Knowledge, actual and potential use of HIV self-sampling testing kits among MSM recruited in eight European countries.

AIM: To describe the knowledge as well as current and potential use of self-sampling kits among men who have sex with men (MSM) and to analyse their preferred biological sample and result communication method. METHODS: We analyse data of MSM of HIV negative or unknown serostatus from an online survey conducted in eight countries (Belgium, Denmark, Germany, Greece, Portugal, Romania, Slovenia and Spain) between April and December 2016. It was advertised mainly in gay dating websites. We conduct a descriptive analysis of the main characteristics of the participants, and present data on indicators of knowledge, use and potential use of HIV self-sampling as well as their preferences regarding blood or saliva sample and face or non-face-to-face result communication by country of residence. RESULTS: A total of 8.226 participants of HIV negative or unknown serostatus were included in the analysis. Overall, 25.5% of participants knew about self-sampling (range: 18.8-47.2%) and 1.1% had used it in the past (range: 0.3-8.9%). Potential use was high, with 66.6% of all participants reporting that they would have already used it if available in the past (range: 62.1-82.1%). Most (78.6%) reported that they would prefer using a blood-based kit, and receiving the result of the test through a non-face-to-face-method (70.8%), even in the case of receiving a reactive result. CONCLUSION: The high potential use reported by MSM recruited in eight different European countries suggests that self-sampling kits are a highly acceptable testing methodology that could contribute to the promotion of HIV testing in this population.

Kidney retransplantation in the ipsilateral iliac fossa: a surgical challenge.

The aim of this study is to review the surgical outcome of kidney retransplantation in the ipsilateral iliac fossa in comparison to first kidney transplants. The database was screened for retransplantations between 1995 and 2013. Each study patient was matched with 3 patients with a first kidney transplantation. Just for graft and patient survival analyses, we added an extra control group including all patients receiving a second transplantation in the contralateral iliac fossa. We identified 99 patients who received a retransplantation in the ipsilateral iliac fossa. There was significantly more blood loss and longer operative time in the retransplantation group. The rate of vascular complications and graft nephrectomies within 1 year was significantly higher in the study group. The graft survival rates at 1 year and 3, 5, and 10 years were 76%, 67%, 61%, and 47% in the study group versus 94%, 88%, 77%, and 67% (p < 0.001) in the first control group versus 91%, 86%, 78%, and 57% (p = 0.008) in the second control group. Patient survival did not differ significantly between the groups. Kidney retransplantation in ipsilateral iliac fossa is surgically challenging and associated with more vascular complications and graft loss within the first year after transplantation. Whenever feasible, the second renal transplant (first retransplant) should be performed contralateral to the prior failed one.

Collaboration of general practitioners and exercise providers in promotion of physical activity a written survey among general practitioners.

BACKGROUND: General practitioners have an ideal position to motivate inactive patients to increase their physical activity. Most patients are able to exercise in regular local facilities outside the health care setting. The purpose of this study was to get insight into general practitioners perceptions and current practices regarding referral of patients to local exercise facilities. Furthermore, collaboration with exercise providers in the community was investigated, and motivators and barriers for referral. METHODS: A written questionnaire sent to a representative random sample of 800 Dutch general practitioners. Descriptive statistics and Chi(2) tests were used. RESULTS: All responding general practitioners (340) recommend their patients to take more exercise when necessary and 87 % say to refer patients sometimes. Limited motivation of the patient (44 %) and reduced health status (34 %) are the most mentioned barriers for advising patients to increase physical activity. When referred, most patients are send to a physical therapist (69 %) but also local exercise facilities were mentioned (54 %). The most important barrier for referring patients to local exercise activities are patients limited financial possibilities (46 %). Restricted knowledge of local exercise- or sport facilities was an additional barrier (19 %). There is little structural collaboration between general practitioners and exercise providers, but when collaboration exists general practitioners refer more often. Positive experiences of patients (67 %), affordable offers (59 %) and information of local exercise facilities (46 %) are seen as important promoting factors for referral. Although 32 % of the general practitioners think that good collaboration would be stimulating, regular meetings with sports and exercise providers were considered the least important for increasing referral (3 %). CONCLUSIONS: Dutch physicians have a positive attitude towards stimulating physical activity but referral to local exercise facilities is low. Referral is partly hindered by restricted knowledge of local exercise facilities. Although general practitioners think that collaboration is important for physical activity promotion, it should not cost them much extra time. A coordinator with knowledge of the local situation can facilitate contacts between GP practices and sports providers.

