MHC-B Diversity and Signs of Respiratory Illness in Wild, East African Chimpanzees ( Pan troglodytes schweinfurthii ).

Many traits, intrinsic and extrinsic to an organism, contribute to interindividual variation in immunity in wild habitats. The vertebrate Major Histocompatibility Complex (MHC) includes genes encoding antigen-presenting molecules that are highly variable, and that variation often predicts susceptibility/resistance to and recovery from pathogen infection. I compare MHC-B variation at two long-term chimpanzee research sites, Kibale National Park in Uganda and Gombe National Park in Tanzania. Using decades of respiratory health data available for these chimpanzees, I test hypotheses associated with maintenance of diversity at MHC loci, including heterozygote, divergent allele, and rare allele advantage hypotheses, and predictions for unique function of MHC-B in great apes. I found, despite confirmation of recent shared ancestry between Kibale and Gombe chimpanzees, including an overlapping MHC-B allele repertoire and similar MHC-B phenotype compositions, chimpanzees from the two research sites experienced differences in the occurrence of respiratory signs and had different associations of MHC-B diversity with signs of respiratory illness. Kibale chimpanzees with heterozygous genotypes and different peptide-binding supertypes were observed less often with respiratory signs than those homozygous or possessing the same supertypes, but this same association was not observed among Gombe chimpanzees. Gombe chimpanzees with specific MHC-B phenotypes that enable engagement of Natural Killer (NK) cells were observed more often with respiratory signs than chimpanzees with other phenotypes, but this was not observed at Kanyawara. This study emphasizes local adaptation in shaping genetic and phenotypic traits in different infectious disease contexts, even among close genetic relatives of the same subspecies, and highlights utility for continued and simultaneous tracking of host immune genes and specific pathogens in wild species.

At-vessel mortality of skates (Rajidae) taken in coastal fisheries and evidence of longer-term survival.

Data on the vigour and at-vessel mortality (AVM) of 6798 skates (comprising Raja clavata n = 6295; R. brachyura n = 208; R. undulata n = 185, R. montagui n = 98 and R. microocellata n = 12) captured by commercial fishing vessels in the inshore waters of the southern North Sea and English Channel were recorded. AVM in longline fisheries averaged 0·44% across five vessels (0-1·47%), although skates were usually unhooked manually and did not usually pass through a bait-stripper. AVM in otter trawls averaged 0·76% (0-2·35%), from four vessels fishing with tow durations of <1·5 h (southern North Sea) or 1-4 h (English Channel). No AVM was noted for skates taken as a by-catch in drift trammel nets (soak times <4 h). Anchored tangle nets resulted in an overall AVM of 2·0-2·7%, but increased from 1·47% (13-28 h soak time) to 6·16% (42-53 h soak time). There were significant differences in the vigour of skates between gears, with R. clavata caught by longline and tangle nets in better condition than those captured by otter trawl or drift trammel net. Similarly, R. undulata caught by tangle net were in better condition than those caught by otter trawl. The vigour of R. undulata was also found to be higher than other skate species for both trawl and tangle net. In total, 5283 skates were tagged with Petersen discs and released, with recapture rates for the various combinations of vessel and gear ranging up to 24·8% for R. clavata. Whilst confirming a degree of post-release survival, quantitative estimates of post-release mortality for skates remain unknown.

A review of capture and post-release mortality of elasmobranchs.

There is a need to better understand the survivorship of discarded fishes, both for commercial stocks and species of conservation concern. Within European waters, the landing obligations that are currently being phased in as part of the European Union's reformed common fisheries policy means that an increasing number of fish stocks, with certain exceptions, should not be discarded unless it can be demonstrated that there is a high probability of survival. This study reviews the various approaches that have been used to examine the discard survival of elasmobranchs, both in terms of at-vessel mortality (AVM) and post-release mortality (PRM), with relevant findings summarized for both the main types of fishing gear used and by taxonomic group. Discard survival varies with a range of biological attributes (species, size, sex and mode of gill ventilation) as well as the range of factors associated with capture (e.g. gear type, soak time, catch mass and composition, handling practices and the degree of exposure to air and any associated change in ambient temperature). In general, demersal species with buccal-pump ventilation have a higher survival than obligate ram ventilators. Several studies have indicated that females may have a higher survival than males. Certain taxa (including hammerhead sharks Sphyrna spp. and thresher sharks Alopias spp.) may be particularly prone to higher rates of mortality when caught.

