Biomedical subjects
Publications and source records attributed to G Evans.
Methods of collection and freezing of semen of some deer species and aspects of controlled reproduction associated with the use of frozen-thawed semen by artificial insemination (AI) are discussed.
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The distribution of neurotensin receptor 1 immunoreactivity in the rat brain was studied using an antibody against the amino-terminal of the receptor expressed as a fusion protein with glutathione-S transferase. Affinity purified antibodies detected the fusion protein and the complete neurotensin receptor sequence expressed in Escherichia coli. The immunostaining was abolished by preabsorption with the amino-terminal fusion protein. Immunoreactive neurotensin receptor 1 immunoreactivity was detected on cell bodies and their processes in a number of CNS regions. In agreement with previous binding studies neurotensin receptor 1 immunoreactivity was particularly localised in cell bodies in the basal forebrain, nucleus basalis and substantia nigra. At the electron microscope level immunoreactivity was found both in axonal bouton and dendrites and spines in the basal forebrain indicating that neurotensin may act both pre- and post-synaptically. There were several regions such as the substantia gelatinosa, ventral caudate-putamen and the lateral reticular nucleus where the neurotensin receptor 1 positive cells had not previously been reported, indicating that distribution of this receptor is widespread.
Few studies have determined whether carotid artery intima-media thickness (IMT) is associated prospectively with risk of first ischemic stroke. In the Atherosclerosis Risk in Communities Study, carotid IMT, an index of generalized atherosclerosis, was defined as the mean of IMT measured by B-mode ultrasonography at six sites of the carotid arteries. The authors assessed the relation of mean IMT to stroke incidence over 6-9 years' follow-up (1987-1995) among 7,865 women and 6,349 men aged 45-64 years without prior stroke at baseline in four US communities. There were 90 incident ischemic stroke events for women and 109 for men. In sex-specific Cox proportional hazards models adjusting only for age, race, and community, the hazard rate ratios comparing extreme mean IMT values (> or =1 mm) to values less than 0.6 mm were 8.5 for women (95% confidence interval: 3.5, 20.7) and 3.6 for men (95% confidence interval: 1.5, 9.2). The relation was graded, and models with cubic splines indicated significant nonlinearity, with hazards increasing more rapidly at lower IMTs than at higher IMTs. Thus, models using linear IMT values substantially underestimate the strength of the association at lower IMTs. The strength of the association was reduced by the inclusion of putative stroke risk factors, but it remained elevated at higher IMTs. Hence, mean carotid IMT is a noninvasive predictor of future ischemic stroke incidence.
The Structural Biology Center beamline, 19ID, has been designed to take full advantage of the highly intense undulator radiation and very low source emittance available at the Advanced Photon Source. In order to keep the X-ray beam focused onto the pre-sample slits, a novel position-sensitive PIN diode array has been developed. The array consists of four PIN diodes positioned upstream of a 0.5 microm-thick metal foil placed in the X-ray beam. Using conventional difference-over-the-sum techniques, two-dimensional position information is obtained from the metal foil fluorescence. Because the full X-ray beam passes through the metal foil, the true beam center-of-mass is measured. The device is compact, inexpensive to construct, operates in a vacuum and has a working range of 8 mm x 10 mm that can be expanded with design modifications. Measured position sensitivity is 1-2 microm. Although optimized for use in the 5-25 keV energy range, the upper limit can be extended by changing metals or adjusting foil thickness.
