Endothelial-Specific Reduction in Arf6 Impairs Insulin-Stimulated Vasodilation and Skeletal Muscle Blood Flow Resulting in Systemic Insulin Resistance in Mice.

BACKGROUND: Much of what we know about insulin resistance is based on studies from metabolically active tissues such as the liver, adipose tissue, and skeletal muscle. Emerging evidence suggests that the vascular endothelium plays a crucial role in systemic insulin resistance; however, the underlying mechanisms remain incompletely understood. Arf6 (ADP ribosylation factor 6) is a small GTPase that plays a critical role in endothelial cell function. Here, we tested the hypothesis that the deletion of endothelial Arf6 will result in systemic insulin resistance. METHODS: We used mouse models of constitutive endothelial cell-specific Arf6 deletion (Arf6f/- Tie2Cre+) and tamoxifen-inducible Arf6 knockout (Arf6f/f Cdh5CreER+). Endothelium-dependent vasodilation was assessed using pressure myography. Metabolic function was assessed using a battery of metabolic assessments including glucose and insulin tolerance tests and hyperinsulinemic-euglycemic clamps. We used a fluorescence microsphere-based technique to measure tissue blood flow. Skeletal muscle capillary density was assessed using intravital microscopy. RESULTS: Endothelial Arf6 deletion impaired insulin-stimulated vasodilation in white adipose tissue and skeletal muscle feed arteries. The impairment in vasodilation was primarily due to attenuated insulin-stimulated nitric oxide bioavailability but independent of altered acetylcholine-mediated or sodium nitroprusside-mediated vasodilation. Endothelial cell-specific deletion of Arf6 also resulted in systematic insulin resistance in normal chow-fed mice and glucose intolerance in high-fat diet-fed obese mice. The underlying mechanisms of glucose intolerance were reductions in insulin-stimulated blood flow and glucose uptake in the skeletal muscle and were independent of changes in capillary density or vascular permeability. CONCLUSIONS: Results from this study support the conclusion that endothelial Arf6 signaling is essential for maintaining insulin sensitivity. Reduced expression of endothelial Arf6 impairs insulin-mediated vasodilation and results in systemic insulin resistance. These results have therapeutic implications for diseases that are associated with endothelial cell dysfunction and insulin resistance such as diabetes.

Endothelial cell-specific reduction in mTOR ameliorates age-related arterial and metabolic dysfunction.

Systemic inhibition of the mammalian target of rapamycin (mTOR) delays aging and many age-related conditions including arterial and metabolic dysfunction. However, the mechanisms and tissues involved in these beneficial effects remain largely unknown. Here, we demonstrate that activation of S6K, a downstream target of mTOR, is increased in arteries with advancing age, and that this occurs preferentially in the endothelium compared with the vascular smooth muscle. Induced endothelial cell-specific deletion of mTOR reduced protein expression by 60-70%. Although this did not significantly alter arterial and metabolic function in young mice, endothelial mTOR reduction reversed arterial stiffening and improved endothelium-dependent dilation (EDD) in old mice, indicating an improvement in age-related arterial dysfunction. Improvement in arterial function in old mice was concomitant with reductions in arterial cellular senescence, inflammation, and oxidative stress. The reduction in endothelial mTOR also improved glucose tolerance in old mice, and this was associated with attenuated hepatic gluconeogenesis and improved lipid tolerance, but was independent of alterations in peripheral insulin sensitivity, pancreatic beta cell function, or fasted plasma lipids in old mice. Lastly, we found that endothelial mTOR reduction suppressed gene expression of senescence and inflammatory markers in endothelial-rich (i.e., lung) and metabolically active organs (i.e., liver and adipose tissue), which may have contributed to the improvement in metabolic function in old mice. This is the first evidence demonstrating that reducing endothelial mTOR in old age improves arterial and metabolic function. These findings have implications for future drug development.

Reduction of double-strand DNA break repair exacerbates vascular aging.