Reproductive characteristics and life-history relationships of starry smooth-hound Mustelus asterias in British waters.

The reproductive biology and other life-history parameters were investigated for Mustelus asterias in British waters, with specimens caught from both commercial fisheries and research-vessel surveys. In total, 504 specimens [238 males, 24-99 cm total length (LT) and 266 females, 28-124 cm LT] were examined, with further information collected from 238 uterine pups. The lengths at 50% maturity were estimated as 70·4 and 81·9 cm LT for males and females, respectively. Ovarian fecundity ranged from one to 28, and uterine fecundity from four to 20. The number, mass and LT of pups were positively correlated with maternal LT. Full-term pups ranged from 205 to 329 mm LT, and the smallest free-living fish caught was 24 cm LT . Parturition occurred in February in the western English Channel and during June to July in the eastern English Channel and southern North Sea, indicating either protracted spawning or asynchronous parturition for the stock as a whole. The reproductive cycle is thought to extend beyond 1 year. Developmental abnormalities observed included atresia in oocytes, uterine eggs that failed to develop, a partly developed pup and an abnormal male with a single aberrant clasper. Data relating to conversion factors, oocyte numbers and diameter and gonado-somatic and hepato-somatic indices are presented, and the seasonality of the reproductive cycle is discussed.

Neural differentiation regulated by biomimetic surfaces presenting motifs of extracellular matrix proteins.

The interaction between cells and the extracellular matrix (ECM) is essential during development. To elucidate the function of ECM proteins on cell differentiation, we developed biomimetic surfaces that display specific ECM peptide motifs in a controlled manner. Presentation of ECM domains for collagen, fibronectin, and laminin influenced the formation of neurites by differentiating PC12 cells. The effect of these peptide sequences was also tested on the development of adult neural stem/progenitor cells. In this system, collagen I and fibronectin induced the formation of beta-III-tubulin positive cells, whereas collagen IV reduced such differentiation. Biomimetic surfaces composed of multiple peptide types enabled the combinatorial effects of various ECM motifs to be studied. Surfaces displaying combined motifs were often predictable as a result of the synergistic effects of ECM peptides studied in isolation. For example, the additive effects of fibronectin and laminin resulted in greater expression of beta-III-tubulin positive cells, whereas the negative effect of the collagen IV domain was canceled out by coexpression of collagen I. However, simultaneous expression of certain ECM domains was less predictable. These data highlight the complexity of the cellular response to combined ECM signals and the need to study the function of ECM domains individually and in combination.

Enhanced cell attachment using a novel cell culture surface presenting functional domains from extracellular matrix proteins.

Many factors contribute to the creation and maintenance of a realistic environment for cell growth in vitro, e.g. the consistency of the growth medium, the addition of supplements, and the surface on which the cells grow. The nature of the surface on which cells are cultured plays an important role in their ability to attach, proliferate, migrate and function. Components of the extracellular matrix (ECM) are often used to coat glass or plastic surfaces to enhance cell attachment in vitro. Fragments of ECM molecules can be immobilised on surfaces in order to mimic the effects seen by whole molecules. In this study we evaluate the application of a novel technology for the immobilisation of functional domains of known ECM proteins in a controlled manner on a surface. By examining the adherence of cultured PC12 cells to alternative growth surfaces, we show that surfaces coated with motifs from collagen I, collagen IV, fibronectin and laminin can mimic surfaces coated with the corresponding whole molecules. Furthermore, we show that the adherence of cells can be controlled by modifying the hydropathic properties of the surface to either enhance or inhibit cell attachment. Collectively, these data demonstrate the application of a new technology to enable optimisation of cell growth in the tissue culture laboratory.

Zinc transporters in the mouse placenta show a coordinated regulatory response to changes in dietary zinc intake.

