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Hemocyte Differentiation Mediates The Mosquito Late-Phase Immune Response Against Plasmodium In Anopheles Gambiae, Ryan C. Smith, Carolina Barillas-Mury, Marcelo Jacobs-Lorena Jun 2015

Hemocyte Differentiation Mediates The Mosquito Late-Phase Immune Response Against Plasmodium In Anopheles Gambiae, Ryan C. Smith, Carolina Barillas-Mury, Marcelo Jacobs-Lorena

Ryan C. Smith

Plasmodium parasites must complete development in the mosquito vector for transmission to occur. The mosquito innate immune response is remarkably efficient in limiting parasite numbers. Previous work has identified a LPS-induced TNFα transcription factor (LITAF)-like transcription factor, LITAF-like 3 (LL3), which significantly influences parasite numbers. Here, we demonstrate that LL3 does not influence invasion of the mosquito midgut epithelium or ookinete-to-oocyst differentiation but mediates a late-phase immune response that decreases oocyst survival. LL3 expression in the midgut and hemocytes is activated by ookinete midgut invasion and is independent of the mosquito microbiota, suggesting that LL3 may be a component of …


Repeated Stressors In Adulthood Increase The Rate Of Biological Ageing, Michaela Hau, Mark F. Haussmann, Timothy J. Greives, Christa Matlack, David Costantini, Michael Quetting, James S. Adelman, Ana C. Miranda, Jesko Partecke Jan 2015

Repeated Stressors In Adulthood Increase The Rate Of Biological Ageing, Michaela Hau, Mark F. Haussmann, Timothy J. Greives, Christa Matlack, David Costantini, Michael Quetting, James S. Adelman, Ana C. Miranda, Jesko Partecke

James S. Adelman

Individuals of the same age can differ substantially in the degree to which they have accumulated tissue damage, akin to bodily wear and tear, from past experiences. This accumulated tissue damage reflects the individual’s biological age and may better predict physiological and behavioural performance than the individual‘s chronological age. However, at present it remains unclear how to reliably assess biological age in individual wild vertebrates. We exposed hand-raised adult Eurasian blackbirds (Turdus merula) to a combination of repeated immune and disturbance stressors for over one year to determine the effects of chronic stress on potential biomarkers of biological ageing including …


The Plasmodium Bottleneck: Malaria Parasite Losses In The Mosquito Vector, Ryan C. Smith, Joel Vega-Rodríguez, Marcelo Jacobs-Lorena Aug 2014

The Plasmodium Bottleneck: Malaria Parasite Losses In The Mosquito Vector, Ryan C. Smith, Joel Vega-Rodríguez, Marcelo Jacobs-Lorena

Ryan C. Smith

Nearly one million people are killed every year by the malaria parasite Plasmodium. Although the disease-causing forms of the parasite exist only in the human blood, mosquitoes of the genus Anopheles are the obligate vector for transmission. Here, we review the parasite life cycle in the vector and highlight the human and mosquito contributions that limit malaria parasite development in the mosquito host. We address parasite killing in its mosquito host and bottlenecks in parasite numbers that might guide intervention strategies to prevent transmission.


Immunization Against A Merozoite Sheddase Promotes Multiple Invasion Of Red Blood Cells And Attenuates Plasmodium Infection In Mice, Ryan C. Smith, Daisy D. Colón-López, Jürgen Bosch Aug 2014

Immunization Against A Merozoite Sheddase Promotes Multiple Invasion Of Red Blood Cells And Attenuates Plasmodium Infection In Mice, Ryan C. Smith, Daisy D. Colón-López, Jürgen Bosch

Ryan C. Smith

Subtilisin-like protease 2 (SUB2) is a conserved serine protease utilized by Plasmodium parasites as a surface sheddase required for successful merozoite invasion of host red blood cells and has been implicated in ookinete invasion of the mosquito midgut. To determine if SUB2 is a suitable vaccine target to interfere with malaria parasite development, the effects of SUB2-immunization on the Plasmodium life cycle were examined in its vertebrate and invertebrate hosts. Swiss Webster mice were immunized with SUB2 peptides conjugated to Keyhole limpet hemocyanin (KLH) or KLH alone, and then challenged with Plasmodium berghei. To determine the effects of immunization on …


