Ethanol inhibits dopamine uptake via organic cation transporter 3: Implications for ethanol and cocaine co-abuse.

Concurrent cocaine and alcohol use is among the most frequent drug combination, and among the most dangerous in terms of deleterious outcomes. Cocaine increases extracellular monoamines by blocking dopamine (DA), norepinephrine (NE) and serotonin (5-HT) transporters (DAT, NET and SERT, respectively). Likewise, ethanol also increases extracellular monoamines, however evidence suggests that ethanol does so independently of DAT, NET and SERT. Organic cation transporter 3 (OCT3) is an emergent key player in the regulation of monoamine signaling. Using a battery of in vitro, in vivo electrochemical, and behavioral approaches, as well as wild-type and constitutive OCT3 knockout mice, we show that ethanol's actions to inhibit monoamine uptake are dependent on OCT3. These findings provide a novel mechanistic basis whereby ethanol enhances the neurochemical and behavioral effects of cocaine and encourage further research into OCT3 as a target for therapeutic intervention in the treatment of ethanol and ethanol/cocaine use disorders.

Distal Transradial Access for Diagnostic Cerebral Angiography and Neurointervention: Systematic Review and Meta-analysis.

BACKGROUND: Radial artery access for cerebral angiography is traditionally performed in the wrist. Distal transradial access in the anatomic snuffbox is an alternative with several advantages. PURPOSE: Our aim was to review the safety and efficacy of distal transradial access for diagnostic cerebral angiography and neurointerventions. DATA SOURCES: We performed a comprehensive search of the literature using PubMed, Scopus, and EMBASE. STUDY SELECTION: The study included all case series of at least 10 patients describing outcomes associated with distal transradial access for diagnostic cerebral angiography or a neurointervention. DATA ANALYSIS: Random-effects models were used to obtain pooled rates of procedural success and complications. DATA SYNTHESIS: A total of 7 studies comprising 348 (75.8%) diagnostic cerebral angiograms and 111 (24.2%) interventions met the inclusion criteria. The pooled success rate was 95% (95% CI, 91%-98%; I2 = 74.33). The pooled minor complication rate was 2% (95% CI, 1%-4%; I2 = 0. No major complications were reported. For diagnostic procedures, the combined mean fluoroscopy time was 13.53 [SD, 8.82] minutes and the mean contrast dose was 74.9 [SD, 35.6] mL. LIMITATIONS: A small number of studies met the inclusion criteria, all of them were retrospective, and none compared outcomes with proximal transradial or femoral access. CONCLUSIONS: Early experience with distal transradial access suggests that it is a safe and effective alternative to proximal radial and femoral access for performing diagnostic cerebral angiography and interventions. Additional studies are needed to establish its efficacy and compare it with other access sites.

Giri-nya-la-nha (talk together) to explore acceptability of targeted smoking cessation resources with Australian Aboriginal women.

OBJECTIVES: To engage with health providers and Aboriginal women to understand what educational resources they want and need to support quit smoking attempts during pregnancy in order to develop a comprehensive evidence-based intervention. STUDY DESIGN: Resources were developed in partnership with Aboriginal people, communities and academics with the aim to be inclusive of diverse communities. We then recruited Aboriginal women of various ages for yarning circles (focus groups) held in three Australian states to explore the acceptability of the resources and seeking further guidance as to the needs of Aboriginal women to support smoking cessation during pregnancy. METHODS: Yarning circles were recorded and transcribed, and data were analysed independently by two researchers. Responses were coded using predetermined themes and further general inductive analysis for emergent themes. RESULTS: Twenty-four Aboriginal women reflected on the resources they included: one pregnant woman, 15 mothers and eight elders. Predetermined themes of attraction, comprehension, cultural acceptability, graphics and layout, persuasion and self-efficacy were explored. Women suggested the following: resources need to be visually attractive and interactive to enhance self-efficacy; additional scientific content on health consequences of smoking and combining with non-pharmacological approaches to quitting. CONCLUSION: Indigenous peoples prefer culturally targeted messages. However, developing effective Aboriginal health promotion requires more than a 'culturally appropriate' adaptation of mainstream resources. Consideration needs to be given to the diversity of Aboriginal communities when developing effective, evidence-based interventions. Aboriginal women are calling for innovative and interactive resources that enhance self-efficacy; the use of videos to explain medical and informational brochure content is well received. Requests for non-pharmacological cessation options were reported in New South Wales and Queensland and should be further explored.