The aim of the study was to determine if the expression of zinc transporters in the mouse placenta is regulated by dietary zinc, commensurate with regulating the supply of zinc to the fetus. Mice were fed diets differing only in the concentration of zinc (moderately zinc-restricted (ZnR)--15 mg Zn/kg; zinc-adequate (ZnA)--50 mg Zn/kg; zinc-supplemented (ZnS)--150 mg Zn/kg) from the onset of pregnancy until collection of tissue at day 17. Compared with mice fed the other diets, fetal weight was reduced in the ZnR group and total non-embryonic weight gain was reduced in mice fed the ZnS diet. Transcript levels of metallothionein and the zinc transporters ZnT1, ZnT4 and ZIP1 were reduced in the placenta of mice fed both the ZnR and ZnS diets compared with mice fed the ZnA diet. Placental ZnT7 and fetal liver metallothionein transcript levels did not differ significantly between mice fed the three diets and placental ZnT5 was reduced in mice fed the ZnS compared with the ZnA diet but did not differ significantly between the ZnA and ZnR diets. The pattern of mRNA expression in placenta was reflected at the protein level for ZnT1. Levels of ZnT5 protein were also highest in mice fed the ZnA diet. Both ZnT1 and ZnT5 were detected in the human villous syncytiotrophoblast by immunohistochemistry. The data indicate that the expression of zinc transporters in mouse placenta is responsive to dietary zinc supply but this modulation of expression is insufficient to maintain optimum fetal nutrition at even a modest level of dietary zinc restriction.

Creation of a CT Image Library for the Lung Screening Study of the National Lung Screening Trial.

The CT Image Library (CTIL) of the Lung Screening Study (LSS) network of the National Lung Screening Trial (NLST) consists of up to three annual screens using CT imaging from each of 17,308 participants with a significant history of smoking but no evidence of cancer at trial enrollment (Fall 2002-Spring 2004). Screens performed at numerous medical centers associated with 10 LSS-NLST screening centers are deidentified of protected health information and delivered to the CTIL via DVD, external hard disk, or Internet/Virtual Private Network transmission. The collection will be completed in late 2006. The CTIL is of potential interest to clinical researchers and software developers of nodule detection algorithms. Its attractiveness lies in its very specific, well-defined patient population, scanned via a common CT protocol, and in its collection of evenly spaced serial screens. In this work, we describe the technical details of the CTIL collection process from screening center retrieval through library storage.

Homeostatic regulation of zinc transporters in the human small intestine by dietary zinc supplementation.

BACKGROUND: The role of intestinal transporter regulation in optimising nutrient absorption has been studied extensively in rodent and cell line models but not in human subjects. AIMS: The aim of the present study was to investigate the response in vivo of zinc transporters in the human enterocyte to dietary zinc supplementation. SUBJECTS: Eighteen patients who had previously undergone ileostomy, all free of any symptoms of inflammatory bowel disease. METHODS: Subjects took a daily zinc supplement of 25 mg for 14 days in a double blind, placebo controlled, crossover trial. The effect of the supplement on expression in ileal biopsies of the zinc transporters SLC30A1, SLC30A4, SLC30A5, SLC39A1, SLC39A4, and metallothionein was measured by reverse transcription-polymerase chain reaction RT-PCR. Expression of SLC30A1, SLC30A5, and SLC39A4 was also examined by immunoblotting. RESULTS: The zinc supplement reduced SLC30A1 mRNA (1.4-fold) together with SLC30A1, SLC30A5, and SLC39A4 protein (1.8-fold, 3.7-fold, and to undetectable levels, respectively) in ileal mucosa and increased metallothionein mRNA (1.7-fold). The supplement had no effect on expression of SLC30A4 or SLC39A1 mRNA. Localisation of SLC30A5 at the apical human enterocyte/colonocyte membrane and also at the apical membrane of Caco-2 cells was demonstrated by immunohistochemistry. Commensurate with these observations in zinc supplemented human subjects, SLC30A1, SLC30A5, and SLC39A4 mRNA and protein were reduced in Caco-2 cells cultured at 200 muM compared with 100 muM zinc. CONCLUSIONS: These observations indicate that, in response to variations in dietary zinc intakes, regulated expression of plasma membrane zinc transporters in the human intestine contributes to maintenance of zinc status.

National Prescribing Service: creating an implementation arm for national medicines policy.