Linking Social Behavior And Stress Physiology In Feral Mares (Equus Caballus): Group Transfers Elevate Fecal Cortisol Levels, Cassandra M.V. Nuñez, James S. Adelman, Jessica Smith, Laurence Gesquiere, Daniel I. Rubenstein Jan 2014

Linking Social Behavior And Stress Physiology In Feral Mares (Equus Caballus): Group Transfers Elevate Fecal Cortisol Levels, Cassandra M.V. Nuñez, James S. Adelman, Jessica Smith, Laurence Gesquiere, Daniel I. Rubenstein

Cassandra M.V. Nuñez

Feral horses (Equus caballus) have a complex social structure, the stability of which is important to their overall health. Behavioral and demographic research has shown that decreases in group (or band) stability reduce female fitness, but the potential effects on the physiological stress response have not been demonstrated. To fully understand how band stability affects group-member fitness, we need to understand not only behavioral and demographic, but also physiological consequences of decreases to that stability. We studied group changes in feral mares (an activity that induces instability, including both male and female aggression) on Shackleford Banks, NC. We found that …


A Free-Ranging, Feral Mare Equus Caballus Affords Similar Maternal Care To Her Genetic And Adopted Offspring, Cassandra M.V. Nuñez, James S. Adelman, Daniel I. Rubenstein Nov 2013

A Free-Ranging, Feral Mare Equus Caballus Affords Similar Maternal Care To Her Genetic And Adopted Offspring, Cassandra M.V. Nuñez, James S. Adelman, Daniel I. Rubenstein

Cassandra M.V. Nuñez

Adoption of nongenetic offspring occurs in a variety of species but is rare in equids. We report a case of adoption by a free-ranging, feral mare Equus caballus and compare the maternal care received by her genetic offspring (born 1995) to that of her adopted offspring (born 1996) for the first 30 weeks of development. We compare five measures of care: (1) total time spent suckling, (2) mare aggression during suckling, (3) number of mare-terminated suckling bouts, (4) contact maintenance, and (5) mare-foal distance. For most behaviors, we detected no difference in the mare’s treatment of the two foals; however, …


Transgenic Mosquitoes Expressing A Phospholipase A(2) Gene Have A Fitness Advantage When Fed Plasmodium Falciparum-Infected Blood, Ryan C. Smith, Christopher Kizito, Jason L. Rasgon, Marcelo Jacobs-Lorena Oct 2013

Transgenic Mosquitoes Expressing A Phospholipase A(2) Gene Have A Fitness Advantage When Fed Plasmodium Falciparum-Infected Blood, Ryan C. Smith, Christopher Kizito, Jason L. Rasgon, Marcelo Jacobs-Lorena

Ryan C. Smith

Genetically modified mosquitoes have been proposed as an alternative strategy to reduce the heavy burden of malaria. In recent years, several proof-of-principle experiments have been performed that validate the idea that mosquitoes can be genetically modified to become refractory to malaria parasite development. We have created two transgenic lines of Anophelesstephensi, a natural vector of Plasmodium falciparum, which constitutively secrete a catalytically inactive phospholipase A2 (mPLA2) into the midgut lumen to interfere with Plasmodium ookinete invasion. Our experiments show that both transgenic lines expressing mPLA2 significantly impair the development of rodent malaria parasites, but only one line impairs the development …


The Genome And Developmental Transcriptome Of The Strongylid Nematode Haemonchus Contortus, Erich M. Schwarz, Pasi K. Korhonen, Bronwyn E. Campbell, Neil D. Young, Aaron R. Jex, Abdul Jabbar, Ross S. Hall, Alinda Mondal, Adina C. Howe, Jason Pell, Andreas Hofmann, Peter R. Boag, Xing-Quan Zhu, T. Ryan Gregory, Alex Loukas, Brian A. Williams, Igor Antoshechkin, C. Titus Brown, Paul W. Sternberg, Robin B. Gasser Aug 2013

