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  • Biochemistry and Biotechnology  (3,415)
  • Engineering  (2,990)
  • Inorganic Chemistry
  • 2015-2019  (34)
  • 1995-1999  (8,153)
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  • 1
    Publication Date: 2021-05-19
    Description: Aleksey Sergeevich Vinnov (25.05.1958 – 04.06.2019) was a Candidate of Sciences (Engineering), an Associate Professor, a talented lecturer, a rector of the Kerch Maritime Technological Institute (KMTI) in 2000–2005, a Senior Researcher in the FSBSI “Southern Scientific Research Institute of Marine Fisheries and Oceanography” (YugNIRO), and a Deputy Head for Operation and Quality of the Limited Liability Company under the Laws of Russian Federation “Aquamarine”. A.S. Vinnov made a great contribution into the development of education and science in the field of fisheries. Aleksey Sergeevich began his professional life as a Junior Researcher in the Astrakhan Technical Institute of Fishing Industry and Economy and defended his Candidate's Thesis in 1988. He dedicated the major part of his working career to the Kerch State Maritime Technological University, where he had been employed for 21 years, taking positions from a senior lecturer of the Department of Fish Processing Technology, an Associate Professor, the Dean of the Technological Faculty, the vice-rector for academic affairs, to the rector of the University. A.S. Vinnov was known for his rational thinking and professional integrity; he was diplomatic and considerate, and proved to be a competent leader. Vinnov's research works were dedicated to the issues of development of production technology for canned fish and fish protein mass, of kinetics of enzymatic hydrolysis, and many other subjects. A.S. Vinnov was honored with several badges of distinction due to his high professional expertise and personal contribution to the development of fisheries field, particularly education.
    Description: Алексей Сергеевич Виннов (25.05.1958 – 04.06.2019 гг.) — кандидат технических наук, доцент, талантливый преподаватель, ректор Керченского морского технологического института с 2000 по 2005 г. (КМТИ), старший научный сотрудник ФГБНУ «Южный научно-исследовательский институт рыбного хозяйства и океанографии» (ЮгНИРО), заместитель директора по производству и качеству ООО «Аквамарин». А.С. Виннов внес огромный вклад в развитие образования и науки рыбной отрасли. Свою трудовую деятельность Алексей Сергеевич начал в должности младшего научного сотрудника Астраханского технического института рыбной промышленности и хозяйства, успешно защитив в 1988 г. кандидатскую диссертацию. Большую часть своей трудовой деятельности он посвятил Керченскому государственному морскому технологическому университету, проработав здесь 21 год в должностях от старшего преподавателя кафедры технологии рыбных продуктов, доцента, декана технологического факультета, проректора по учебной работе до ректора университета. А.С. Виннов обладал рациональным мышлением, был очень грамотным, тактичным человеком, умелым руководителем. Научная деятельность Алексея Сергеевича была посвящена изучению вопросов усовершенствования технологии рыбных консервов, рыбных белковых масс, кинетики ферментативного гидролиза, а также многим другим. За личный вклад в развитие образования и рыбной отрасли, за высокий профессионализм А.С. Виннов был удостоен ряда почетных знаков отличия.
    Description: Published
    Keywords: Researchers ; Engineering ; Fish processing ; Canned food ; Rector ; Proteins ; Hydrolysis
    Repository Name: AquaDocs
    Type: Journal Contribution , Refereed
    Format: pp.91-95
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  • 2
    Publication Date: 2021-05-19
    Description: Наряду с исследованиями по определению сырьевой базы Азово-Черноморского бассейна, при создании института АзЧерНИРО в 1933 г. была сразу организована научно-исследовательская лаборатория техники лова рыбы. За все время функционирования этого подразделения такие специалисты, как С.С. Виннов, О.И. Саковец, С.Я. Наместников, Е.Е. Шапунов, В.М. Кириллов, В.Г. Герасимов, В.С. Долбиш, Н.Г. Думин, В.Г. Васильев, А.А. Яковлев, Е.С. Деньгин, В.И. Абакаров, В.Н. Миронов, В.В. Стрельцов, А.С. Вайнерман, В.К. Яшкин, Ю.В. Шишов, А.М. Стафикопуло в течение многих лет вносили огромный вклад в развитие рыболовства в Азово-Черноморском бассейне и Мировом океане. До 1950 г. траловый промысел в Черном море отсутствовал, хотя были известны попытки внедрить этот вид лова в черноморское рыболовство. Так, в начале 1909 г. в северо-западной части Черного моря работал траулер «Федя». К концу 1911 г. количество траулеров, которые работали в данном регионе моря, увеличилось до 9 судов. Уловы состояли на 98-99 % из осетровых и на 1-2 % из камбалы. Ввиду очень большого вылова молоди осетровых рыб траловый промысел вскоре был запрещен. В 1932 г. из Мурманска для выяснения эффективности тралового лова в Черном море был послан рыболовный траулер «Абрек». Почти у всего черноморского побережья пробовали ловить отечественным промысловым тралом, применяемым в Баренцевом море для добычи трески. При этом отмечалось, что уловы камбалы были очень низкими. Очевидно, одной из причин была неприспособленность самой конструкции трала для лова этого вида рыб. В конце 1933 г. траулер вернулся в Мурманск. В 1949 г. Черноморская научно-промысловая экспедиция возобновила работы по освоению тралового лова в Черном море. Перед экспедицией стояла задача всесторонне изучить ихтиофауну моря и выяснить возможности применения ряда поисковых орудий лова, в том числе донного трала. С этой целью было разработано и испытано несколько типов донных тралов. С конца 1949 г. были начаты исследования по разработке конструкции разноглубинного трала для Черного моря. К тому времени инженерами М.К. Кокоревым, В.Ф. Шушпановым и А.Н. Потехиным была разработана специальная подъемно-распорная система. В соответствии с ее техническими данными в дальнейшем проектировались сетные части трала. Первый проект опытного рыболовного разноглубинного морского трала разрабатывался для лова мелких черноморских пелагических рыб в толще воды, на глубинах от 0 до 100 м, с одного судна. Большой вклад в развитие тралового промысла в Черном море внес А.Н. Самарянов.
    Description: Published
    Keywords: Engineering ; Fishing gear ; Fishing fleet ; Fishing vessels ; Trawlers ; Anchovy fisheries ; Sprat fisheries ; Purse fishing ; Commercial species ; Mullets
    Repository Name: AquaDocs
    Type: Conference Material , Non Refereed
    Format: pp.247-254
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  • 3
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2018-08-17
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 4
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2018-08-17
    Description: Polyamide thickness and roughness have been identified as critical properties that affect thin-film composite membrane performance for reverse osmosis. Conventional formation methodologies lack the ability to control these properties independently with high resolution or precision. An additive approach is presented that uses electrospraying to deposit monomers directly onto a substrate, where they react to form polyamide. The small droplet size coupled with low monomer concentrations result in polyamide films that are smoother and thinner than conventional polyamides, while the additive nature of the approach allows for control of thickness and roughness. Polyamide films are formed with a thickness that is controllable down to 4-nanometer increments and a roughness as low as 2 nanometers while still exhibiting good permselectivity relative to a commercial benchmarking membrane.
    Keywords: Engineering
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  • 5
    Publication Date: 2018-09-07
    Description: Zeolitic imidazolate framework (ZIF) membranes are emerging as a promising energy-efficient separation technology. However, their reliable and scalable manufacturing remains a challenge. We demonstrate the fabrication of ZIF nanocomposite membranes by means of an all-vapor-phase processing method based on atomic layer deposition (ALD) of ZnO in a porous support followed by ligand-vapor treatment. After ALD, the obtained nanocomposite exhibits low flux and is not selective, whereas after ligand-vapor (2-methylimidazole) treatment, it is partially transformed to ZIF and shows stable performance with high mixture separation factor for propylene over propane (an energy-intensive high-volume separation) and high propylene flux. Membrane synthesis through ligand-induced permselectivation of a nonselective and impermeable deposit is shown to be simple and highly reproducible and holds promise for scalability.
    Keywords: Engineering
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 6
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2018-09-14
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 7
    Publication Date: 2018-09-14
    Description: Insects are among the most agile natural flyers. Hypotheses on their flight control cannot always be validated by experiments with animals or tethered robots. To this end, we developed a programmable and agile autonomous free-flying robot controlled through bio-inspired motion changes of its flapping wings. Despite being 55 times the size of a fruit fly, the robot can accurately mimic the rapid escape maneuvers of flies, including a correcting yaw rotation toward the escape heading. Because the robot’s yaw control was turned off, we showed that these yaw rotations result from passive, translation-induced aerodynamic coupling between the yaw torque and the roll and pitch torques produced throughout the maneuver. The robot enables new methods for studying animal flight, and its flight characteristics allow for real-world flight missions.
    Keywords: Engineering
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 8
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2018-04-27
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 9
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2018-09-14
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 10
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2018-03-09
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 11
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2018-01-19
    Keywords: Engineering
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    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 12
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-06-23
    Keywords: Engineering
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    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 13
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-06-23
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 14
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-06-16
    Keywords: Engineering
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 15
    Publication Date: 2017-06-23
    Description: Exoskeletons and active prostheses promise to enhance human mobility, but few have succeeded. Optimizing device characteristics on the basis of measured human performance could lead to improved designs. We have developed a method for identifying the exoskeleton assistance that minimizes human energy cost during walking. Optimized torque patterns from an exoskeleton worn on one ankle reduced metabolic energy consumption by 24.2 ± 7.4% compared to no torque. The approach was effective with exoskeletons worn on one or both ankles, during a variety of walking conditions, during running, and when optimizing muscle activity. Finding a good generic assistance pattern, customizing it to individual needs, and helping users learn to take advantage of the device all contributed to improved economy. Optimization methods with these features can substantially improve performance.
