Two variants of the rat brain sodium-driven chloride bicarbonate exchanger (NCBE): developmental expression and addition of a PDZ motif

Regulation of pH in the central nervous system is critical to normal brain function and response to pathophysiological conditions. Here we identify two novel variants of the sodium-driven chloride bicarbonate exchanger (NCBE) from brain. The developmental expression pattern seen by in situ hybridization for the 90-bp containing insert (insert A) reveals strong expression in spinal cord and brain beginning in embryonic development. High-level expression is seen in cerebellar Purkinje cells and principal cells in hippocampus. The variant missing a 39-bp insert at the 3' end (insert B) encodes a protein in which the deduced carboxyterminal three amino acids are replaced with a unique 21 amino acid stretch terminating in a PDZ motif. rb2NCBE, the PDZ motif-encoding variant, is more highly expressed in astrocytes than is rb1NCBE. Both variants are expressed at similar levels in neurons. Expression varies with age and cell type. The FLAG epitope was fused in-frame at the amino terminus and each variant was expressed using a retroviral vector to study subcellular localization. Both variants were associated with the plasma membrane, but rb2NCBE colocalized with actin filaments to a greater extent, suggesting the PDZ form may interact with the cytoskeleton, whereas rb1NCBE was more often seen in intracellular vesicles. The PDZ motif-containing variant was much more active in pH regulation, with the expected ionic dependence on Na+, HCO3- and Cl- when expressed in 3T3 cells. These results are a first step towards understanding the regulation of expression and activity of this transporter in the brain.     

Entorhinal cortex disruption causes memory deficit in early Alzheimer's disease as shown by PET

Voxel-based mapping of the correlations between cognitive scores and resting-state brain glucose utilization measured by PET has recently emerged as a novel way to reveal in living patients with Alzheimer's disease (AD) the neural systems whose disruption underlies particular neuropsychological, especially mnemonic, deficits. We have now applied this approach using a novel cognitive paradigm designed to selectively assess verbal episodic memory, and show that in early AD disruption of the left entorhinal cortex underlies this memory deficit, consistent with post mortem data showing that this brain area is affected earliest and most severely by tau pathology in AD.     

Accumulation of prion protein in the brain that is not associated with transmissible disease

Prion diseases or transmissible spongiform encephalopathies are characterized histopathologically by the accumulation of prion protein (PrP) ranging from diffuse deposits to amyloid plaques. Moreover, pathologic PrP isoforms (PrP(Sc)) are detected by immunoblot analysis and used both as diagnostic markers of disease and as indicators of the presence of infectivity in tissues. It is not known which forms of PrP are associated with infectivity. To address this question, we performed bioassays using human brain extracts from two cases with phenotypically distinct forms of familial prion disease (Gerstmann-Str?ussler-Scheinker P102L). Both cases had PrP accumulations in the brain, but each had different PrP(Sc) isoforms. Only one of the brains had spongiform degeneration. Tissue from this case transmitted disease efficiently to transgenic mice (Tg PrP101LL), resulting in spongiform encephalopathy. In contrast, inoculation of tissue from the case with no spongiform degeneration resulted in almost complete absence of disease transmission but elicited striking PrP-amyloid deposition in several recipient mouse brains. Brains of these mice failed to transmit any neurological disease on passage, but PrP-amyloid deposition was again observed in the brains of recipient mice. These data suggest the possible isolation of an infectious agent that promotes PrP amyloidogenesis in the absence of a spongiform encephalopathy. Alternatively, the infectious agent may be rendered nonpathogenic by sequestration in amyloid plaques, or PrP amyloid can seed amyloid accumulation in the brain, causing a proteinopathy that is unrelated to prion disease. Formation of PrP amyloid may therefore not necessarily be a reliable marker of transmissible spongiform encephalopathy infectivity.     

Geographical and socioeconomic variation in the prevalence of asthma symptoms in English and Scottish children.

