Psychotic disorders in young adults with perinatally acquired HIV: a UK case series.

BACKGROUND: Increasing numbers of children with perinatally acquired HIV (PaHIV) are transitioning into adult care. People living with behaviourally acquired HIV are known to be at more risk of psychosis than uninfected peers. Young adults living with PaHIV face numerous risk factors; biological: lifelong exposure to a neurotrophic virus, antiretroviral medication and immune dysfunction during brain development, and environmental; social deprivation, ethnicity-related discrimination, and migration-related issues. To date, there is little published data on the prevalence of psychotic illness in young people growing up with PaHIV. METHODS: We conducted a retrospective case note review of all individuals with PaHIV aged over 18 years registered for follow up at a dedicated clinic in the UK (n = 184). RESULTS: In total, 12/184 (6.5%), median age 23 years (interquartile range 21-26), had experienced at least one psychotic episode. The presentation and course of the psychotic episodes experienced by our cohort varied from short-lived symptoms to long term illness and nine (75%) appear to have developed a severe and enduring mental illness requiring long term care. CONCLUSION: The prevalence of psychosis in our cohort was clearly above the lifetime prevalence of psychosis in UK individuals aged 16-34 years, which has been reported to be 0.5-1.0%. This highlights the importance of clinical vigilance regarding the mental health of young people growing up with PaHIV and the need to integrate direct access to mental health services within the HIV centres providing medical care.

Accuracy of colposcopy-directed punch biopsies: a systematic review and meta-analysis.

BACKGROUND: The colposcopy-directed punch biopsy is widely used in the management of women with abnormal cervical cytology; however, its accuracy compared with definitive histology from an excision biopsy is not well established. OBJECTIVES: To assess the accuracy of the colposcopy-directed punch biopsy to diagnose high-grade cervical intraepithelial neoplasia (CIN) by performing a systematic review and meta-analysis. SEARCH STRATEGY: A systematic search of MEDLINE, EMBASE and the Cochrane Library was performed. SELECTION CRITERIA: Articles that compared the colposcopically directed cervical punch biopsy with definitive histology from an excisional cervical biopsy or hysterectomy. DATA COLLECTION AND ANALYSIS: Random effects and hierarchical summary receiver operating characteristic regression models were used to compute the pooled sensitivity and specificity applying different test cut-offs for outcomes of high-grade CIN. MAIN RESULTS: Thirty-two papers comprising 7873 paired punch/definitive histology results were identified. The pooled sensitivity for a punch biopsy defined as test cut-off CIN1+ to diagnose CIN2+ disease was 91.3% (95% CI 85.3-94.9%) and the specificity was 24.6% (95% CI 16.0-35.9%). In most of the studies, the majority of enrolled women had positive punch biopsies. Pooling of the four studies where the excision biopsy was performed immediately after the punch biopsy, and where the rate of positive punch biopsies was considerably lower, yielded a sensitivity of 81.4% and specificity of 63.3%. AUTHOR'S CONCLUSION: The observed high sensitivity of the punch biopsy derived from all studies is probably the result of verification bias.

Non-viral transfection of goat germline stem cells by nucleofection results in production of transgenic sperm after germ cell transplantation.

Germline stem cells (GSCs) can be used for large animal transgenesis, in which GSCs that are genetically manipulated in vitro are transplanted into a recipient testis to generate donor-derived transgenic sperm. The objectives of this study were to explore a non-viral approach for transgene delivery into goat GSCs and to investigate the efficiency of nucleofection in producing transgenic sperm. Four recipient goats received fractionated irradiation at 8 weeks of age to deplete endogenous GSCs. Germ cell transplantations were performed 8-9 weeks post-irradiation. Donor cells were collected from testes of 9-week-old goats, enriched for GSCs by Staput velocity sedimentation, and transfected by nucleofection with a transgene construct harboring the human growth hormone gene under the control of the goat beta-casein promoter (GBC) and a chicken beta-globin insulator (CBGI) sequence upstream of the promoter. For each recipient, transfected cells from 10 nucleofection reactions were pooled, mixed with non-transfected cells to a total of 1.5 × 10(8) cells in 3 ml, and transplanted into one testis (n = 4 recipients) by ultrasound-guided cannulation of the rete testis. The second testis of each recipient was removed. Semen was collected, starting at 9 months after transplantation, for a period of over a year (a total of 62 ejaculates from four recipients). Nested genomic PCR for hGH and CBGI sequences demonstrated that 31.3% ± 12.6% of ejaculates were positive for both hGH and CBGI. This study provides proof-of-concept that non-viral transfection (nucleofection) of primary goat germ cells followed by germ cell transplantation results in transgene transmission to sperm in recipient goats.