    Keywords: Engineering
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  • 16
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-11-24
    Keywords: Engineering
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 17
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-12-01
    Description: Kim et al . (Reports, 28 April 2017, p. 430) presented results for the solar-driven harvesting of water from air via metal-organic frameworks as a prodigious potential advance toward remedying global water shortages. Basic thermodynamics and a survey of multiple off-the-shelf technologies show that their approach is vastly inferior in efficiency (and thereby in feasibility) to available alternatives.
    Keywords: Engineering
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  • 18
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-12-01
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 19
    Publication Date: 2017-12-01
    Description: In their comment, Bui et al . argue that the approach we described in our report is vastly inferior in efficiency to alternative off-the-shelf technologies. Their conclusion is invalid, as they compare efficiencies in completely different operating conditions. Here, using heat transfer and thermodynamics principles, we show how Bui et al .’s conclusions about the efficiencies of off-the-shelf technologies are fundamentally flawed and inaccurate for the operating conditions described in our study.
    Keywords: Engineering
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  • 20
    Publication Date: 2017-05-12
    Description: Bacteria within communities can interact to organize their behavior. It has been unclear whether such interactions can extend beyond a single community to coordinate the behavior of distant populations. We discovered that two Bacillus subtilis biofilm communities undergoing metabolic oscillations can become coupled through electrical signaling and synchronize their growth dynamics. Coupling increases competition by also synchronizing demand for limited nutrients. As predicted by mathematical modeling, we confirm that biofilms resolve this conflict by switching from in-phase to antiphase oscillations. This results in time-sharing behavior, where each community takes turns consuming nutrients. Time-sharing enables biofilms to counterintuitively increase growth under reduced nutrient supply. Distant biofilms can thus coordinate their behavior to resolve nutrient competition through time-sharing, a strategy used in engineered systems to allocate limited resources.
    Keywords: Engineering
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  • 21
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-05-12
    Keywords: Engineering
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    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 22
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-02-10
    Description: Author: Jake Yeston
    Keywords: Inorganic Chemistry
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 23
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-01-27
    Description: Author: Jake Yeston
    Keywords: Inorganic Chemistry
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 24
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-02-17
    Description: Engineering has an image problem. The phrase "engineering disaster" rolls off the tongue, while great technical achievements are more often heralded as "scientific miracles." Enter Dream Big. Sponsored by the American Society of Civil Engineers with support from Bechtel Corporation, the film sets out to reframe engineering as a force for good and a profession in service to people and the planet. Author: Donna Riley
    Keywords: Engineering
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  • 25
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-07-21
    Keywords: Engineering
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 26
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-07-21
    Keywords: Engineering
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    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 27
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-05-26
    Keywords: Engineering
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  • 28
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-11-10
    Keywords: Engineering
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  • 29
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-01-27
    Description: Polynitrogens have the potential for ultrahigh-performing explosives or propellants because singly or doubly bonded polynitrogens can decompose to triply bonded dinitrogen (N2) with an extraordinarily large energy release. The large energy content and relatively low activation energy toward decomposition makes the synthesis of a stable polynitrogen allotrope an extraordinary challenge. Many elements exist in different forms (allotropes)—for example, carbon can exist as graphite, diamond, buckyballs, or graphene. However, no stable neutral allotropes are known for nitrogen, and only two stable homonuclear polynitrogen ions had been isolated until now—namely, the N3− anion (1) and the N5+ cation (2). On page 374 of this issue, Zhang et al. (3) report the synthesis and characterization of the first stable salt of the cyclo-N5− anion, only the third stable homonuclear polynitrogen ion ever isolated. Author: Karl O. Christe
    Keywords: Inorganic Chemistry
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  • 30
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2016-07-08
    Description: The Internet connects billions of computational platforms of various sizes, from supercomputers to smart phones. However, the same types of data transmission can connect computational resources to much simpler sensors “at the edge of the net” that collect, analyze, and transmit data, as well as controllers that receive instructions. Devices deployed in the environment, homes and offices, and even our bodies would expand the number of connected devices to the trillions. This “Internet of Things” (IoT) underlies the vision of smart homes and buildings that could sense and transmit their status and respond appropriately (1), or track and report on the state of objects (vehicles, goods, or even animals) in the environment. However, the practical implementation of the IoT has been relatively slow, in part because all of these edge devices must draw electrical power from their local environment. We analyze the use of photovoltaics (PV) to power devices and help bring the IoT to fruition. Authors: Richard Haight, Wilfried Haensch, Daniel Friedman
    Keywords: Engineering
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  • 31
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2016-04-22
    Description: So prized by the ancient Romans were Egyptian obelisks that, at one time, more of them stood in Rome than in Egypt. In the 19th century, France, Britain, and the United States—inspired by Napoleon Bonaparte's expedition to Egypt in 1798— acquired their own major obelisks from Alexandria and Luxor. Cleopatra's Needles, by Egyptologist Bob Brier, explores the engineering challenges associated with building and erecting these massive monuments. Author: Andrew Robinson
    Keywords: Engineering
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  • 32
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2016-10-21
    Description: A physicist reveals the engineering marvels that underlie the modern metropolis Author: Sybil Derrible
    Keywords: Engineering
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2016-09-09
    Description: Author: Jake Yeston
    Keywords: Inorganic Chemistry
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2016-11-11
    Description: Author: Jake Yeston
    Keywords: Inorganic Chemistry
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 131-131 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: No Abstract
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 123-130 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: This paper presents the electrode separation method for the boundary condition of a-Si TFT mixed-level simulation. The Poisson equation and the continuity equation are formulated into equivalent circuits. So, a circuit simulator can be used to handle the two-dimensional numerical simulation of a-Si TFT. The boundary condition problem between a semiconductor and an external circuit is solved by the electrode separation method. An electrode is separated into two nodes to fit Kirchhoff's current law and the semiconductor equations, respectively. A simple a-Si TFT/LCD circuit is taken as an example for the electrode separation method. For mixed-level simulation this technique is very useful. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 133-151 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: Most traditional theories of speech production are currently based on plane waves. However, it is well known that, for acoustic waveguides, higher acoustical modes start to propagate and can become predominant above cut-on frequencies. This paper thus presents the transmission line matrix method, a numerical method initially designed for electromagnetic waves, and its adaptation to acoustic waveguides. The method, and in particular the representation of boundary conditions, is validated by comparison with known analytical theories. It is then used to show the dramatic effect of higher order modes upon the radiation characteristics of uniform ducts, as well as the importance of source location. Finally, first applications to bent and bifurcating rectangular ducts are presented, and the transfer function of a vowel [a] is shown to display frequency patterns typical of those measured on human subjects and that cannot be explained by one-dimensional propagation only. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 189-205 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: The atomic mixing model that forms the basis of the IMPETUS software is described in detail. The model simulates the mixing and particle emission that occurs when a solid is bombarded with energetic particles, such as in SIMS or SNMS. The methods employed for computing the deposition of the bombarding particles and their energies along with the modelling of the particle yield and the surface recession speed are described.The material volume concentrations are governed by a set of partial differential equations. A description of the finite element method that is employed for their solution is given. Results from the application of IMPETUS II to a number of typical structures are given. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 3-19 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: This paper presents an efficient space domain integral formulation of planar dielectric structures based on the concepts of multiresolution analysis (MRA) theory. Battle-Lemarie multiresolution expansions are utilized in the moment method solution of a volume integral equation for the unknown electric field. The generation of sparse moment matrices is explained in view of the cancellation property of wavelet basis functions. The formulation is then applied to two- and three-dimensional dielectric structures including waveguides and resonators, and the numerical results and consequences of matrix sparsity are discussed in detail. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 21-40 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: An efficient wavelet-packet-based time-frequency technique is proposed in this paper. This technique has found applications in inverse synthetic aperture radar (ISAR). It combines the Wigner-Ville distribution (WVD) of each sub-band signal partitioned from an input signal by the wavelet-packet transform (WPT). These distributions are used to estimate the parameters in Doppler processing. Similar parametric compensation is carried out in range processing so that a specific scatterer is focused in both range and cross-range. The end result uses the Fourier imaging method to obtain a high-resolution radar image. This technique is primarily based on estimation and compensation of the Doppler scattering parameters of the dominant scatterers for improved focus. It can be of great value to the scattering-oriented ISAR imaging. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 273-274 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 255-257 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: We re-examine the potential drop method to give a direct estimation of the absolute error in transmission-line matrix (TLM) diffusion modelling. In particular, the open-circuit boundary is taken into account. Promising results are presented with a structure relevant to thermal analysis of semiconductor devices. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 259-271 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: There is a class of material processing simulators which require solution of the diffusion equation over a surface which is evolving due to some physical or chemical process related to the concentration of the diffusing species. Because of its explicit and unconditionally stable nature, the transmission line matrix (TLM) method is well suited to efficiently solve the diffusion equation in these instances. However, the methodology for using TLM on a dynamic problem space is not well established.This paper describes the development of algorithms to handle the insertion, deletion and motion of TLM nodes along the one-dimensional surface of a two-dimensional thin film process simulator called GROFILMS. These routines are completely compatible with concurrent time scaling to increase computational efficiency. Central to this development is the use of an asymmetric TLM cell which provides more flexibility for representation and minimizes the disturbance effects of these nodal operations. The scaling routines apply conservation of charge and continuity of current into the affected region to recompute the scattered or the incident pulses as a result of the evolving network. Verification of these results through comparison with known analytic solutions is achieved where possible. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 41-54 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: The local support and vanishing moment property of wavelet bases have been recently used to obtain a sparse matrix representation of integral equations in the spatial domain. In this paper, an application of the cubic spline and the corresponding semi-orthogonal wavelets in the spectral domain is proposed for the evaluation of the reflection coefficient for open/short transmission lines. Because of the nearly optimal time (space)-frequency (wavenumber)-window product of the cubic spline and wavelet, the double spectral integrals appearing in the formulation can be computed more efficiently than with the commonly used piecewise sinusoidal (PWS) or triangular basis functions. It is shown that the time-frequency-window product of the triangular and PWS function are close to each other, whereas those of the cubic spline/wavelet are close to 0·5, the lowest possible value corresponding to functions of Gaussian class. Both the PWS and wavelet bases are applied to microstrip and coplanar waveguides with isotropic and anisotropic substrates, and the results are compared with published theoretical and experimental data. It is observed that even though the number of splines/wavelets required for an accurate representation of the current distribution in the transmission line is almost twice as high as the number of PWS functions, the overall computation time decreases significantly in the former case. