Fluorescence in situ hybridization mapping of translocations and deletions involving the short arm of human chromosome 12 in malignant hematologic diseases

Translocations and deletions of the short arm of chromosome 12 [t(12p) and del(12p)] are common recurring abnormalities in a broad spectrum of hematologic malignant diseases. We studied 20 patients and one cell line whose cells contained 12p13 translocations and/or 12p deletions using fluorescence in situ hybridization (FISH) with phage, plasmid, and cosmid probes that we previously mapped and ordered on 12p12-13. FISH analysis showed that the 12p13 translocation breakpoints were clustered between two cosmids, D12S133 and D12S142, in 11 of 12 patients and in one cell line. FISH analysis of 11 patients with deletions demonstrated that the deletions were interstitial rather than terminal and that the distal part of 12p12, including the GDI-D4 gene and D12S54 marker, was deleted in all 11 patients. Moreover, FISH analysis showed that cells from 3 of these patients contained both a del(12p) and a 12p13 translocation and that the affected regions of these rearrangements appeared to overlap. We identified three yeast artificial chromosome (YAC) clones that span all the 12p13 translocation breakpoints mapped between D12S133 and D12S142. They have inserts of human DNA between 1.39 and 1.67 Mb. Because the region between D12S133 and D12S142 also represents the telomeric border of the smallest commonly deleted region of 12p, we also studied patients with a del(12p) using these YACs. The smallest YAC, 964c10, was deleted in 8 of 9 patients studied. In the other patient, the YAC labeled the del(12p) chromosome more weakly than the normal chromosome 12, suggesting that a part of the YAC was deleted. Thus, most 12p13 translocation breakpoints were clustered within the sequences contained in the 1.39 Mb YAC and this YAC appears to include the telomeric border of the smallest commonly deleted region. Whether the same gene is involved in both the translocations and deletions is presently unknown.     

Improvement in glucose tolerance after one year of follow-up in a Hindu community in Africa

Oral glucose tolerance was studied following a 75 g glucose load in 108 (82.4%) of 131 male and 110 (79.1%) of 139 female members of a Hindu subcommunity aged 15 years and over in Dar es Salaam. One year later, the glucose tolerance tests were repeated in 93 (86.1%) and 93 (84.5%) of the 108 male, and 110 of the female subjects, respectively. In the first survey, 25 (26.9%) of the 93 male and 24 (25.8%) of the 93 female subjects had impaired glucose tolerance (IGT), 6 (6.4%) and 15 (16.1%), respectively, had diabetes mellitus; and 62 (66.7%) and 54 (58.1%), respectively, had normal glucose tolerance. In the repeat survey, of the 93 male and 93 female subjects, 8 (8.6%) and 7 (7.5%) had IGT, 4 (4.3%) and 10 (10.8%) had diabetes; and 81 (87.1%) and 76 (81.7%) were normal, respectively. Of the 21 subjects diagnosed as having diabetes in the first survey, 13 (61.9%) continued to show diabetic glucose tolerance; 4 (19%) IGT and 4 (19%) had normal glucose tolerance with no gender difference. One (1.6%) of the 62 male subjects and none of the 54 females with normal glucose tolerance in the first survey progressed to IGT, while the remainder retained normal glucose tolerance. Diabetes and IGT rates in both surveys were higher for the older than the younger persons. A significant fall in mean fasting and post-75 g blood glucose levels, and in mean systolic and diastolic pressure levels was observed between the first and second surveys in both genders. There was, however, no significant difference in body mass index (BMI), serum total cholesterol and triglyceride levels between surveys, suggesting that major dietary changes had not taken place. Male subjects who showed persistent IGT had significantly lower mean level of body mass index (kg/m²) than subjects who reverted to normal, whilst for the whole group those who had persistent IGT were older. It is tempting to speculate that these changes were due to community action. However, in view of the lack of change in weight and lipids and similar results in other communities in Tanzania when retested at 1 week, further studies are needed to establish the significance of the findings.     

Two pathways of exocytosis of cytoplasmic granule contents and target cell killing by cytokine-induced CD3+ CD56+ killer cells

Cytokine-induced killer (CIK) cells are non-major histocompatibility complex-restricted cytotoxic cells generated by incubation of peripheral blood lymphocytes with anti-CD3 monoclonal antibody (MoAb), interleukin-2 (IL-2), IL-1, and interferon-gamma. Cells with the greatest effector function in CIK cultures coexpress CD3 and CD56 surface molecules. CIK cell cytotoxicity can be blocked by MoAbs directed against the cell surface protein leukocyte function associated antigen-1 but not by anti-CD3 MoAbs. CIK cells undergo release of cytoplasmic cytotoxic granule contents to the extracellular space upon stimulation with anti-CD3 MoAbs or susceptible target cells. Maximal granule release was observed from the CD3+ CD56+ subset of effector cells. The cytoplasmic granule contents are lytic to target cells. Treatment of the effector cells with a cell-permeable analog of cyclic adenosine monophosphate (cAMP) inhibited anti-CD3 MoAb and target cell- induced degranulation and cytotoxicity of CIK cells. The immunosuppressive drugs cyclosporin (CsA) and FK506 inhibited anti-CD3- mediated degranulation, but did not affect cytotoxicity of CIK cells against tumor target cells. In addition, degranulation induced by target cells was unaffected by CsA and FK506. Our results indicate that two mechanisms of cytoplasmic granule release are operative in the CD3+ CD56+ killer cells; however, cytotoxicity proceeds through a cAMP- sensitive, CsA- and FK506-insensitive pathway triggered by yet-to-be- identified target cell surface molecules.     

