Role of capsid sequence and immature nucleocapsid proteins p9 and p15 in Human Immunodeficiency Virus type 1 genomic RNA dimerization

HIV-1 genomic RNA (gRNA) dimerization is important for viral infectivity and is regulated by proteolytic processing of the Gag precursor protein (Pr55gag) under the direction of the viral protease. The processing occurs in successive steps and, to date, the step associated with formation of a wild-type (WT) level of gRNA dimers has not been identified. The primary cleavage divides Pr55gag into two proteins. The C-terminal polypeptide is termed NCp15 (NCp7-p1-p6) because it contains the nucleocapsid protein (NC), a key determinant of gRNA dimerization and packaging. To examine the importance of precursor polypeptides NCp15 and NCp9 (NCp7-p1), we introduced mutations that prevented the proteolytic cleavages responsible for the appearance of NCp9 or NCp7. Using native Northern blot analysis, we show that gRNA dimerization was impaired when both the secondary (p1-p6) and tertiary (p7-p1) cleavage sites of NCp15 were abolished, but unaffected when only one or the other site was abolished. Though processing to NCp9 therefore suffices for a WT level of gRNA dimerization, we also show that preventing cleavage at the p7-p1 site abolished HIV-1 replication. To identify the minimum level of protease activity compatible with a WT level of gRNA dimers, we introduced mutations Thr26Ser and Ala28Ser in the viral protease to partially inactivate it, and we prepared composite HIV-1 resulting from the cotransfection of various ratios of WT and protease-inactive proviral DNAs. The results reveal that a 30% processing of Pr55gag into mature capsid proteins (CA/CA-p2) yielded a WT level of gRNA dimers, while a 10% Pr55gag processing hardly increased gRNA dimerization above the level seen in protease-inactive virions. We found that full gRNA dimerization required less than 50% WT NC in complementation asssays. Finally, we show that if we destroy alpha helix 1 of the capsid protein (CA), gRNA dimerization is impaired to the same extent as when the viral protease is inactivated. Cotransfection studies show that this CA mutation, in contrast to the NC-disabling mutations, has a dominant negative effect on HIV-1 RNA dimerization, viral core formation, and viral replication. This represents the first evidence that a capsid mutation can affect HIV-1 RNA dimerization.     

Pre‐medication and renal pre‐conditioning: a role for alprazolam,atropine, morphine and promethazine

Four pre‐medication drugs are used to relieve pain, allay anxiety, reduce secretion and enhance hypnosis, were evaluated for their effects on ischemia reperfusion (I/R) injury which is one of the major complications of vascular and transplantation surgery. Right kidney was removed from female rats (210–250 g) 3 weeks before surgical procedure. Different doses of morphine (0.5, 2 and 5 mg/kg), promethazine (1, 2 and 5 mg/kg), atropine (0.1, 0.3 and 0.5 mg/kg) and alprazolam (0.08, 0.32 and 0.64 mg/kg) were administered subcutaneously 30 min before left renal artery occlusion and 6 h reperfusion. Left kidneys were processed for histological evaluations. Creatinine and BUN were measured in serum samples. Morphine, promethazine, atropine and alprazolam at all evaluated doses significantly decreased serum creatinine and BUN levels and histopathological scores. The effects of promethazine (1 mg/kg) and all doses of alprazolam were more potent than other pre‐medication drugs and doses. This study suggested a protective effect of these pre‐medication drugs on I/R injury. Although obvious studies are required, these findings may lead to effective therapies against I/R injury.     

Does electronic clinical microbiology results reporting influence medical decision making: a pre- and post-interview study of medical specialists

Background  

Clinicians view the accuracy of test results and the turnaround time as the two most important service aspects of the clinical microbiology laboratory. Because of the time needed for the culturing of infectious agents, final hardcopy culture results will often be available too late to have a significant impact on early antimicrobial therapy decisions, vital in infectious disease management. The clinical microbiologist therefore reports to the clinician clinically relevant preliminary results at any moment during the diagnostic process, mostly by telephone. Telephone reporting is error prone, however. Electronic reporting of culture results instead of reporting on paper may shorten the turnaround time and may ensure correct communication of results. The purpose of this study was to assess the impact of the implementation of electronic reporting of final microbiology results on medical decision making.     

Material deprivation and health-related dysfunction in older Dutch people: findings from the SMILE study.