Health and reproductive profiles of malaria antigen-producing transgenic goats derived by somatic cell nuclear transfer.

Nuclear transfer (NT) using transfected primary cells is an efficient approach for the generation of transgenic goats. However, reprogramming abnormalities associated with this process might result in compromised animals. We examined the health, reproductive performance, and milk production of four transgenic does derived from somatic cell NT. Goats were derived from two fetal cell lines, each transfected with a transgene expressing a different version of the MSP-1(42) malaria antigen, either glycosylated or non-glycosylated. Two female kids were produced per cell line. Health and growth of these NT animals were monitored and compared with four age-matched control does. There were no differences in birth and weaning weights between NT and control animals. The NT does were bred and produced a total of nine kids. The control does delivered five kids. The NT does expressing the glycosylated antigen lactated only briefly, probably as a result of over-expression of the MSP-1(42) protein. However, NT does expressing the non-glycosylated antigen had normal milk yields and produced the recombinant protein. These data demonstrated that the production of healthy transgenic founder goats by somatic cell NT is readily achievable and that these animals can be used successfully for the production of a candidate Malaria vaccine.

Perimenopausal women in estrogen vasomotor trials: contribution to placebo effect and efficacy outcome.

OBJECTIVE: To determine the difference, if any, in the placebo response when both perimenopausal and postmenopausal women are enrolled, compared with postmenopausal women alone, in a study assessing the efficacy of synthetic conjugated estrogen tablets on moderate-to-severe vasomotor symptoms (MSVS). METHODS: A total of 120 healthy women (72 active; 48 placebo) complaining of moderate-to-severe vasomotor symptoms were enrolled in a randomized, placebo-controlled, double-blind, multicenter clinical trial. In all, 109 patients completed treatment to week 12. Women were enrolled using minimal inclusion and exclusion criteria, and included perimenopausal women (n = 34; 0 to 6 months since last menses) as well as menopausal women (n = 79; > 12 months since last menses). RESULTS: Changes in MSVS in the intent-to-treat (ITT) population showed differences between the active and placebo treatments at weeks 4 (p < 0.022), 8 (p < 0.010) and 12 (p < 0.010). At week 12, the mean percentage reduction in MSVS was 81% for the active treatment group and 58% in the placebo treatment group. To examine the placebo response, the ITT population was broken down into groups defined by the time since their last menses. The perimenopausal group (0 to 6 months since last menses) demonstrated a consistently higher placebo response than that of the postmenopausal groups (> 12 months since last menses). CONCLUSIONS: Perimenopausal women contributed to a higher placebo response, compared with the rate of response previously reported in clinical studies of estrogen replacement in postmenopausal women. Including perimenopausal women in future vasomotor symptom trials will require study populations of sufficient size to maintain the statistical power to demonstrate a difference between therapeutic response to active or placebo treatment.

Estrogen levels and lipid peroxidation following exercise.

Recent large-scale clinical trials have confirmed that estrogens are powerful antioxidants that effectively prevent lipid peroxidation. In particular, estrogen decreases oxidative modification of low-density lipoprotein both in vitro and in vivo. Estrogen levels fluctuate during the menstrual cycle and during perimenopause, and they are permanently depressed at menopause. Thus, it is of interest to evaluate the lipid peroxidation response under conditions of low estrogen status. It is of additional interest to evaluate lipid peroxidation under such conditions in response to situations that augment free radical production, such as exercise. Aerobic exercise, and more recently anaerobic exercise regimens, are reported to variably affect free radical production and lipid peroxidation. In addition to the exercising woman's estrogen status and menstrual history, other factors to consider when evaluating an exercising woman's risk for increased lipid peroxidation include the intensity of exercise and/or the woman's fitness level as well as her current dietary antioxidant status. (c) 2001 by CHF, Inc.