Medicines make an essential contribution to the health of the community, but rapidly rising drug budgets have caused governments to seek ways of ensuring this expenditure results in value for money. The National Prescribing Service was established against this background to implement a quality use of medicines service as part of the National Medicines Policy. A range of programmes that attempt to use evidence-based strategies to deliver evidence-based messages have been established. These use multifaceted interventions, such as newsletters, prescriber feedback, clinical audit and educational visiting, that are provided both centrally, through the national office, and locally, through Divisions of General Practice. The work is underpinned by an evaluation strategy that incorporates strong qualitative elements as well as an emphasis on time-series analyses for changes in drug utilization. Some 80% of Australian general practitioners have voluntarily participated in activities such as educational visiting and clinical audit within the National Prescribing Service programmes. New programmes for the community and consumers will be coordinated with the work that has become well established within general practice.

Aggressive fibromatosis of the abdominal wall, limbs and limb girdles.

BACKGROUND: Aggressive fibromatosis (AF) is a rare soft-tissue tumour. It is histologically benign but locally aggressive and destructive. Surgery is the mainstay of treatment. This was a review of patients treated at the Royal Marsden Hospital between 1986 and 2003. METHODS: Records of patients with abdominal wall (23) and limb or limb girdle (86) AF were studied to determine patient, tumour and treatment characteristics and outcome. Children and patients with intra-abdominal AF were excluded. The data were analysed by Cox regression to calculate univariate hazard ratios. RESULTS: Twenty-one (19.3 per cent) of 109 patients developed recurrence after a median follow-up of 39 months. Recurrence did not correlate with surgical margin or any other variable. CONCLUSION: Wide excisional, function-preserving surgery is the goal in treatment of AF. Surgery for recurrent disease is often curative. Tumours are frequently irresectable at presentation and an initial period of expectant observation is advisable because growth arrest is a common feature of the disease.

Expression, candidate gene, and population studies of the melanocortin 5 receptor.

In mouse the melanocortin 5 receptor is known to regulate sebaceous gland function. To clarify its role in man, we have studied melanocortin 5 receptor expression in skin, and allelic variation at the melanocortin 5 receptor locus in diverse human populations and candidate disease groups. Melanocortin 5 receptor protein and mRNA expression were studied by immunohistochemistry and reverse transcriptase polymerase chain reaction. Melanocortin 5 receptor mRNA was detected in normal skin and cultured keratinocytes but not in cultured fibroblasts or melanocytes. Immunohistochemistry revealed melanocortin 5 receptor immunoreactivity in the epithelium and appendages, including the sebaceous gland, eccrine glands, and apocrine glands, as well as low level expression in the interfollciular epidermis. In order to screen for genetic diversity in the melanocortin 5 receptor that might be useful for allelic association studies we sequenced the entire melanocortin 5 receptor coding region in a range of human populations. One nonsynonymous change (Phe209Leu) and four synonymous changes (Ala81Ala, Asp108Asp, Ser125Ser, and Thr248Thr) were identified. Similar results were found in each of the populations except for the Inuit in which only the Asp108Asp variant was seen. The apparent "global distribution" of melanocortin 5 receptor variants may indicate that they are old in evolutionary terms. Variation of melanocortin 5 receptor was examined in patients with acne (n = 21), hidradenitis supprativa (n = 4), and sebaceous gland lesions comprising sebaceous nevi, adenomas, and hyperplasia (n = 13). No additional mutations were found. In order to determine the functional status of the Phe209Leu change, increase in cAMP in response to stimulation with alpha-melanocyte-stimulating hormone was measured in HEK-293 cells transfected with either wild-type or the Phe209Leu variant. The variant melanocortin 5 receptor was shown to act in a concentration-dependent manner, which did not differ from that of wild type. We have therefore found no evidence of a causative role for melanocortin 5 receptor in sebaceous gland dysfunction, and in the absence of any association between variation at the locus and disease group, the pathophysiologic role of the melanocortin 5 receptor in man requires further study.

Loss of function mutations of the human melanocortin 1 receptor are common and are associated with red hair.

