Inequalities in HIV testing uptake and needs among men who have sex with men living in Ireland: findings from an internet survey.

OBJECTIVES: HIV disproportionately affects men who have sex with men (MSM) in Ireland. The aim of this study was to improve understanding of HIV testing among MSM living in Ireland to inform prevention and testing initiatives. METHODS: We used data from the MSM Internet Survey Ireland 2015 (MISI 2015), a cross-sectional survey of MSM living in Ireland. We identified factors associated with never having tested for HIV using univariable and multivariable logistic regression. We identified preferred sites for future tests and examined the relationships between unmet HIV testing needs and socio-demographic groups. RESULTS: More than one-third (n = 1006; 36%) of MSM had never tested for HIV. Multivariable logistic regression showed that untested men were more likely to be aged 18-24 years, live outside Dublin, have a lower level of education, be born in Ireland, identify as bisexual, be out to fewer people, and not have had sex with a man in the previous 12 months. The same groups of men also had the least knowledge about HIV and were least confident in accessing an HIV test. Men who had never tested for HIV were more likely to prefer testing by their general practitioner (GP) or using home sampling HIV kits and less likely to prefer testing in a sexual health clinic. CONCLUSIONS: HIV prevention and testing programmes for MSM should be targeted towards younger men, those living outside Dublin and those with lower levels of education. We recommend increased promotion and availability of free HIV testing services in a range of clinical and nonclinical settings (including self-sampling and home testing).

Knowledge about folic acid supplementation in women presenting for antenatal care.

BACKGROUND/OBJECTIVES: The incidence of neural tube defects (NTDs) in Ireland has increased in recent years. This study examines knowledge about folic acid (FA) supplementation for the prevention of NTDs among women presenting for antenatal care. SUBJECTS/METHODS: Women were recruited at their convenience in the first trimester after sonographic confirmation of an ongoing singleton pregnancy. A detailed questionnaire was completed under the supervision of a research dietitian. Clinical and socio-demographic details were collected. RESULTS: Of the 587 women studied, 96% took FA during early pregnancy. Of these, 56.4% cited brain/spinal development or the prevention of brain/spinal defects, spina bifida or NTDs as the reason for taking FA. Multivariate analysis showed that women who were experiencing material deprivation or who were living in Ireland <5 years were least likely to be knowledgeable about the benefits of FA supplementation (P<0.05 for both). Over half (57.1%) of the women did not take FA preconceptionally. The main reason reported for not supplementing preconceptionally was that the woman did not expect to get pregnant (76.4%). Over one-third of women (35%), however, reported that they did not know they needed to take FA before becoming pregnant. CONCLUSIONS: These results highlight the need for a renewed public health campaign in Ireland about the importance of FA. As well as focusing on women who have recently come to live in Ireland, this campaign needs focus on women living in deprivation, as these are the women most at risk of having inadequate knowledge about the importance of FA in improving pregnancy outcomes.

A review of European guidelines on periconceptional folic acid supplementation.

Strong evidence that folic acid (FA) prevents the majority of cases of neural tube defects (NTDs) has led to national organisations developing guidelines for women concerning periconceptional supplementation. In Europe, there is evidence of national variations in the incidence of NTDs, with a recent Irish study reporting an increase in the rate. This review compares the periconceptional FA supplementation guidelines between the different countries in Europe. An online search of country-specific guidelines produced before 2015 concerning periconceptional FA supplementation was conducted. If an English version was not available directly, the EUROCAT register was searched for the English version of the recommendations. We identified national guidelines from 20 European countries. Over half recommended that FA supplements be taken by women planning a pregnancy, but three recommended that they should be taken by all women of child-bearing age. Four guidelines recommended starting FA at least 4 weeks preconceptionally, but no country recommended starting FA at least 12 weeks preconceptionally as suggested by recently published studies. There is a need for further consideration of the duration of preconceptional FA supplementation specifically. The latest scientific evidence in this area should inform the development of European guidelines on FA, as there is wide variation in current recommendations. Overall, the wide variation in national guidelines concerning periconceptional FA supplementation may in part explain the differences in national rates of NTDs reported by EUROCAT. National guidelines on FA supplementation should be standardised across European countries.