The Genome And Developmental Transcriptome Of The Strongylid Nematode Haemonchus Contortus, Erich M. Schwarz, Pasi K. Korhonen, Bronwyn E. Campbell, Neil D. Young, Aaron R. Jex, Abdul Jabbar, Ross S. Hall, Alinda Mondal, Adina C. Howe, Jason Pell, Andreas Hofmann, Peter R. Boag, Xing-Quan Zhu, T. Ryan Gregory, Alex Loukas, Brian A. Williams, Igor Antoshechkin, C. Titus Brown, Paul W. Sternberg, Robin B. Gasser

Adina Howe

Background The barber's pole worm, Haemonchus contortus, is one of the most economically important parasites of small ruminants worldwide. Although this parasite can be controlled using anthelmintic drugs, resistance against most drugs in common use has become a widespread problem. We provide a draft of the genome and the transcriptomes of all key developmental stages of H. contortus to support biological and biotechnological research areas of this and related parasites. Results The draft genome of H. contortus is 320 Mb in size and encodes 23,610 protein-coding genes. On a fundamental level, we elucidate transcriptional alterations taking place throughout the life …


House Finch Populations Differ In Early Inflammatory Signaling And Pathogen Tolerance At The Peak Of Mycoplasma Gallisepticum Infection, James S. Adelman, Laila Kirkpatrick, Jessica L. Grodio, Dana M. Hawley May 2013

House Finch Populations Differ In Early Inflammatory Signaling And Pathogen Tolerance At The Peak Of Mycoplasma Gallisepticum Infection, James S. Adelman, Laila Kirkpatrick, Jessica L. Grodio, Dana M. Hawley

James S. Adelman

Host individuals and populations often vary in their responses to infection, with direct consequences for pathogen spread and evolution. While considerable work has focused on the mechanisms underlying differences in resistance—the ability to kill pathogens— we know little about the mechanisms underlying tolerance— the ability to minimize fitness losses per unit pathogen. Here, we examine patterns and mechanisms of tolerance between two populations of house finches (Haemorhous [formerly Carpodacus] mexicanus) with different histories with the bacterial pathogen Mycoplasma gallisepticum (MG). After infection in a common environment, we assessed two metrics of pathology, mass loss and eye lesion severity, as proxies …


Enterobacter-Activated Mosquito Immune Responses To Plasmodium Involve Activation Of Srpn6 In Anopheles Stephensi, Ryan C. Smith, Abraham G. Eappen, Marcelo Jacobs-Lorena May 2013

Enterobacter-Activated Mosquito Immune Responses To Plasmodium Involve Activation Of Srpn6 In Anopheles Stephensi, Ryan C. Smith, Abraham G. Eappen, Marcelo Jacobs-Lorena

Ryan C. Smith

Successful development of Plasmodium in the mosquito is essential for the transmission of malaria. A major bottleneck in parasite numbers occurs during midgut invasion, partly as a consequence of the complex interactions between the endogenous microbiota and the mosquito immune response. We previously identified SRPN6 as an immune component which restricts Plasmodium berghei development in the mosquito. Here we demonstrate that SRPN6 is differentially activated by bacteria in Anopheles stephensi, but only when bacteria exposure occurs on the lumenal surface of the midgut epithelium. Our data indicate that AsSRPN6 is strongly induced following exposure to Enterobacter cloacae, a common component …


45-Year Trends In Women's Use Of Time And Household Management Energy Expenditure., Edward Archer, Robin P. Shook, Diana M. Thomas, Timothy S. Church, Peter T. Katzmarzyk, James R. Hebert, Kerry L. Mciver, Gregory A. Hand, Carl J. Lavie, Steven N. Blair Feb 2013

45-Year Trends In Women's Use Of Time And Household Management Energy Expenditure., Edward Archer, Robin P. Shook, Diana M. Thomas, Timothy S. Church, Peter T. Katzmarzyk, James R. Hebert, Kerry L. Mciver, Gregory A. Hand, Carl J. Lavie, Steven N. Blair

Robin Shook

Context: Relationships between socio-environmental factors and obesity are poorly understood due to a dearth of longitudinal population-level research. The objective of this analysis was to examine 45-year trends in time-use, household management (HM) and energy expenditure in women. Design and Participants: Using national time-use data from women 19–64 years of age, we quantified time allocation and household management energy expenditure (HMEE) from 1965 to 2010. HM was defined as the sum of time spent in food preparation, post-meal cleaning activities (e.g., dish-washing), clothing maintenance (e.g., laundry), and general housework. HMEE was calculated using body weights from national surveys and metabolic …