The melanocortin 1 receptor is a G-protein-coupled receptor that acts as a control point for control of the eumelanin/phaeomelanin ratio in mouse hair. MC1 receptor loss of function mutations lead to an increase in the ratio of phaeomelanin/eumelanin in many mammals resulting in yellow or red coat colours. We have previously shown that several common point mutations in the human MC1 receptor are overrepresented in North European redheads and in individuals with pale skin. In order to determine the functional significance of these changes we have carried out transfection and binding studies. Expression of the Val60Leu, Arg142His, Arg151Cys, Arg160Trp, and Asp294His receptors in COS 1 cells revealed that these receptors were unable to stimulate cAMP production as strongly as the wild type receptor in response to alpha-melanocyte-stimulating hormone stimulation. None of the mutant receptors displayed complete loss of alphaMSH binding, with only the Arg142His and Asp294His displaying a slight reduction in binding affinity.

Turning research into policy: a survey on adolescent condom use.

PIP: Results from a 1991-92 survey of the condom use beliefs, attitudes, and practices of 373 US inner-city high school students reveal educational policy implications which would further the goal of encouraging sexually active youth to use condoms. First, the issue of sexual desire must be treated realistically. Many teenage girls, as well as boys, want to fulfill their desires by having sexual intercourse. The standard educational approach to teenage boys also assumes that they are sexual exploiters, but this may also simply be a cultural construct rather than a reality. Addressing physical pleasure issues may be particularly important in encouraging condom use, but it would be inappropriate for such discussions to encourage sexual activity. Adolescent ambivalence towards risk taking and mortality must also be considered. Standard definitions of risk (which exclude the health risk of pregnancy) contain the common perception that adolescent males are greater risk-takers than females. When such definitions exclude certain behaviors, the classification of risk becomes a social construct. Adolescents are not completely unaware of the dangers of risky behavior and may even overestimate their chances of getting pregnant or contracting HIV. Adolescents may find the risk of offending a partner to be more important than the risk of contracting a disease in 10 years. When teenagers believe themselves to be in love, they are less likely to insist on condom use. Educators must deconstruct risks and address each one specifically. High levels of knowledge about HIV transmission fail to predict previous, current, or intended condom use. Thus, while facts are important, teenagers also need to learn the social skills surrounding condom use. Students have easy accessibility to condoms, but embarrassment may pose a barrier to acquisition. This embarrassment, however, showed no correlation to actual or intended condom use. By working within the context of the adolescent mind and world, where condom use is seen as a strategy complicated by sexual desire, feelings of immortality, and negative trade-offs, educators may be able to learn how to maximize condom use and, thus, help young people protect themselves against pregnancy, sexually transmitted diseases, and HIV.^ieng

Medical sociology and health policy: where are the connections?

The attempt to assess the impact of medical sociology on health policy encounters operational difficulties regarding what should be counted. After reviewing these difficulties, we argue that there is potential interest among policymakers for sociological contributions to policy debates, but that policy impact has been limited by sociologists' ambivalence, academic career considerations, and by health research becoming a distinct field of research. Suggestions for increasing sociologists' policy impact include attending to policy relevance in both the design of studies and the dissemination of results and by sociologists' becoming more oriented to the field of health policy research.

Design, expression, and crystallization of recombinant lectin from the garden pea (Pisum sativum).

The propeptide form of the lectin from the garden pea (Pisum sativum agglutinin) has been expressed in Escherichia coli by attaching its cDNA to an inducible promoter. By a number of criteria, including the ability to form dimers, hemagglutination titer, Western blot, and enzyme-linked immunosorbent assay, the resulting propeptide molecule is virtually indistinguishable from the mature proteolytically processed lectin isolated from peas. Preliminary crystallization experiments using the recombinant propeptide lectin yield crystals in space group P2(1)2(1)2(1) with a = 64.8 A, b = 73.8 A, and c = 109.0 A (1 A = 0.1 nm) that diffract to 2.8-A resolution. This unit cell size is quite similar to the unit cell determined for native pea lectin, suggesting that the overall structure of the recombinant prolectin is virtually identical.

Fluorescence quenching studies of the structures of calf gamma-II, III, and IV crystallins.