The cognitive and behavioural phenotype of Roifman syndrome.

BACKGROUND: Roifman syndrome (OMIM 300258) is a multi-system disorder with a physical phenotype that includes Beta-cell immunodeficiency, intra-uterine and postnatal growth retardation, spondyloepiphyseal dysplasia, retinal dystrophy and characteristic facial dysmorphism. So far, six cases, all boys, have been reported in the literature. Roifman postulated that the syndrome may be due to a mutation in an X-linked gene or an autosomal gene giving rise to a sex-limited trait, but the definitive pathogenetic mechanism has still not been elucidated. Very little is known about the cognitive and behavioural phenotype of Roifman syndrome and no standardized measures of cognitive abilities have been reported. METHODS: We report the seventh case of a boy with Roifman syndrome and present the first systematic documentation of the cognitive and behavioural phenotype of an individual with the syndrome. RESULTS: In spite of having been reported as appearing intellectually 'able', formal evaluation showed very significant intellectual disability and neuropsychological impairment across cognitive domains. CONCLUSIONS: The findings suggest that Roifman syndrome may be an example of an X-linked mental retardation syndrome (XLMRS).

Estrus synchronization and pregnancy rates in beef cattle given CIDR-B, prostaglandin and estradiol, or GnRH.

Two experiments were conducted to determine estrous response and pregnancy rate in beef cattle given a controlled internal drug release (CIDR-B) device plus prostaglandin F2 alpha (PGF) at CIDR-B removal, and estradiol or gonadotropin releasing hormone (GnRH). In Experiment I, crossbred beef heifers received a CIDR-B device and 1 mg estradiol benzoate (EB), plus 100 mg progesterone (E + P group; n = 41), 100 micrograms gonadotropin releasing hormone (GnRH group; n = 42), or no further treatment (Control group; n = 42), on Day 0. On Day 7, CIDR-B devices were removed and heifers were treated with PGF. Heifers in the E + P group were given 1 mg EB, 24 h after PGF, and then inseminated 30 h later. Heifers in the GnRH group were given 100 micrograms GnRH, 54 h after PGF, and concurrently inseminated. Control heifers were inseminated 12 h after onset of estrus. The estrous rate was lower (P < 0.01) in the GnRH group (55%) than in either the E + P (100%) or Control (83%) groups. The mean interval from CIDR-B removal to estrus was shorter (P < 0.01) and less variable (P < 0.01) in the E + P group than in the GnRH or Control groups. Pregnancy rate in the E + P group (76%) was higher (P < 0.01) than in the GnRH (48%) or Control (38%) groups. In Experiment II, 84 cows were treated similarly to the E + P group in Experiment I. Cows received 100 mg progesterone and either 1 mg EB or 5 mg estradiol-17 beta (E-17 beta) on Day 0 and either 1 mg of EB or 1 mg of E-17 beta on Day 8 (24 h after CIDR-B removal), in a 2 x 2 factorial design, and were inseminated 30 h later. There were no differences among groups for estrous rates or conception rates. The mean interval from CIDR-B removal to estrus was 44.2 h, s = 11.2. Conception rates were 67%, 62%, 52%, and 71% in Groups E-17 beta/E-17 beta, E-17 beta/EB, EB/E-17 beta, and EB/EB, respectively. In cattle given a CIDR-B device and estradiol plus progesterone, treatment with either EB or E-17 beta effectively synchronized estrus and resulted in acceptable conception rates to fixed-time artificial insemination.

Cluster of diplopia cases after periocular anesthesia without hyaluronidase.

