Upregulation of the Coagulation Factor VII Gene during Glucose Deprivation Is Mediated by Activating Transcription Factor 4

Background: Constitutive production of blood coagulation proteins by hepatocytes is necessary for hemostasis. Stressful conditions trigger adaptive cellular responses and delay processing of most proteins, potentially affecting plasma levels of proteins secreted exclusively by hepatocytes. We examined the effect of glucose deprivation on expression of coagulation proteins by the human hepatoma cell line, HepG2. Methodology/Principal Findings: Expression of coagulation factor VII, which is required for initiation of blood coagulation, was elevated by glucose deprivation, while expression of other coagulation proteins decreased. Realtime PCR and ELISA demonstrated that the relative percentage expression +/− SD of steady-state F7 mRNA and secreted factor VII antigen were significantly increased (from 100+/−15% to 188+/−27% and 100+/−8.8% to 176.3+/−17.3% respectively, p<0.001) at 24 hr of treatment. The integrated stress response was induced, as indicated by upregulation of transcription factor ATF4 and of additional stress-responsive genes. Small interfering RNAs directed against ATF4 potently reduced basal F7 expression, and prevented F7 upregulation by glucose deprivation. The response of the endogenous F7 gene was replicated in reporter gene assays, which further indicated that ATF4 effects were mediated via interaction with an amino acid response element in the F7 promoter. Conclusions/Significance: Our data indicated that glucose deprivation enhanced F7 expression in a mechanism reliant on prior ATF4 upregulation primarily due to increased transcription from the ATF4 gene. Of five coagulation protein genes examined, only F7 was upregulated, suggesting that its functions may be important in a systemic response to glucose deprivation stress

Privacy guidelines for telemedicine developers

This presentation was given at the Med-e-tel 2005 Conference in Luxembourg on April 8th, 2005.Ethical Issues in eHealth: This presentation provides an analysis of privacy for telemedicine based on common international privacy practices and a recently published privacy framework for information systems development. Some practical privacy guidelines are ultimately provided for telemedicine developers.ISOL Research Centre, Waterford Institute of Technology, Waterford, Irelan

Myosin Va Plays a Role in Nitrergic Smooth Muscle Relaxation in Gastric Fundus and Corpora Cavernosa of Penis

The intracellular motor protein myosin Va is involved in nitrergic neurotransmission possibly by trafficking of neuronal nitric oxide synthase (nNOS) within the nerve terminals. In this study, we examined the role of myosin Va in the stomach and penis, proto-typical smooth muscle organs in which nitric oxide (NO) mediated relaxation is critical for function. We used confocal microscopy and co-immunoprecipitation of tissue from the gastric fundus (GF) and penile corpus cavernosum (CCP) to localize myosin Va with nNOS and demonstrate their molecular interaction. We utilized in vitro mechanical studies to test whether smooth muscle relaxations during nitrergic neuromuscular neurotransmission is altered in DBA (dilute, brown, non-agouti) mice which lack functional myosin Va. Myosin Va was localized in nNOS-positive nerve terminals and was co-immunoprecipitated with nNOS in both GF and CCP. In comparison to C57BL/6J wild type (WT) mice, electrical field stimulation (EFS) of precontracted smooth muscles of GF and CCP from DBA animals showed significant impairment of nitrergic relaxation. An NO donor, Sodium nitroprusside (SNP), caused comparable levels of relaxation in smooth muscles of WT and DBA mice. These normal postjunctional responses to SNP in DBA tissues suggest that impairment of smooth muscle relaxation resulted from inhibition of NO synthesis in prejunctional nerve terminals. Our results suggest that normal physiological processes of relaxation of gastric and cavernosal smooth muscles that facilitate food accommodation and penile erection, respectively, may be disrupted under conditions of myosin Va deficiency, resulting in complications like gastroparesis and erectile dysfunction

Incidence of and risks associated with Giardia infections in herds on dairy farms in the New York City Watershed

