Increased monocyte activation with age among HIV-infected long term non-progressor children: implications for early treatment initiation.

OBJECTIVES: The key to newer therapeutic and eradication approaches often lies in understanding slow disease progression in HIV infection. The paediatric population has been poorly studied in this regard. We aimed to describe a cohort of perinatally infected long-term nonprogressor (LTNP) children living with HIV in India and to evaluate the immune biomarkers of disease progression. METHODS: LTNPs (ART-naïve, with a CD4 count ≥ 500 cells/µL at age ≥ 7 years) among the cohort of HIV-infected children were identified and monitored longitudinally, and their CD4 T-cell counts and plasma viral loads were measured every 6 months. The plasma monocyte/macrophage activation markers, namely soluble CD14 (sCD14), soluble CD163 (sCD163) and interferon-inducible protein-10 (IP-10) were measured by enzyme-linked immunosorbent assay (ELISA) in LTNPs and progressors. The Mann-Whitney U-test was used to compare the two groups and P values < 0.05 were considered statistically significant. Spearman's rank or Pearson's correlation coefficient (r) was calculated to determine the associations between variables. RESULTS: Among 378 children living with HIV-1 surveyed in our cohort, 40 (10.6%) were LTNPs. Longitudinal analysis of the LTNP data showed that both CD4 count and viral load declined significantly with age (P < 0.0001 for both). Plasma sCD14 levels were significantly (P < 0.005) higher in progressors and sCD163 levels were significantly (P < 0.0001) higher in LTNPs. CONCLUSIONS: The prevalence of LTNPs in our cohort of perinatally infected children living with HIV was 10.6%. We observed a trend for associations between the increasing sCD163 monocyte/macrophage activation marker levels, declining CD4 counts and the gradual loss of nonprogressor status with age in the LTNPs. These findings underscore the need for early antiretroviral therapy in those children with proven slow disease progression.

Bioanalytical method development, validation and quantification of dorsomorphin in rat plasma by LC-MS/MS.

The present investigation describes the development and validation of a sensitive liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method for the estimation of dorsomorphin in rat plasma. A sensitive LC-MS/MS method was developed using multiple reaction monitoring mode, with the transition of m/z (Q1/Q3) 400.2/289.3 for dorsomorphin and m/z (Q1/Q3) 306.2/236.3 for zaleplon. Chromatographic separation was achieved on a reverse phase Agilent XDB C18 column (100 × 4.6 mm, 5 µm). The mobile phase consisted of acetonitrile and 5 mm ammonium acetate buffer (pH 6.0) 90:10 v/v, at a flow rate of 0.8 mL/min. The effluence was ionized in positive ion mode by electrospray ionization (ESI) and quantitated by mass spectrometry. The retention times of dorsomorphin and internal standard were found to be 2.13 and 1.13 min, respectively. Mean extraction recovery of dorsomorphin and internal standard in rat plasma was above 80%. Dorsomorphin calibration curve in rat plasma was linear (r(2) ≥ 0.99) ranging from 0.005 to 10 µg/mL. Inter-day and intra-day precision and accuracy were found to be within 85-115% (coefficient of variation). This method was successfully applied for evaluation of the oral pharmacokinetic profile of dorsomorphin in male Wistar rats.

CpG methylation patterns and decitabine treatment response in acute myeloid leukemia cells and normal hematopoietic precursors.

The DNA hypomethylating drug decitabine maintains normal hematopoietic stem cell (HSC) self-renewal but induces terminal differentiation in acute myeloid leukemia (AML) cells. The basis for these contrasting cell fates, and for selective CpG hypomethylation by decitabine, is poorly understood. Promoter CpGs, with methylation measured by microarray, were classified by the direction of methylation change with normal myeloid maturation. In AML cells, the methylation pattern at maturation-responsive CpGs suggested at least partial maturation. Consistent with partial maturation, in gene expression analyses, AML cells expressed high levels of the key lineage-specifying factor CEBPA, but relatively low levels of the key late-differentiation driver CEBPE. In methylation analysis by mass spectrometry, CEBPE promoter CpGs that are usually hypomethylated during granulocyte maturation were significantly hypermethylated in AML cells. Decitabine-induced hypomethylation was greatest at these and other promoter CpGs that are usually hypomethylated with myeloid maturation, accompanied by cellular differentiation of AML cells. In contrast, decitabine-treated normal HSCs retained immature morphology, and methylation significantly decreased at CpGs that are less methylated in immature cells. High expression of lineage-specifying factor and aberrant epigenetic repression of some key late-differentiation driver genes distinguishes AML cells from normal HSCs, and could explain the contrasting differentiation and methylation responses to decitabine.