Macrophage activation has been recognized as playing a central role in chronic inflammatory diseases in general and, more specifically, in the vascular wall during the progression of atherosclerotic lesions. Macrophage-activating factors present within the atherosclerotic lesion include the colony-stimulating factors and gamma interferon (IFNgamma). In the present study, the effects of IFNgamma on macrophage binding and uptake of fluorochrome-labeled high density lipoprotein (HDL) were investigated by flow cytometry and by measuring the amount of the type B scavenger receptors CD36 and scavenger receptor type B (SR-BI) by Northern blot analysis. IFNgamma-, but not granulocyte macrophage colony-stimulating factor (GM-CSF)-treated murine peritoneal macrophages displayed a two- to threefold decrease in Dil-labeled HDL uptake. This effect was observed in the absence of a comparable decrease in SR-BI message and protein or CD36 message. This decrease in both HDL binding and uptake was reversed by the peroxisome proliferator-activated receptor gamma (PPARgamma) agonist, 15-deoxy-delta12,4-prostaglandin J2 (15d-PGJ2), which also inhibited the IFNgamma induction of the beta2 integrin CD11a. Furthermore, 15d-PGJ2 increased the expression of SR-BI and CD36 message and SR-BI protein which was reflected in an increase in HDL binding and uptake. These results suggest a role for PPARgamma agonists in modulating the IFNgamma-mediated macrophage effector functions relevant to atherosclerotic disease progression.
The purpose of this study was to examine the independent and interactive effects of HIV-1 serostatus and cocaine on neuropsychological (NP) performance in a sample of 237 gay and bisexual urban-dwelling African American men. Consistent with current evidence, it was expected that the greatest neuropsychological performance deficits would be evident (1) in the symptomatic seropositives (SSPs), especially in domains affected by HIV (i.e., memory and psychomotor speed), and on tests that are sensitive to subtle slowing; (2) in those who are recent and frequent cocaine abusers; and (3) in those who are both HIV seropositive and cocaine abusers. Multivariate analyses controlling for age and alcohol use confirmed expectations, with symptomatic seropositives (SSPs) evidencing significantly poorer psychomotor speed than the seronegatives (SNs), and slower reaction time and poorer nonverbal memory than the asymptomatic seropositives (ASPs). Moderate to heavy recent cocaine use was associated with slower psychomotor speed. However, contrary to expectations, no interaction of serostatus and cocaine was noted for any NP domain, and the expected serostatus and cocaine effects on verbal memory and frontal systems were not obtained. Level of alcohol consumption exacerbated the detrimental effects of HIV-1 on a computerized reaction time test which is especially sensitive to subtle slowing. This study provides one of the first descriptions of the neuropsychological effects of HIV-AIDS in a non-injection drug-using community sample of gay and bisexual African American men.
The survival to term of 414 sheep embryos, thawed and transferred after conventional cryopreservation and storage for 13 years, was evaluated. A concurrent experiment involving the transfer of vitrified embryos to 91 ewes and artificial insemination of 51 ewes with frozen-thawed semen from sires of the long-term cryopreserved embryos provided forms of control treatments. The donor ewes had a mean ovulation rate of 10.9, and 7.1 embryos per ewe were cryopreserved. Each recipient ewe received two embryos and pregnancy was assessed at Day 18, Day 54 and term. The pregnancy rate was lower in the long-term embryo group than the artificial insemination group at Day 18 (P<0.01) and Day 54 (P<0.05), although the difference at term (31% v. 49%) was not significant, with the vitrified embryo group being similar to the long-term group. Embryo survival to birth was 21%, with the majority of loss (80%) occurring by Day 18. The later stage of development and higher grade of transferred embryos and the older age of donor ewes resulted in a significantly higher (P<0.01) pregnancy rate at Day 54 and term, and percentage of lambs born and weaned. Other effects of donor ewes (genotype, superovulation treatment, number of ovulations and embryos cryopreserved) were not significant. Implications for the design of genetic evaluation and germplasm conservation programmes using embryo cryopreservation technology are discussed.
