. This study is consistent with previous population genetic studies wherein African-derived samples have a greater level of genetic diversity compared to Caucasians. These data also demonstrate that patterns of LD are not consistent across the entire TRBV locus.

Notes: Matrix metalloproteinase-7 (MMP-7) generates soluble Fas Ligand (FasL), which is involved in the apoptotic loss of CD4+ T cells during HIV infection. We evaluated whether two polymorphisms in MMP-7 promoter could influence CD4+ recover in response to antiretroviral therapy, and found that these polymorphisms are ineffective.

Notes: The objective of this study was to identify single nucleotide polymorphisms (SNPs) within four functionally related immune response genes in the horse, and to develop genotyping techniques that could be useful for future genomic studies of horse infectious and allergic diseases. The genes analysed were: the lipopolysaccharide (LPS) receptor gene CD14, the toll-like receptor 4 gene TLR4, the gene Cɛ encoding the IgE heavy chain molecule and the gene FcɛR1 alpha coding for the alpha subunit of the IgE receptor molecule. Horse-specific primers amplifying selected gene regions were designed and SNPs were searched by selective resequencing and/or by PCR-SSCP (polymerase chain reaction-sequence specific conformational polymorphism) or PCR-RFLP (PCR-restriction fragment length polymorphism). Gene expression was analysed by RT-PCR (reverse transcriptase-PCR) of all four genes examined. For CD14, the cDNA sequence was determined and a novel sequence of the 5′UTR region was identified. The protein-coding sequence was identical to that previously deposited in GenBank. 5′UTR, intronic and both synonymous and non-synonymous exonic SNPs were identified. Three SNPs were found in the CD14 gene, four in the TLR4 gene; two SNPs were identified in the Cɛ gene, and one SNP was found in the FcɛR1 alpha gene. PCR-RFLP was developed for genotyping eight of the SNPs identified. The RT-PCR assay showed that all the SNPs reported here are parts of expressed genes. The results showed that important immunity-related genes in horses are polymorphic and that even non-synonymous SNPs with potential functional impact may occur. The methods developed for genotyping and haplotyping the SNPs identified represent, along with markers described previously, a potentially useful tool for genomic analysis of the function and role of these genes in immunity and in mechanisms of disease.

Notes: Recent studies have reported the association of a pro-inflammatory profile of genetic polymorphisms in IL-1B, IL-1RN, TNF-A, and IL-10 genes with an increased risk of non-cardia gastric cancer. Because gastric cancer and duodenal ulcer are mutually exclusive outcomes of Helicobacter pylori infection, we aimed to investigate possible allelic variant associations of several functional polymorphisms in the IL-1B, IL-1RN, TNFA, and LTA genes in the susceptibility to duodenal ulcer. Genomic DNA from 118 patients with duodenal ulcer and 97 healthy controls was typed for the IL-1B polymorphisms at positions −511, −31, and +3954, the VNTR polymorphism in intron 2 of the IL-1RN gene, the TNFA−308, TNFA −238, and the NcoI and BsI LTA polymorphisms by PCR, SSCP and TaqMan assays. H. pylori infection and non-steroidal anti-inflammatory drugs (NSAIDs) use was investigated in patients and controls. Logistic regression analysis identified H. pylori infection (OR: 12.86; 95%CI: 3.85–43), NSAID use (OR: 11.95; 95%CI: 4.19–34.05), and family history-ulcer (OR: 3.79; 95%CI: 1.68–8.54) as independent risk factors for duodenal ulcer. When the effect of the combinations of IL-1 and TNF genotypes was studied we found that the distribution of all possible combinations of these eight polymorphisms was similar in duodenal ulcer patients and controls. The simultaneous carriage of alleles IL-1RN*2/IL-1B −31T/IL-1B −511C/IL-IB +3954C/TNF-HaplotypeE negative (termed in some studies as ‘low-producing’ alleles) was increased in H. pylori-positive duodenal ulcer patients compared to H. pylori-infected healthy controls (10.5% vs. 5.9%) although the difference did not reach statistical significance (OR: 1.85; 95%CI: 0.57–5.99, P = 0.41). Moreover, no differences were found with respect to H. pylori status, NSAID use, age, gender, smoking habit, type of complication, recurrence of the ulcer, and need for surgical treatment. Our data show no association between allelic variants of IL-1 and TNF gene polymorphisms in the susceptibility to and final outcome of duodenal ulcer.

Notes: A novel DRB1*15 allele, DRB1*1516, has been identified in a Guangdong Han individual. Its sequence was confirmed by sequencing of polymerase chain reaction products and clones. This allele differed by one nucleotide from DRB1*150101 at position 220 (G→A), resulting in an amino acid substitution from Gly to Arg at codon 45.

Notes: The contribution of the tumour necrosis factor (TNF) B + 252 (TNFB) dimorphism and microsatellite polymorphisms of TNFa and TNFb to the pathogenesis of systemic lupus erythematosus (SLE) was studied in Japanese patients. The TNFB dimorphism was determined using the restriction fragment length polymorphism (RFLP) method with NcoI digestion followed by specific polymerase chain reaction (PCR) amplification. TNFa and TNFb microsatellite polymorphisms were determined using the DNA sequencer and GeneScan program (Applera Corporation, Foster City, CA) followed by specific PCR amplification. HLA-DRB1*15 typing was carried out by the PCR-sequence specific conformational polymorphism (SSCP) method. In SLE, the allele frequency of TNFB*2 significantly increased (68.9%, P 〈 0.05) and the genotype frequency of TNFB*2/2 also increased (52.8%, P 〈 0.05). TNFB*2 showed no significant linkage disequilibrium with HLA-DRB1*1501. The prevalence of TNFa13 and TNFb4 showed very slight increases, but these increases were not significant. An association analysis indicated that TNFB*2/2 conferred greater, or at least equal, susceptibility to SLE in Japanese patients in comparison with HLA-DRB1*1501. The TNFB*2/2 genotype may contribute additively with DRB1*1501 to SLE in Japanese patients. No association was observed between auto-antibodies and TNF. TNFB*2 is a genetic marker for SLE in Japanese patients, while TNFa and TNFb microsatellites are not associated with SLE.