Advanced age is the greatest risk factor for cardiovascular disease (CVD), the leading cause of death. Arterial function is impaired in advanced age which contributes to the development of CVD. One underexplored hypothesis is that DNA damage within arteries leads to this dysfunction, yet evidence demonstrating the incidence and physiological consequences of DNA damage in arteries, and in particular, in the microvasculature, in advanced age is limited. In the present study, we began by assessing the abundance of DNA damage in human and mouse lung microvascular endothelial cells and found that aging increases the percentage of cells with DNA damage. To explore the physiological consequences of increases in arterial DNA damage, we evaluated measures of endothelial function, microvascular and glycocalyx properties, and arterial stiffness in mice that were lacking or heterozygous for the double-strand DNA break repair protein ATM kinase. Surprisingly, in young mice, vascular function remained unchanged which led us to rationalize that perhaps aging is required to accumulate DNA damage. Indeed, in comparison to wild type littermate controls, mice heterozygous for ATM that were aged to ~18 mo (Old ATM +/-) displayed an accelerated vascular aging phenotype characterized by increases in arterial DNA damage, senescence signaling, and impairments in endothelium-dependent dilation due to elevated oxidative stress. Furthermore, old ATM +/- mice had reduced microvascular density and glycocalyx thickness as well as increased arterial stiffness. Collectively, these data demonstrate that DNA damage that accumulates in arteries in advanced age contributes to arterial dysfunction that is known to drive CVD.

Endothelial cell telomere dysfunction induces senescence and results in vascular and metabolic impairments.

In advanced age, increases in oxidative stress and inflammation impair endothelial function, which contributes to the development of cardiovascular disease (CVD). One plausible source of this oxidative stress and inflammation is an increase in the abundance of senescent endothelial cells. Cellular senescence is a cell cycle arrest that occurs in response to various damaging stimuli. In the present study, we tested the hypothesis that advanced age results in endothelial cell telomere dysfunction that induces senescence. In both human and mouse endothelial cells, advanced age resulted in an increased abundance of dysfunctional telomeres, characterized by activation of DNA damage signaling at telomeric DNA. To test whether this results in senescence, we selectively reduced the telomere shelterin protein telomere repeat binding factor 2 (Trf2) from endothelial cells of young mice. Trf2 reduction increased endothelial cell telomere dysfunction and resulted in cellular senescence. Furthermore, induction of endothelial cell telomere dysfunction increased inflammatory signaling and oxidative stress, resulting in impairments in endothelial function. Finally, we demonstrate that endothelial cell telomere dysfunction-induced senescence impairs glucose tolerance. This likely occurs through increases in inflammatory signaling in the liver and adipose tissue, as well as reductions in microvascular density and vasodilation to metabolic stimuli. Cumulatively, the findings of the present study identify age-related telomere dysfunction as a mechanism that leads to endothelial cell senescence. Furthermore, these data provide compelling evidence that senescent endothelial cells contribute to age-related increases in oxidative stress and inflammation that impair arterial and metabolic function.

Endothelial specific reduction in Arf6 impairs insulin-stimulated vasodilation and skeletal muscle blood flow resulting in systemic insulin resistance.

Background: Much of what we know about insulin resistance is based on studies from metabolically active tissues such as liver, adipose tissue, and skeletal muscle. Emerging evidence suggests that the vascular endothelium plays a crucial role in systemic insulin resistance, however, the underlying mechanisms remain incompletely understood. ADP ribosylation factor 6 (Arf6) is a small GTPase that plays a critical role in endothelial cell (EC) function. Here, we tested the hypothesis that the deletion of endothelial Arf6 will result in systemic insulin resistance. Methods: We used mouse models of constitutive EC-specific Arf6 deletion (Arf6 f/- Tie2Cre) and tamoxifen inducible Arf6 knockout (Arf6 f/f Cdh5Cre). Endothelium-dependent vasodilation was assessed using pressure myography. Metabolic function was assessed using a battery of metabolic assessments including glucose- and insulin-tolerance tests and hyperinsulinemic-euglycemic clamps. A fluorescence microsphere-based technique was used to measure tissue blood flow. Intravital microscopy was used to assess skeletal muscle capillary density. Results: Endothelial Arf6 deletion impaired insulin-stimulated vasodilation in white adipose tissue (WAT) and skeletal muscle feed arteries. The impairment in vasodilation was primarily due to attenuated insulin-stimulated nitric oxide (NO) bioavailability but independent of altered acetylcholine- or sodium nitroprusside-mediated vasodilation. In vitro Arf6 inhibition resulted in suppressed insulin stimulated phosphorylation of Akt and endothelial NO synthase. Endothelial cell-specific deletion of Arf6 also resulted in systematic insulin resistance in normal chow fed mice and glucose intolerance in high fat diet fed obese mice. The underlying mechanisms of glucose intolerance were reductions in insulin-stimulated blood flow and glucose uptake in the skeletal muscle and were independent of changes in capillary density or vascular permeability. Conclusion: Results from this study support the conclusion that endothelial Arf6 signaling is essential for maintaining insulin sensitivity. Reduced expression of endothelial Arf6 impairs insulin-mediated vasodilation and results in systemic insulin resistance. These results have therapeutic implications for diseases that are associated with endothelial cell dysfunction and insulin resistance such as diabetes.