Regional distribution of gonadotropin-releasing hormone-like, beta-endorphin-like, and methionine-enkephalin-like immunoreactivities in the central nervous system of the goat.

Regional distribution of gonadotropin-releasing hormone (GnRH)-like-, beta-endorphin (beta-end)-like-, and methionine-enkephalin (met-enk)-like-immunoreactivity was quantified across various regions of the central nervous system (CNS) of male and female goats by using highly specific radioimmunoassays. All the animals were sacrificed during the months of March through June (non-breeding season). Although the distribution of these three neuropeptides was similar to other mammalian species, species-specific gender differences in the levels of neuropeptides were noticed in the goat CNS. Highest levels of GnRH-like immunoreactivities were found in the hypothalamus. The hypothalamus of male goats exhibited significantly higher levels of GnRH-like immunoreactivities compared to female goats. Other regions exhibiting GnRH-like immunoreactivities included olfactory bulbs, preoptic and supraoptic regions, and mamillary bodies. Both beta-end- and met-enk immunoreactivities were detected in all selected regions of goat CNS, but highest levels of these opioid peptide-like immunoreactivities were limited to the forebrain regions of the goat. The supraoptic area of the female goats contained significantly higher levels of beta-end-like immunoreactivities than that of the male goats. Met-enk-peptide-like immunoreactivity also exhibited gender-specific differences in its content in some regions of the CNS. The male goats exhibited significantly higher levels of met-enk-like immunoreactivity in both the striatal and hypothalamic regions of the brain.

Parent-offspring trios: a resource to facilitate the identification of type 2 diabetes genes.

The transmission disequilibrium test with use of trios (an affected proband with both parents) is a robust method for assessing the role of gene variants in disease that avoids the problem of population stratification that may confound conventional case/control studies and allows the detection of parent-of-origin effects. Trios have played a major role in defining genes in a number of polygenic conditions, including type 1 diabetes. We assessed the prevalence, clinical characteristics, and suitability for defining type 2 susceptibility genes of European type 2 diabetes trios. In a Caucasian population in the U.K., only 2.5% of type 2 patients had both parents alive. Using a nationwide strategy, we collected 182 trios defined by strict clinical criteria. Immunological and genetic testing resulted in the exclusion of 25 trios as a result of latent autoimmune diabetes (n = 13), inconsistent family relationships (n = 7), and maternally inherited diabetes and deafness (n = 5). The 157 remaining probands had similar treatment requirements to familial type 2 diabetic subjects but presented at a younger age, were more obese, and more frequently had affected parents. Using this resource, we have not found any evidence for linkage disequilibrium between type 2 diabetes and the glucokinase gene markers GCK1 and GCK2 and the chromosome 20 marker D20S197. We conclude that European type 2 diabetes trios are difficult to collect but provide an important additional approach to dissecting the genetics of type 2 diabetes.

Missense mutations in the insulin promoter factor-1 gene predispose to type 2 diabetes.

The transcription factor insulin promoter factor-1 (IPF-1) plays a central role in both the development of the pancreas and the regulation of insulin gene expression in the mature pancreatic beta cell. A dominant-negative frameshift mutation in the IPF-l gene was identified in a single family and shown to cause pancreatic agenesis when homozygous and maturity-onset diabetes of the young (MODY) when heterozygous. We studied the role of IPF-1 in Caucasian diabetic and nondiabetic subjects from the United Kingdom. Three novel IPF-1 missense mutations (C18R, D76N, and R197H) were identified in patients with type 2 diabetes. Functional analyses of these mutations demonstrated decreased binding activity to the human insulin gene promoter and reduced activation of the insulin gene in response to hyperglycemia in the human beta-cell line Nes2y. These mutations are present in 1% of the population and predisposed the subject to type 2 diabetes with a relative risk of 3.0. They were not highly penetrant MODY mutations, as there were nondiabetic mutation carriers 25-53 years of age. We conclude that mutations in the IPF-1 gene may predispose to type 2 diabetes and are a rare cause of MODY and pancreatic agenesis, with the phenotype depending upon the severity of the mutation.