<p>Abstract</p> <p>Background</p> <p>The primary aims of this study were to determine the incidence of <it>Giardia </it>infections in dairy herds on farms in the New York City Watershed region and to evaluate risk factors associated with infections. Because co-infections of <it>Giardia </it>and <it>Cryptosporidium </it>spp. are common in this population, we also evaluated the effect of herd infection status on <it>Giardia </it>infections.</p> <p>Methods</p> <p>Farms were grouped into three cohorts based on their prior infection status with <it>Giardia </it>and/or <it>Cryptosporidium </it>spp. The sampling plan included collecting fecal samples from all calves below 30 days of age and proportional sampling of calves, young stock, and adults. A total of 10,672 fecal samples were collected and analyzed for the presence of <it>Giardia </it>cysts using zinc sulfate flotation. Herds enrolled in the study were sampled seasonally for a study period of two years. The probability of shedding cysts past a certain age and the factors that influenced the likelihood of shedding were evaluated using survival analysis. Linear regression was used to evaluate factors that were associated with the intensity of shedding.</p> <p>Results</p> <p>The majority of <it>Giardia </it>infections occurred in calves within their first 180 days of age, with the most number of calves shedding <it>Giardia </it>cysts between 11 and 20 days of age. The incidence of shedding of <it>Giardia </it>cysts ranged from 0.0004 per animal day for cattle in the low risk cohort to 0.0011 per animal day for cattle in the high risk cohort. The likelihood of shedding was influenced by the prior infection status of the herd and the season of collection. Infected animals shed on average 9,658 cysts/gram and the intensity of shedding <it>Giardia </it>cysts varied significantly with the age (p < 0.0001) and the season of collection (p = 0.0151 for Spring).</p> <p>Conclusion</p> <p><it>Giardia </it>infections are common in dairy herds in the New York City watershed, particularly in calves less than 6 months of age. Seasonality may be an important factor in the perpetuation of infections based on changes in management practices corresponding to weather patterns of a particular season. A dairy herd's prior infection status with <it>Cryptosporidium </it>influences the likelihood of infection with <it>Giardia</it>.</p

Deletion of Irs2 causes reduced kidney size in mice: role for inhibition of GSK3β?

<p>Abstract</p> <p>Background</p> <p>Male <it>Irs2</it>^-/-mice develop fatal type 2 diabetes at 13-14 weeks. Defects in neuronal proliferation, pituitary development and photoreceptor cell survival manifest in <it>Irs2</it>^-/-mice. We identify retarded renal growth in male and female <it>Irs2</it>^-/-mice, independent of diabetes.</p> <p>Results</p> <p>Kidney size and kidney:body weight ratio were reduced by approximately 20% in <it>Irs2</it>^-/-mice at postnatal day 5 and was maintained in maturity. Reduced glomerular number but similar glomerular density was detected in <it>Irs2</it>^-/-kidney compared to wild-type, suggesting intact global kidney structure. Analysis of insulin signalling revealed renal-specific upregulation of PKBβ/Akt2, hyperphosphorylation of GSK3β and concomitant accumulation of β-catenin in <it>Irs2</it>^-/-kidney. Despite this, no significant upregulation of β-catenin targets was detected. Kidney-specific increases in Yes-associated protein (YAP), a key driver of organ size were also detected in the absence of <it>Irs2</it>. YAP phosphorylation on its inhibitory site Ser127 was also increased, with no change in the levels of YAP-regulated genes, suggesting that overall YAP activity was not increased in <it>Irs2</it>^-/-kidney.</p> <p>Conclusions</p> <p>In summary, deletion of <it>Irs2 </it>causes reduced kidney size early in mouse development. Compensatory mechanisms such as increased β-catenin and YAP levels failed to overcome this developmental defect. These data point to <it>Irs2 </it>as an important novel mediator of kidney size.</p

Analysis of dietary patterns and cross-sectional and longitudinal associations with hypertension, high BMI and type 2 diabetes in Peru

OBJECTIVE: To determine if specific dietary patterns are associated with risk of hypertension, type 2 diabetes mellitus (T2DM) and high BMI in four sites in Peru. DESIGN: We analysed dietary patterns from a cohort of Peruvian adults in four geographical settings using latent class analysis. Associations with prevalence and incidence of hypertension, T2DM and high BMI were assessed using Poisson regression and generalised linear models, adjusted for potential confounders. SETTING: Four sites in Peru varying in degree of urbanisation. PARTICIPANTS: Adults aged ≥35 years (n 3280). RESULTS: We identified four distinct dietary patterns corresponding to different stages of the Peruvian nutrition transition, reflected by the foods frequently consumed in each pattern. Participants consuming the 'stage 3' diet, characterised by high proportional consumption of processed foods, animal products and low consumption of vegetables, mostly consumed in the semi-urban setting, showed the highest prevalence of all health outcomes (hypertension 32·1 %; T2DM 10·7 %; high BMI 75·1 %). Those with a more traditional 'stage 1' diet characterised by potato and vegetables, mostly consumed in the rural setting, had lower prevalence of hypertension (prevalence ratio; 95 CI: 0·57; 0·43, 0·75), T2DM (0·36; 0·16, 0·86) and high BMI (0·55; 0·48, 0·63) compared with the 'stage 3' diet. Incidence of hypertension was highest among individuals consuming the 'stage 3' diet (63·75 per 1000 person-years; 95 % CI 52·40, 77·55). CONCLUSIONS: The study found more traditional diets were associated with a lower prevalence of three common chronic diseases, while prevalence of these diseases was higher with a diet high in processed foods and low in vegetables