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 69-84 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: The use of wavelet expansions in numerical solutions of electromagnetic frequency-domain integral equation formulations is steadily growing. In this paper we review the recently suggested impedance matrix compression (IMC) method for a more effective integration of wavelet-based transforms into existing numerical solvers. The difference between the IMC method and the previous approaches to applying wavelets in computational electromagnetics is twofold. Firstly, the transformation is effected by means of a digital filtering approach. This approach renders the transform algorithm adaptive and facilitates the derivation of a basis which best suits the problem at hand. Secondly, the conventional thresholding procedure applied to the impedance matrix is substituted for by a compression process in which only the significant terms in the expansion of the (yet-unknown) current are retained and hence a substantially smaller number of coefficients has to be determined. A few numerical results are included to demonstrate the advantages of the presented method over the currently used ones. The feasibility of ensuring a slow growth in the number of unknowns even when there is a rapid increase in the problem complexity is shown by an illustrative example. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 87-103 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: An efficient novel algorithm is introduced for ground wave propagation problems. First, ground wave propagation characteristics for a vertically polarized short electric dipole over a smooth spherical earth are reviewed, reducing the vector electromagnetic problem for the three-dimensional spherical geometry to an equivalent two-dimensional rectilinear scalar potential problem which is solved by spectral analysis and synthesis. Alternative evaluations of the spectral integral yield ray optical and normal mode solutions, which are conventionally referred to as the Norton and Wait formulations, respectively. Combining these formulations in an efficient manner yields a hybrid algorithm which is constructed so as to account adaptively for the characteristics of ground wave propagation in interference, intermediate and diffraction regions (including mixed paths) for various source and/or receiver heights. Numerical comparisons are made with reference results obtained via the parabolic equation (PE) method, in parametric ranges where PE is reliable; this permits assessment of the effectiveness of the hybrid approach. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 183-184 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: No Abstract
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 153-166 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: Most methods for the numerical calculation of inverse Laplace transformations f(t) = L-1[F(s)] have serious limitations concerning the class of functions F(s) that can be inverted or the achievable accuracy. The procedures described in the paper can be used to invert rational as well as irrational or transcendental functions of the complex variable s. The required accuracy of the results can be enhanced without changing the algorithm, only at the cost of a longer computation time. The described methods were verified with many examples including transients in lumped/distributed systems with sections of lossy multiconductor transmission lines or with distributed RC elements. © 1998 John Wiley & Sons, Ltd.
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    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 167-181 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: In this study, a software package is described which has been developed for the simulation of detection and tracking in surface wave high frequency (SW-HF) radars. The aim is to investigate the problems related to detection and tracking of surface targets at beyond the horizon ranges. In SW-HF radars target detection and tracking involves stochastic features such as target RCS fluctuations, atmospheric, galactic and/or man-made ambient interference components, and the sea clutter with the dominant resonant Bragg returns which affect target detection are all incorporated in the package. In the model, first, terrain data are fed into the simulator by means of a specially designed graphical user interface. Then, a scenario is prepared where the radar's location, coverage and operational parameters can be defined, together with different targets and their sailing routes. The radial propagation paths for angular resolution cells are extracted from the terrain data with the lengths of (possible) sea-island transitions. The surface wave path losses are calculated over the smooth spherical earth's surface with finite conductivity. Surface roughness and mixed-path propagation effects are also included in these calculations. The target detection is performed in the frequency domain after calculation of the noise floor, signal to noise (SNR) and clutter to noise (CNR) ratios. Since SW-HF radars yield coarse range and azimuth but accurate velocity measurements, different Kalman filter techniques are applied for the target tracking, and algorithms are added to improve the track-measurement data correlation. Various simulation tests are performed and the results are presented. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 187-187 
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    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 277-287 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: The solution of Maxwell's equations is considered using the finite difference time domain (FDTD) method and the client-server software paradigm on a heterogeneous network of distributed computers. The performance of compilers is evaluated and the PVM 3.3 message-passing harness, in both its conventional and TCP mode of operation, is compared with direct calls to the lesser known TCP/IP routines on the operating system. An example is given showing the impact of using different floating-point representations, in each subdomain, when computing field updates. As far as we are aware to date, there has been no discussion of the actual content of the communication between concurrent FDTD processes; accordingly we have defined one such protocol. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 307-316 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: This paper presents an FE modelling approach for the calculation of transient eddy currents in thin conductive layers, where the complexity of the geometry prohibits both a detailed FE modelling and the use of analytical tools. The method allows an estimation of the maximum values of eddy currents at a highly reduced modelling effort and the use of a commercial FE software package. A prerequisite is a slowly varying magnetic field that can be assumed to be unaffected by the eddy currents. It is shown that neither the source of the magnetic field nor the insulating environment has to be modelled. The model is built up exclusively from 2D elements and is excited via the magnetic vector potential. The spatial distribution of the latter is calculated separately in a magnetostatic calculation based on Biot-Savart's law. It is then applied in time-varying form as a dynamic boundary condition at every node of the model. The method was applied to a simple problem for which the results of a detailed FE calculation were available, to document its validity. Further numerical results are presented for the plasma vessel and the heat radiation shield of the Wendelstein 7-X fusion experiment in the case of an emergency discharge. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 55-68 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: Multiresolution time domain (MRTD) analysis is applied directly to Maxwell's equations to model inhomogeneous dielectric material. In our approach, scaling and wavelet functions are used as a complete basis for the method of moments. The MRTD scheme is used to analyze different types of resonant cavity structures with varying dielectric perturbations in one, two and three dimensions. The results presented here agree very well with those obtained by FDTD, FEM and integral equation methods. MRTD allows for considerable savings in memory and computation time in comparison to FDTD, while maintaining the same accuracy of the results. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 1-1 
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    Keywords: Engineering ; Numerical Methods and Modeling
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    Topics: Electrical Engineering, Measurement and Control Technology
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 105-121 
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    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: This paper describes a computationally efficient diakoptic algorithm for the solution of closed-mesh electrical network equations. The algorithm is designed for use with time-varying and/or discontinuous equations describing limited size power supply systems, allowing them to be torn into smaller subdivisions in a manner that is independent of their structure. The efficiency of the algorithm is demonstrated by comparing its performance with that obtained when using a more conventional sparse matrix technique, on the basis of the number of floating point operations required for the solution of a range of typical network problems. Mention is made of the parallel processes inherent in the resultant equation structure. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 207-219 
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    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: The IMPETUS II code simulates the atomic mixing and particle emission that occurs when a solid is bombarded by energetic particles (as in SIMS or SNMS). The underlying model consists of a system of partial differential equations that are solved by a finite difference method (FDM). Special techniques are also employed to model thin layers and sharp interfaces, to deal efficiently with wide homogeneous layers (when the solution is tending to a steady state), to model linear diffusion in order to smooth the sharp interfaces before they enter to domain of the FDM.In this paper the special techniques are described in detail. Results from test problems, demonstrating these techniques, are shown. An algorithm that describes the way the IMPETUS II code is structured is given. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 221-229 
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    Keywords: Engineering ; Numerical Methods and Modeling
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    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: This paper presents a physical-mathematical model for abrupt heterojunction transistors and its solution using numerical methods with application to InP/InGaAs HBTs. The physical model is based on the combination of the drift-diffusion transport model in the bulk with thermionic emission and tunnelling transmission through the emitter-base interface. Fermi-Dirac statistics and bandgap narrowing distribution between the valence and conduction bands are considered in the model. A compact formulation is used that makes it easy to take into account other effects such as the non-parabolic nature of the bands or the presence of various subbands in the conduction process. The simulator has been implemented for distributed memory multicomputers, making use of the MPI message-passing standard library. In order to accelerate the solution process of the linear system, iterative methods with parallel incomplete factorization-based preconditioners have been used. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 273-273 
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    Keywords: Engineering ; Numerical Methods and Modeling
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 233-242 
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    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: The propagation factors of buried microstrip lines are considered. The associated eigenvalue problems are solved by the edge-based vectorial finite element method (VFEM). To improve the higher-order VFEM solutions, the problem infinite domain is truncated by second-order absorbing boundary conditions (ABCs). This reduces the number of unknowns and, consequently, the numerical effort. Owing to the many advantages of both the edge-based VFEM and the second-order ABCs, this full-wave method is implemented by a general computer program for solving unbounded domain problems involving complex geometries and inhomogeneous media. The examples presented show the validity of this technique. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 243-254 
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    Keywords: Engineering ; Numerical Methods and Modeling
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    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: Various local absorbing boundary conditions are implemented in the TLM method and their corresponding reflections are compared as a waveguide termination. The instability in different ABCs is discussed and reflections from the terminating walls are computed. An improvement for the matched termination wall is proposed and reflection of this absorbing condition is compared with ordinary matched termination. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 317-318 
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    Keywords: Engineering ; Numerical Methods and Modeling
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 289-298 
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    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: Oscillatory flow present in the melt during InSb single crystal growth using an RF-heating Czochralski method has been numerically investigated by means of the finite difference method using the HSMAC algorithm. The thermal boundary conditions required for the numerical simulation model were obtained experimentally by measuring the temperature profile along the crucible of a Czochralski system by means of thermocouples mounted in the crucible. Results of numerical simulations showed that the use of a third-order upwind discretization scheme was necessary to catch the oscillatory behaviour of the fluid flow in the melt. It was shown that this oscillatory behaviour strongly depends on the crystal rotation rate. Indeed, the oscillation period increases when the crystal rotation rate is above a critical rotation rate. In order to avoid such oscillations, crystal rotation rates lower than this critical value of crystal rotation rate must be selected for the growth of high quality crystals free of striations. © 1998 John Wiley & Sons, Ltd.