Regulation Of Anti-Plasmodium Immunity By A Litaf-Like Transcription Factor In The Malaria Vector Anopheles Gambiae, Ryan C. Smith, Abraham G. Eappen, Andrea J. Radtke, Marcelo Jacobs-Lorena Oct 2012

Regulation Of Anti-Plasmodium Immunity By A Litaf-Like Transcription Factor In The Malaria Vector Anopheles Gambiae, Ryan C. Smith, Abraham G. Eappen, Andrea J. Radtke, Marcelo Jacobs-Lorena

Ryan C. Smith

The mosquito is the obligate vector for malaria transmission. To complete its development within the mosquito, the malaria parasite Plasmodium must overcome the protective action of the mosquito innate immune system. Here we report on the involvement of the Anopheles gambiae orthologue of a conserved component of the vertebrate immune system, LPS-induced TNFα transcription factor (LITAF), and its role in mosquito anti-Plasmodium immunity. An. gambiae LITAF-like 3 (LL3) expression is up-regulated in response to midgut invasion by both rodent and human malaria parasites. Silencing of LL3 expression greatly increases parasite survival, indicating that LL3 is part of an anti-Plasmodium defense …


Bean And Rice Meals Reduce Postprandial Glycemic Response In Adults With Type 2 Diabetes: A Cross-Over Study, Sharon V. Thompson, Donna M. Winham, Andrea M. Hutchins Apr 2012

Bean And Rice Meals Reduce Postprandial Glycemic Response In Adults With Type 2 Diabetes: A Cross-Over Study, Sharon V. Thompson, Donna M. Winham, Andrea M. Hutchins

Donna Winham

Background: Around the world, beans and rice are commonly consumed together as a meal. With type 2 diabetes increasing, the effect of this traditional diet pattern on glycemic response has not been studied fully. Methods: We evaluated the glycemic response of bean and rice traditional meals compared to rice alone in adults with type 2 diabetes. Seventeen men and women with type 2 diabetes controlled by metformin (n = 14) or diet/exercise (n=3) aged 35–70 years participated in the randomized 4 × 4 crossover trial. The white long grain rice control, pinto beans/rice, black beans/rice, red kidney beans/rice test meals, …


What Is The Risk For Metabolic Syndrome In Police Officers?, Warren D. Franke Jan 2012

What Is The Risk For Metabolic Syndrome In Police Officers?, Warren D. Franke

Warren D Franke

The report by Jayakrishnan and colleagues in this Journal is noteworthy since it provides an assessment of the presence of the metabolic syndrome and related cardiovascular risk factors in one of the largest, if not the largest, cohort of police officers to date. Two aspects of this research especially warrant comment.


Quantitative Normative Gait Data In A Large Cohort Of Ambulatory Persons With Parkinson’S Disease, Chris J. Hass, Paul Malczak, Joe Nocera, Elizabeth L. Stegemoller, Aparna Wagle Shukala, Irene Malaty, Charles E. Jacobson Iv, Michael S. Okun, Nick Mcfarland Jan 2012

Quantitative Normative Gait Data In A Large Cohort Of Ambulatory Persons With Parkinson’S Disease, Chris J. Hass, Paul Malczak, Joe Nocera, Elizabeth L. Stegemoller, Aparna Wagle Shukala, Irene Malaty, Charles E. Jacobson Iv, Michael S. Okun, Nick Mcfarland

Elizabeth L. Stegemoller

Background: Gait performance is widely evaluated to assess health status in older adult populations. While several investigators have presented normative values for spatiotemporal gait parameters drawn from older adult populations, the literature has been void of large-scale cohort studies, which are needed in order to provide quantitative, normative gait data in persons with Parkinson’s disease. The aim of this investigation was to provide reference values for clinically important gait characteristics in a large sample of ambulatory persons with Parkinson’s disease to aid both clinicians and researchers in their evaluations and treatments of gait impairment. Methodology/Principal Findings: Gait performance was collected …