The structures of the calf lens crystallin fractions gamma-II, gamma-III, and gamma-IV have been investigated using the fluorescence quenching method. The three crystallin fractions showed very large differences in the quenching rates of their fluorescent tryptophan residues, for quenching by acrylamide or iodide in pH 7.5 phosphate buffer solutions. The experimentally measured quenching rate constants were kq(II) = 3.2 x 10(8), kq(III) = 9.9 x 10(8), and kq(IV) = 1.8 x 10(9) M-1 sec-1. Smaller rate constants were obtained for iodide quenching of the three crystallins, but the values were in approximately the same ratios as the ones found for acrylamide quenching. The conclusion is that the tryptophan residues in gamma-II crystallin are 6-10 times less easily quenched than those of gamma-IV crystallin and 3-6 times less easily quenched than those of gamma-III. These conclusions are in accord with those reached by Mandal et. al. based on fluorescence and CD data, who found the following order of Trp hydrophobicities: gamma-II greater than gamma-III greater than gamma-IV. The significance of these structural differences for lens function and stability remains to be elucidated.

Acrylamide and iodide fluorescence quenching as a structural probe of tryptophan microenvironment in bovine lens crystallins.

Fluorescence quenching using acrylamide and iodide quenchers has been used to investigate the microenvironments of tryptophan residues in bovine alpha-, beta-, and gamma-crystallin fractions. Acrylamide quenching is very sensitive to the degree of tryptophan accessibility to the solvent containing the acrylamide. Since acrylamide is able to diffuse into the interior of the protein, accessibility to acrylamide may result from Trp residues lying at the surface of the protein or from the existence of channels leading to the interior of the protein. Iodide ion is hydrated and is limited by its large size and charge to quenching of tryptophan residues lying at or near the surface of proteins. Tryptophan residues in the lens crystallin fractions were found to be highly accessible to acrylamide, yet the rate of quenching by acrylamide was very low, indicating that the tryptophan residues of the lens crystallin fractions occupy predominately hydrophobic environments. The high accessibility to acrylamide likely results from diffusion of acrylamide into the interior of the protein. Accessibility to iodide was much lower, as was the rate of quenching by iodide, adding support to the conclusions from acrylamide quenching.

Tyrosine-to-tryptophan energy transfer and the structure of calf gamma-II crystallin.

The efficiency of electronic energy transfer from tyrosine to tryptophan residues in the lens protein calf gamma II crystallin has been determined from measurements of fluorescence excitation spectra at 25 degrees C and independently from phosphorescence spectroscopy at 77 K. The total transfer efficiency from the fifteen tyrosines to the four tryptophans in native calf gamma II was found to be t = 78 +/- 10%, at 25 degrees C. The expected value based on Förster theory and the X-ray structure of the protein is t = 83%, in good agreement with the present experimental result. The transfer efficiency measured in denatured calf gamma II at 25 degrees C was t = 20 +/- 10%, in good agreement with the value t = 25% expected for the completely denatured protein, based on Förster theory and the known amino acid sequence. These results are of interest for several reasons. First, energy transfer can provide a simple , experimental confirmation of lens protein structures determined from X-ray data or from computer modeling studies. Second, the present studies show that energy transfer measurements can be used to monitor the effect of denaturants on lens protein structure, an aspect not readily investigated by X-ray crystallography. Third, the present electronic energy transfer studies may be relevant to understanding the mechanism of UV photodamage in lens proteins and hence in whole lenses.

Paralysis of accommodation in infectious mononucleosis.

A case report of a 22-year-old patient with accommodative paralysis is presented including (1) the five-year history beginning with infectious mononucleosis; (2) recent clinical examination showing accomodative paralysis and reduced pupilary responses to light and near; (3) objective recordings confirming both the absence of any accommodation and the presence of pupillary responses to monocular and binocular near stimuli and to light, the latter with pupillary escape; and finally (4) neuropharmacological tests showing 7-diopter accommodative responses to pilocarpine (an acetylcholine substitute acting directly on the ciliary muscle receptor sites) and absent responses to demecarium bromide (a cholinesterase blocking agent which potentiates neurally released acetylcholine). Infectious mononucleosis includes ocular signs and symptoms. In young persons with accommodative difficulties, infectious mononucleosis should be suspected.