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    International Journal of Numerical Modelling: Electronic Networks, Devices and Fields 11 (1998), S. 299-306 
    ISSN: 0894-3370
    Keywords: Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Electrical Engineering, Measurement and Control Technology
    Notes: The application of the FDTD algorithm on generalized non-orthogonal meshes, following the basic ideas of Holland (1983), has been investigated by many authors for several years now, and detailed dispersion analysis as well as convergence studies have been published. Already in 1992 also a general stability criterion was given for the time integration using the standard leap-frog scheme (Lee et al.). Many authors, however, still propose some damped time stepping algorithms to work around unexpected instabilities in the discretization method. In this paper the origin of this type of instability is revealed, and a technique to obtain a stable discretization of Maxwell's equations on non-orthogonal grids is proposed. To obtain more insight into the stability properties of the method, it is reformulated according to the matrix-vector notation of the Finite Integration Technique. © 1998 John Wiley & Sons, Ltd.
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    Biotechnology and Bioengineering 57 (1998), S. 46-54 
    ISSN: 0006-3592
    Keywords: smooth muscle ; polyglycolic acid ; biodegradable ; tissue engineering ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The engineering of functional smooth muscle (SM) tissue is critical if one hopes to successfully replace the large number of tissues containing an SM component with engineered equivalents. This study reports on the effects of SM cell (SMC) seeding and culture conditions on the cellularity and composition of SM tissues engineered using biodegradable matrices (5 × 5 mm, 2-mm thick) of polyglycolic acid (PGA) fibers. Cells were seeded by injecting a cell suspension into polymer matrices in tissue culture dishes (static seeding), by stirring polymer matrices and a cell suspension in spinner flasks (stirred seeding), or by agitating polymer matrices and a cell suspension in tubes with an orbital shaker (agitated seeding). The density of SMCs adherent to these matrices was a function of cell concentration in the seeding solution, but under all conditions a larger number (approximately 1 order of magnitude) and more uniform distribution of SMCs adherent to the matrices were obtained with dynamic versus static seeding methods. The dynamic seeding methods, as compared to the static method, also ultimately resulted in new tissues that had a higher cellularity, more uniform cell distribution, and greater elastin deposition. The effects of culture conditions were next studied by culturing cell-polymer constructs in a stirred bioreactor versus static culture conditions. The stirred culture of SMC-seeded polymer matrices resulted in tissues with a cell density of 6.4 ± 0.8 × 108 cells/cm3 after 5 weeks, compared to 2.0 ± 1.1 × 108 cells/cm3 with static culture. The elastin and collagen synthesis rates and deposition within the engineered tissues were also increased by culture in the bioreactors. The elastin content after 5-week culture in the stirred bioreactor was 24 ± 3%, and both the elastin content and the cellularity of these tissues are comparable to those of native SM tissue. New tissues were also created in vivo when dynamically seeded polymer matrices were implanted in rats for various times. In summary, the system defined by these studies shows promise for engineering a tissue comparable in many respects to native SM. This engineered tissue may find clinical applications and provide a tool to study molecular mechanisms in vascular development. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 46-54, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 518-528 
    ISSN: 0006-3592
    Keywords: ammonium ; UDP-GlcNAc ; N -glycosylation ; BHK-21 cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of different ammonium concentrations and glucosamine on baby hamster kidney (BHK)-21 cell cultures grown in continuously perfused double membrane bioreactors was investigated with respect to the final carbohydrate structures of a secretory recombinant glycoprotein. The human interleukin-2 (IL-2) mutant glycoprotein variant IL-Mu6, which bears a novel N-glycosylation site (created by a single amino acid exchange of Gln100 to Asn), was produced under different defined protein-free culture conditions in the presence or absence of either glutamine, NH4Cl, or glucosamine. Recombinant glycoprotein products were purified and characterized by amino acid sequencing and carbohydrate structural analysis using matrix-assisted laser desorption ionization time of flight mass spectrometry, high-pH anion-exchange chromatography with pulsed amperometric detection, and methylation analysis. In the absence of glutamine, cells secreted glycoprotein forms with preponderantly biantennary, proximal fucosylated carbohydrate chains (85%) with a higher NeuAc content (58%). Under standard conditions in the presence of 7.5 mM glutamine, complex-type N-glycans were found to be mainly biantennary (68%) and triantennary structures (33%) with about 50% containing proximal α1-6-linked fucose; 37% of the antenna were found to be substituted with terminal α2-3-linked N-acetylneuraminic acid. In the presence of 15 mM exogenously added NH4Cl, a significant and reproducible increase in tri- and tetraantennary oligosaccharides (45% of total) was detected in the secretion product. In glutamin-free cultures supplemented with glucosamine, an intermediate amount of high antennary glycans was detected. The increase in complexity of N-linked oligosaccharides is considered to be brought about by the increased levels of intracellular uridine diphosphate-GlcNAc/GalNAc. These nucleotide sugar pools were found to be significantly elevated in the presence of high NH3/NH4+ and glucosamine concentrations. ©1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 518-528, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 557-570 
    ISSN: 0006-3592
    Keywords: Alcaligenes eutrophus ; polyhydroxyalkanoates ; metabolic engineering ; mathematical modeling ; enzyme kinetics ; regulation of metabolism ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model describing intracellular polyhydroxybutyrate (PHB) synthesis in Alcaligenes eutrophus has been constructed. The model allows investigation of issues such as the existence of rate-limiting enzymatic steps, possible regulatory mechanisms in PHB synthesis, and the effects different types of rate expressions have on model behavior. Simulations with the model indicate that activities of all PHB pathway enzymes influence overall PHB flux and that no single enzymatic step can easily be identified as rate limiting. Simulations also support regulatory roles for both thiolase and reductase, mediated through AcCoA/CoASH and NADPH/NADP+ ratios, respectively. To make the model more realistic, complex rate expressions for enzyme-catalyzed reactions were used which reflect both the reversibility of the reactions and the reaction mechanisms. Use of the complex kinetic expressions dramatically changed the behavior of the system compared to a simple model containing only Michaelis-Menten kinetic expressions; the more complicated model displayed different responses to changes in enzyme activities as well as inhibition of flux by the reaction products CoASH and NADP+. These effects can be attributed to reversible rate expressions, which allow prediction of reaction rates under conditions both near and far from equilibrium. ©1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 557-570, 1998.
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  • 68
    ISSN: 0006-3592
    Keywords: rhG-CSF ; fusion protein ; secretion efficiency ; glycosylation ; multimer ; conformation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The synthesis and secretion of recombinant human granulocyte colony-stimulating factor (rhG-CSF) are investigated in fed-batch cultures at high cell concentration of recombinant Saccharomyces cerevisiae, and some important characteristics of the secreted rhG-CSF are demonstrated. Transcription of the recombinant gene is regulated by a GAL1-10 upstream activating sequence (UASG), and the rhG-CSF is expressed in a hybrid fusion protein consisting of signal sequence of Kluyveromyces lactis killer toxin and N-terminal 24 amino acids of human interleukin 1β. The intracellular KEX2 cleavage leads to excretion of mature rhG-CSF into extracellular culture broth, and the cleavage process seems to be highly efficient. In spite of relatively low copy number the plasmid propagation is stably maintained even at nonselective culture conditions. The rhG-CSF synthesis does not depend on galactose level, whereas the production of extracellular rhG-CSF was significantly enhanced by increasing the inducer concentration above a certain level and also by supplementing the nonionic surfactant to the culture medium, which is notably due to the enhanced secretion efficiency. Various immunoblotting analyses demonstrate that none of the rhG-CSF is accumulated in the cell wall fraction and that a significant amount of intracellular rhG-CSF antibody-specific immunoreactive proteins is located in the ER. A core N-glycosylation at fused IL-1β fragment is likely to play a critical role in directing the high-level secretion of rhG-CSF, and the O-glycosylation of secreted rhG-CSF seems nearly negligible. Also the extracellular rhG-CSF is observed to exist as various multimers, and the nature of molecular interaction is evidently not the covalent disulfide bridges. The CD spectra of purified rhG-CSF and Escherichia coli-derived standard show that the conformations of both are similar and are almost identical to that reported for natural hG-CSF. ©1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 600-609, 1998.