Does Immunocontraception In Feral Horses (Equus Caballus) Extend Reproductive Cycling Beyond The Normal Breeding Season?, Cassandra M.V. Nuñez, James S. Adelman, Daniel I. Rubenstein Jan 2012

Does Immunocontraception In Feral Horses (Equus Caballus) Extend Reproductive Cycling Beyond The Normal Breeding Season?, Cassandra M.V. Nuñez, James S. Adelman, Daniel I. Rubenstein

Cassandra M.V. Nuñez

Although the physiological effects of immunocontraceptive treatment with porcine zona pellucida (PZP) have been well studied, little is known about PZP’s effects on the scheduling of reproductive cycling. Recent behavioral research has suggested that recipients of PZP extend the receptive breeding period into what is normally the non-breeding season. To determine if this is the case, we compiled foaling data from wild horses (Equus caballus) living on Shackleford Banks, North Carolina for 4 years pre- and 8 years post-contraception management with PZP (pre-contraception, n = 65 births from 45 mares; post-contraception, n = 97 births from 46 mares). Gestation lasts …


Phaseolus Beans: Impact On Glycaemic Response And Chronic Disease Risk In Human Subjects, Andrea M. Hutchins, Donna M. Winham, Sharon V. Thompson Dec 2011

Phaseolus Beans: Impact On Glycaemic Response And Chronic Disease Risk In Human Subjects, Andrea M. Hutchins, Donna M. Winham, Sharon V. Thompson

Donna Winham

Consumption of Phaseolus vulgaris bean species such as pinto, black, navy or kidney may be beneficial in the prevention and treatment of chronic diseases. In particular, conditions that are promoted by increased glycaemic stress (hyperglycaemia and hyperinsulinaemia) including diabetes, CVD and cancer seem to be reduced in individuals who eat more of these beans. The present paper discusses the influence of P. vulgaris species on glycaemic response and the impact that relationship may have on the risk of developing diabetes, CVD and cancer.


Perceptions Of Flatulence From Bean Consumption Among Adults In 3 Feeding Studies, Donna M. Winham, Andrea M. Hutchins Nov 2011

Perceptions Of Flatulence From Bean Consumption Among Adults In 3 Feeding Studies, Donna M. Winham, Andrea M. Hutchins

Donna Winham

Background: Many consumers avoid eating beans because they believe legume consumption will cause excessive intestinal gas or flatulence. An increasing body of research and the 2010 Dietary Guidelines for Americans supports the benefits of a plant-based diet, and legumes specifically, in the reduction of chronic disease risks. The purpose of the current research was to investigate the perception of increased flatulence and gastrointestinal discomfort among participants who consumed a ½ cup of beans daily for 8 or 12 weeks. Methods: Participants in three studies to test the effects of beans on heart disease biomarkers completed the same weekly questionnaire to …


Immunocontraception In Wild Horses (Equus Caballus) Extends Reproductive Cycling Beyond The Normal Breeding Season, Cassandra M.V. Nuñez, James S. Adelman, Daniel I. Rubenstein Oct 2010

Immunocontraception In Wild Horses (Equus Caballus) Extends Reproductive Cycling Beyond The Normal Breeding Season, Cassandra M.V. Nuñez, James S. Adelman, Daniel I. Rubenstein

Cassandra M.V. Nuñez

Although the physiological effects of immunocontraceptive treatment with porcine zona pellucida (PZP) have been well studied, little is known about PZP's effects on the scheduling of reproductive cycling. Recent behavioral research has suggested that recipients of PZP extend the receptive breeding period into what is normally the non-breeding season. To determine if this is the case, we compiled foaling data from wild horses (Equus caballus) living on Shackleford Banks, North Carolina for 4 years pre- and 8 years post-contraception management with PZP (pre-contraception, n = 65 births from 45 mares; post-contraception, n = 97 births from 46 mares). Gestation lasts …


A Survey Of Lymphoedema Practitioners Across The Us., Jane M. Armer, Electra D. Paskett, Mei R. Fu, Joseph L. Feldman, Robin Shook, Melanie K. Schneider, Bob R. Stewart, Janice N. Cormier Jan 2010