PURPOSE: To describe a cluster of cases of iatrogenic diplopia after cataract surgery that occurred in 1998, when hyaluronidase was unavailable for use in periocular anesthetic regimens. SETTING: The clinical practices of the authors. METHODS: This study comprised a retrospective chart review. RESULTS: Twenty-five cases of transient or permanent diplopia were reported. Of these, 13 eyes had retrobulbar and 10 had peribulbar injections; in 2 cases the injection technique was unknown. The inferior rectus was affected in 19 eyes; of these, 1 had a temporary palsy and 18 had permanent restriction. Temporary paresis developed in the lateral rectus in 5 cases and the superior rectus in 2. Eleven cases were submitted by 4 anterior segment surgeons, who collectively had a zero incidence of iatrogenic postoperative diplopia in the preceding 4 to 11 years of practice (approximately 6900 cases). CONCLUSION: Hyaluronidase may be more important than previously suspected in preventing anesthetic-related damage to the extraocular muscles. The inferior rectus muscle is particularly vulnerable, presumably because of the injection technique.

Heterotrimeric and small molecular mass GTP-binding proteins of rat brain neuronal and glial nuclei.

Heterotrimeric and small molecular mass guanine nucleotide binding (GTP-binding) proteins were found in neuronal and glial nuclei isolated from rat brain. Neuronal nuclei bound 0.213 +/- 0.055 pmoles of GTP/microg protein (n = 8) and glial nuclei bound 0.145 +/- 0.038 pmoles of GTP/microg protein (n = 8). The intrinsic GTPase activity of neuronal and glial nuclei was 0.0019 +/- 0.0005 pmoles GTP hydrolyzed/min/microg protein (n = 10) and 0.0022 +/- 0.0006 pmoles GTP hydrolyzed/min/microg protein (n = 10), respectively. Western blot analysis was carried out using a peptide-specific antibody that recognizes a common sequence in the alpha-subunit of the various heterotrimeric G-proteins. The antibody revealed the presence of a polypeptide of molecular mass of 40 kDa only in neuronal nuclei. Small molecular mass GTP-binding proteins were detected by incubating nitrocellulose blots with [alpha-32P]GTP. The results demonstrated the presence of 25-26 kDa GTP-binding proteins in both populations of nuclei. However, the binding of [alpha-32P]GTP to neuronal nuclei was approximately 3-fold greater than to the glial nuclei. Further analysis by two-dimensional polyacrylamide gel electrophoresis resolved the neuronal nuclei 26 kDa protein into three forms (a-c) with the most acidic form (c) being the major species. The neuronal 25 kDa protein was resolved into two forms that were present in approximately equal concentration. In glial nuclei, only the 26 kDa (c) and a small amount of the 25 kDa proteins were detected. However, both populations of nuclei contained the small molecular mass GTP-binding protein, ran. Differential association of non-ran small molecular mass GTP-binding proteins and heterotrimeric G-proteins with neuronal nuclei suggests a potential role for these guanine nucleotide binding proteins in the function of this cell type.

Intradermal immunization with a bovine herpesvirus-1 DNA vaccine induces protective immunity in cattle.

Although intramuscular (i.m.) injection of DNA encoding glycoprotein D (gD) of bovine herpesvirus-1 (BHV-1) induces immune responses in cattle, this route of delivery is inefficient. Here we assessed three parameters that may enhance the efficacy of a gD DNA vaccine in cattle. First, the immune response generated by i.m. injected plasmid expressing a secreted form of gD (tgD) was determined and found to be very similar in magnitude to the response induced by gD-expressing plasmid. Secondly, gD- and tgD-expressing plasmids were administered by intradermal (i.d.) immunization, which resulted in a superior immune response to the secreted form, but no improvement in the response to the membrane-associated form. However, the form of gD used for immunization did not influence the immunoglobulin subtype, the ratio of antigen-specific IgG1 to IgG2 being approximately 4:1. Finally, the effect of promoter strength was assessed by replacing the Rous sarcoma virus (RSV) promoter, which was used in the original experiments, with the human cytomegalovirus immediate early promoter and first intron A (HCMV/IA). Although upon transfection in vitro the HCMV/IA promoter appeared to be stronger than the RSV promoter, there was only a 2-fold higher antibody response in vivo upon i.d. injection of cattle. Protection against virus challenge was obtained in the calves immunized i.d. with tgD-encoding plasmid, as shown by a significant reduction in weight loss, virus excretion, temperature response and clinical disease. No significant protection was observed in the animals vaccinated i.d. with the gD-expressing plasmid, which correlates with the lower level of immunity pre-challenge.