Anesthetic challenges of decompressive craniotomy for the right frontal lobe abscess in a patient with Ebstein's anomaly: A rare case report.

Ebstein's anomaly is a rare congenital heart disease characterized by downward displacement of the tricuspid valve, atrialization of right ventricle, enlarged right atrium with tricuspid regurgitation, or stenosis. It is often associated with atrial septal defect (ASD) or patent foramen ovale with right to left shunt. It has a varied clinical presentation and can manifest as cyanosis, ventricular, or supraventricular arrhythmias and recurrent episodes of congestive heart failure. We describe the anesthetic management of a female patient with Ebstein's anomaly who had right frontal lobe abscess due to paradoxical embolism for decompressive craniotomy.

MDA-7/IL-24, a novel tumor suppressor/cytokine is ubiquitinated and regulated by the ubiquitin-proteasome system, and inhibition of MDA-7/IL-24 degradation enhances the antitumor activity.

Steady-state protein levels are determined by the balance between protein synthesis and degradation. Protein half-lives are determined primarily by degradation, and the major degradation pathways involve either lysosomal destruction or an ATP-dependent process involving ubiquitination to target proteins to the proteosome. Studies have shown that multiple tumor-suppressor proteins are ubiquitinated and degraded by the 26S proteasome. In the present study, we investigated whether the tumor suppressor/cytokine melanoma differentiation-associated gene-7/interleukin-24 gene (MDA-7/IL-24) protein is ubiquitinated and its degradation controlled by the proteasome. Treatment of ovarian (2008) and lung (H1299) tumor cells with adenoviral delivery of mda-7 (Ad-mda7) or Ad-mda7 plus the proteosome inhibitor MG132 showed that MDA-7 protein expression was dependent upon proteosome activity. Western blot and immunoprecipitation analyses verified that the MDA-7 protein was ubiquitinated and that ubiquitinated-MDA-7 levels were increased in MG132-treated cells. These results were confirmed using small interfering RNA (siRNA)-mediated knockdown of ubiquitin. Furthermore, ubiquitinated MDA-7 protein was degraded by the 26S proteasome, as MDA-7 accumulation was observed only when cells were treated with MG132 but not with lysosome or protease inhibitors. Inhibition of the catalytic beta-5 subunit of the 20S proteasome using siRNA resulted in MDA-7 protein accumulation. Finally, treatment of tumor cells with Ad-mda7 plus the proteasome inhibitor bortezomib resulted in increased tumor cell killing. Our results show that MDA-7/IL-24 is ubiquitinated and degraded by the 26S proteasome. Furthermore, inhibition of MDA-7 degradation results in enhanced tumor killing, identifying a novel anticancer strategy.

Survey for tuberculosis in a tribal population in North Arcot District.

SETTING: Tribal villages in the jungles of the Jawadhu hills, South India. OBJECTIVE: To estimate the prevalence of tuberculosis (TB) infection and disease in a remote tribal population. DESIGN: A cross-sectional survey with two-stage screening for identification of cases. A stratified probability proportional sample with the hamlet as the unit. METHODS: Among 56 revenue divisions with a population of about 66,000, 24 revenue divisions were selected. Among 26,320 persons registered, children < 10 years were tuberculin tested and reactions were read after 72 hours. Those over 15 were X-rayed, and tuberculosis symptoms were investigated. Sputum was collected from those with abnormal X-ray or symptoms and examined for smear and culture positivity and sensitivity. RESULTS: Of the 6952 children tested and read, 5% had BCG scars and the prevalence of infection was 5%. The annual risk of infection was 1.1. Among adults, the prevalence of bacillary cases was 8/1000 and X-ray cases 29/1000. The prevalence of bacillary disease was higher among males, particularly with increasing age. Thirty symptomatic cases had normal X-rays and 63 X-ray cases had no symptoms. Thus prevalence would have been underestimated if either method had been used alone for screening. Isoniazid resistance was seen in 12% of patients, two of whom also had rifampicin resistance (2.6%). CONCLUSIONS: The prevalence and pattern of tuberculosis in this tribal group is similar to that observed in non-tribal areas.