BACKGROUND: There has been controversy over the relation between poverty and asthma in the community. The aim of this analysis was to disentangle geographical and socioeconomic variation in asthma symptoms. METHODS: The analysis is based on parental reports of symptoms from data collected in 1990 and 1991. Children aged 5-11 years from three populations (English representative sample, Scottish representative sample, and an English inner city sample) were included. Of 17 677 eligible children, between 14 490 (82.0%) and 15 562 (88.0%) children were available for analysis according to symptom group. RESULTS: Wheezy symptoms were less prevalent in the Scottish sample than in the English samples and asthma attacks were most prevalent in the English representative sample. Asthma attacks were less prevalent in inner city areas than in the English representative sample (OR 0.79, 95% CI 0.66 to 0.95), but persistent wheeze and other respiratory symptoms were more prevalent (OR 1.95, 95% CI 1.65 to 2.32 and OR 1.67, 95% CI 1.52 to 1.84, respectively). The prevalence of persistent wheeze was higher in children whose father's social class was low and in those living in areas with a high Townsend score (an index of poverty) than in other children (p<0.001). Of the 14 areas with the highest Townsend score, 13 had an OR above 1 and six had an OR significantly higher than the reference area. CONCLUSIONS: Persistent wheeze is more prevalent in poor areas than in less deprived areas. This may indicate that poverty is associated with severe asthma or that a high percentage of persistent asthma symptoms in inner city areas are unrecognised and untreated.     

Motivation to reduce alcohol consumption and subsequent attempts at reduction and changes in consumption in increasing and higher‐risk drinkers in England: a prospective population survey

Aims

To assess how far motivation to reduce alcohol consumption in increasing and higher‐risk drinkers in England predicts self‐reported attempts to reduce alcohol consumption and changes in alcohol intake during the following 6 months.

Methods

This study used self‐reported data from 2928 higher‐risk drinkers in the Alcohol Toolkit Study (ATS): a series of monthly cross‐sectional household surveys of adults aged 16+ years of age in England. Alcohol consumption was measured in an initial survey and in a 6‐month telephone follow‐up interview using the Alcohol Use Disorders Identification Test (AUDIT)‐C questionnaire. Motivation was measured in the initial survey using the Motivation to Reduce Alcohol Consumption (MRAC) scale. Attempts to reduce alcohol consumption during the past 6 months were recorded at follow‐up. Data were analysed using repeated‐measures difference‐in‐differences and logistic regression models.

Results

Participants with higher initial motivation to reduce alcohol consumption were more likely to report that they had made an attempt to reduce consumption at follow‐up [adjusted odds ratio (OR_adj) = 2.39, 95% confidence interval (CI) = 1.75–3.29]. There was an overall reduction in alcohol consumption between initial survey and follow‐up (OR_adj = 0.72, 95% CI = 0.65–0.79), but there was insufficient evidence of an additional effect of motivation to reduce consumption on subsequent changes in alcohol consumption, with the difference‐in‐differences effect instead suggesting an average increase (OR_adj = 1.37, 95% CI = 1.00–1.88).

Conclusions

Increasing and higher‐risk drinkers in England who report greater motivation to reduce their consumption are more likely to report making an attempt to reduce during the next 6 months, but this may not be associated with a reduction in alcohol consumption.     

Exposure to environmental microbiota explains persistent abdominal pain and irritable bowel syndrome after a major flood

Background

After an environmental disaster, the affected community is at increased risk for persistent abdominal pain but mechanisms are unclear. Therefore, our study aimed to determine association between abdominal pain and poor water, sanitation and hygiene (WaSH) practices, and if small intestinal bacterial overgrowth (SIBO) and/or gut dysbiosis explain IBS, impaired quality of life (QOL), anxiety and/or depression after a major flood.

Results

New onset abdominal pain, IBS based on the Rome III criteria, WaSH practices, QOL, anxiety and/or depression, SIBO (hydrogen breath testing) and stools for metagenomic sequencing were assessed in flood victims. Of 211 participants, 37.9% (n = 80) had abdominal pain and 17% (n = 36) with IBS subtyped diarrhea and/or mixed type (n = 27 or 12.8%) being the most common. Poor WaSH practices and impaired quality of life during flood were significantly associated with IBS. Using linear discriminant analysis effect size method, gut dysbiosis was observed in those with anxiety (Bacteroidetes and Proteobacteria, effect size 4.8), abdominal pain (Fusobacteria, Staphylococcus, Megamonas and Plesiomonas, effect size 4.0) and IBS (Plesiomonas and Trabulsiella, effect size 3.0).