In order to investigate the interaction of fresh and frozen-thawed spermatozoa with oviduct epithelial cells, spermatozoa were co-incubated with ovine oviduct epithelial cell monolayers (OECM) derived from either complete oviducts, at any stage of the oestrous cycle (Experiments 1 and 2), or from different regions of the oviduct at different stages of the cycle (Experiment 3). Fresh and frozen-thawed spermatozoa displayed different patterns of binding to, and release from, the OECM. Frozen-thawed spermatozoa immediately bound to the complete oviduct OECM and were released after 2 h. A small proportion of fresh spermatozoa bound immediately, increasing to a maximum after 2 h, and were gradually released thereafter. When only the cells that were released from the OECM were observed by chlortetracycline staining in Experiment 2, it was found that the presence of an OECM increased the number of capacitated fresh spermatozoa while decreasing the number of capacitated frozen-thawed spermatozoa. Overall, the OECM advanced the membrane state of both types of spermatozoa from uncapacitated to acrosome-reacted. Fresh and frozen-thawed spermatozoa bound to OECM derived from the cells of the isthmus and the ampulla in similar proportions. However, more spermatozoa were capacitated when incubated with OECM derived from isthmic rather than ampullary cells. Higher proportions of fresh spermatozoa bound to, and were acrosome-reacted following incubation with OECM derived from post- rather than pre-ovulatory tracts. Such differences were not observed for frozen-thawed spermatozoa. The findings reported in this study show that fresh and frozen-thawed spermatozoa behave differently when in contact with oviduct cells in vitro. This may be a consequence of the more advanced membrane state of the frozen spermatozoa upon thawing.
This study investigated the effects of a reduction in gonadotrophins, by means of differently timed active and passive gonadotrophin-releasing hormone (GnRH) immunization at various stages, on the maintenance of early pregnancy in pigs. In the first experiment crossbred sows (n = 11) were immunized against GnRH using a commercial vaccine on the day of farrowing, mated at the first oestrus, and a booster immunization was administered 10 days (n = 7) or 20 days (n = 4) after mating. Plasma samples were collected every second day and assayed for GnRH antibodies and progesterone. Pregnancy testing was carried out by real time ultrasound. None of the sows receiving the booster immunization 10 days after mating were pregnant on Day 18 after mating. All sows receiving the booster on Day 20 after mating aborted, with a mean vaccination-to-abortion interval of 10.0 +/- 1.5 days. In the second experiment, crossbred gilts (n = 6) were passively immunized by infusing (i.v.) GnRH immune pig serum on Day 12 after mating. Luteinizing hormone profiles were determined on the day before immunization and one day afterwards. Daily plasma samples were assayed for GnRH antibodies and progesterone. None of the gilts were pregnant 18 days after mating, compared with 5 of 6 non-immunized controls. Booster immunization 10 days after mating resulted in failure of embryonic development and establishment of pregnancy before the corpora lutea (CL) regressed, according to progesterone profiles, whereas immunization 20 days after mating resulted in regression of CL followed by abortion. Passive immunization 12 days after mating had a similar effect to the active immunization 10 days after mating. These results demonstrate two different outcomes of active GnRH immunization depending on the timing of immunization, and indicate that loss of pregnancy between Days 12 and 18 may occur due a reduction in progesterone rather than complete failure of the CL, as occurs at later stages. The findings may provide an explanation for the reduced fertility of pigs in the summer-autumn period.
In this paper we examine new empirical evidence on the coherence and magnitude of the main classes in the Goldthorpe class schema. Particular attention is paid to issues that have recently been a source of academic dispute: the coherence and size of the service class and the distinction between the service class and intermediate classes. Using recently available British data collected by the Office for National Statistics we examine: (i) the extent to which measures of class-relevant job characteristics are empirically discriminated by the categories of the schema; (ii) the structure of a 'contract type' dimension of employment relations conceived of as a categorical latent variable; and (iii) the association between this latent variable and both the Goldthorpe class schema and a related measure socio-economic group (SEG). We find that the data are consistent with the existence of a three category latent 'contract type' variable largely corresponding to the notions of service, intermediate and wage-labour contracts explicit in discussions of the theoretical rationale for the Goldthorpe schema. We further find a substantial degree of fit between the latent 'contract types' and the schema. However, the service class fault line appears to lie within class I and II of the schema rather than between them and the intermediate classes which suggests a revised, smaller service class would better capture the reality of the contemporary British occupational structure.