A Survey Of Lymphoedema Practitioners Across The Us., Jane M. Armer, Electra D. Paskett, Mei R. Fu, Joseph L. Feldman, Robin Shook, Melanie K. Schneider, Bob R. Stewart, Janice N. Cormier

Robin Shook

An online survey was developed by the American Lymphedema Framework Project (ALFP) executive and research committees to assess the current scope of practice in lymphoedema treatment in the US. The analysis included responses from 415 therapists from 46 states. Respondents typically had a background in physical or occupational therapy. Nearly all reported having completed a 135-hour training course on lymphoedema treatment and management. Treatment options most often reported were comprehensive decongestive therapy (CDT), exercise/movement, risk-reduction education, and skin care.


30-Month Post-Breast Cancer Treatment Lymphoedema, Jane M. Armer, Bob R. Stewart, Robin Shook Apr 2009

30-Month Post-Breast Cancer Treatment Lymphoedema, Jane M. Armer, Bob R. Stewart, Robin Shook

Robin Shook

Background—Quantification of lymphoedema (LE) has been problematic, and the reported incidence of LE varies greatly among women treated with surgery and radiation for breast cancer. Aims—This study aims to describe LE occurrence over time among breast cancer survivors using four diagnostic criteria based on three measurement techniques. Methods—Limb volume and symptom assessment data were followed after surgery every three months for 12 months, then every six months for 30 months. Limb volume changes (LVC) were measured by circumferences and by perometry, and by symptom experience via interview. Standard survival analysis methods identified when the criteria indicating LE were met. Results—Trends …


Alfp: Identifying Issues In Lymphoedema In Hte Us, Jane Armer, Joseph Feldman, Mei Fu, Nicole Stout, Bonnie Lasinski, Catherine Tuppo, Marcia Beck, Julie Rodrick, Robin Shook, Pierre Gonon, Constance Brooks, Christine Moffatt, Bob Stewart, Janice Cormier Jan 2009

Alfp: Identifying Issues In Lymphoedema In Hte Us, Jane Armer, Joseph Feldman, Mei Fu, Nicole Stout, Bonnie Lasinski, Catherine Tuppo, Marcia Beck, Julie Rodrick, Robin Shook, Pierre Gonon, Constance Brooks, Christine Moffatt, Bob Stewart, Janice Cormier

Robin Shook

The American Lymphedema Framework Project (ALFP) is a national initiative introduced in 2008 to improve the management of lymphoedema (LE) and related disorders in the United States. A National Stakeholders Conference was held in March 2009 to share perspectives on the current state of LE, establish priority issues, and recommend actions to move the field forward.


Protection Of Retinal Cells From Ischemia By A Novel Gap Junction Inhibitor, Satyabrata Das, Dingo Lin, Snehalata Jena, Aibin Shi, Srinivas Battina, Duy H. Hua, Rachel A. Allbaugh, Dolores J. Takemoto Sep 2008

Protection Of Retinal Cells From Ischemia By A Novel Gap Junction Inhibitor, Satyabrata Das, Dingo Lin, Snehalata Jena, Aibin Shi, Srinivas Battina, Duy H. Hua, Rachel A. Allbaugh, Dolores J. Takemoto

Rachel A. Allbaugh

Retinal cells which become ischemic will pass apoptotic signal to adjacent cells, resulting in the spread of damage. This occurs through open gap junctions. A class of novel drugs, based on primaquine (PQ), was tested for binding to connexin 43 using simulated docking studies. A novel drug has been synthesized and tested for inhibition of gap junction activity using R28 neuro-retinal cells in culture. Four drugs were initially compared to mefloquine, a known gap junction inhibitor. The drug with optimal inhibitory activity, PQ1, was tested for inhibition and was found to inhibit dye transfer by 70% at 10 μM. Retinal …


Behavioral Effects Of Contraception Management: The Use Of Porcine Zona Pellucida On Wild Horses, Cassandra M.V. Nuñez Jan 2008

Behavioral Effects Of Contraception Management: The Use Of Porcine Zona Pellucida On Wild Horses, Cassandra M.V. Nuñez