Wula (Voices) of Aboriginal women on barriers to accepting smoking cessation support during pregnancy: Findings from a qualitative study.

AIM: To gather Aboriginal women's stories of smoking and becoming pregnant to identify the barriers in accepting smoking cessation support during pregnancy. METHODS: Qualitative data were collected through use of yarning methodology between August 2015 and January 2016 by an Aboriginal Researcher with experience in social and community services. A short on-line survey was used to collect quantitative data. Interviews only recorded the therapeutic yarning process, which ranged from 9 to 45min duration, averaging 30min. Audio-recorded interviews were transcribed and independently coded. A general inductive analysis was used to determine emergent themes. RESULTS: Twenty Aboriginal women between 17-38 years of age, who were pregnant or recently given birth, living in the Hunter New England (HNE) area took part. Eleven women were still smoking; nine had quit. Most were highly aware of the implications of smoking for their babies. Major themes identified for accepting support were: ambivalence towards a need for support, health professional advice, reduction in smoking, and attitudes to Nicotine Replacement Therapy (NRT). Women reported being advised to cut down, rather than to quit; reducing consumption may be a barrier to accepting NRT. Women recommended enhanced clinical support and Aboriginal community engagement in cessation care. DISCUSSION/CONCLUSIONS: Aboriginal women in the HNE area reported quitting or reducing their cigarette intake during pregnancy. Health Professionals working with Aboriginal women during pregnancy should give consistent messages to quit smoking completely, and offer increased, ongoing and extensive smoking cessation support to Aboriginal mothers. Clinical practices could partner with Aboriginal communities to support the delivery of smoking cessation services.

Development and initial validation of the Bristol Impact of Hypermobility questionnaire.

OBJECTIVES: Stage 1 - to identify the impact of joint hypermobility syndrome (JHS) on adults; Stage 2 - to develop a questionnaire to assess the impact of JHS; and Stage 3 - to undertake item reduction and establish the questionnaire's concurrent validity. DESIGN: A mixed methods study employing qualitative focus groups and interviews (Stage 1); a working group of patients, clinicians and researchers, and 'think aloud' interviews (Stage 2); and quantitative analysis of questionnaire responses (Stage 3). SETTING: Stages 1 and 2 took place in one secondary care hospital in the UK. Members of a UK-wide patient organisation were recruited in Stage 3. PARTICIPANTS: In total, 15, four and 615 participants took part in Stages 1, 2 and 3, respectively. Inclusion criteria were: age ≥18 years; diagnosis of JHS; no other conditions affecting physical function; able to give informed consent; and able to understand and communicate in English. INTERVENTIONS: None. MAIN OUTCOME MEASURES: The development of a questionnaire to assess the impact of JHS. RESULTS: Stage 1 identified a wide range of impairments, activity limitations and participation restrictions In Stage 2, a draft questionnaire was developed and refined following 'think aloud' analysis, leaving 94 scored items. In Stage 3, items were removed on the basis of low severity and/or high correlation with other items. The final Bristol Impact of Hypermobility (BIoH) questionnaire had 55 scored items, and correlated well with the physical component score of the Short Form 36 health questionnaire (r=-0.725). CONCLUSIONS: The BIoH questionnaire demonstrated good concurrent validity. Further psychometric properties need to be established.

Spontaneous delayed migration/shortening of the pipeline embolization device: report of 5 cases.

Five patients were found to have spontaneous delayed migration/shortening of their Pipeline Embolization Devices on follow-up angiography. The device migrated proximally in 4 patients and distally in 1 patient. One patient had a subarachnoid hemorrhage and died as a result of migration of the Pipeline Embolization Device, and another patient presented with complete MCA occlusion and was left severely disabled. Mismatch in arterial diameter between inflow and outflow vessels was a constant finding. Migration of the Pipeline Embolization Device was managed conservatively, with additional placement of the device, or with parent vessel occlusion. Obtaining complete expansion of the embolization device by using a longer device, increasing vessel coverage, using adjunctive aneurysm coiling, and avoiding dragging and stretching of the device are important preventive measures. Neurointerventionalists should be aware of this potentially fatal complication and take all necessary preventive measures.