Pyridoxamine treatment ameliorates large artery stiffening and cerebral artery endothelial dysfunction in old mice.

Age-related increases in large artery stiffness are associated with cerebrovascular dysfunction and cognitive impairment. Pyridoxamine treatment prevents large artery stiffening with advancing age, but the effects of pyridoxamine treatment on the cerebral vasculature or cognition is unknown. The purpose of this study was to investigate the effects of pyridoxamine on blood pressure, large artery stiffness, cerebral artery function, and cognitive function in old mice. Old male C57BL/6 mice consumed either pyridoxamine (2 g/L) or vehicle control in drinking water for ∼7.5 months and were compared with young male C57BL/6 mice. From pre- to post-treatment, systolic blood pressure increased in old control mice, but was maintained in pyridoxamine treated mice. Large artery stiffness decreased in pyridoxamine-treated mice but was unaffected in control mice. Pyridoxamine-treated mice had greater cerebral artery endothelium-dependent dilation compared with old control mice, and not different from young mice. Old control mice had impaired cognitive function; however, pyridoxamine only partially preserved cognitive function in old mice. In summary, pyridoxamine treatment in old mice prevented age-related increases in blood pressure, reduced large artery stiffness, preserved cerebral artery endothelial function, and partially preserved cognitive function. Taken together, these results suggest that pyridoxamine treatment may limit vascular aging.

Effects of hatch window and nutrient access in the hatcher on performance and processing yield of broiler chicks reared according to time of hatch.

The effects of hatch window and hatching basket nutrient availability on organ weights, performance, and processing yield of broilers were investigated. Eggs were hatched in illuminated hatchers. At the end of each hatch window period (HWP), hatched chicks were placed into control (CTL) hatching baskets with no nutrients or baskets providing access to feed and water (FAW). This resulted in 6 treatments in a factorial arrangement of 3 HWP (early, middle, or late) and 2 basket types (CTL or FAW). Chicks remained in experimental baskets until 504 h and were then subjected to a 4 h holding period at the hatchery without nutrient access. Subsequently, 1,500 hatched chicks were reared in floor pens for 42 d with 5 replicate pens per treatment. Common diets and water were provided ad libitum. Bird weights and feed consumption were recorded weekly. Individual bird weights were taken at 21 and 42 d. At 43 d, 14 males from each pen were processed. There was an interaction between HWP and basket type on placement BW (P = 0.028) and BW change in the hatcher (P < 0.001). The HWP influenced BW at hatch (P = 0.007), 7 d (P < 0.001), and 14 d (P < 0.001) and FI at 7 d (P < 0.001) and 14 d (P = 0.002). Chicks from FAW baskets were heavier (P < 0.001) than those from CTL baskets at 7 d; afterward, they were similar (P > 0.05) in BW. Yolk and liver weights were similar (P > 0.05) between basket treatments at 3 d posthatch. No differences (P > 0.05) in FCR, mortality, or processing were observed between basket treatments. Interestingly, early hatching chicks were lightest at hatch but subsequently had higher FI and BWG. These findings indicate that hatcher nutrient access may reduce weight loss in the hatcher, especially for early hatching chicks, but had no influence on subsequent performance or processing yields beyond 7 d.