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    Biotechnology and Bioengineering 58 (1998), S. 101-116 
    ISSN: 0006-3592
    Keywords: biofilm ; structure ; shape ; surface ; cellular automata ; discrete ; modeling ; roughness ; fractal ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A hybrid differential-discrete mathematical model has been used to simulate biofilm structures (surface shape, roughness, porosity) as a result of microbial growth in different environmental conditions. In this study, quantitative two- and three-dimensional models were evaluated by introducing statistical measures to characterize the complete biofilm structure, both the surface structure and volume structure. The surface enlargement, coefficient of roughness, fractal dimension of surface, biofilm compactness, and solids hold-up were found to be good measures of biofilm structure complexity. Among many possible factors affecting the biofilm structure, the influence of biomass growth in relation to the diffusive substrate transport was investigated. Porous biofilms, with many channels and voids between the “finger-like” or “mushroom” outgrowth, were obtained in a substrate-transport-limited regime. Conversely, compact and dense biofilms occurred in systems limited by the biomass growth rate and not by the substrate transfer rate. The surface complexity measures (enlargement, roughness, fractal dimension) all increased with increased transport limitation, whereas the volume measures (compactness, solid hold-up) decreased, showing the change from a compact and dense to a highly porous and open biofilm. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:101-116, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 155-163 
    ISSN: 0006-3592
    Keywords: endogenous respiration ; activated sludge ; multi-time scales ; identifiability ; observability ; model reduction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this article, an autonomous four-compartment model that describes the endogenous respiration in an aerobic biodegradation process is proposed and analyzed theoretically. First, the multi-time scale of the system's behavior, to be taken into account in subsequent analyses, is emphasized. Then, an identifiability and observability study, given measurements of MLVSS (mixed liquor volatile suspended solids) and respiration rate, is performed for use under practical circumstances, such as in state and parameter estimation. It appears that the process is observable, but not fully identifiable. Hence, for the identification of some of the model parameters, additional measurements or experiments, also indicated here, have to be performed. Furthermore, it is shown that, under quasi-steady state conditions which, in general, appear shortly after initialization of an endogenous respiration experiment, the model can be reduced significantly. Finally, results of parameter estimation from available data are presented and discussed. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 155-163, 1998.
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  • 71
    ISSN: 0006-3592
    Keywords: hybridoma ; futile cycling ; hollow fiber bioreactor ; glutamine ; NMR ; C-13 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of changes in extracellular glutamine level on metabolism of a murine hybridoma was examined with in vivo nuclear magnetic resonance (NMR) spectroscopy. Cells were cultured in a hollow-fiber bioreactor at high cell density to allow intracellular metabolite levels to be determined on a metabolically relevant time scale. Steady infusions of [1-13C] glucose were used to label glycolytic and tricarboxylic acid cycle intermediates, which permitted continuous monitoring with NMR spectroscopy during changes in environmental glutamine level. Samples of the extracellular medium were also analyzed to determine the effect of glutamine on other metabolites associated with primary and secondary metabolism. The changes in glutamine concentration had several effects on primary and secondary metabolism, depending on the rate the changes were made. For a brief reduction in feed glutamine concentration from 4 to 0 mM (which produced a rapid change from 0.67 to ∼0 mM in residual glutamine), large changes were observed in the rate of consumption of metabolites normally associated with energy production. Antibody synthesis was strongly stimulated and nitrogen metabolism was significantly altered. For a more prolonged reduction from 2.4 to 1.2 mM (which produced a slower reduction from 0.30 to 0.08 mM in residual glutamine), much smaller changes were observed even though the concentration of glutamine at the reduced feed level was very low. Energy metabolism did not appear to be limited by glutamine at 0.08 mM, which suggests that significant futile cycling may occur in energy producing pathways when excess glucose and glutamine are available. However, this concentration of extracellular glutamine appeared to affect some anabolic pathways, which require amino groups from glutamine. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 172-186, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 211-215 
    ISSN: 0006-3592
    Keywords: protein ; conformational memory ; organic solvent ; molecular imprinting ; enzyme ; catalysis ; transition state analogue ; bovine serum albumin ; β-lactoglobulin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The induction of catalytic activity in proteins by lyophilization in the presence of a transition state analogue (biomolecular imprinting) has been attempted. It was shown that proteins which were freeze-dried with n-isopropyl-4-nitrobenzyl-amine (a transition state analogue for the reaction of dehydrofluorination of 4-fluoro-4-[p-nitrophenyl] butan-2-one) displayed higher β-elimination activity as compared to their-non-imprinted counterparts. It was also found that native bovine serum albumin has a high dehydrofluorination activity towards the above substrate with kinetic parameters rather similar to those of a catalytic antibody prepared by Shokat et al. (1989). A comparison of the kinetic parameters determined in this study with those obtained for analogous catalytic antibodies and imprinted polymers was made. © 1998 John Wiley & Sons, Inc. Biotechnol. Bioeng. 57: 211-215, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 272-279 
    ISSN: 0006-3592
    Keywords: biofilm ; plasmid transfer ; conjugation ; retrotransfer ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A strain of Pseudomonas putida harboring plasmids RK2 and pDLB101 was exposed to a pure culture biofilm of Bacillus azotoformans grown in a rotating annular reactor under three different concentrations of the limiting nutrient, succinate. Experimental results demonstrated that the broad host range RSF1010 derivative pDLB101 was transferred to and expressed by B. azotoformans. At the lower concentrations, donor mediated plasmid transfer increased with increasing nutrient levels, but the highest nutrient concentration yielded the lowest rate of donor to recipient plasmid transfer. For transconjugant initiated transfer, the rate of transfer increased with increasing nutrient concentrations for all cases. At the lower nutrient concentrations, the frequency of plasmid transfer was higher between donors and recipients than between transconjugants and recipients. The reverse was true at the highest succinate concentration. The rates and frequencies of plasmid transfer by mobilization were compared to gene exchange by retrotransfer. The initial rate of retrotransfer was slower than mobilization, but then increased dramatically. Retrotransfer produced a plasmid transfer frequency more than an order of magnitude higher than simple mobilization. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 272-279, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 280-286 
    ISSN: 0006-3592
    Keywords: biofilm ; plasmid transfer ; conjugation ; mathematical models ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A strain of Pseudomonas putida that harbors plasmids RK2 and pDLB101 was exposed to a pure culture biofilm of Bacillus azotoformans grown in a rotating annular reactor. Transfer of the RK2 mobilizable pDLB101 plasmid to B. azotoformans was monitored over a 4-day period. Experimental results demonstrated that the broad host range, RSF1010 derivative pDLB101 was transferred to and expressed by B. azotoformans. In the companion article to this work, the rate of plasmid transfer was quantified as a function of the limiting nutrient, succinate, and as a function of the mechanism of transfer. A biofilm process simulation program (AQUASIM) was modified to analyze resultant experimental data. Although the AQUASIM package was not designed to simulate or predict genetic events in biofilms, modification of the rate process dynamics allowed successful modeling of plasmid transfer. For the narrow range of substrate concentrations used in these experiments, nutrient level had only a slight effect on the rate and extent of plasmid transfer in biofilms. However, further simulations using AQUASIM revealed that under nutrient poor conditions, the number of transconjugants appearing in the biofilm was limited. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 280-286, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 314-320 
    ISSN: 0006-3592
    Keywords: Phaffia rhodozyma ; chemostat ; continuous fermentation ; astaxanthin ; peat ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Response surface methodology was applied to optimize the growth of the yeast Phaffia rhodozyma in continuous fermentation using peat hydrolysates as substrate. A second-order, complete, factorial design of the experiments was used to develop empirical models providing a quantitative interpretation of the relationships between the two variables studied, dilution rate and pH. Maximum biomass concentration in the fermentor was obtained by employing the following predicted optimum fermentation conditions: a dilution rate of 0.017/h and a pH level of 7.19. A verification experiment, conducted at previously optimized conditions for maximum biomass volumetric productivity (a dilution rate of 0.022/h, and a pH level of 6.90), produced values for biomass concentration, residual substrate concentration, biomass yield, and biomass volumetric productivity that were very close to the predicted values, indicating the reliability of the empirical model. The concentration of the pigment astaxanthin produced by the yeast under the optimized growth conditions was found to be 544 mg astaxanthin/kg dry cell biomass. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 314-320, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 330-341 
    ISSN: 0006-3592
    Keywords: brewers' yeast ; collision theory ; flocculation ; modeling ; surface erosion ; floc splitting ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Flocculation of yeast cells occurs during the fermentation of beer. Partway through the fermentation the cells become flocculent and start to form flocs. If the environmental conditions, such as medium composition and fluid velocities in the tank, are optimal, the flocs will grow in size large enough to settle. After settling of the main part of the yeast the green beer is left, containing only a small amount of yeast necessary for rest conversions during the next process step, the lagering. The physical process of flocculation is a dynamic equilibrium of floc formation and floc breakup resulting in a bimodal size distribution containing single cells and flocs.The floc size distribution and the single cell amount were measured under the different conditions that occur during full scale fermentation. Influences on flocculation such as floc strength, specific power input, and total number of yeast cells in suspension were studied. A flocculation model was developed, and the measured data used for validation. Yeast floc formation can be described with the collision theory assuming a constant collision efficiency. The breakup of flocs appears to occur mainly via two mechanisms, the splitting of flocs and the erosion of yeast cells from the floc surface. The splitting rate determines the average floc size and the erosion rate determines the number of single cells. Regarding the size of the flocs with respect to the scale of turbulence, only the viscous subrange needs to be considered. With the model, the floc size distribution and the number of single cells can be predicted at a certain point during the fermentation. For this, the bond strength between the cells, the fractal dimension of the yeast, the specific power input in the tank and the number of yeast cells that are in suspension in the tank have to be known. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 330-341, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 367-379 
    ISSN: 0006-3592
    Keywords: fluidized-bed ; consecutive reaction kinetics ; distributed fraction of methanogens ; rate-limiting ; parametric sensitivity ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A kinetic model involving the distributed fractions of acidogens and methanogens is proposed. To determine the fluxes and biochemical reaction rates of the substrate sucrose and its intermediates, volatile fatty acids (VFAs) in bulk liquid and within the biofilm, a kinetic model was developed by combining the solid-phase model with the liquid-phase model. The predicted substrate removal efficiencies of the conventional and tapered fluidized-bed bioreactors (CFB, TFBs) are in good agreement with the experimental results. The biofilm thickness in TFBs are thicker than that in CFB, resulting in performance enhancement with TFBs. The simulated results obtained from the kinetic model show that methanogenesis is the rate-limiting step of degradation of the simple organic compound (sucrose), and the chemical oxygen demand (COD) concentration in the effluent is mainly contributed by the intermediates VFAs. The distributed fractions of acidogens and methanogens determined experimentally are 0.4 and 0.6, respectively. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 367-379, 1998.