Visual social preferences of lone zebrafish in a novel environment: strain and anxiolytic effects.

Zebrafish (Danio rerio) have an innate tendency to join shoals. Based on this, we refined visual choice tests to focus on social interaction and novelty preference. Our design follows mouse three-chamber sociability protocols, except testing is conducted under 940 Lux fluorescent lighting. Initially, we compared performance among zebrafish strains: inbred (AB) or wild-crossbred (WIK) from Zebrafish International Resource Center, to golden and short-fin from Petco stores. AB fish exhibited a preference for shoaling; they dwelled longest near transparent boxes containing zebrafish, while short fin favored blue boxes without fish. AB and golden exhibited a strong preference for social novelty, not evident in short-fin or WIK fish. Serotonin and cannabinoids shape mammalian social behavior, and equivalents of both receptor types are expressed in the zebrafish brain. We examined the effects of the cannabinoid receptor agonist WIN 55,212-2 (1 mg/l), or serotonin 5-HT(1A) receptor agonist buspirone (10 mg/l) on Petco short-fin social choice. Fish were bath exposed to test compounds for 10 min, under these conditions [(3) H]CP55,940 (4 nm) bound to brain with a concentration of 1.9-6.4 fmol/mg 5-30 min afterward. Social approach was measured 20 min after acclimation to the test arena. WIN 55,212-2 and buspirone increased dwelling near boxed zebrafish. In zebrafish whole-brain homogenates, buspirone displaced [(3) H] 8-hydroxy-N,N-dipropylaminotetralin (dissociation constant, K(D) = 16 ± 1.2 nm) with an inhibition constant (K(i) ) of 1.8 ± 1.0 nm lower than that of WAY 100,635 (K(i) ∼1000 nm). These fish social choice tests may enhance social behavior research, and are useful for studying the effects of genetic manipulations, pharmaceuticals or environmental toxins.

National survey of College Tutors in the UK regarding training in medical education.

BACKGROUND: College Tutors are responsible for the organization of training and should possess the pre-requisite knowledge and skills to facilitate this. METHODS: An anonymized survey of anaesthetic College Tutors in the UK was conducted with regard to training in medical education. RESULTS: A response rate of 65.54% was achieved. Around 16% had a formal postgraduate teaching qualification and another 27% were interested in attaining one. However, 84% were of the opinion that formal teaching qualifications were not essential for College Tutors. The more recently appointed College Tutors (<2 yr experience) had more formal qualifications and thought these were important. Appraisal and assessment courses were considered the most valuable for professional development of the role of the College Tutor, and were identified as challenging. CONCLUSIONS: This survey highlights that training in medical education for College Tutors is inadequate. It is the responsibility of the Colleges and the Postgraduate Deans to ensure College Tutors are supported appropriately to develop the knowledge and skills required for the role.

History of science--spores.