Prevalence of gestational diabetes mellitus in urban and rural Tanzania

Aim

To estimate prevalence of gestational diabetes mellitus (GDM) and associated determinants in urban and rural Tanzania.

Methods

A cross-sectional study was conducted from 2011 through 2012 in selected urban and rural communities. Pregnant women (609 urban, 301 rural), who were not previously known to have diabetes, participated during usual ante-natal clinic visits. Capillary blood samples were collected at fasting and 2 h after 75 g glucose load and were measured using HemoCue. Diagnosis of GDM was made using 1999 World Health Organization (WHO) criteria.

Results

Women in rural areas were younger (26.6 years) than in urban areas (27.5 years). Mean gestational age, height, and mid-upper arm circumference (MUAC) were similar for the two areas. Overall prevalence of GDM averaged 5.9%, with 8.4% in urban area and 1.0% in rural area. Prevalence of GDM was higher for women who had a previous stillbirth (OR 2.8, 95% CI 1.5–5.4), family history of type 2 diabetes (OR 2.1, 95% CI 1.1–4.2), and MUAC above 28 cm (OR 1.9, 95% CI 1.1–3.3), and lower for women with normal hemoglobin compared with anemia (OR 0.45, 95% CI 0.22–0.93).

Conclusions

Prevalence of GDM is higher than expected in urban areas in Tanzania, indicating an increasing population who are at risk for delivery complications and type 2 diabetes in Sub-Saharan Africa.     

Redox modulatory protective effects of ω-3 fatty acids rich fish oil against experimental colitis

Ulcerative colitis (UC), a form of inflammatory bowel disease (IBD), is an immune-modulated disorder characterized by chronic and recurring inflammatory episodes. Oxidative stress and COX pathway of prostaglandin (PG) biosynthesis are indispensable to pathogenesis of UC. Any imbalance between PGs can compromise the mucosal homeostasis, leading to mucosal damage and chronic inflammation. However, blocking these PGs using classical Cox inhibitors such as non-steroidal anti-inflammatory drugs (NSAIDs) can instead aggravate signs of IBD. Therefore, realizing the need for safer and well tolerable alterative treatment approaches, currently, we evaluated the efficacy of n-3 fatty acids rich fish oil (FO) in the resolution of UC. Using a dextran sodium sulfate (DSS) model of experimental colitis, we have demonstrated that supplementation of FO containing 180?mg EPA and 120?mg DHA for 1 month relieved the signs (diarrhea, bloody stools, weight loss) of colitis-associated inflammation. To understand the biophysical changes associated with FO mediated inflammatory regulation, impedance measurement and Fourier transform infrared spectroscopy (FTIR) were done. These changes were also correlated with oxidative stress through markers such as GST, glutathione peroxidase (GPx), LPO, catalase, protein carbonyl content, GR, etc. in colonic mucosa. The modulation of COX mediated pathways in UC-associated inflammation was observed by protein expressions of various pro-inflammatory cytokines such as TNF-α and enzymes of PG synthesis such as COX-2, PGES, TXAS, and anti-inflammatory PGDS. Refuting the earlier reports that suggested the contradictory effects of FO, in the current study, we evidently demonstrated that the protective effects of FO are mediated through molecular mechanisms involving the redox-regulation of metabolism of key lipid metabolites.     

Efficacy of a new model for delivering integrated TB and HIV services for people living with HIV/AIDS in Delhi – case for a paradigm shift in national HIV/TB cross-referral strategy

Under National TB/HIV framework, all TB patients are referred by Revised National Tuberculosis Programme (RNTCP) service providers to Integrated Counseling and Testing Centers (ICTCs) for voluntary counseling and testing (C&T) and ICTC “TB-suspects” are referred to RNTCP facilities for TB diagnosis and treatment. HIV–TB coinfected patients are then referred to Anti Retroviral Treatment (ART) center for initiation of ART between two weeks and two months of initiating TB treatment. During the third phase of National AIDS Control Programme (NACP-III, April 2007–April 2012), 30749/130503 (23.6%) TB/HIV cross-referrals were lost to follow up (LTFU) and there was missed opportunity for 940/1884 (49.9%) HIV–TB coinfected patients for initiation of ART during TB treatment. This motivated Delhi State AIDS Control Society (DSACS) and State TB Cell (STC) to revise existing cross-referral strategy. The new strategy was launched in May 2012, wherein HIV–TB coinfected and HIV-positive “TB-suspects” were referred to nearest ART center for HIV care and investigations of TB at Chest Clinic/Designated Microscopy Centre (DMC) located within the same hospital instead of referral to area RNTCP facility. Outcome of the strategy was evaluated in March 2013. The new HIV–TB cross-referral strategy in Delhi has shown advantage over national strategy: first, improved retention of coinfected clients in HIV care; second, ensured timely initiation of TB-treatment and ART; and third, significantly improved survival of HIV–TB coinfected patients.