The SH2-containing inositol polyphosphate 5-phosphatase, SHIP-2, binds filamin and regulates submembraneous actin.

SHIP-2 is a phosphoinositidylinositol 3,4,5 trisphosphate (PtdIns[3,4,5]P3) 5-phosphatase that contains an NH2-terminal SH2 domain, a central 5-phosphatase domain, and a COOH-terminal proline-rich domain. SHIP-2 negatively regulates insulin signaling. In unstimulated cells, SHIP-2 localized in a perinuclear cytosolic distribution and at the leading edge of the cell. Endogenous and recombinant SHIP-2 localized to membrane ruffles, which were mediated by the COOH-terminal proline-rich domain. To identify proteins that bind to the SHIP-2 proline-rich domain, yeast two-hybrid screening was performed, which isolated actin-binding protein filamin C. In addition, both filamin A and B specifically interacted with SHIP-2 in this assay. SHIP-2 coimmunoprecipitated with filamin from COS-7 cells, and association between these species did not change after epidermal growth factor stimulation. SHIP-2 colocalized with filamin at Z-lines and the sarcolemma in striated muscle sections and at membrane ruffles in COS-7 cells, although the membrane ruffling response was reduced in cells overexpressing SHIP-2. SHIP-2 membrane ruffle localization was dependent on filamin binding, as SHIP-2 was expressed exclusively in the cytosol of filamin-deficient cells. Recombinant SHIP-2 regulated PtdIns(3,4,5)P3 levels and submembraneous actin at membrane ruffles after growth factor stimulation, dependent on SHIP-2 catalytic activity. Collectively these studies demonstrate that filamin-dependent SHIP-2 localization critically regulates phosphatidylinositol 3 kinase signaling to the actin cytoskeleton.

The yeast inositol polyphosphate 5-phosphatases inp52p and inp53p translocate to actin patches following hyperosmotic stress: mechanism for regulating phosphatidylinositol 4,5-bisphosphate at plasma membrane invaginations.

The Saccharomyces cerevisiae inositol polyphosphate 5-phosphatases (Inp51p, Inp52p, and Inp53p) each contain an N-terminal Sac1 domain, followed by a 5-phosphatase domain and a C-terminal proline-rich domain. Disruption of any two of these 5-phosphatases results in abnormal vacuolar and plasma membrane morphology. We have cloned and characterized the Sac1-containing 5-phosphatases Inp52p and Inp53p. Purified recombinant Inp52p lacking the Sac1 domain hydrolyzed phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)] and PtdIns(3, 5)P(2). Inp52p and Inp53p were expressed in yeast as N-terminal fusion proteins with green fluorescent protein (GFP). In resting cells recombinant GFP-tagged 5-phosphatases were expressed diffusely throughout the cell but were excluded from the nucleus. Following hyperosmotic stress the GFP-tagged 5-phosphatases rapidly and transiently associated with actin patches, independent of actin, in both the mother and daughter cells of budding yeast as demonstrated by colocalization with rhodamine phalloidin. Both the Sac1 domain and proline-rich domains were able to independently mediate translocation of Inp52p to actin patches, following hyperosmotic stress, while the Inp53p proline-rich domain alone was sufficient for stress-mediated localization. Overexpression of Inp52p or Inp53p, but not catalytically inactive Inp52p, which lacked PtdIns(4,5)P(2) 5-phosphatase activity, resulted in a dramatic reduction in the repolarization time of actin patches following hyperosmotic stress. We propose that the osmotic-stress-induced translocation of Inp52p and Inp53p results in the localized regulation of PtdIns(3,5)P(2) and PtdIns(4,5)P(2) at actin patches and associated plasma membrane invaginations. This may provide a mechanism for regulating actin polymerization and cell growth as an acute adaptive response to hyperosmotic stress.