Bacterial endospores were first studied 130 years ago by Cohn in 1876 and independently by Koch in the same year. Although spore dormancy and resistance have been much studied since then, questions still remain concerning the basic mechanisms and the kinetics of heat inactivation in particular. Likewise, the extreme dormancy and longevity of spores was recognized early on and later greatly extended but still evade complete understanding. Evidence has accumulated for the involvement of specific spore components such as calcium, dipicolinic acid, small acid soluble proteins in the core and peptidoglycan in the cortex. Involvement of physical factors too, such as the relative dehydration of the core, maybe in a high-viscosity state or even in a glassy state, has added to appreciation of the multicomponent nature of dormancy and resistance. Spore-former morphology formed the basis for early classification systems of sporeformers from about 1880 and consolidated in the mid-1900s, well prior to the use of modern genetic procedures. With respect to sporulation, groundbreaking sequence studies in the 1950s provided the basis for later elucidation of the genetic control widely relevant to many cell differentiation mechanisms. With respect to the breaking of dormancy (activation and germination), the elucidation of mechanisms began in the 1940s following the observations of Hills at Porton who identified specific amino acid and riboside 'germinants', and laid the basis for the later genetic analyses, the identification of germinant receptor genes and the elucidation of key germination reactions. The nonexponential nature of germination kinetics has thwarted the development of practical Tyndallization-like processing. So inactivation by heat remains the premier method of spore control, the basis of a huge worldwide industry, and still relying on the basic kinetics of inactivation of Clostridium botulinum spores, and the reasoning regarding safety first evolved by Bigelow et al. in 1920 and Esty and Meyer in 1922. 'Newer' processes such as treatment with ionizing radiation (first proposed in 1905) and high hydrostatic pressure (first proposed in 1899) may be introduced if consumer resistance and some remaining technical barriers could be overcome.

Chicken recombinant antibodies against infectious bursal disease virus are able to form antibody-virus immune complex.

Virus particles exposed to specific anti-virus antibodies result in the formation of immune complexes (Icx). Recent vaccination strategies have employed this feature, and an infectious bursal disease virus (IBDV) vaccine based on Icx has been released and is expected to replace conventional IBDV vaccines. We evaluated whether chicken recombinant antibodies (rAb) specific for IBDV, rather than conventional chicken anti-IBDV sera, could be used to generate Icx. Out of 14 rAb expressed as soluble single-chain variable fragments (scFv), nine were able to completely neutralize Bursavac, a live IBDV vaccine, when tested in ovo. When these rAb were mixed with IBDV and inoculated into either 18-day-old embryos, or 1-day-old or 2-week-old specific pathogen free chicks, a rAb.IBDV complex was formed. These Icx were similar to those produced by polyclonal chick anti-IBDV sera and IBDV. Following inoculation of the rAb.IBDV complex, the virus was rendered non-infectious for 5 to 7 days. After this time virus was released from the Icx, resulting in infection of the inoculated chicks and subsequent induction of an immune response and protection against virulent IBDV challenge. The results indicated that genetically derived antibodies can replace polyclonal sera in the formulation of Icx vaccines.

Chicken recombinant antibodies specific for very virulent infectious bursal disease virus.

A phage-displayed single chain variable fragment (scFv) antibody library was constructed from the immune spleen cells of chickens immunized with very virulent infectious bursal disease virus (vvIBDV) strain CS89. A library consisting of around 9.2 x 10(7) clones was subjected to 3 rounds of panning against captured CS89 virus. Analysis of individual clones by nucleotide sequencing revealed at least 22 unique scFv antibodies binding to vvIBDV in ELISA. Testing of the scFv antibody panel in ELISA against classical, variant or vaccine strains and a wide variety of vvIBDV isolates from the UK, China, France, Belgium, Africa, Brazil, Indonesia and the Netherlands identified one antibody, termed chicken recombinant antibody 88 (CRAb 88) that was specific for vvIBDV. CRAb 88 was capable of recognizing all vvIBDV strains tested regardless of their country of origin and showed no reactivity with classical, variant or vaccine strains, lending support to the use of this scFv as a powerful diagnostic tool for the differentiation of vvIBDV strains. Immunoprecipitation studies revealed that CRAb 88 was directed towards a highly conformational epitope located within the major neutralizing protein VP2. Sequence analysis of the hypervariable region of VP2 of the IBDV strains tested indicate that Ile(256) and Ile(294) may play roles in binding of CRAb 88. This is the first reagent of its type capable of positively distinguishing vvIBDV from other IBDV strains.

Isolation of a variant infectious bronchitis virus in Australia that further illustrates diversity among emerging strains.