Conclusion

Disturbed gut microbiota because of environmentally-derived organisms may explain persistent abdominal pain and IBS after a major environmental disaster in the presence of poor WaSH practices.

     

FOLFCIS Treatment and Genomic Correlates of Response in Advanced Anal Squamous Cell Cancer

Background

Treatment of advanced anal squamous cell cancer (SCC) is usually with the combination of cisplatin and 5-fluorouracil, which is associated with heterogeneous responses across patients and significant toxicity. We examined the safety and efficacy of a modified schedule, FOLFCIS (leucovorin, fluorouracil, and cisplatin), and performed an integrated clinical and genomic analysis of anal SCC.

The influence of genetic counselling in the era of DNA testing on knowledge, reproductive intentions and psychological wellbeing

Subjects of reproductive age at risk of having an affected child with a severe single gene disorder such as Duchenne muscular dystrophy (DMD) or cystic fibrosis (CF) were surveyed to ascertain: their views on genetic counselling and antenatal testing; their knowledge of their risk of having an affected child; and their psychological wellbeing. Questionnaires were posted to 209 individuals at 130 addresses; a 65% response rate was achieved. The majority of those surveyed were under 40 years of age (91%), half of them had received genetic counselling only once and for 47% the first encounter was after the diagnosis of their affected child. Most patients expressed their intention to use prenatal testing. However, less than 50% of those counselled knew their risk of having an affected child. Knowledge of risk was associated with the type of disease in the family (p<0.001) (inheritance of DMD was poorly understood by relevant subjects) and was positively associated with the participant's level of education (p<0.05). We did not detect a significant association between the number of intended children and the risk of having an affected child. In terms of family relations, genetic counselling appears to be beneficial for the nuclear family, the couple and their children, but some counsellees reported a detericration in relations with other relatives. The results indicate that couples at risk of having a child with a severe genetic disorder value the counselling provided, but many of them do not remember important facts in relation to their risk status.     

Overexpression of mitochondrial Hsp70/Hsp75 protects astrocytes against ischemic injury in vitro.

Mitochondrial heat shock protein 70 (mtHsp70/Hsp75/Grp75/mortalin/TRAP-1/PBP74) is an essential mitochondrial chaperone and a member of the heat shock protein 70 (HSP70) family. Although many studies have shown the protective properties of overexpression of the cytosolic inducible member of the HSP70 family, Hsp72, few studies have investigated the protective potential of Hsp75 against ischemic injury. Mitochondria are one of the primary targets of ischemic injury in astrocytes. In this study, we analyzed the effects of Hsp75 overexpression on cellular levels of reactive oxygen species (ROS), mitochondrial membrane potential, ATP levels, and viability during the ischemia-like conditions of oxygen-glucose deprivation (OGD) or glucose deprivation (GD) in primary astrocytic cultures. We show that Hsp75 overexpression decreases ROS production and preserves mitochondrial membrane potential during GD, and preserves ATP levels and cell viability during OGD. These findings indicate that Hsp75 can provide protection against ischemia-like in vitro injury and suggest that it should be further studied as a potential candidate for protection against ischemic injury.     

Designing a statewide agenda for nursing research

This article describes the background, development, and beginning implementation of a nursing research action plan for New York state. Working together over the past two years and building on past collaborative efforts, the New York State Nurses Association (NYSNA) and the Foundation of the NYSNA created an agenda for nursing research in New York state. The agenda contains five major goals, subgoals, an action plan for achieving these goals, and desired outcomes. Several implementation activities, including initiation of Web-based activities, strategies to bring research to the clinical nurse, and the development of a proposal to create evidence-based practice teams, are under way. The process of agenda development may serve as a model for other associations or organizations.