Differential SKIP expression in PTEN-deficient glioblastoma regulates cellular proliferation and migration.

Glioblastoma is the most common and lethal primary malignant brain tumor in adults. The tumor suppressor gene PTEN is deleted, mutated or hypermethylated in more than 60% of glioblastoma cases resulting in hyperactivation of the phosphoinositide 3-kinase pathway, which leads to sustained PI(3,4,5)P3 signaling, and thereby hyperactivation of Akt and other effectors. PI(3,4,5)P3 is also hydrolyzed to PI(3,4)P2 by inositol polyphosphate 5-phosphatases such as SKIP, but the role this pathway has in glioblastoma is unknown. Microarray expression profiling of SKIP in human glioblastoma has revealed both increased and decreased SKIP gene expression. Here we have screened PTEN-deficient glioblastoma for SKIP protein expression by immunohistochemistry and report that SKIP expression is increased in some cases or decreased relative to normal brain. Using the U-87MG PTEN-deficient cell line we show that SKIP knockdown did not further enhance cell proliferation or survival. However, SKIP overexpression in U-87MG cells suppressed anchorage-independent cell growth and growth factor-induced PI(3,4,5)P3/Akt signaling. Although, SKIP knockdown did not affect cell proliferation or survival, cell migration was significantly retarded, associated with significantly increased PI(4,5)P2 signals, and decreased phosphorylation of the actin-regulatory protein cofilin, a PI(4,5)P2-binding protein. Notably, overexpression of SKIP also inhibited migration of U-87MG cells to a similar degree as observed with PTEN reconstitution, however, via distinct mechanisms. PTEN reconstitution promoted sustained lamellipodia generation and focal adhesion formation. In contrast, SKIP overexpression reduced sustained lamellipodia formation, talin incorporation into focal adhesions and recruitment of PI(4,5)P2-binding proteins to the plasma membrane. Notably, analysis of two independent ONCOMINE microarray data sets revealed a significant correlation between increased SKIP mRNA expression in glioblastoma and improved long-term survival. Therefore, SKIP expression in glioblastoma may affect the local invasion of PTEN-deficient tumors.

Simple ion chromatographic method for the determination of chlormequat residues in pears.

Current methods for quantitative determination of chlormequat residues in food crops are characterized by rather low recoveries and the need for derivatization (in case of gas chromatography, GC), or by high capital investment (in case of liquid chromatography-mass spectrometry, LC-MS). We propose a cation-exchange chromatography method for the analysis of chlormequat in pears. The method is based on extraction of the target compound with 40 mM HCl, followed by centrifugation and filtration. The filtrate is directly injected into an ion chromatograph equipped with a commercially available cation-exchange column and a suppressed conductivity detection system. While the limit of detection (LOD) (0.5 mg/kg) may not be small enough to allow dietary analysis, the method meets all validation requirements and is an alternative for the existing GC and LC-MS methods in quality control.

R 8110, a new short-acting hypnotic in dogs.

The clinical, respiratory and cardiovascular effects of intravenous injections of R 8110, a fluor analogue of etomidate, were studied in unpremedicated dogs. Clinical observations were carried out after intravenous injections of 3 and 4 mg kg-1 of R 8110. Cardiovascular studies were conducted after an intravenous injection of 3 mg kg-1. The drug proved to be a safe and reliable agent for induction and produced a short-lasting hypnosis and some analgesia. Both induction and recovery were smooth and rapid. Heart rate and systolic and diastolic blood pressure decreased significantly (P less than or equal to 0.05) 10 minutes after injection; the influence on arterial blood parameters was minimal.

Alleviation of excessive gas accumulation in the ruminant stomach by ritanserin.