Auditory perception in individuals with Friedreich’s Ataxia

INTRODUCTION: Friedreich's ataxia (FRDA) is an inherited ataxia with a range of progressive features including axonal degeneration of sensory nerves. The aim of this study was to investigate auditory perception in affected individuals. METHODS: Fourteen subjects with genetically defined FRDA participated. Two control groups, one consisting of healthy, normally hearing individuals and another comprised of subjects with sensorineural hearing loss, were also assessed. Auditory processing was evaluated using structured tasks designed to reveal the listeners' ability to perceive temporal and spectral cues. Findings were then correlated with open-set speech understanding. RESULTS: Nine of 14 individuals with FRDA showed evidence of auditory processing disorder. Gap and amplitude modulation detection levels in these subjects were significantly elevated, indicating impaired encoding of rapid signal changes. Electrophysiologic findings (auditory brainstem response, ABR) also reflected disrupted neural activity. Speech understanding was significantly affected in these listeners and the degree of disruption was related to temporal processing ability. Speech analyses indicated that timing cues (notably consonant voice onset time and vowel duration) were most affected. CONCLUSION: The results suggest that auditory pathway abnormality is a relatively common consequence of FRDA. Regular auditory evaluation should therefore be part of the management regime for all affected individuals. This assessment should include both ABR testing, which can provide insights into the degree to which auditory neural activity is disrupted, and some functional measure of hearing capacity such as speech perception assessment, which can quantify the disorder and provide a basis for interventio

Heteromeric TRPV4/TRPC1 channels mediate calcium-sensing receptor-induced relaxations and nitric oxide production in mesenteric arteries: comparative study using wild-type and TRPC1−/- mice

We have previously provided pharmacological evidence that stimulation of calcium-sensing receptors (CaSR) induces endothelium-dependent relaxations of rabbit mesenteric arteries through activation of heteromeric TRPV4/TRPC1 channels and nitric oxide (NO) production. The present study further investigates the role of heteromeric TRPV4/TRPC1 channels in these CaSR-induced vascular responses by comparing responses in mesenteric arteries from wild-type (WT) and TRPC1-/- mice. In WT mice, stimulation of CaSR induced endothelium-dependent relaxations of pre-contracted tone and NO generation in endothelial cells (ECs), which were inhibited by the TRPV4 channel blocker RN1734 and the TRPC1 blocking antibody T1E3. In addition, TRPV4 and TRPC1 proteins were colocalised at, or close to, the plasma membrane of endothelial cells (ECs) from WT mice. In contrast, in TRPC1-/- mice, CaSR-mediated vasorelaxations and NO generation were greatly reduced, unaffected by T1E3, but blocked by RN1734. In addition, the TRPV4 agonist GSK1016790A (GSK) induced endothelium-dependent vasorelaxations which were blocked by RN1734 and T1E3 in WT mice, but only by RN1734 in TRPC1-/- mice. Moreover, GSK activated cation channel activity with a 6pS conductance in WT ECs but with a 52 pS conductance in TRPC1-/- ECs. These results indicate that stimulation of CaSR activates heteromeric TRPV4/TRPC1 channels and NO production in ECs, which are responsible for endothelium-dependent vasorelaxations. This study also suggests that heteromeric TRPV4-TRPC1 channels may form the predominant TRPV4-containing channels in mouse mesenteric artery ECs. Together, our data further implicates CaSR-induced pathways and heteromeric TRPV4/TRPC1 channels in the regulation of vascular tone

Middle Holocene environmental change and archaeology in coastal wetlands: further implications for our understanding of the history of Taxus woodland

A radiocarbon-dated multi-proxy palaeoenvironmental record from Beckton in the Lower Thames Valley, Southern England, has permitted a detailed reconstruction of human activities and environmental change during the middle-Holocene. Peat accumulation occurred over river terrace gravels from ca. 7200 to 6600 until at least 3450–3240 cal. BP, and in the later period a trackway and platform structure provide unequivocal evidence for human exploitation of the floodplain environment during the Bronze Age. The site is unique in offering the first certain evidence of the utilisation of Taxus in the construction of a wooden prehistoric platform. Across north-west Europe during the middle-Holocene, the colonisation of Taxus on peat is well documented; at Beckton, it occurred between ca. 5220–4940 and 4410–4220 cal. BP. This research provides important insights into the former distribution of Taxus, and reasons for its expansion and decline during the Holocene, which has relevance to present-day concerns over the conservation and management of Taxus woodland. Abandonment of the site occurred in response to environmental change to wetter conditions. The study employed multi-proxy analyses, including pollen, plant and wood macrofossils, and uniquely Coleoptera; Coleopteran analysis has significant potential to enhance understandings of environmental change and human–environment interactions in coastal wetland research