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  • 78
    ISSN: 0006-3592
    Keywords: depolymerization ; kinetics ; endo -enzymes ; theoretical equation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Monitoring the time evolution of the concentration of a selected range of molecular weights of substrate, referred to as “detectable” substrate, has been used to determine endo-enzymic activities in polysaccharide depolymerizing processes. In the methodologies based on the use of dye-labeled substrates, the “detectable” substrate extends from a given molecular weight threshold downward. On the contrary, in the fluorescent probe-flow injection analysis methodology, initially developed to determine (1 → 3)-(1 → 4)-β-d-glucanase activities, the “detectable” substrate extends from a given molecular weight threshold upward. Assuming that the time evolution of the molecular weight distribution of the substrate follows the most probable distribution (the enzymic attack is random and its mechanism is single attack), a theoretical equation describing the time evolution of the concentration of “detectable” substrate (from a given molecular weight threshold upward or downward) has been deduced. This equation, Wd = Wo · (1 + αt) · e-αt, where Wd is the concentration of “detectable” substrate, Wo is the initial concentration of the substrate, t is the depolymerization time, and α is a parameter correlated through a hyperbola with the initial concentrations of enzyme and substrate and the Michaelis-Menten constant, Km, has been tested against different (1 → 3)-(1 → 4)-β-d-glucan/(1 → 3)-(1 → 4)-β-d-glucanase systems using the fluorescent probe-flow injection analysis methodology and Calcofluor as the fluorescent probe. The most important predictions of the theoretical equation, which allow accurate determination of both endo-enzymic activities and kinetic constants, have been experimentally confirmed. ©1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 387-393, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 438-446 
    ISSN: 0006-3592
    Keywords: DNA ; alginate ; encapsulation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Alginate gels produced by an external or internal gelation technique were studied so as to determine the optimal bead matrix within which DNA can be immobilized for in vivo application. Alginates were characterized for guluronic/mannuronic acid (G/M) content and average molecular weight using 1H-NMR and LALLS analysis, respectively. Nonhomogeneous calcium, alginate, and DNA distributions were found within gels made by the external gelation method because of the external calcium source used. In contrast, the internal gelation method produces more uniform gels. Sodium was determined to exchange for calcium ions at a ratio of 2:1 and the levels of calcium complexation with alginate appears related to bead strength and integrity. The encapsulation yield of double-stranded DNA was over 97% and 80%, respectively, for beads formed using external and internal calcium gelation methods, regardless of the composition of alginate. Homogeneous gels formed by internal gelation absorbed half as much DNAse as compared with heterogeneous gels formed by external gelation. Testing of bead weight changes during formation, storage, and simulated gastrointestinal (GI) conditions (pH 1.2 and 7.0) showed that high alginate concentration, high G content, and homogeneous gels (internal gelation) result in the lowest bead shrinkage and alginate leakage. These characteristics appear best suited for stabilizing DNA during GI transit. ©1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 438-446, 1998.
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    Biotechnology and Bioengineering 57 (1998), S. 454-461 
    ISSN: 0006-3592
    Keywords: propionic acid ; extractive fermentation ; solvent ; partition ; acid recovery ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Solvent selection for extractive fermentation for propionic acid was conducted with three systems: Alamine 304-1 (trilaurylamine) in 2-octanol, 1-dodecanol, and Witcohol 85 NF (oleyl alcohol). Among them, the solvent containing 2-octanol exhibited the highest partition coefficient in acid extraction, but it was also toxic to propionibacteria. The most solvent-resistant strain among five strains of the microorganism was selected. Solvent toxicity was eliminated via two strategies: entrapment of dissolved toxic solvent in the culture growth medium with vegetable oils such as corn, olive, or soybean oils; or replacement of the toxic 2-octanol with nontoxic Witcohol 85 NF. The complete recovery of acids from the Alamine 304-1/Witcohol 85 NF was also realized with vacuum distillation. ©1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 454-461, 1998.
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    Biotechnology and Bioengineering 58 (1998), S. 258-262 
    ISSN: 0006-3592
    Keywords: mass balance ; metabolic flux ; 13C tracer ; NMR spectroscopy ; mass spectroscopy ; hybridoma ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The estimation of intracellular fluxes of mammalian cells using only mass balances of the relevant metabolites is not possible because the set of linear equations defined by these mass balances is underdetermined. In order to quantify fluxes in cyclic pathways the mass balance equations can be complemented with several constraints: (1) the mass balances of co-metabolites, such as ATP or NAD(P)H, (2) linear objective functions, (3) flux data obtained by isotopic-tracer experiments. Here, these three methods are compared for the analysis of fluxes in the primary metabolism of continuously cultured hybridoma cells. The significance of different theoretical constraints and different objective functions is discussed after comparing their resulting flux distributions to the fluxes determined using 13CO2 and 13C-lactate measurements of 1 - 13C-glucose-fed hybridoma cells. Metabolic fluxes estimated using the objective functions “maximize ATP” and “maximize NADH” are relatively similar to the experimentally determined fluxes. This is consistent with the observation that cancer cells, such as hybridomas, are metabolically hyperactive, and produce ATP and NADH regardless of the need for these cofactors. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:258-262, 1998.
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  • 82
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    Biotechnology and Bioengineering 58 (1998), S. 263-266 
    ISSN: 0006-3592
    Keywords: Streptomyces lividans ; simple structured modeling ; cybernetic modeling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The growth of Streptomyces lividans in defined media was modeled using a simple structured growth model. Conventional unstructured models like Monod kinetics, substrate inhibition kinetics, and the logistic equation were also used in an attempt to fit the data, but the results were all unsatisfactory. The main reason for failure in applying simple unstructured models is that they cannot describe the long lag phases sometimes observed during growth of S. lividans. The simple structured growth model was derived along similar principles to cybernetic growth models. This model quite accurately describes the growth of S. lividans. It assumes that the rate of assimilation of a substrate depends on the concentration of a specific key enzyme. This key enzyme is only produced in the presence of the substrate, and it is broken down at a steady rate. An enzyme synthesis allocation variable, w, similar to the cybernetic variable, u, described in cybernetic growth models, is proposed to control enzyme synthesis. Until the key enzyme concentration approaches its maximum level, very little substrate is consumed. And consequently, the lag phase is sustained. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:263-266, 1998.
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  • 83
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    Biotechnology and Bioengineering 58 (1998), S. 267-271 
    ISSN: 0006-3592
    Keywords: metabolic engineering ; mathematical programming ; mixed integer nonlinear programming (MINLP) ; metabolic control analysis ; Dictyostelium discoideum ; tricarboxylic acid cycle ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The design of new generation bioprocessing plants is increasingly dependent on the design of process-compatible microorganisms. The latter, whether through genetic or physiological manipulations, can be greatly assisted by metabolic engineering. An emerging powerful tool in metabolic engineering research is computer-assisted cell design using mathematical programming. In this work, the problem of optimizing cellular metabolic networks has been formulated as a Mixed Integer Nonlinear Programming (MINLP) model. The model can assist genetic engineers to identify which cellular enzymes should be modified, and the new levels of activity required to produce an optimal network. Results are presented from the tricarboxylic acid cycle in Dictyostelium discoideum. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:267-271, 1998.
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  • 84
    ISSN: 0006-3592
    Keywords: metabolic flux analysis ; 13C tracer experiments ; fractional enrichment ; NADH ; NADPH ; pentose phosphate pathway ; Aspergillus oryzae ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Conventional metabolic flux analysis uses the information gained from determination of measurable fluxes and a steady-state assumption for intracellular metabolites to calculate the metabolic fluxes in a given metabolic network. The determination of intracellular fluxes depends heavily on the correctness of the assumed stoichiometry including the presence of all reactions with a noticeable impact on the model metabolite balances. Determination of fluxes in complex metabolic networks often requires the inclusion of NADH and NADPH balances, which are subject to controversial debate. Transhydrogenation reactions that transfer reduction equivalents from NADH to NADPH or vice versa can usually not be included in the stoichiometric model, because they result in singularities in the stoichiometric matrix. However, it is the NADPH balance that, to a large extent, determines the calculated flux through the pentose phosphate pathway. Hence, wrong assumptions on the presence or activity of transhydrogenation reactions will result in wrong estimations of the intracellular flux distribution. Using 13C tracer experiments and NMR analysis, flux analysis can be performed on the basis of only well established stoichiometric equations and measurements of the labeling state of intracellular metabolites. Neither NADH/NADPH balancing nor assumptions on energy yields need to be included to determine the intracellular fluxes. Because metabolite balancing methods and the use of 13C labeling measurements are two different approaches to the determination of intracellular fluxes, both methods can be used to verify each other or to discuss the origin and significance of deviations in the results. Flux analysis based entirely on metabolite balancing and flux analysis, including labeling information, have been performed independently for a wild-type strain of Aspergillus oryzae producing α-amylase. Two different nitrogen sources, NH4+ and NO3-, have been used to investigate the influence of the NADPH requirements on the intracellular flux distribution. The two different approaches to the calculation of fluxes are compared and deviations in the results are discussed. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:254-257, 1998.