Phospholipid metabolism in Alzheimer's disease and in a human cholinergic cell.

There is evidence available suggesting that membrane alterations occur in Alzheimer's disease including the metabolism of membrane phospholipids. We have quantitated in vitro the phospholipase D activity of homogenates from Alzheimer's disease brain tissue. There was a significant increase of this enzyme activity as compared to controls. Amyloid beta protein is the predominant protein of the characteristic senile plaques found in Alzheimer's disease. Treatment of LA-N-2 cells, a human cholinergic neuroblastoma clone, with amyloid beta protein results in an activation of phospholipases A, C and D.

Phospholipase D activity of rat brain neuronal nuclei.

Phospholipase D activity of rat brain neuronal nuclei, measured with exogenous phosphatidylcholine as substrate, was characterized. The measured activity of neuronal nuclei was at least 36-fold greater than the activity in glia nuclei. The pH optimum was 6.5, and unsaturated but not saturated fatty acids stimulated the enzyme. The optimal concentration of sodium oleate for stimulation of the enzyme activity was 1.2 mM in the presence of 0.75 mM phosphatidylcholine. This phospholipase D activity was cation independent. In the absence of NaF, used as a phosphatidic acid phosphatase inhibitor, the principal product was diglyceride; whereas in the presence of NaF, the principal product was phosphatidic acid. The phospholipase D, in addition to having hydrolytic activity, was able to catalyze a transphosphatidylation reaction. Maximum phosphatidylethanol formation was seen with 0.2-0.3 M ethanol. GTPgammaS, ATPgammaS, BeF2, AIF3, phosphatidic acid, and phosphatidylethanol inhibited the neuronal nuclei phospholipase D activity. The addition of the cytosolic fraction of brain, liver, kidney, spleen, and heart to the incubation mixtures resulted in inhibition of the phospholipase D activity. Phospholipase D activity was detectable in nuclei prepared from rat kidney, spleen, heart, and liver.

Acidic phospholipids inhibit the phospholipase D activity of rat brain neuronal nuclei.

An oleate dependent form of phospholipase D is present in rat brain neuronal nuclei and both the hydrolytic and transphosphatidylation activities measured. Several acidic phospholipids were found to inhibit this activity in a dose dependent manner. The IC50 values varied from 3.5 microM for PIP2 to 200 microM for phosphatidic acid. The hydrolysis of PIP2 by phospholipase C would be expected to result in the disinhibition of the oleate dependent phospholipase D activity.

Amyloid beta protein (25-35) stimulation of phospholipases A, C and D activities of LA-N-2 cells.

[3H]Myristic acid prelabeled LA-N-2 cells were exposed to varying concentrations of amyloid beta protein (25-35), from 20 to 250 micrograms/ml, and the activation of phospholipases A and D estimated. A progressive increase in phosphatidylethanol formation, a measure of phospholipase D activity, and of free fatty acid release, a measure of phospholipase A activity, was observed over a time-course of 60 min. [3H]Inositol prelabeled LA-N-2 cells were exposed to varying concentrations of A beta P, from 20 to 125 micrograms/ml, and phospholipase C activation was measured. There was an increased release of inositol phosphates in the presence of amyloid beta protein as a function of incubation time. The effects of adrenergic, metabotropic amino acid and bombesin antagonists on the A beta P mediated stimulation of phospholipase C activity was investigated. Propranolol, a beta adrenergic antagonist, 7-chloro-kynurenic acid, a metabotropic amino acid antagonist, and [Tyr4-D-Phe12]bombesin, a bombesin antagonist, blunted the A beta P stimulation of phospholipase C activity in [3H]inositol prelabeled LA-N-2 cells. This suggests that amyloid beta protein activation of phospholipase C may be receptor mediated. The phospholipase C inhibitor U 71322 prevented the activation of phospholipase C by A beta P. However, this activation was not effected by tocopherol, propylgallate, or vitamin C.