Cassandra M.V. Nuñez

The advent of immunocontraception with porcine zona pellucida (PZP) has all but revolutionized wild horse management, providing a more humane method of population control than earlier strategies. Early studies on Assateague Island National Seashore have described it as an ideal form of fertility control in that it reduces the chance of conception to below 10%, can be delivered remotely, is reversible (after short-term use), lacks debilitating physiological side effects, cannot pass through the food chain, and shows minimal effects on social behaviors. However, recent research in other populations has revealed behavioral and physiological side effects of long-term PZP use. These …


Bifunctional Abietadiene Synthase: Mutual Structural Dependence Of The Active Sites For Protonation-Initiated And Ionization-Initiated Cyclizations, Reuben J. Peters, Ora A. Carter, Yan Zhang, Brian W. Matthews, Rodney B. Croteau Mar 2003

Bifunctional Abietadiene Synthase: Mutual Structural Dependence Of The Active Sites For Protonation-Initiated And Ionization-Initiated Cyclizations, Reuben J. Peters, Ora A. Carter, Yan Zhang, Brian W. Matthews, Rodney B. Croteau

Reuben J. Peters

Abietadiene synthase from grand fir catalyzes two sequential, mechanistically distinct cyclizations, of geranylgeranyl diphosphate and of copalyl diphosphate, in the formation of a mixture of abietadiene isomers as the committed step of diterpenoid resin acid biosynthesis. Each reaction is independently conducted at a separate active site residing in what were considered to be structurally distinct domains typical of terpene cyclases. Despite the presence of an unusual 250-residue N-terminal insertional element, a tandem pair of charged residues distal to the insertion was shown to form a functional part of the C-terminal active site. Because abietadiene synthase resembles the ancestral plant terpene …


Mechanism Of Abietadiene Synthase Catalysis: Stereochemistry And Stabilization Of The Cryptic Pimarenyl Carbocation Intermediates, Reuben J. Peters, Matthew M. Ravn, Robert M. Coates, Rodney Croteau Jun 2002

Mechanism Of Abietadiene Synthase Catalysis: Stereochemistry And Stabilization Of The Cryptic Pimarenyl Carbocation Intermediates, Reuben J. Peters, Matthew M. Ravn, Robert M. Coates, Rodney Croteau

Reuben J. Peters

Abietadiene synthase (AS) catalyzes the complex cyclization-rearrangement of (E,E,E)-geranylgeranyl diphosphate (8, GGPP) to a mixture of abietadiene (1a), double bond isomers 2a-4a and pimaradienes 5a-7a as a key step in the biosynthesis of the abietane resin acid constituents (1b-4b) of conifer oleoresin. The reaction proceeds at two active sites by way of the intermediate, copalyl diphosphate (9). In the second site, a putative tricyclic pimaradiene or pimarenyl(+) carbocation intermediate of undefined C13 stereochemistry and annular double bond position is formed. Three 8-oxy-17-nor analogues of 9 (17 and 19a,b) and three isomeric 15,16-bisnorpimarenyl-N-methylamines (26a-c) were synthesized and evaluated as alternative substrates …


Abietadiene Synthase Catalysis:  Conserved Residues Involved In Protonation-Initiated Cyclization Of Geranylgeranyl Diphosphate To (+)-Copalyl Diphosphate, Reuben J. Peters, Rodney B. Croteau Feb 2002

Abietadiene Synthase Catalysis:  Conserved Residues Involved In Protonation-Initiated Cyclization Of Geranylgeranyl Diphosphate To (+)-Copalyl Diphosphate, Reuben J. Peters, Rodney B. Croteau

Reuben J. Peters

Abietadiene synthase catalyzes two sequential, mechanistically distinct cyclization reactions in the formation of a mixture of abietadiene double bond isomers as the committed step in resin acid biosynthesis. Each reaction is carried out at a separate active site residing in a structurally distinct domain, and the reactions are kinetically separable. The first cyclization reaction is initiated by protonation of the terminal double bond of the universal diterpene precursor, geranylgeranyl diphosphate. The pH dependence of the overall reaction is consistent with an acid-base catalytic mechanism, and a divalent metal ion plays a role in this reaction probably by binding the diphosphate …