The relationships between forestomach motility and eructation rate were studied in sheep and cattle. Three ewes and 2 heifers were implanted with strain gauges on the reticulo-rumen and fitted with a cannula in the dorsal sac of the rumen. Studies were performed in sheep after induction of hypocalcemia by Na2EDTA infusion and cattle were studied after ruminal distension. Experiments were performed by measuring the rate and volume of eructated ruminal gases, using a technique by which the trachea is transected. The frequency of reticulo-ruminal contractions decreased 40% within 30 minutes of Na2EDTA infusion to the sheep. The volume of eructated gas (for 30-minute periods) decreased from 10.7 L to 5.5 L at the end of the 60-minute infusion period. Pretreatment with ritanserin (0.1 mg/kg, subcutaneously) not only prevented bloating during the ruminal stasis induced by hypocalcemia, but also significantly increased the eructated volume of gas. In cattle, ritanserin given at the same dose level (0.1 mg/kg, subcutaneously) significantly increased the volume of eructated gas after ruminal distension. This study supports the hypothesis that the caudal esophageal sphincter has a role in the rate of ruminal gas eructation and indicates that its relaxation may be due to a 5-hydroxytryptamine antagonist.

Treatment of tetanus in the horse by injections of tetanus antitoxin into the subarachnoid space.

In 40 horses with tetanus, large doses of tetanus antitoxin (TAT) were injected into the subarachnoid space. In all the horses that recovered, the disease stabilized immediately after the injection. The results (77.5% recovery) were much better than in a previous series of horses with tetanus (50% recovery), in which TAT was injected either intravenously, intramuscularly, or in the epidural space.

Pathogenesis and pharmacology of diarrhea.

The etiological factors involved in diarrhea are multiple. Also the mechanisms and mediators involved are multiple: intracellular mediators (Ca, cAMP, cGMP, calmodulin, phospholipids), extracellular mediators (hormones, neurotransmitters, prostaglandins, enterotoxins...), intramural blood flow and oxygen, intestinal motility (local- and peristaltic motility). Till now, antidiarrheals are not so versatile that they provide a solution to all types of diarrhea. The mechanisms of action of fluid replacement therapy, loperamide, alpha 2 agonist and some nonsteroidal anti-inflammatory substances are reviewed.

BHV4 (bovine herpes virus 4) related disorders in Belgian cattle: a study of two problem herds.

Two cattle farms, with a ten year history of BHV4 related postpartum metritis accompanied by fertility problems, were monitored during the winter season 1985-1986. BHV4 was isolated from the lochia from 55% of the animals on farm A and 66% of those on farm B. Respectively 59% and 30% of the animals presented postpartum metritis. In some animals virus multiplication was followed by severe leucopenia lasting several weeks. Indirect immunofluorescence (IIF) BHV4 seropositive as well as IIF seronegative animals were affected. The latter responded with a rapid or late IIF antibody reaction. No BHV4 seroneutralizing antibodies could be detected. The authors also suggest a possible role of BHV4 in the respiratory problems observed during the study.

Alterations in intestinal fluid movement.

Fluid loss in diarrhoea is caused by alterations in the basic mechanisms of transport both in the small intestine and in the colon. The role of blood flow is discussed with reference to anatomy and blood flow distribution, fluid transport, luminal pressure and intestinal motility. The control of intestinal fluid transport and of absorption and secretion processes by intracellular and extracellular mediators is reviewed. Treatment of disorders of fluid transport consist of fluid replacement therapy or the use of antidiarrhoeal agents with high specificity and safety, such as loperamide. The effects of interactions between loperamide and bacterial toxins, theophylline and prostaglandins on the antisecretory activity of loperamide are detailed.

Mechanisms of action of loperamide.

The pharmacology of loperamide is discussed, with particular reference to the drug's antidiarrhoeal properties. Its absorption, distribution, metabolism and excretion are considered and rapid concentration in the small intestine is described. The mechanism of action of loperamide on fluid and electrolyte transport is examined with reference to opiate agonism, calcium-channel blocking, calmodulin inhibition, and paracellular permeability. The colon appears to be the main site of action of loperamide on gut motility and transit. In the proximal colon, the drug accentuates segmenting motor activity. Work relating to the activity of loperamide on bacterial toxins and other secretory agents is outlined. Diarrhoea induced by prostaglandins can be prevented by pretreatment with the drug. Details are given of clinical applications of loperamide for diarrhoea control in children and adults. The drug has no significant side-effects and presents no serious threat of abuse.

