High-frequency reactivation of X-linked genes in chinese hamster × human hybrid cells

Three genes on the human inactive X chromosome retained in the Chinese hamster x human hybrid cell line X8/6T2 have been reactivated using the demethylating agent, 5-azacytidine (5-aza-CR). Pulse-labeling and histochemical methods permitted detection and measurement of reactivation rates of the hypoxanthine phosphoribosyltransferase (Hpt) and glucose-6-phosphate dehydrogenase (G6pd) genes within 48 h of treatment. About 50% of the cells became active for these genes, which represents a reactivation rate some 30-fold greater than previously reported in similar systems. The phosphoglycerate kinase (Pgk) gene was not reactivated as frequently as the Hptor G6pdgenes. Segregation analysis of progeny of treated cells showed that enzyme-positive and enzyme-negative cells were produced in proportions supporting the notion that 5-aza-CR causes demethylation by replicative loss and that demethylation leads to reactivation.     

Clinical impact of a 2-week psychotropic medication washout in unipolar depressed inpatients

BACKGROUND: Short-term discontinuation of psychiatric medications is required in many types of research studies. Yet there are few studies of the clinical impact of psychotropic discontinuation. We studied the impact of a short-term medication washout in unipolar depressed patients consecutively admitted to hospital for neuroimaging and cerebrospinal fluid (CSF) studies. METHOD: Patients (n=51) with unipolar depression who were taking antidepressant or mood stabilizing medication at or within 1 week of admission, and who had not been responding satisfactorily, were assessed for severity of psychopathology within 1 week of hospital admission and 41 of the group were reassessed following an approximately 2-week medication washout. RESULTS: On average, patients remained stable during the washout or improved on clinical measures. No serious adverse clinical changes were observed. LIMITATIONS: Potential sample bias, small sample size. CONCLUSION: The results suggest that similar studies can be conducted without causing undue worsening of symptoms. The benefit of medication washout may be related to the fact that many of the patients had been responding poorly to the medication they were taking. There is a need for further research on the effects of medication washout, for example in outpatients or those who are responding well to treatment, but have intolerable side-effects.     

Infection of immunodeficient horses with Sarcocystis neurona does not result in neurologic disease

Equine protozoal myeloencephalitis is a progressive neurologic disease of horses most commonly caused by infection with the apicomplexan parasite Sarcocystis neurona. Factors affecting neuroinvasion and neurovirulence have not been determined. We investigated the pathogenesis of infection with S. neurona in horses with severe combined immune deficiency (SCID). Two immunocompetent (IC) Arabian horses and two Arabian horses with SCID were infected orally with 5 x 10(5) sporocysts of S. neurona. Four IC horses and one SCID horse were infected intravenously (i.v.) with 5 x 10(8) merozoites of the WSU-1 isolate of S. neurona. Despite prolonged parasitemia and persistent infection of visceral tissues (skeletal muscle, cardiac muscle, lung, liver, and spleen) as demonstrated by PCR and culture, SCID horses did not develop neurologic signs after oral or i.v. infection. S. neurona was undetectable in the neuronal tissues of SCID horses by either PCR, immunohistochemistry, or culture. In contrast, although parasitemia was undetectable in orally infected IC horses and of only short duration in i.v. infected IC horses, four of six IC horses developed neurologic signs. S. neurona was detectable by PCR and/or culture of neural tissue but not visceral tissue of IC horses with neurologic disease. Infected SCID horses are unable to clear S. neurona from visceral tissues, but the infection does not result in neurologic signs; in contrast, IC horses rapidly control parasitemia and infection of visceral tissues but frequently experience neuroinvasion and exhibit clinical signs of neurologic disease.     

Magnesium intake and serum C-reactive protein levels in children.

The objective of this study was to determine whether magnesium consumption is associated with inflammation (C-reactive protein [CRP]) in children. The study was an analysis of child (age 6-17 years) participants in the cross-sectional, nationally representative National Health and Nutrition Examination Survey (NHANES). Children consuming less than 75% of RDA were 1.94 times more likely (p < 0.05) to have elevated serum CRP levels than children consuming above the RDA. In adjusted analyses controlling for demographics, cardiovascular risk factors, and BMI, children with consumption of less than 75% RDA were 58% more likely to have elevated CRP (OR 1.58, 95% CI 1.07-infinity). Children with intakes below the RDA are more likely to have elevated CRP levels.     