Australian infectious bronchitis viruses (IBV) have undergone a separate evolution due to geographic isolation. Consequently, changes occurring in Australian IBV illustrate, independently from other countries, types of variability that could occur in emerging IBV strains. Previously, we have identified two distinct genetic groups of IBV, designated subgroups 1 and 2. IBV strains of subgroup 1 have S1 and N proteins that share a high degree of amino acid identity, 81 to 98% in S1 and 91 to 99% in N. Subgroup 2 strains possess S1 and N proteins that share a low level of identity with subgroup 1 strains: 54 to 62% in S1 and 60 to 62% in N. This paper describes the isolation and characterisation of a third, previously undetected genetic group of IBV in Australia. The subgroup 3 strains, represented by isolate chicken/Australia/N2/04, had an S1 protein that shared a low level of identity with both subgroups 1 and 2: 61 to 63% and 56 to 59%, respectively. However, the N protein and the 3' untranslated region were similar to subgroup 1: 90 to 97% identical with the N protein of subgroup 1 strains. This N4/02 subgroup 3 of IBV is reminiscent of two other strains, D1466 and DE072, isolated in the Netherlands and in the USA, respectively. The emergence of the subgroup 3 viruses in Australia, as well as the emergence of subgroup 2 in 1988, could not be explained by any of the mechanisms that are currently considered to be involved in generation of IBV variants.

Membrane traffic in cytokinesis.

A crucial facet of mammalian cell division is the separation of two daughter cells by a process known as cytokinesis. An early event in cytokinesis is the formation of an actomyosis contractile ring, which functions like a purse string in the constriction of the forming furrow between the cells. Far less well characterized are the membrane-trafficking steps which deliver new membrane to the cell surface during the plasma membrane expansion known to accompany furrow formation. It is now clearly established that the plasma membrane at the cleavage furrow of mammalian cells has a distinct lipid and protein composition from the rest of the plasma membrane. This may reflect a requirement for both increased surface area during furrowing and for the co-ordinated delivery of intracellular signalling or membrane re-modelling activities to the correct spatial coordinates during cleavage. In this review, we discuss recent work within the area of membrane traffic and cytokinesis.

Insect renal tubules constitute a cell-autonomous immune system that protects the organism against bacterial infection.

Innate immunity is a widespread and important defence against microbial attack, which in insects is thought to originate mainly in the fat body. Here we demonstrate that the fluid-transporting Malpighian (renal) tubule of Drosophila melanogaster constitutes an autonomous immune-sensing tissue utilising the nitric oxide (NO) signalling pathway. Reverse transcriptase PCR (RT-PCR) shows that tubules express those genes encoding components of the Imd pathway. Furthermore, isolated tubules bind and respond to lipopolysaccharide (LPS), by upregulating anti-microbial peptide (diptericin) gene expression and increased bacterial killing. Excised, LPS-challenged tubules, as well as tubules from LPS-infected flies, display increased NO synthase (NOS) activity upon immune challenge. Targetted expression of a Drosophila NOS (dNOS) transgene to only principal cells of the tubule main segment using the GAL4/UAS system increases diptericin expression. In live flies, such targetted over-expression of dNOS to tubule principal cells confers increased survival of the whole animal upon E. coli challenge. Thus, we describe a novel role of Malpighian tubules in immune sensing and insect survival.

Reduced insulin-stimulated GLUT4 bioavailability in stroke-prone spontaneously hypertensive rats.

AIMS/HYPOTHESIS: Insulin-stimulated glucose transport is impaired in a genetic model of hypertension, the stroke-prone spontaneously hypertensive rat (SHRSP), yet the molecular mechanisms that underlie this defect in the animals remain unclear. METHODS: We examined the effects of insulin on the trafficking of the insulin-responsive glucose transporter GLUT4 to the plasma membrane in isolated adipocytes from SHRSP and normotensive control Wistar-Kyoto (WKY) rats. RESULTS: Treatment of isolated adipocytes with insulin resulted in trafficking of GLUT4 to the plasma membrane. There was no significant difference in the magnitude of insulin-stimulated GLUT4 trafficking from intracellular membranes to the plasma membrane between strains. In contrast, we demonstrated that there is a significant reduction in GLUT4 accessible to the glucose photolabel Bio-LC-ATB-BGPA at the plasma membrane of SHRSP adipocytes compared with control rats. CONCLUSIONS/INTERPRETATION: We propose that a large proportion of GLUT4 translocated to the plasma membrane in response to insulin is not able to bind substrate and catalyse transport in the SHRSP. Therefore, there is a reduction in bioavailable GLUT4 in SHRSP animals that is likely to account, at least in part, for the reduced insulin-stimulated glucose uptake.