Until modelling is complete, macromolecular structures are refined in the absence of a model for some of the atoms in the crystal. Techniques for defining positional probability distributions of atoms, and using them to model the missing part of a macromolecular crystal structure and the bulk solvent, are described. The starting information may consist of either a tentative structural model for the missing atoms or an electron-density map. During structure completion and refinement, the use of probability distributions enables the retention of low-resolution phase information while avoiding premature commitment to uncertain higher resolution features. Homographic exponential modelling is proposed as a flexible, compact and robust parametrization that proves to be superior to a traditional Fourier expansion in approximating a model protein envelope. The homographic exponential model also has potential applications to ab initio phasing of Fourier amplitudes associated with macromolecular envelopes.
It is demonstrated that standard in-house protein crystal X-ray diffraction apparatus can be used to measure very low resolution reflections with only a few modifications. The apparatus and modifications are described in detail and tested on two different macromolecular crystal samples: lysozyme and the 30S ribosomal subunit. Contrast-variation measurements on tetragonal hen egg-white lysozyme demonstrate the potential usefulness of the apparatus in providing accurate data for the determination of macromolecular envelopes. In contrast, the measurement of very low resolution diffraction from crystals of the 30S ribosome subunit illustrates how in-house facilities can provide data from small weakly diffracting crystals of a very large macromolecule.
BACKGROUND: Juvenile polyposis syndrome (JPS) is characterised by gastrointestinal (GI) hamartomatous polyposis and an increased risk of GI malignancy. Juvenile polyps also occur in the Cowden (CS), Bannayan-Ruvalcaba-Riley (BRRS) and Gorlin (GS) syndromes. Diagnosing JPS can be problematic because it relies on exclusion of CS, BRRS, and GS. Germline mutations in the PTCH, PTEN and DPC4 (SMAD4) genes can cause GS, CS/BRRS, and JPS, respectively. AIMS: To examine the contribution of mutations in PTCH, PTEN, and DPC4 (SMAD4) to JPS. METHODS: Forty seven individuals from 15 families and nine apparently sporadic cases with JPS were screened for germline mutations in DPC4, PTEN, and PTCH. RESULTS: No patient had a mutation in PTEN or PTCH. Five different germline mutations were detected in DPC4; three of these were deletions, one a single base substitution creating a stop codon, and one a missense change. None of these patients had distinguishing clinical features. CONCLUSIONS: Mutations in PTEN and PTCH are unlikely to cause juvenile polyposis in the absence of clinical features indicative of CS, BRRS, or GS. A proportion of JPS patients harbour DPC4 mutations (21% in this study) but there remains uncharacterized genetic heterogeneity in JPS.
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The objective of this study was to describe associations of retinal arteriolar abnormalities with clinical and subclinical manifestations of atherosclerosis and a broad group of risk factors for vascular disease. A biracial population of 8772 adults (aged 48 to 72 years) living in 4 communities was examined from 1993 to 1995 were studied for that purpose. Retinal arteriovenous nicking and focal arteriolar narrowing were determined by light-box grading of a 45 degrees fundus photograph by use of a standardized protocol. Diameters of arterioles and venules were measured in digitized photographs, and a summary arteriolar-to-venular ratio was derived as an index of generalized arteriolar narrowing. Focal arteriolar narrowing was associated only with hypertension. Generalized arteriolar narrowing was associated with carotid plaque but not with any other evidence of atherosclerosis, either clinical (cardiovascular disease or stroke) or subclinical (carotid or popliteal artery thickness or lower limb obstructive disease), or with plasma cholesterol. It was also associated with smoking, with inflammatory markers (white blood cell count, fibrinogen, and reduced albumin), and with the triglyceride and high density lipoprotein cholesterol changes associated with inflammation. Arteriovenous nicking was inconsistently associated with subclinical atherosclerosis. It was not associated with cardiovascular disease, stroke, or plasma cholesterol. Arteriovenous nicking was associated with markers of inflammation and vascular endothelial dysfunction (von Willebrand factor and factor VIII). Arteriolar narrowing and nicking appear to be related to hypertension and inflammatory factors. Nicking may also be related to endothelial dysfunction. Results suggest that these arteriolar changes are pathologically distinct from atherosclerosis. Including their measurement in population studies may permit evaluation of the independent contribution of arteriolar disease to various ischemic diseases of the heart, brain, and other organs.