T cells mediate cell non-autonomous arterial ageing in mice.

KEY POINTS: Increased large artery stiffness and impaired endothelium-dependent dilatation occur with advanced age. We sought to determine whether T cells mechanistically contribute to age-related arterial dysfunction. We found that old mice exhibited greater proinflammatory T cell accumulation around both the aorta and mesenteric arteries. Pharmacologic depletion or genetic deletion of T cells in old mice resulted in ameliorated large artery stiffness and greater endothelium-dependent dilatation compared with mice with T cells intact. ABSTRACT: Ageing of the arteries is characterized by increased large artery stiffness and impaired endothelium-dependent dilatation. T cells contribute to hypertension in acute rodent models but whether they contribute to chronic age-related arterial dysfunction is unknown. To determine whether T cells directly mediate age-related arterial dysfunction, we examined large elastic artery and resistance artery function in young (4-6 months) and old (22-24 months) wild-type mice treated with anti-CD3 F(ab'2) fragments to deplete T cells (150 µg, i.p. every 7 days for 28 days) or isotype control fragments. Old mice exhibited greater numbers of T cells in both aorta and mesenteric vasculature when compared with young mice. Old mice treated with anti-CD3 fragments exhibited depletion of T cells in blood, spleen, aorta and mesenteric vasculature. Old mice also exhibited greater numbers of aortic and mesenteric IFN-γ and TNF-α-producing T cells when compared with young mice. Old control mice exhibited greater large artery stiffness and impaired resistance artery endothelium-dependent dilatation in comparison with young mice. In old mice, large artery stiffness was ameliorated with anti-CD3 treatment. Anti-CD3-treated old mice also exhibited greater endothelium-dependent dilatation than age-matched controls. We also examined arterial function in young and old Rag-1-/- mice, which lack lymphocytes. Rag-1-/- mice exhibited blunted increases in large artery stiffness with age compared with wild-type mice. Old Rag-1-/- mice also exhibited greater endothelium-dependent dilatation compared with old wild-type mice. Collectively, these results demonstrate that T cells play an important role in age-related arterial dysfunction.

Effects of Hatch Window and Nutrient Access in the Hatcher on Performance and Processing Yields of Broilers Reared with Equal Hatch Window Representation.

The objective of this experiment was to investigate the effects of feed and water availability in hatching baskets on broiler performance, processing yield, and organ weights while considering the influence of hatch window. Cobb 500 eggs were transferred into illuminated hatchers with two hatching basket types [control (CTL) hatching baskets with no nutrients provided or baskets containing feed and water (FAW)]. Chicks were pulled sequentially to establish four hatch window periods (HWP): early, pre-peak, post-peak, or late. Chicks were then held for 4 h at the hatchery without nutrient access and subsequently reared in 26 floor pens designated as CTL (n = 13) or FAW (n = 13), with 13 chicks from each of the 4 HWP per pen (52 chicks per pen). At 43 d, 16 males from each pen were processed. Chicks from FAW baskets were 1 g heavier (p < 0.001) than those from CTL baskets at placement and were heavier through 28 d (p = 0.003) but similar (p > 0.05) in body weight (BW) for the remainder of the 42 d. No differences (p > 0.05) in feed conversion ratio, mortality, or processing data were observed between CTL and FAW groups. Early-hatching chicks were lighter (p < 0.001) than those from all other HWP at placement, but were only lighter (p < 0.001) than the post-peak group by 42 d. In summary, it was found that hatching basket nutrient access increased the BW of broilers during the first 4 wk of growth, with no other effects on performance or yield. Also, earlier-hatching chicks were generally able to compensate for a lighter placement BW.

Aging differentially impacts vasodilation and angiogenesis in arteries from the white and brown adipose tissues.