An ELISA for detection of infectious bursal disease virus and differentiation of very virulent strains based on single chain recombinant chicken antibodies.

Two chicken single-chain variable antibody fragments (scFv) designated scFv154 and scFv88, previously shown to react with either all or very virulent (vv) infectious bursal disease virus (IBDV) strains, respectively, were evaluated for use in an enzyme-linked immunosorbent assay (ELISA) for differentiation of vvIBDV. Specificity and sensitivity of the vvIBDV ELISA was assessed when scFv154 and scFv88 were expressed as soluble antibodies (sAb), phage antibodies (pAb) or hyper-phage antibodies (hpAb). The highest test sensitivity and specificity was obtained using hpAb154 to detect all IBDV and pAb88 to differentiate vvIBDV strains. Such an ELISA was eight to 16 times more sensitive for IBDV antigen detection than the mouse monoclonal antibody ELISA. Using field samples, the scFv ELISA was able to differentiate between flocks infected with vvIBDV and those infected with classical or variant IBDV. In one instance IBDV was detected in a flock found to be negative by the monoclonal antibody ELISA. The results showed that scFv can be utilized as highly specific and sensitive ELISA reagents for the detection and discrimination of avian pathogens.

Recurrent chylothorax in a patient with non-Hodgkins lymphoma: case report.

Spontaneous chylothorax could arise as a complication of Iymphoma. There are no reports on the frequency of it's occurrence. It is associated with a high mortality rate. This is mainly due to severe nutritional deficiencies and wasting. This case describes a patient with non-Hodgkins Iymphoma who developed recurrent bilateral chylothorax requiring repeated pleural aspirations and eventually talc pleurodesis which failed.

Virus strains from a flock exhibiting unusually high mortality due to infectious bursal disease.

OBJECTIVE: To characterise infectious bursal disease viruses (IBDVs) isolated from commercial broiler flocks exhibiting unusually high mortality due to infectious bursal disease (IBD). DESIGN: An IBD outbreak occurred in mid 1999 on two broilers farms (A and B) in northern New South Wales amongst chickens 28 to 38 days of age, with a sharp rise in mortality of 2.5%. Initial histopathological diagnosis indicated acute IBD. Since acute IBD caused by classical pathogenic and very virulent (vv) IBDVs is exotic to Australia, samples from both farms A and B were obtained and used for virus characterisation. METHOD: Tissue homogenates were made from six bursae collected from farm B. One histological sample from farm A was also used. Nucleotide sequencing of the hypervariable region (HVR) within the VP2 gene of IBDVs was determined and the deduced amino acid sequences compared with previously characterised Australian and overseas IBDVs. The phylogenetic relationship between IBDVs from farm B and IBDVs from Australia and overseas was then determined. Pathogenicity of one isolate, N2/99 from farm B, was compared with 3 other local IBDVs, as well as with three pathogenic overseas strains in 3-week-old specific pathogen-free (SPF) chickens. RESULTS: Initial histopathological characterisation of a sample of bursa from a bird on farm A showed widespread acute lymphoid necrosis, follicular haemorrhage and stromal oedema, indicative of acute IBD. Subsequent analysis using reverse transcriptase polymerase chain reaction (RT-PCR), followed by nucleotide sequencing of the same bursal sample, as well as 6 samples from nearby farm B, showed that the IBDVs involved were similar in sequence to Australian vaccine strains and not to classical pathogenic or vvIBDVs. One isolate, N2/99 from farm B, was only marginally more pathogenic than other local IBDVs. It induced mild clinical signs in 30% of chicks and no mortality. In comparison, vvIBDV CS89 and classical pathogenic 52/70 strains induced severe clinical signs in 100% and 80% of chickens, respectively with mortalities of 27% and 12%, respectively. CONCLUSIONS: The results illustrated the value of nucleotide sequencing as a method for discrimination of local and exotic types of IBDV.