BACKGROUND: Even in generally wealthy Western countries material deprivation and poverty are not uncommon. There is, however, little data on the prevalence of material deprivation and its associations with health-related dysfunction in older people. METHODS: Cross-sectional data from the SMILE study were used to examine the prevalence of material deprivation and the associations between material deprivation and health-related dysfunction in persons aged 55 years and older (n > 4000). Material deprivation was measured with a comprehensive questionnaire assessing seven subdomains referring to current and anticipated financial problems and poverty in childhood. Health-related dysfunction was measured using the SF36-based physical and mental components. In addition, self-reported heart disease was examined as an indicator of health-related dysfunction as well. RESULTS: Almost 29% of subjects experienced at least one financial problem. Those reporting material deprivation had more than twice the risk of physical (OR = 2.22; 95% CI: 1.72-2.86) and mental (OR = 2.34; 95% CI: 1.84-2.97) dysfunction compared with non-deprived persons. A slightly weaker association was found when self-reported heart disease was used as an outcome variable (OR = 1.74; 95% CI: 1.40-2.15). Although odds ratios were generally higher for diseased older persons, no significant interaction effect between chronic disease and material deprivation subscales was found. CONCLUSIONS: Material deprivation in the Netherlands is not uncommon and is strongly related to both mental and physical dysfunction, and therefore needs further attention in public health policy. Longitudinal research is necessary to clarify the causal nature of our results and to develop appropriate interventions.     

Socioeconomic profile and nutritional status of children in rubber smallholdings

This paper will present the socioeconomic profile and nutritional status of children aged 1-6 years in the rubber smallholdings of Peninsula Malaysia. A total of 323 households were involved in this study. The sociodemographic data were obtained through interviews with heads of households using a set of questionnaires. Anthropometric measurements were taken from 506 children aged 1-6 years from these households. The weight and height of the children were compared with the reference values of the National Center for Health Statistics (NCHS) and the nutritional status was classified based on the recommendations of WHO. The average age of the fathers was 39.9+/-8.6 years and 34.4+/-7.0 years for the mothers. The mean household size was 6.67+/-2.27. The majority (49.7%) of the heads of households received 4-6 years of formal education and 7.9% received no formal education. Based on the monthly per capita income, 24.0% were found to be in the hardcore poor category, 38.3% fall into the poor category and 37.7% in the above poverty income group. The prevalence of stunting and underweight among children between the ages of 1-6 years were highest among children from the hardcore poor, followed by the poor category and above the poverty line income group. Wasting was present in all income groups, with a prevalence of 4.2% found among the hardcore poor, 9.4% among the poor group and 8.4% in the above poverty income group. The Pearson Product Moment Correlation showed significant relationships between household total income and height-for-age (r = 0.131, P = 0.05) and weight-for-age (r = 0.127, P = 0.05). There were also significant correlations between monthly per capita income with height-for-age (r = 0.16, P < 0.01) and weight-for-age (r = 0.13, P < 0.05). The acreage of land utilised was correlated with height-for-age (r = 0.11, P < 0.05), weight-for-age (r = 0.17, P < 0.05) and weight-for-height (r = 0.16, P < 0.05). However, stepwise multiple regression analysis indicated that the predictor of height-for-age was monthly per capita income (R2 = 0.03, P < 0.01) and acreage of land utilised was a predictor for weight-for-age (R2 = 0.03, P < 0.01) and weight-for-height (R2 = 0.01, P < 0.01). Because income and acreage of land utilised have been shown to be associated with nutritional status, it is recommended that intervention programs that focus on generation of income and diversification of land utilisation should be undertaken. A multidiscipline approach involving the family, community and government agencies should be applied to any type of intervention program.     

Genetic effects on age-dependent onset and islet cell autoantibody markers in type 1 diabetes

Age-dependent associations between type 1 diabetes risk genes HLA, INS VNTR, and CTLA-4 and autoantibodies to GAD65 (GADAs), ICA512/IA-2, insulin, and islet cells were determined by logistic regression analysis in 971 incident patients with type 1 diabetes and 702 control subjects aged 0-34 years. GADAs were associated with HLA-DQ2 in young but not in older patients (P = 0.009). Autoantibodies to insulin were negatively associated with age (P < 0.0001) but positively associated with DQ8 (P = 0.03) and with INS VNTR (P = 0.04), supporting possible immune tolerance induction. ICA512/IA-2 were negatively associated with age (P < 0.0001) and with DQ2 (P < 0.0001) but positively associated with DQ8 (P = 0.04). Males were more likely than females to be negative for GADA (P < 0.0001), autoantibodies to islet cells (P = 0.04), and all four autoantibody markers (P = 0.004). The CTLA-4 3' end microsatellite marker was not associated with any of the autoantibodies. We conclude that age and genetic factors such as HLA-DQ and INS VNTR need to be combined with islet autoantibody markers when evaluating the risk for type 1 diabetes development.