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  • 85
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    Biotechnology and Bioengineering 58 (1998), S. 272-281 
    ISSN: 0006-3592
    Keywords: gene transfer ; retrovirus ; cell cycle ; intracellular stability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Recombinant retroviruses are currently used as gene delivery vehicles for the purpose of gene therapy. It is generally believed that the efficiency of retroviral transduction depends on the cell cycle status of the target cells. However, it has been reported that this is not the case for the transduction of human and murine fibroblasts, in contrast to other cell types such as lymphocytes. The predictions of a mathematical model that we constructed, offer an explanation of this contradiction, based on the dynamics of the underlying processes of target cell growth and the intracellular decay of retroviral vectors. The model suggests that the utility of synchronization experiments, that are usually employed to study cell cycle specificity, is severely limited when the time scales of the above kinetic events are comparable to each other. The predictions of the model also suggest the use of retroviral vectors as cell cycle markers, as an alternative way to detect cell cycle dependence of retroviral transduction. This method obviates the need for cell synchronization and therefore, it does not perturb the cell cycle or interfere with the life cycle of retroviral vectors. Moreover, it does not depend on the intracellular stability of retroviral vectors. Our results show that in contrast to previously reported results, transduction of murine fibroblasts is cell cycle dependent, and they are consistent with the current notion that mitosis is the phase that confers transduction susceptibility. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:272-281, 1998.
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  • 86
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    Biotechnology and Bioengineering 58 (1998), S. 282-291 
    ISSN: 0006-3592
    Keywords: cybernetic models ; metabolic engineering ; storage pathways ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A cybernetic model is proposed to examine generic features of storage pathways. This model is capable of describing synthesis of carbon and non-carbon storage polymers. The effect of environmental conditions is evaluated using storage polymer level as a fraction of total biomass as a gauge of pathway performance. The base wild-type pathway is then analyzed to determine the effect of genetic alterations upon system performance. Proposed modifications are tested using the cybernetic model as a diagnostic tool to ascertain the ramifications of potential genetic alterations. A methodology is developed within the cybernetic framework to describe alterations of enzyme activity and over-expression of pathway enzymes. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:282-291, 1998.
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  • 87
    ISSN: 0006-3592
    Keywords: metabolic engineering ; carbon metabolism ; Escherichia coli mutants ; microbial growth ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Escherichia coli strains devoid of one or both of the two pyruvate kinase isoenzymes (PKA and PKF), were grown on minimal media in batch fermentations. The strain lacking both PKs showed a 28% decrease on its specific growth rate when compared to the wild type. However, protein and CO2 yields did not change. Using radioactive 1-C14 glucose and collecting the CO2 produced by the cultures, it was found that the mutant lacking both pyruvate kinases, metabolized glucose mainly through the pentose pathway (PP). The increased participation of the PP in glucose metabolism in this strain, was also reflected on the levels of the glucose-6-phosphate and 6-phosphogluconate dehydrogenases.© 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:292-295, 1998.
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  • 88
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    Biotechnology and Bioengineering 58 (1998), S. 296-298 
    ISSN: 0006-3592
    Keywords: lac promoter ; galactose ; galactokinase mutant ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In Escherichia coli, strong overexpression of a recombinant protein has been shown to be deleterious due to a heavy metabolic burden on the host cell, which may completely cease cell growth before maximum product accumulation has occurred.Aiming at a reduction of very high product formation rates, we engineered E. coli strains by mutating the Leloir pathway for galactose metabolization, so that galactose can be utilized to induce lac derived promoters. The induction with galactose was effective in every strain and expression construct tested, and it reduced the metabolic burden on a highly overproducing clone so that cell growth and product accumulation could be maintained for several generations. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:296-298, 1998.
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  • 89
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    Biotechnology and Bioengineering 58 (1998), S. 299-302 
    ISSN: 0006-3592
    Keywords: phosphoglucomutase ; site-directed mutagenesis ; kinetic constants ; Pm promoter ; metabolic engineering ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Mutants of Escherichia coli deficient in phosphoglucomutase accumulate amylose when the cells are grown on maltose or galactose as carbon source. In the presence of physiological levels of phosphoglucomutase, most of the sugar is catabolized, leading to strongly reduced levels of amylose accumulation. By varying the expression level of heterologous phosphoglucomutase, we show that the minimum level needed to block amylose accumulation corresponds to a phosphoglucomutase activity of 150-600 nmole substrate transformed per min per mg of total soluble protein. Mutant phosphoglucomutases with strongly reduced Vmax values and increased Km values for the substrate glucose-1-phosphate or the co-substrate glucose-1,6-diphosphate, could also reduce amylose accumulation, but much higher enzyme expression levels were required. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58:299-302, 1998.
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  • 90
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    Biotechnology and Bioengineering 57 (1998), S. 718-731 
    ISSN: 0006-3592
    Keywords: biofilm ; modeling ; reaction-diffusion-growth ; cellular automata ; immobilized cells ; structure ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The theoretical basis and quantitative evaluation of a new approach for modeling biofilm growth are presented here. Soluble components (e.g., substrates) are represented in a continuous field, whereas discrete mapping is used for solid components (e.g., biomass). The spatial distribution of substrate is calculated by applying relaxation methods to the reaction-diffusion mass balance. A biomass density map is determined from direct integration in each grid cell of a substrate-limited growth equation. Spreading and distribution of biomass is modeled by a discrete cellular automaton algorithm. The ability of this model to represent diffusion-reaction-microbial growth systems was tested for a well-characterized system: immobilized cells growing in spherical gel beads. Good quantitative agreement with data for global oxygen consumption rate was found. The calculated concentration profiles of substrate and biomass in gel beads corresponded to those measured. Moreover, it was possible, using the discrete spreading algorithm, to predict the spatial two- and three-dimensional distribution of microorganisms in relation to, for example, substrate flux and inoculation density. The new technique looks promising for modeling diffusion-reaction-microbial growth processes in heterogeneous systems as they occur in biofilms. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 57: 718-731, 1998
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  • 91
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    Biotechnology and Bioengineering 58 (1998), S. 561-571 
    ISSN: 0006-3592
    Keywords: metabolism analysis ; AB fermentation equations ; on-line physiological state diagnosis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fermentation equations for acetone-butanol (AB) were applied in a metabolic analysis of the reaction network under various conditions; that is, at different pHs and a high NADH2 turnover rate using methyl viologen, in a Clostridium acetobutylicum culture. The results disclosed variations in the pattern of rate changes that reflected changes in the physiological state. A linear relationship was found to exist between NADH2 generation and butanol production rate. By coupling an automated measurement system with the fermentation model, on-line estimation of the culture state was accomplished. Based on the AB fermentation model, new parameters were defined for on-line diagnosis of the physiological state and determination of the best timing for amplifying NADH2 generation by the addition of methyl viologen to obtain a high level of butanol productivity. A potential means of achieving optimal control for a high level of solvent production, involving the correlation of certain rates, is proposed. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 58: 561-571, 1998.
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  • 92
    ISSN: 0006-3592
    Keywords: aqueous two-phase systems ; immobilized enzymes ; continuous extraction of product ; penicillin G acylase ; synthesis of antibiotics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Yields of kinetically controlled synthesis of antibiotics catalyzed by penicillin G acylase from Escherichia coli (PGA) have been greatly increased by continuous extraction of water soluble products (cephalexin) away from the surroundings of the enzyme. In this way its very rapid enzymatic hydrolysis has been avoided. Enzymes covalently immobilized inside porous supports acting in aqueous two-phase systems have been used to achieve such improvements of synthetic yields. Before the reaction is started, the porous structure of the biocatalyst can be washed and filled with one selected phase. In this way, when the pre-equilibrated biocatalyst is mixed with the second phase (where the reaction product will be extracted), the immobilized enzyme remains in the first selected phase in spite of its possibly different natural trend.Partition coefficients (K) of cephalexin in very different aqueous two-phase systems were firstly evaluated. High K values were obtained under drastic conditions. The best K value for cephalexin (23) was found in 100% PEG 600-3 M ammonium sulfate where cephalexin was extracted to the PEG phase. Pre-incubation of immobilized PGA derivatives in ammonium sulfate and further suspension with 100% PEG 600 allowed us to obtain a 90% synthetic yield of cephalexin from 150 mM phenylglycine methyl ester and 100 mM 7-amino desacetoxicephalosporanic acid (7-ADCA). In this reaction system, the immobilized enzyme remains in the ammonium sulfate phase and hydrolysis of the antibiotic becomes suppressed because of its continuous extraction to the PEG phase. On the contrary, synthetic yields of a similar process carried out in monophasic systems were much lower (55%) because of a rapid enzymatic hydrolysis of cephalexin. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:73-79, 1998.
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  • 93
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    Biotechnology and Bioengineering 59 (1998), S. 99-107 
    ISSN: 0006-3592
    Keywords: Monod kinetics ; mixed substrate growth ; continuous culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In natural environments, heterotrophic microorganisms encounter complex mixtures of carbon sources, each of which is present only at very low concentrations. Under such conditions no significant growth could be expected if cells utilized only one of the available carbon compounds as suggested by the principle of diauxic growth. Indeed, there is much evidence that microbial cells utilize many carbon sources simultaneously. In order to predict bacterial growth under such conditions we developed a model describing the specific growth rate as a function of the individual concentrations of several simultaneously utilized carbon substrates. Together with multisubstrate models previously published, this model was evaluated for its ability to describe growth of Escherichia coli during the simultaneous utilization of mixtures of sugars in carbon-limited continuous culture. Using the μmax and Ks constants determined for single substrate growth with six different sugars, the model was able for most experiments to adequately describe the specific growth rate of the culture, i.e., the experimentally set dilution rate, from the measured concentrations of the individual sugars. The model provides an explanation why bacteria can still grow relatively fast under environmental conditions where the concentrations of carbon substrates are usually extremely low. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:99-107, 1998.