Proximal femoral diaphysectomy in cerebral palsy.

A study was conducted to evaluate the outcome of massive proximal femoral shortening in the cerebral palsy patient with severe spastic quadriplegia and hip instability. A retrospective review of 13 children (age range: three to 19 years of age) representing 18 hips treated with massive shortening of the proximal femur was conducted. Bilateral procedures were performed in five patients. All procedures were performed between February 1986 and March 1990. Radiographs were evaluated for preoperative and postoperative migration percentage (MP) and femoral neck-shaft angle (NSA). Charts were reviewed for complications and clinical results. All femoral osteotomies healed without difficulty. Clinical follow-up averaged 27.6 months. Satisfactory results occurred in all but one hip. Radiographs taken an average of 19.5 months postoperatively showed improved MP in all but one hip. The average preoperative MP was 70% and postoperative MP was 18%. Femoral NSA also was improved. Heterotopic bone formed in 13 hips but caused no significant problems. Other complications included postoperative seizure, urinary tract infection, cast sores, transient arm weakness, weight loss, pin protrusion through skin, and femur fracture after cast removal. Based on the good results and minimal complications in this series, massive femoral shortening appears to be a superior alternative to proximal femoral resection in these difficult patients.

Dual camera sequencer for microsurgical documentation.

All currently available documentary formats have inherent compromises. The use of both video and 35-mm still photography seems to be a practical solution but introduces image quality loss (when using a beam splitter type of dual camera adapter) or increased complexity of operation and resultant surgeon distraction (when using a movable mirror type of dual camera adapter). An electronic sequencer was designed to simplify the operation of a movable mirror type of dual camera adapter, permitting the efficient production of high image quality video recordings and 35-mm slides.

Enzymatic activities during differentiation of the human neuroblastoma cells, LA-N-1 and LA-N-2.

The presence of 10(-5) M retinoic acid (RA) in the culture medium of LA-N-1, a catecholaminergic cell line, and LA-N-2, a cholinergic cell line, enhanced their morphological differentiation. Tyrosine hydroxylase (TH) activity of the LA-N-1 cells was increased in the RA-treated cells compared with control cultures at day 4 and remained elevated. Choline acetyltransferase (ChAT) activity in the LA-N-2 cells gradually increased until 8 days in vitro (DIV) both in the untreated control and the RA treated cultures. This activity in control and treated cells decreased gradually to a constant level of activity. The ChAT activity at 8 DIV of RA-treated LA-N-2 cells was increased 2.1-fold (P less than 0.001) as compared to the control cultures. This increase in ChAT activity was accompanied by a 73% decrease of acetylcholinesterase (AChE) activity in LA-N-2 cells by 8 DIV. AChE activity of LA-N-1 cells was unchanged during the time course of the experiment. Phospholipase-A2 (PL-A2) activity in RA-treated LA-N-2 cells was increased at day 4 as compared with the control cultures. There were no differences observed in phospholipase-D (PL-D), choline kinase and GPC-phosphodiesterases activities in RA-treated and -untreated LA-N-1 and LA-N-2 cells.

Regulation of the choline, ethanolamine and serine base exchange enzyme activities of rat brain microsomes by phosphorylation and dephosphorylation.

The base exchange enzyme activities of rat brain microsomes were estimated subsequent to preincubations under conditions for either protein phosphorylation or dephosphorylation. Quantitatively the choline base exchange activity was most affected by these treatments. Exposure of the microsomes to alkaline phosphatase resulted in a decrease of all three base exchange activities. Pretreatment with a cAMP-dependent protein kinase resulted in increases of all 3 enzyme activities. Conditions favoring protein kinase C phosphorylation resulted in stimulation of the choline base exchange activity.