Selective blockade of the responses of reticulo-ruminal muscle to 5-HT in sheep.

The effects of intravenous administration of 5-HT on the patterns of reticulo-rumen contractions were studied in conscious sheep. Contractions were measured by strain gauges applied to the external wall of the reticulum and the ventral and/or dorsal sac of the rumen. The responses to 5-HT comprised (i) a short-lived contraction followed by (ii) a sustained increase in muscle tone and (iii) a concomitant inhibition of the extrinsic reticulo-rumen contractions. The corresponding blockades produced by (i) atropine, (ii) 5-HT antagonists and (iii) chemical sympathectomy suggest involvement of a peripheral cholinergic mechanism in the initial contractile response and a central adrenergic mechanism in the reflex inhibition of extrinsic reticulo-rumen contractions.

Clinical evaluation of R 51163, a new sedative in cattle.

R 51163, a purine alkyl piperidine derivative, has been shown to produce reliable sedation in cattle of varying age and breed in three different types of experiments: transport and regrouping of cattle which had only been handled occasionally; manipulations and minor interventions; minor and major surgery with or without supplementary local anaesthesia. With dose rates of R 51163 at 0.05 mg/kg i.v. and of 0.10-0.15 mg/kg i.m. sedation became prominent within 10-15 min and lasted at least 60 min. Caesarean sections were performed under adequate sedation with R 51163 with dose ranging from 0.018 to 0.05 mg/kg i.v. and from 0.075 to 0.10 mg/kg i.m. The data suggest possible increased responsiveness to the drug at the time of calving.

Clinical, cardiovascular and respiratory effects of R8110 in premedicated dogs.

The clinical, cardiovascular and respiratory effects after i.v. administration of R8110, a fluoro analogue of etomidate (Fig. 1), were studied in pre-medicated dogs. The clinical observations were made at doses of 3 and 4 mg/kg body weight (BW) injected slowly i.v., whereas cardiovascular and respiratory studies were carried out at a dose rate of 3 mg/kg R8110 i.v. Induction and recovery were smooth and no significant side-effects were observed. The cardiovascular system was slightly influenced, but respiration was hardly affected. The effect of pre-medication on respiration and the cardiovascular system was hardly potentiated by R8110. Although there were significant changes in cardiovascular and biochemical parameters, all values remained within physiological limits. R8110 appeared to be a safe and reliable induction agent.

Comparison of the effects of etomidate and its fluoro analogue, R 8110, on plasma cortisol, 11 beta-deoxycortisol, 17 alpha-hydroxyprogesterone and testosterone concentrations in dogs.

R 8110, an imidazole derivative, was shown to be clinically superior to etomidate for induction and maintenance of anaesthesia in dogs. The present study compared the effects of intravenous (i.v.) R 8110, etomidate and Ringer solution on cortisol biosynthesis by the adrenal gland in seven male labradors. A tetracosactide challenge was carried out 30 min after the i.v. injection of 3 mg/kg of both drugs and after i.v. Ringer solution (1 ml/kg). Etomidate and R 8110 both suppressed the cortisol response to tetracosactide almost completely and increased the plasma 11 beta-deoxycortisol levels more than 20 fold. Maximal 11 beta-deoxycortisol values were reached 120 min after R 8110, and not less than 300 min after etomidate. Plasma 17 alpha-hydroxyprogesterone and testosterone concentrations did not differ between placebo and R 8110 treatment, but they decreased after etomidate. These results indicate that the effects of R 8110 on steroid biosynthesis in dogs are less pronounced than those of etomidate and are largely limited to a temporary inhibition of the 11 beta-hydroxylase in the adrenal gland.