Aging adipose tissues (ATs) manifest reduced vascularity and increased hypoxia and inflammation that contribute to local and systemic metabolic dysfunction. However, the mechanisms that underlie these age-related changes are incompletely understood. In this study, we sought to examine insulin-stimulated vasodilation and angiogenesis in the arterial vasculature from three major AT depots, perigonadal white (pgWAT), subcutaneous white (scWAT) and brown (BAT) from young and old mice. Here, we demonstrate that in young mice, insulin-stimulated vasodilation is lower in feed arteries from pgWAT compared to scWAT (p < 0.05), but no differences were found between feed arteries in other AT depots (p > 0.05). Insulin-stimulated vasodilation was lower in old compared to young feed arteries from all three AT depots (p < 0.05 for all). In the presence of endothelial nitric oxide synthase inhibitor, L-NAME, insulin-stimulated vasodilation was decreased in young (p < 0.05), but was unaffected in old (p > 0.05) from all AT depots. We also observed no age-related differences in endothelium-independent dilation, as assessed by sodium nitroprusside (p > 0.05). We next investigated angiogenic capacity of the vasculature in these AT depots. In young mice, BAT vasculature demonstrated the highest angiogenic potential, followed by pgWAT and scWAT. We found that aging decreased angiogenic sprout formation in pgWAT and BAT (both p < 0.05), but increased angiogenic potential in scWAT (p < 0.05), indicating dissimilar impact of aging on angiogenesis in different AT depots. Collectively, these data suggest that aging leads to a consistent impairment in insulin-stimulated vasodilation and reduction in NO bioavailability in all three AT, although aging differentially impacts angiogenic capacity across different AT depots.

Impact of high-fat diet on vasoconstrictor reactivity of white and brown adipose tissue resistance arteries.

Blood flow regulation is a critical factor for tissue oxygenation and substrate supply. Increased reactivity of arteries to vasoconstrictors may increase vascular resistance, resulting in reduced blood flow. We aimed to investigate the effect of a high-fat (HF) diet on stiffness and vasoconstrictor reactivity of white adipose tissue (WAT) and brown adipose tissue (BAT) resistance arteries and also investigated the interconversion of both adipose depots in the setting of a HF diet. Vasoconstrictor reactivity and passive morphology and mechanical properties of arteries from B6D2F1 mice (5 mo old) fed normal chow (NC) or a HF diet (8 wk) were measured using pressure myography. Receptor gene expression in WAT and BAT arteries and markers of WAT and BAT were assessed in whole tissue lysates by real-time RT-PCR. Despite greater receptor-independent vasoconstriction (in response to KCl, P < 0.01), vasoconstriction in response to angiotensin II ( P < 0.01) was lower in NC-BAT than NC-WAT arteries and similar in response to endothelin-1 ( P = 0.07) and norepinephrine ( P = 0.11) in NC-BAT and NC-WAT arteries. With the exception of BAT artery reactivity to endothelin-1 and angiotensin II, the HF diet tended to attenuate reactivity in arteries from both adipose depots and increased expression of adipose markers in BAT. No significant differences in morphology or passive mechanical properties were found between adipose types or diet conditions. Alterations in gene expression of adipose markers after the HF diet suggest beiging of BAT. An increase in brown adipocytes in the absence of increased BAT mass may be a compensatory mechanism to dissipate excess energy from a HF diet. NEW & NOTEWORTHY Despite no differences in passive mechanical properties and greater receptor-independent vasoconstriction, receptor-mediated vasoconstriction was either lower in brown than white adipose tissue arteries or similar in brown and white adipose tissue arteries. A high-fat diet has a greater impact on vasoconstrictor responses in white adipose tissue but leads to altered adipose tissue gene expression consistent with beiging of the brown adipose tissue.

The impact of ageing on adipose structure, function and vasculature in the B6D2F1 mouse: evidence of significant multisystem dysfunction.