Surgical treatment of tracheal collapse using pliable total ring prostheses: results in one experimental and 4 clinical cases.

Pliable total ring prostheses were created from the polyvinyl chloride drip chambers of intravenous administration sets. The total ring prostheses were placed in one clinically normal research dog and in 4 client-owned dogs diagnosed with tracheal collapse. The research dog was euthanized one month after placement of the prostheses. Histopathological analysis of the trachea adjacent to the prostheses revealed a mild inflammatory response. The follow-up period for the clinical cases was from 4 months to 11 years. Radiographs taken and fluoroscopy performed 1 day to 5 months after surgery revealed improvement or resolution of the tracheal collapse. One dog was asymptomatic 28 weeks following surgery. Two dogs died 7 and 9 years after surgery, with one requiring intermittent medical management for coughing. They were euthanized for nonrespiratory illness. One dog had a persistent nonproductive cough, due to collapse of the mainstem bronchi, when last evaluated 4 months postoperatively. Pliable total ring prostheses provided adequate stability to the trachea and had the advantage of conforming to the trachea and being easy to create, place, and suture.

Production of goats by somatic cell nuclear transfer.

In this study, we demonstrate the production of transgenic goats by nuclear transfer of fetal somatic cells. Donor karyoplasts were obtained from a primary fetal somatic cell line derived from a 40-day transgenic female fetus produced by artificial insemination of a nontransgenic adult female with semen from a transgenic male. Live offspring were produced with two nuclear transfer procedures. In one protocol, oocytes at the arrested metaphase II stage were enucleated, electrofused with donor somatic cells, and simultaneously activated. In the second protocol, activated in vivo oocytes were enucleated at the telophase II stage, electrofused with donor somatic cells, and simultaneously activated a second time to induce genome reactivation. Three healthy identical female offspring were born. Genotypic analyses confirmed that all cloned offspring were derived from the donor cell line. Analysis of the milk of one of the transgenic cloned animals showed high-level production of human antithrombin III, similar to the parental transgenic line.

Mechanisms involved in the protective effect of estradiol-17beta on lipid peroxidation and DNA damage.

Previous studies from our laboratory have shown that estrogens can protect against lipoprotein peroxidation and DNA damage. In this study, the mechanism of estradiol-17beta (E2) action was investigated by comparing E2 with selective scavengers of reactive oxygen species (ROS) in terms of inhibition of 1) human low-density lipoprotein (LDL) peroxidation (measured by the diene conjugation method) and 2) DNA damage (measured by the formation of strand breaks in supercoiled OX-174 RFI DNA). In addition, the direct effect of E2 on the generation of individual ROS was also measured. By use of ROS scavengers, it was determined that lipoprotein peroxidation was predominantly due to superoxide (39%), with some contributions from hydrogen peroxide (23%) and peroxy (38%) radicals. E2 was a more effective inhibitor of peroxidation than all the ROS scavengers combined. In DNA damage, scavengers of hydrogen peroxide, hydroxyl, and superoxide radical offered significant protection (49-65%). E2 alone offered a similar degree of protection, and no additional effect was evident when it was combined with ROS scavengers. E2 caused a significant reduction (37%) in the production of superoxide radical by bovine heart endothelial cells in culture but had no effect on the formation of either hydrogen peroxide or hydroxyl radicals. These studies show that 1) the protection offered by E2 in terms of lipid peroxidation could be due to its ability to inhibit generation of superoxide radical and prevent further chain propagation, and 2) in DNA damage protection, E2 mainly appears to inhibit chain propagation.

Exercised-induced increase in lipid peroxidation parameters in amenorrheic female athletes.