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  • 94
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    Biotechnology and Bioengineering 59 (1998), S. 635-639 
    ISSN: 0006-3592
    Keywords: Spathoglottis plicata ; orchid ; encapsulation ; two-coat systems ; complex coacervation ; artificial seed ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Complex coacervation of alginate-chitosan and alginate-gelatin were used to develop two-coat systems for the encapsulation of Spathoglottis plicata seeds and protocorms (top-shaped structures formed after seed germination of orchids). Both the seeds and the protocorms could withstand the encapsulation treatments with high viability. About 54% of seeds and 40% of large protocorms (1.6-2.0 mm) were able to tolerate a 6-h desiccation treatment. However, viability of the small protocorms (0.7-0.9 mm) was greatly reduced if they were desiccated before encapsulation. Encapsulation after desiccation significantly increased the percentage viability of seeds and protocorms. Treatment with abscisic acid (ABA, 10-5 M) before desiccation and encapsulation resulted in high percentage viability in seeds and large protocorms whereas the small protocorms were found to be less tolerant to the treatments. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:635-639, 1998.
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  • 95
    ISSN: 0006-3592
    Keywords: Trichoderma reesei ; cellulase ; cellobiohydrolase ; endoglucanase ; microcrystalline cellulose ; cellulose hydrolysis ; adsorption isotherm ; synergism ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Microcrystalline cellulose (10 g/L Avicel) was hydrolysed by two major cellulases, cellobiohydrolase I (CBH I) and endoglucanase II (EG II), of Trichoderma reesei. Two types of experiments were performed, and in both cases the enzymes were added alone and together, in equimolar mixtures. In time course studies the reaction time was varied between 3 min and 48 h at constant temperature (40°C) and enzyme loading (0.16 μmol/g Avicel). In isotherm studies the enzyme loading was varied in the range of 0.08-2.56 μmol/g at 4°C and 90 min. Adsorption of the enzymes and production of soluble sugars were followed by FPLC and HPLC, respectively. Adsorption started quickly (50% of maximum achieved after 3 min) but was not completed before 60-90 min. For CBH I a linear relationship was observed between the production of soluble sugars and adsorption, showing that the average activity of the bound CBH I molecules does not change with increasing saturation. For EG II the corresponding curve levelled off which is explained by initial hydrolysis of loose ends on Avicel. The enzymes competed for binding sites, binding of EG II was considerably affected by CBH I, especially at high concentration. CBH I produced more soluble sugars than EG II, except at conversions below 1%. At 40°C when the enzymes were added together they produced 27-45% more soluble sugars than the sum of what they produced alone, i.e. synergistic action was observed (the final conversion after 48 h of hydrolysis was 3, 6, and 13% for EG II, CBH I, and their mixture, respectively). At 4°C, on the other hand, when the conversion was below 2.5%, almost no synergism could be observed. Molar proportions of the produced sugars were rather stable for CBH I (11-15%, 82-89%, and 〈6% for glucose, cellobiose, and cellotriose, respectively), while it varied considerably with both time and enzyme concentration for EG II. The observed stable but high glucose to cellobiose ratio for CBH I indicates that the processivity for this enzyme is not perfect. EG II produced significant amounts of glucose, cellobiose, and cellotriose, which are not the expected products of a typical endoglucanase activity on a solid substrate. We explain this by hypothesizing that EG II may show processivity due to its extended substrate binding site and the presence of its cellulose binding domain. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:621-634, 1998.
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  • 96
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    Biotechnology and Bioengineering 59 (1998), S. 647-650 
    ISSN: 0006-3592
    Keywords: biomass separation ; flocculation ; biomass measurement ; yeast ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We introduce the ratio of nonflocculent versus total biomass as a criterion for starting cell separation from the medium. This criterion can be applied for the automation of the process regardless of the process dynamics. Its minimum indicates the optimum period of time for the start of the separation process with regard not only to nonflocculent cell concentration, but also medium attributes. In contrast to the concentration of nonflocculent cells, which has two minima, first at the beginning of the process and another broader one in the period during which maximum flocculation is present, the ratio has a single minimum and can therefore be implemented as a criterion for cell separation. To calculate the ratio value, in addition to an on-line method for nonflocculent biomass measurement described elsewhere, an on-line method for the total biomass of flocculent yeast is proposed. It is based on the absorbency measurement of the cell biomass, previously deflocculated by EDTA. Therefore, it can be applied in bioprocesses with transparent media and yeast that can be deflocculated by EDTA. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:647-650, 1998.
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  • 97
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    Biotechnology and Bioengineering 59 (1998), S. 640-646 
    ISSN: 0006-3592
    Keywords: Vitreoscilla hemoglobin ; metabolic engineering ; fermentation ; acetoin ; 2,3-butanediol ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Serratia marcescens was transformed with plasmid vector pUC8 or pUC8 containing the bacterial (Vitreoscilla) hemoglobin gene (vgb) on either a 2.3-kb fragment (pUC8:15) or 1.4-kb fragment (pUC8:16) of Vitreoscilla DNA. The vgb-bearing strains were compared with the pUC8 transformant and untransformed S. marcescens with respect to growth in Luria-Bertani (LB) broth supplemented with glucose or casein acid hydrolysate. Growth (on a viable cell basis) was similar to that in unsupplemented LB. Total acid excretion (as estimated by medium pH) was similar for all strains in both LB plus 2% casein acid hydrolysate and LB without additions. Acid excretion in LB plus 2% glucose was somewhat greater at up to 10 h in culture for the two vgb-bearing strains; from 10 to 26 h in culture, the pHs of these cultures continued to decrease (to 4.1-4.2), whereas those of the non-vgb-bearing strains returned to near the starting pH (7.4-7.8). Concomitantly, after 26 h of culture in LB plus 2% glucose, the non-vgb-bearing strains had produced about 15 times as much acetoin and about three to four times as much 2,3-butanediol as the vgb-bearing strains. In general, for all strains, much more acetoin and 2,3-butanediol were produced in LB plus 2% glucose than in unsupplemented LB. The exception was acetoin production by the strain bearing vgb on plasmid pUC8:15; after 26 h of culture in LB without supplementation it was between three and four times that of the other strains, and about 50% higher than its level in LB plus 2% glucose. When grown with the 2% casein acid hydrolysate supplement, the strain bearing vgb on plasmid pUC8:15 produced much more acetoin and 2,3-butanediol than the other strains after 26 hours in culture. The results confirm that vgb can significantly alter carbon metabolism and suggest that the use of vgb technology for directed metabolic engineering may be a complicated process, depending in part on medium composition. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:640-646, 1998.
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  • 98
    ISSN: 0006-3592
    Keywords: cellulose ; gel ; fiber ; immobilization ; adsorption ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We prepared a new composite gel fiber by the gel formation of cellulose acetate and titanium iso-propoxide. The fiber is harder than alginate gel; it is also stable in common solvents, phosphate solution, and electrolyte solutions over a wide range of pH from 3 to 10. The fiber shows amphoretic adsorption properties depending on pH, namely, it acts anionic with decreasing pH and cationic with increasing pH. However, the fiber had no adsorption property for a pyrogen endotoxin. The β-galactosidase and α-chymotrypsin not retained in alginate gel were immobilized on the fibers by this method. The pH, temperature, and repeated run stabilities of the immobilized enzyme were compared to those of the native one. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:651-656, 1998.
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  • 99
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    Biotechnology and Bioengineering 59 (1998), S. 659-665 
    ISSN: 0006-3592
    Keywords: F plasmid ; low-copy plasmids ; plasmid stability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A low-copy expression vector has been constructed from a 9 Kbp region of the Escherichia coli F plasmid containing the oriV and oriS origins of replication. This plasmid carries the β-lactamase gene (Apr) and the araBAD promoter/araC regulator for arabinose-inducible gene expression. A derivative which carries a lacZ reporter gene was found to be stably maintained for at least 150 generations. A related multi-copy plasmid was stably maintained in arabinose-free medium, but no plasmid-bearing segregants remained after 60 generations when lacZ expression was induced. Induced expression resulted in 27% (multi-copy) and 12% (low-copy) decreases in growth rate. The uninduced levels of β-galactosidase were 200 units (multi-copy) and 15 units (low-copy). © 1998 John-Wiley & Sons, Inc. Biotechnol Bioeng 59:659-665, 1998.
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  • 100
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    Biotechnology and Bioengineering 59 (1998), S. 666-672 
    ISSN: 0006-3592
    Keywords: mRNA stability ; plasmid copy number ; gene expression ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effects of mRNA stability and plasmid copy number on gene expression in Escherichia coli were evaluated by constructing multicopy (pMB1-based) and low-copy (F-based) plasmids containing an arabinose-inducible promoter system, the lacZ reporter gene, and mRNA-stabilizing 5′ hairpin structures. Product formation and cell growth were evaluated under a number of inducer concentrations. The introduction of a 5′ hairpin into the untranslated region of the mRNA resulted in significantly higher gene expression from the multicopy plasmids at low inducer concentrations and increased gene expression from the low-copy plasmids across all inducer concentrations investigated. With high inducer concentrations, expression from high-copy plasmids significantly slowed cell growth, whereas expression from the low-copy plasmids had little effect on growth rate. At inducer concentrations between 1 × 10-4 and 4 × 10-4%, the productivity of low-copy plasmids containing the 5′-hairpin was equal to or greater than that from multicopy plasmids. Together, these two gene expression strategies may find important use in metabolic engineering and heterologous gene expression. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 59:666-672, 1998.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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