The critical influence of the white adipose tissue (WAT) on metabolism is well-appreciated in obesity, but adipose tissue dysfunction as a mechanism underlying age-associated metabolic dysfunction requires elucidation. To explore this possibility, we assessed metabolism and measures of epididymal (e)WAT mitochondria and artery function in young (6.1 ± 0.4 months) and old (29.6 ± 0.2 months) B6D2F1 mice. There were no group differences in average daily oxygen consumption, fasted blood glucose or plasma free fatty acids, but fasted plasma insulin and the homeostatic model assessment of insulin resistance (HOMA-IR%) were higher in the old (∼50-85%, P < 0.05). Tissue mass (P < 0.05) and adipocyte area were lower (∼60%) (P < 0.01) and fibrosis was greater (sevenfold, P < 0.01) in eWAT with older age. The old also exhibited greater liver triglycerides (∼60%, P < 0.05). The mitochondrial respiratory oxygen flux after the addition of glutamate and malate (GM), adenosine diphosphate (d), succinate (S) and octanoyl carnitine (O) were one- to twofold higher in eWAT of old mice (P < 0.05). Despite no change in the respiratory control ratio, substrate control ratios of GMOd/GMd and GMOSd/GMd were ∼30-40% lower in old mice (P < 0.05) and were concomitant with increased nitrotyrosine (P < 0.05) and reduced expression of brown adipose markers (P < 0.05). Ageing reduced vascularity (∼50%, P < 0.01), angiogenic capacity (twofold, P < 0.05) and expression of vascular endothelial growth factor (∼50%, P < 0.05) in eWAT. Finally, endothelium-dependent dilation was lower (P < 0.01) in isolated arteries from eWAT arteries of the old mice. Thus, metabolic dysfunction with advancing age occurs in concert with dysfunction in the adipose tissue characterized by both mitochondrial and arterial dysfunction.

Unmet psychosocial needs in haematological cancer: a systematic review.

PURPOSE: Psychosocial need implies a desire or requirement for support that underlies a person's psychological, social and emotional wellbeing. This is not a new concept in the wider cancer literature, yet remains a relatively unexplored area in relation to haematological malignancies. The well-recognised differences between haematological and other types of cancer diagnosis warrant further investigation to try and highlight the potential differences in the needs of this patient group. METHOD: A systematic review of key online databases and psycho-oncology journals was conducted to identify papers that formally assessed unmet psychosocial needs in adults with a diagnosis of haematological cancer. The breadth of methodologies of included studies made a meta-analytical approach unfeasible, therefore studies were analysed using a narrative synthesis approach. RESULTS: Eighteen studies were found to be relevant and a specific focus was placed on those papers that looked solely at participants with a haematological diagnosis. The key areas of need identified were: psychological need, notably fear of recurrence; information needs; and needs relating to both family and healthcare professionals. Fear of recurrence was the most commonly identified psychosocial need within this literature. CONCLUSIONS: The clinical implications of these findings highlight the need for more widespread access to psychological support for haematology patients and for more to be done to tackle patients' fears and concerns throughout the course of their illness. Assessment and identification of unmet needs is an important step enabling the development of clinical services that support and maintain psychological wellbeing through treatment and into survivorship.

Life-long caloric restriction reduces oxidative stress and preserves nitric oxide bioavailability and function in arteries of old mice.

Aging impairs arterial function through oxidative stress and diminished nitric oxide (NO) bioavailability. Life-long caloric restriction (CR) reduces oxidative stress, but its impact on arterial aging is incompletely understood. We tested the hypothesis that life-long CR attenuates key features of arterial aging. Blood pressure, pulse wave velocity (PWV, arterial stiffness), carotid artery wall thickness and endothelium-dependent dilation (EDD; endothelial function) were assessed in young (Y: 5-7 month), old ad libitum (Old AL: 30-31 month) and life-long 40% CR old (30-31 month) B6D2F1 mice. Blood pressure was elevated with aging (P < 0.05) and was blunted by CR (P < 0.05 vs. Old AL). PWV was 27% greater in old vs. young AL-fed mice (P < 0.05), and CR prevented this increase (P < 0.05 vs. Old AL). Carotid wall thickness was greater with age (P < 0.05), and CR reduced this by 30%. CR effects were associated with amelioration of age-related changes in aortic collagen and elastin. Nitrotyrosine, a marker of cellular oxidative stress, and superoxide production were greater in old AL vs. young (P < 0.05) and CR attenuated these increase. Carotid artery EDD was impaired with age (P < 0.05); CR prevented this by enhancing NO and reducing superoxide-dependent suppression of EDD (Both P < 0.05 vs. Old AL). This was associated with a blunted age-related increase in NADPH oxidase activity and p67 expression, with increases in superoxide dismutase (SOD), total SOD, and catalase activities (All P < 0.05 Old CR vs. Old AL). Lastly, CR normalized age-related changes in the critical nutrient-sensing pathways SIRT-1 and mTOR (P < 0.05 vs. Old AL). Our findings demonstrate that CR is an effective strategy for attenuation of arterial aging.