OBJECTIVE: To determine plasma lipid peroxidation parameters in eumenorrheic and amenorrheic athletes and to evaluate differences in their response to exercise-induced oxidative stress. In female athletes, intense physical exercise has been shown to be associated with an increased occurrence of menstrual dysfunction with lower levels of E2. Recently, a protective role has been demonstrated for estrogens as free radical scavengers. DESIGN: Comparison of eumenorrheic and amenorrheic athletes before and after an acute bout of exercise. SETTING: Academic Research Environment. PATIENT(S): Seven eumenorrheic (normally menstruating) and seven amenorrheic (<3 menses/year) female athletes aged 18 to 35 years participating in regular training. MAIN OUTCOME MEASURE(S): Plasma and low-density lipoprotein oxidation parameters, plasma E2 and vitamin E levels, and creatine kinase activity. RESULT(S): Both the amenorrheic and eumenorrheic athletes demonstrated a significant decrease in the lag time of conjugated diene formation after exercise (P < 0.01), with greater magnitude of change occurring in the amenorrheic athletes (P < 0.05). In addition, postexercise samples from amenorrheic (but not eumenorrhic) athletes showed a significant (P < 0.01) increase in oxysterol formation as compared to baseline values. Amenorrheic athletes also demonstrated a significantly higher baseline creatine kinase activity and a nonsignificant (P = 0.04) trend of an increase in creatine kinase activity after exercise. CONCLUSION(S): The results of this study shows that amenorrheic female athletes demonstrate an increased potential for lipid peroxidation after exercise. This could be related to lower plasma E2 levels in this group, considering the strong free radical scavenging ability of estrogens identified recently.

An interleukin (IL)-13 receptor lacking the cytoplasmic domain fails to transduce IL-13-induced signals and inhibits responses to IL-4.

Interleukin (IL)-13 is a pleiotropic immunoregulatory cytokine that shares many, although not all, of the biological activities of IL-4. The overlapping biological properties of IL-4 and IL-13 appear to be due to the existence of shared components of the receptors, and we and others showed that the IL-4 receptor-alpha is involved in signal transduction paths activated by both. We show here that expression of the IL-13 receptor-alpha in two factor-dependent cell lines, the premyeloid FD5 and the T lymphoid CT4.S, conferred the ability to grow continuously in response to IL-13; to respond to IL-13 with tyrosine phosphorylation of JAK1, Tyk2, IL-4Ralpha, IRS-2, and STAT6; and to respond to IL-4 with tyrosine phosphorylation of Tyk2 in addition to those induced in parental cell lines. Expression of a truncated IL-13 receptor-alpha that lacked the cytoplasmic domain demonstrated that this domain was essential for IL-13-dependent growth and phosphorylation of the above substrates. Expression of this truncated IL-13 receptor also resulted in an inhibition of biochemical and biological responses to IL-4 that was exacerbated by the presence of IL-13. These dominant inhibitory effects indicate that the extracellular domain of the truncated IL-13 receptor competes with gammac for complexes of IL-4 and the IL-4 receptor-alpha, or, when itself bound to IL-13, competes with IL-4 for the IL-4 receptor-alpha.

Cyclosporin treatment of perianal fistulas in dogs.

The purpose of this pilot study was to investigate the efficacy of cyclosporin in treating perianal fistulas (PAF) in dogs. Based on resolution of all fistulas in all dogs with remission times up to > 18 months, we conclude that cyclosporin therapy is the treatment of choice for PAF in dogs.

Estradiol-17beta as an antioxidant: some distinct features when compared with common fat-soluble antioxidants.