Supercritical carbon dioxide extraction of turmeric (Curcuma longa).

Turmeric oil was extracted from turmeric (Curcuma longa) with supercritical carbon dioxide in a semicontinuous-flow extractor. Extraction rate was measured as a function of pressure, temperature, flow rate, and particle size. The extraction rate increased with an increase in CO(2) flow rate and with a reduction of particle size. The effect of pressure and temperature on turmeric extraction suggested the use of higher pressure and lower temperature at which solvent density is greater and thus the solubility of the oil in the solvent is greater in the range of 313-333 K and 20-40 MPa. The major components ( approximately 60%) of the extracted oil were identified as turmerone and ar-turmerone by GC-MS.

Feasibility of involving literate tribal youths in tuberculosis case-finding in a tribal area in Tamil Nadu.

SETTING: Tribal area in South India with a population of 96,000, where the tuberculosis case-finding activity was very poor. OBJECTIVES: To investigate the feasibility of (1) involving literate (who can read and write) tribal youth volunteers for detecting cases of pulmonary tuberculosis (PT) in their respective hamlets; and (2) antituberculosis drug delivery to sputum-positive patients at their homes by village health nurses (VHNs). DESIGN: One volunteer from each of 61 hamlets was selected and trained in the detection of subjects with chest symptoms, sputum collection and transportation to the Primary Health Centre for smear examination. All smear-positive patients were treated with 2RHZ/6TH and the drugs were supplied by VHNs fortnightly at their homes. RESULTS: During a period of 1 year (1992-93), the total population screened was 9383 persons; of these 5755 were aged 15 years and above. A total of 338 symptomatic subjects were identified; 12 sputum-positive cases were detected and started on treatment. Antituberculosis drugs were supplied by VHNs to patients for the first 9 months of the study and by literate youths for the next 3 months. Spot drug checks revealed that 11 of the 12 patients were regular in drug consumption. CONCLUSION: It is feasible to train literate tribal youth volunteers within a short time to detect subjects with chest symptoms in the community and thereby cases of pulmonary tuberculosis. They can serve as an excellent model for community participation in difficult areas.

Critical assessment of smear-positive pulmonary tuberculosis patients after chemotherapy under the district tuberculosis programme.

This is a status report of a retrospectively assembled cohort of 3357 smear-positive patients initiated on anti-tuberculosis chemotherapy in the North Arcot district between April 1986 and March 1988. The patients were contacted once at their homes between November 1988 and June 1989 (6 and 36 months after start of treatment), and information on their status, including death, could be obtained from 76% of them. Regimens were selected by the patients. 2306 (69%) had accepted short course regimens (SCC) and 1051 (31%) had been started on standard chemotherapy (non-SCC), 43% and 35% in SCC and non-SCC respectively had completed 80% or more of their treatment. Overall mortality was 28%. Of those remaining, 31% had active disease and were excreting bacilli, among which 65% of the cultures were resistant to isoniazid and 12% to rifampicin. Combined resistance to isoniazid and rifampicin was seen in 4% and to isoniazid and streptomycin was seen in 19%. A significant finding was that even among those who had taken less than 50% of their treatment, 56% were bacteriologically negative. However, inadequate or irregular chemotherapy resulted in over four times the mortality and about twice the rate of smear positivity as compared with those taking adequate chemotherapy. No comparisons are made between patients on short-course and standard regimens as the patients selected their treatment and the groups are not comparable.