A Comparison Of Iron Absorption In Adults And Infants Consuming Identical Infant Formulas, Richard F. Hurrell, Lena Davidsson, Manju B. Reddy, Peter Kastenmayer, James D. Cook Jan 1998

A Comparison Of Iron Absorption In Adults And Infants Consuming Identical Infant Formulas, Richard F. Hurrell, Lena Davidsson, Manju B. Reddy, Peter Kastenmayer, James D. Cook

Manju B. Reddy

Fe absorption was estimated in adults and infants from the erythrocyte incorporation of Fe isotopes added to infant formula. Fe absorption was measured in adults using radioisotopes, and in infants with a stable-isotope technique. In adults, the geometric mean Fe absorption from a ready-to-feed soya formula with its native phytic acid content was 2.4%. This increased to 6.0 % (P < 0.05) after almost complete dephytinization. In infants, mean Fe absorption values were 3.9 and 8.7% respectively from the same products (P(O.05). In adults, mean Fe absorption from a spray-dried soya formula containing 110 mg ascorbic acid was 4.1 %, increasing to 5.3 % (P < 0.05) when ascorbic acid was doubled to 220 mgfl. In infants, mean Fe absorption values were 5.7 and 9.5 % (P < 0.05) from the same products. Mean Fe absorption from a milk-based formula was 6.5 % in adults compared with 6.7 % in infants. All meals in the adult and infant studies were fed using an identical meal size of 217 g. Increasing the meal size threefold in adults did not change fractional Fe absorption. Mean Fe absorption values for each meal were lower in adults than in infants, but the relative inhibitory effect of phytic acid and the enhancing effect of ascorbic acid were similar. We conclude that Fe absorption studies in adults can be used to assess the influence of enhancers and inhibitors of Fe absorption in infant formulas fed to infants. Further studies, however, are required to extend these findings to weaning foods and complete meals.


Iron Regulatory Protein 1 Is Not Required For The Modulation Of Ferritin And Transferrin Receptor Expression By Iron In A Murine Pro-B Lymphocyte Cell Line, Kevin Schalinske, Kenneth P. Blemings, Daniel W. Steffen, Opal S. Chen, Richard S. Eisenstein Sep 1997

Iron Regulatory Protein 1 Is Not Required For The Modulation Of Ferritin And Transferrin Receptor Expression By Iron In A Murine Pro-B Lymphocyte Cell Line, Kevin Schalinske, Kenneth P. Blemings, Daniel W. Steffen, Opal S. Chen, Richard S. Eisenstein

Kevin Schalinske

Iron regulatory proteins (IRPs) are cytoplasmic RNA binding proteins that are central components of a sensory and regulatory network that modulates vertebrate iron homeostasis. IRPs regulate iron metabolism by binding to iron responsive element(s) (IREs) in the 5* or 3* untranslated region of ferritin or transferrin receptor (TfR) mRNAs. Two IRPs, IRP1 and IRP2, have been identified previously. IRP1 exhibits two mutually exclusive functions as an RNA binding protein or as the cytosolic isoform of aconitase. We demonstrate that the BayF3 family of murine pro-B lymphocytes represents the first example of a mammalian cell line that fails to express IRP1 …


Comparison Of The Effect Of Different Opsonins On The Phagocytosis Of Fluorescein-Labeled Staphylococcal Bacteria By Chicken Heterophils, Claire B. Andreasen, James R. Andreasen Jr., Anita E. Sonn, Julie A. Oughton Jan 1996

Comparison Of The Effect Of Different Opsonins On The Phagocytosis Of Fluorescein-Labeled Staphylococcal Bacteria By Chicken Heterophils, Claire B. Andreasen, James R. Andreasen Jr., Anita E. Sonn, Julie A. Oughton

Claire B. Andreasen

Heterophil phagocytosis of fluorescein-labeled staphylococcal bacteria was analyzed by flow cytometry. Opsonization with two types of normal pooled sera and staphylococcal antisera significantly increased bacterial phagocytosis compared to samples without an opsonin. The staphylococcal antisera did not significantly increase bacterial phagocytosis compared to the normal pooled sera. Opsonization appears to increase bacterial phagocytosis but specific antisera may not increase phagocytosis beyond that caused by pooled normal sera.