Estrogens are potent antioxidants both in vitro and in vivo. In this study the antioxidant affect of estradiol-17beta (estradiol) was compared with those of fat-soluble antioxidants (alpha-tocopherol and beta-carotene) in terms of both fatty acid (thiobarbituric acid-reactive substances and diene conjugation) and cholesterol oxidation (oxysterols). The addition of alpha-tocopherol (54 micromol/L) inhibited low-density lipoprotein (LDL) oxidation by 92.6% and high-density lipoprotein (HDL) oxidation by 76.5%. In similar experiments, estradiol (54 micromol/L) inhibited LDL oxidation by 77.5% but inhibited HDL oxidation by only 55.4%. Beta-carotene had no antioxidant effect. Lag times (diene conjugation method) for alpha-tocopherol and beta-carotene increased by 175% and 125%, respectively. Estradiol markedly reduced the maximum formation of diene conjugates as compared with results with alpha-tocopherol and beta-carotene, and it exhibited a linear curve (no change in lag time). In terms of cholesterol oxidation, estradiol was far more effective than alpha-tocopherol or beta-carotene in inhibiting oxysterol formation (microg/ml plasma) (control = 24.56 +/- 2.31, beta-carotene = 20.59 +/- 3.32, alpha-tocopherol = 20.19 +/- 1.58, estradiol = 14.38 +/- 0.70). This study shows that estradiol is as effective an antioxidant as alpha-tocopherol in terms of fatty acid peroxidation but is far more effective than alpha-tocopherol in terms of cholesterol peroxidation.

Quinacrine noncompetitive inhibitor binding site localized on the Torpedo acetylcholine receptor in the open state.

Open-channel blockers of the nicotinic acetylcholine receptor (nAcChR) are widely thought to act sterically by entering and "plugging" the open channel of the nAcChR. However, quinacrine, a fluorescent open-channel blocker, has been recently shown to bind to the nAcChR at a site near the lipid bilayer while the receptor is in a closed, desensitized state, suggesting that at least one open-channel blocker might act allosterically outside the channel [Valenzuela et al. (1992) J. Biol. Chem. 267, 8238]. To determine whether or not quinacrine also binds near the lipid bilayer when the receptor is in an open state, a short-range lipophilic quencher (5-doxylstearate, 5-SA) was used to assess the proximity of the nAcChR-bound quinacrine to the lipid bilayer while the receptor was transiently open by an agonist. Initial experiments using a stopped-flow instrument established the conditions required to monitor a portion of the changes in quinacrine fluorescence associated with its binding to the receptor in the open state. 5-SA (80 microM) reduced the amplitude of the rapid agonist-induced change in quinacrine emission to 44% +/- 12% of the control value, indicating that the quinacrine was binding to a site proximal to the membrane-partitioned 5-SA. Control experiments established that 5-SA had no effect on the ability of the receptor to undergo agonist-induced conformational changes, suggesting that little, if any, 5-SA distributed into the channel lumen and perturbed the functional activity of the receptor. Together, the results indicate that quinacrine binds to a site on the open receptor that is in contact with the lipid bilayer and not in the channel lumen.

Superior and distinct antioxidant effects of selected estrogen metabolites on lipid peroxidation.

The effect of the estrogen metabolites, 4-hydroxyestrone and 17alpha-dihydroequilin (metabolites of estradiol-17beta and equilin, respectively), were examined for antioxidant effects on plasma and lipoprotein lipid peroxidation . Lipid peroxidation was evaluated by products of both fatty acid (thiobarbituric acid-reactive substances [TBARS]) and cholesterol (oxysterols) oxidation from lipoproteins or whole plasma. Although all estrogens significantly reduced lipid peroxidation, 4-hydroxyestrone was far more potent than either equilin or 17alpha-dihydroequilin in inhibiting TBARS formation in lipoproteins induced by Cu2+. Similar effects were also noted on TBARS formation in THP-l macrophages in culture. However, 17alpha-dihydroequilin (along with equilin) strongly inhibited oxysterol formation, whereas 4-hydroxyestrone was ineffective. These studies suggest that different estrogens might act preferentially on distinct lipid substrates in exhibiting antioxidant effects.

Using INSPIRIT to conduct spiritual assessments.

Notes that pastoral caregivers are increasingly interested in spiritual assessment of patients. Reports results when the Index of Core Spiritual Experiences (INSPIRIT) is used for assessment with medical and surgical outpatients as well as patient family members (N = 371). Reports subscales within the instrument, mean scores, and notes that 80% of all respondents reported at least one experience which they regarded as spiritual.