Structural analysis and properties of dextran produced by Leuconostoc mesenteroides NRRL B-640

A water-soluble dextran was produced by purified dextransucrase from Leuconostoc mesenteroides NRRL B-640. The dextran was purified by alcohol precipitation. The structure of dextran was determined by FT-IR, ¹H NMR, ¹³C NMR and 2-dimensional NMR spectroscopic techniques. NMR techniques (1D ¹H, ¹³C and 2D HMQC) were used to fully assign the ¹H and ¹³C spectra. All the spectral data showed that the dextran contains d-glucose residues in a linear chain with consecutive α(1 → 6) linkages. No branching was observed in the dextran structure. The viscosity of dextran solution decreased with the increase in shear rate exhibiting a typical non-Newtonian pseudoplastic behavior. The surface morphology of dried and powdered dextran studied using Scanning electron microscopy revealed the cubical porous structure.     

Stimulation of fatty acid synthesis in vitro by gonadotrophin-induced testicular ribonucleic acid

RNA from testes of hypophysectomized rats treated with follicle-stimulating hormone and luteinizing hormone markedly stimulates in vitro the incorporation of acetate and malonate (as CoA derivatives) into polyunsaturated fatty acids. The system in vitro contains the components necessary for both protein and fatty acid synthesis. That the RNA is a hormone-induced messenger type that causes enzyme synthesis that then causes fatty acid synthesis is supported by the following observations: (1) the stimulation of RNA synthesis by follicle-stimulating hormone and luteinizing hormone is decreased by injection of the animals with actinomycin D; (2) puromycin in the system in vitro decreases the synthesis of polyunsaturated fatty acids; (3) the activity of the RNA preparation is destroyed by digestion with ribonuclease; in fact, the digest is inhibitory, which is a characteristic of messenger-RNA-mediated protein synthesis; (4) protein that might be denatured enzyme is virtually absent from the effective RNA preparations.     

Role of Community Group Exposure in Reducing Sexually Transmitted Infection-Related Risk among Female Sex Workers in India

Background

Empowering female sex workers (FSWs) to address structural barriers and forming community groups (CGs) through community mobilization are seen as essential components of HIV prevention programs in India. Taking the membership of a CG as an exposure intervention, we hypothesized whether participation in a CG lead to reduced sexually transmitted infections (STIs) and increased treatment-seeking behavior among FSWs in three selected states of India — Andhra Pradesh, Maharashtra and Tamil Nadu.

Methods and Findings

The propensity score matching (PSM) approach examined the effect of CG membership, as against no membership, on STI-related risk, described as selected outcome measures — presence of any STI, self-reported STI symptoms, and treatment-seeking behavior among FSWs. A cross sectional bio-behavioral survey was administered in 2009–2010 and covered 7,806 FSWs through two-stage probability-based conventional and time location cluster sampling in 23 administrative districts of Andhra Pradesh, Maharashtra and Tamil Nadu. Only 2,939 FSWs were reported to be members of a CG and among them 4.5% had any STIs. A majority of FSWs were aged above 24 years (86.4%), had ever been married (73%), operated from a public place for solicitation (81.5%), and had ever received HIV test results (75.6%). The average effect of CG exposure was reduction in STI prevalence by 4%, while self-reported STI symptom treatment-seeking behavior increased by 13.7%.

Conclusion

FSWs who were exposed to a CG were at a substantially lower risk of STIs than those who were unexposed. The FSWs exposed to a CG had a higher chance of seeking STI treatment from public and private health facilities. Collectivization related challenges must be overcome to provide access to tailored STI prevention and care services.     