Retrograde migration of glove powder in the human female genital tract

BACKGROUND: This study in humans was undertaken to evaluate earlier results from animal research showing a retrograde migration of glove powder from the vagina into the intra-abdominal cavity. METHODS: One study group was gynaecologically examined with powdered gloves the day before an abdominal hysterectomy and another group 4 days pre-operatively. There were two control groups similarly examined with powder-free gloves. Cell smears were taken from the peritoneal fluid and during the operation further smears were taken from the Fallopian tubes, uterine cavity and cervical canal. RESULTS: Statistically significant differences were found for large starch particles at all locations between the study and control groups examined 1 day pre-operatively. Considering small starch particles, there were significant differences in cervix (P < 0.001), uterus (P < 0.01) and the Fallopian tubes (P < 0.01). The combined results also show significant differences between both large and small starch particles in cervix, uterus and the Fallopian tubes. There were also differences between the study and control groups examined 4 days pre-operatively, but these were not statistically significant except for small and large starch particles in uterus (P < 0.01, P < 0.05) and cervix (P < 0.05, P < 0.05). CONCLUSIONS: This study has pointed out a retrograde migration of starch also in humans after a gynaecological examination with powdered gloves. Consequently, powder or any other potentially harmful substance that can migrate from the vagina should be avoided.     

Emergence of influenza A H1N2 reassortant viruses in the human population during 2001

A recombinant vaccinia virus encoding rotavirus protein NSP3 driven by an internal ribosome entry site (IRES) from the encephalomyocarditis (EMC) virus was able to abate protein synthesis in BSC1 cells by 25-fold, with as much as 30% of the remaining protein synthesis being NSP3. Hence NSP3 shuts off host cell protein synthesis down to the level seen during rotavirus infection but is unable to prevent translation from EMC IRES-driven genes. This effect was abolished by deletions in the eIF4G-binding (aa 274-313) and the dimerization (aa 150-206) but not the viral mRNA-binding (aa 83-149) domains, supporting that NSP3 functions in vivo as a dimer. Binding of eIF4G by NSP3 has been implicated in interfering with mRNA 5'-3' circularization, hence such circularization is essential for translation in mammalian cells.     

Intracytoplasmic lumina--a useful diagnostic feature of adenocarcinomas

The monoclonal antibody NCRC-11, which has epithelial membrane antigen (EMA)-like immunoreactivity, was used to identify intracytoplasmic lumina in a series of 105 adenocarcinomas from various sites and in 283 breast carcinomas; 55% of the non-breast carcinomas and all breast carcinomas except one of spindle cell type contained intracytoplasmic lumina. The highest frequency (16.4% of tumour cells) was found in invasive lobular carcinoma of the breast. The use of antibodies with EMA reactivity is advocated in the routine investigation of metastatic and undifferentiated tumours.     

A pilot study of the effect of mechanical ventilation and heat exchange on house-dust mites and Der p 1 in New Zealand homes

We have examined the effect of reducing relative humidity (RH), with inbuilt mechanical ventilation and heat-exchange (MVHE) units, on house-dust- mite (HDM) counts and allergen levels, in a pilot study of 10 Wellington dwellings. Recent international prevalence studies in adults and children have confirmed a high prevalence of asthma in New Zealand, Sensitivity to HDM is common among the general population, and HDM is the major allergen associated with asthma. Recent studies of allergen levels have confirmed high concentrations of Der p 1 in the domestic environment. While humidity was significantly reduced in those dwellings fitted with ventilation units, no systematic effect on mites or Der p 1 was observed during the study period. When the reductions in humidity were examined in the context of the time spent below the critical equilibrium humidity (CEH), the intervention led to RH values below the CEH for only 39% of the total of 24-h periods for which measurements were made. Reducing RH by means of MVHE in New Zealand domestic dwellings does not lower humidity sufficiently, or long enough, to have any measurable effect on HDM populations.