Multilevel analysis of environmental Salmonella prevalences and management practices on 49 broiler breeder farms in four south-eastern States, USA.

A two-part serial survey of 49 broiler breeder farms was conducted in four south-eastern states: Arkansas, Alabama, Georgia and North Carolina. Broiler breeder farms from three to five broiler company complexes in each state were visited on two separate occasions to document management practices and perform environmental sampling for Salmonella prevalence estimation. Salmonella was detected in 88% of the broiler breeder houses that were sampled and was identified on all 49 farms enrolled. Many management characteristics were consistent across the different states and companies. Multilevel analysis was used to evaluate management characteristics as risk factors for Salmonella prevalence and to estimate the proportion of variance residing at the different hierarchical sampling levels. Management characteristics associated with increased Salmonella prevalence included treatment of the flock for any disease, having dusty conditions in the house, having dry conditions under the slats and walking through the house more than one time per day to pick-up dead birds. After adjusting for state as a fixed effect, the percentages of variance in Salmonella prevalence occurring at the complex, farm, visit, house and individual sample levels were 5.2%, 6.8%, 11.8%, 2.8% and 73.4%, respectively. The intraclass correlations for samples collected from the same house; for samples from different houses during the same visit; for samples from different visits to the same farm; and for samples from different farms in the same complex were as follows: 0.27, 0.24, 0.12 and 0.05, respectively.

Comparisons of sperm storage tubule distribution and number in 4 strains of mature broiler breeders and in turkey hens before and after the onset of photostimulation.

The biological basis of sustained fertility in broiler and turkey hens is their capacity to store sperm in the oviductal sperm storage tubules (SST) located in the uterovaginal junction. The objectives of this study were to determine if the numbers of SST varied between 4 strains of broiler breeders and determine the number of SST in the turkey before (less than 9 d of photostimulation) and after (up to 22 d of photostimulation and laying) photostimulation. No statistical differences were observed in SST numbers in the 4 strains of broilers examined or in turkey hens before and after the onset of egg production. The mean numbers of SST for broilers and turkeys were 4,893 and 30,566, respectively. We conclude that any differences between the fertility of the 4 broiler breeder strains examined cannot be explained by differences in SST numbers. However, differences in the duration of fertility between broilers and turkeys are, in part, related to their respective numbers of number of SST. Furthermore, we conclude that turkey SST are morphologically differentiated and functional before the onset of photostimulation and while the oviduct is morphologically undeveloped.

Use of a screening questionnaire for post-traumatic stress disorder (PTSD) on a sample of UK ICU patients.

BACKGROUND: Although rates vary across studies, research in recent years shows that prevalence of post-traumatic stress disorder (PTSD) following intensive care unit (ICU) can be high. Presently no screening tool assessing all three PTSD symptom categories has been validated in ICU patients. The aim of the study was to conduct a preliminary validation of such a measure, the UK- Post-Traumatic Stress Syndrome 14-Questions Inventory (UK-PTSS-14). METHODS: A case series cohort study performed at two ICUs in two UK district general hospitals. The UK-PTSS-14 was administered at three time-points (4-14 days, 2 months and 3 months post-ICU discharge). At time-point three participants also completed the Post-traumatic Stress Diagnostic Scale (PDS) and the Impact of Events Scale (IES). RESULTS: Forty-four patients completed the 3-month follow up. The UK-PTSS-14 was internally reliable at all three time-points (Cronbach's alpha=0.89, 0.86 and 0.84, respectively). Test-retest reliability was highest between time-points two and three (ICC=0.90). Concurrent validity at time-point three was high against the PDS (r=0.86) and the IES (r=0.71). Predictive validity was highest at time-point two (r=0.85 with the PDS and r=0.71 with the IES). Receiver operator characteristic curve analysis suggested the highest levels of sensitivity (86%) and specificity (97%) for diagnosis of PTSD were at time-point two, with an optimum decision threshold of 45 points. CONCLUSION: This preliminary validation study suggests that the UK-PTSS-14 could be reliably used as a screening instrument at 2 months post-discharge from the ICU to identify those patients in need of referral to specialist psychological services.

Expectations and experience of labial reduction: a qualitative study.

OBJECTIVE: To understand women's reasons for undergoing labial reduction surgery, their expectations and experiences. DESIGN: A retrospective qualitative study. SETTING: British National Health Service Hospital. SAMPLE: Six women who had experienced surgery for labial reduction. Method Qualitative study using semi-structured interviews. RESULTS: Results relating to 'Normality and defect', 'Sex lives' and 'The process of accessing surgery' are presented in this study. The women had seen their presurgery genital appearance as 'defective' and sought a 'normal' genital appearance. They thought that their presurgery genital appearance impacted on their sex lives, but their expectations of the effects of surgery on their sex lives were not all fulfilled. Information about labial surgery came from both the popular media and the health services. An emphasis on, for example, physical discomfort rather than appearance may have been used to legitimise a request for surgery. The process of accessing surgery had exposed them to potentially conflicting messages about their genital appearance. CONCLUSIONS: Women presenting for labial reduction may have unrealistic expectations of surgery, but their perceptions and expectations are long-standing and seem to be based on strong cultural norms. The gynaecologist is also meeting those women who have already negotiated the referral process. As demand for this surgery appears to be increasing, further research is needed. These findings may add to the case for the potential value of specialist staff to provide psychosocial interventions within gynaecology services.

Families' views on ward rounds in neonatal units.

OBJECTIVE: To discover parental preferences about visiting during ward rounds. DESIGN: Survey using a short structured interview SETTING AND PARTICIPANTS: Families of babies cared for in a regional neonatal intensive care unit. RESULTS: Eighty six respondents, no refusals. Sixty three had visited during a ward round, and 13 had come in especially for the round. About half had overheard conversations about other babies or thought discussions about their baby had been overheard. Concerns about these experiences were only expressed by respondents who had actually experienced overhearing. Parents and families had little information about the ward round, held diverse views, and expressed different priorities. They described a mixture of concerns about communication, practicalities, and issues of ethics and principle. Confidentiality was a matter of concern for some, but many parents expected some sharing of information between families on the unit. CONCLUSIONS: Units should consider: the information they have for parents about ward rounds; the possibility that consultations may be overheard; the opportunities for parents to communicate with the clinical team.

An exploration of midwives' and obstetricians' knowledge of genetic screening in pregnancy and their perception of appropriate counselling.

OBJECTIVE: to describe levels of knowledge relevant to genetic screening in a sample of midwives and obstetricians in the late 1990s, and to compare these with those found by Smith et al. in London and Wales, and reported in 1994; to describe health professionals' perceptions of an appropriate counselling process relating to genetic screening in comparison with recognised good practice; and to consider links between knowledge and perceptions of the counselling process. DESIGN: a questionnaire study, including responses to a counselling scenario. PARTICIPANTS AND SETTING: responses were obtained from 81 midwives and obstetricians from maternity services in the North West of England. FINDINGS: knowledge about procedures was very good, but knowledge about the probability of genetic abnormality was relatively poor, and respondents overestimated the efficacy and usefulness of tests. These findings were similar to those of Smith et al. (1994), suggesting that they are a good reflection of the national picture. In terms of reported information-giving and counselling practice, there were some respondents who would not check the woman's understanding of her baby's risk of a genetic abnormality, or the risk of a false result. Some respondents would give information based on their own (necessarily limited) experience, rather than national statistics, and give advice based on the choices they themselves would make. These findings reflect earlier, basic research on people's understanding of probabilistic information. IMPLICATIONS: the findings of this study, together with those of earlier work cited, suggest a need for education and training which includes a specific focus on biases in understanding this type of probabilistic information. They also raise the question as to why tests which provide probabilistic data have been introduced without consideration of the known problems in understanding such information.