Neem Leaf Glycoprotein Activates CD8+ T Cells to Promote Therapeutic Anti-Tumor Immunity Inhibiting the Growth of Mouse Sarcoma

In spite of sufficient data on Neem Leaf Glycoprotein (NLGP) as a prophylactic vaccine, little knowledge currently exists to support the use of NLGP as a therapeutic vaccine. Treatment of mice bearing established sarcomas with NLGP (25 µg/mice/week subcutaneously for 4 weeks) resulted in tumor regression or dormancy (Tumor free/Regressor, 13/24 (NLGP), 4/24 (PBS)). Evaluation of CD8⁺ T cell status in blood, spleen, TDLN, VDLN and tumor revealed increase in cellular number. Elevated expression of CD69, CD44 and Ki67 on CD8⁺ T cells revealed their state of activation and proliferation by NLGP. Depletion of CD8⁺ T cells in mice at the time of NLGP treatment resulted in partial termination of tumor regression. An expansion of CXCR3⁺ and CCR5⁺ T cells was observed in the TDLN and tumor, along with their corresponding ligands. NLGP treatment enhances type 1 polarized T-bet expressing T cells with downregulation of GATA3. Treg cell population was almost unchanged. However, T∶Treg ratios significantly increased with NLGP. Enhanced secretion/expression of IFNγ was noted after NLGP therapy. In vitro culture of T cells with IL-2 and sarcoma antigen resulted in significant enhancement in cytotoxic efficacy. Consistently higher expression of CD107a was also observed in CD8⁺ T cells from tumors. Reinoculation of sarcoma cells in tumor regressed NLGP-treated mice maintained tumor free status in majority. This is correlated with the increment of CD44^hiCD62L^hi central memory T cells. Collectively, these findings support a paradigm in which NLGP dynamically orchestrates the activation, expansion, and recruitment of CD8⁺ T cells into established tumors to operate significant tumor cell lysis.     

Subcellular location of enzymes involved in oxidation of n-alkane by Cladosporium resinae

More than 70% of n-hexadecane-grown cells of Cladosporium resinae ATCC 22711 were converted to spheroplasts when they were treated with chitinase and lytic enzyme from Trichoderma harziamum. The light mitochondrial fraction, containing microbodies, mitochondria and vacuoles, was isolated from spheroplasts. Vacuoles in cells were demonstrated by the inability of acridine orange to stain organelles previously treated with 2.5 μM Bafilomycin A₁, a vacuolar ATPase inhibitor. Microbodies, mitochondria and vacuoles were separated from the light mitochondrial fraction by self-generated density-gradient ultracentrifugation using iodixanol as gradient medium. NADH-dependent n-alkane monooxygenase activity and fatty alcohol oxidase activity were located in the cytoplasm and mitochondrial fractions respectively. Received: 21 September 1998 / Received revision: 21 January 1999 / Accepted: 31 January 1999     

A comparison of time-lapse cinemicrography and flow cytometry for the study of accelerated cell-cycle transit

Abstract. Two methods for the study of cell-cycle progression, time-lapse cinemicrography (TLCM) and flow cytometry (FCM), were compared for their ability to measure the shortening of cell-cycle transit time induced by temporary inhibition of DNA synthesis. DNA synthesis was reversibly inhibited by aphidicolin (APH) in synchronized HeLa cells obtained by mitotic collection. TLCM directly measured intermitotic time intervals and thereby directly obtained the cell-cycle transit time distribution. In contrast, FCM measured time dependent changes in the fractions of cells in the cell-cycle phases from which the distribution of cells traversing a cell-cycle boundary, such as that between G₁ and S phase, was determined. Nevertheless, both methods provided equivalent measures of the cell-cycle transit time and its dispersion. However, TLCM apeared to provide a better measure of skewness of the transit time distribution than did FCM. Further, both methods were able to detect changes in the cell cycle transit on the order of 1 h or less. The TLCM data showed a greater precision (due to a larger number of data points) than that from FCM. However, FCM was able to directly measure changes in the transit of G₁ phase whereas TLCM would require two different experiments to make a similar determination. The results obtained in this study show that FCM can replace TLCM to study most aspects of cell-cycle progression.     

Polyphenol Oxidase Activity and Phenolic Content Pattern During Shoot Development of Grape (Vitis vinifera L.) in Different Growing Seasons

The PPO activity and content of phenolic compounds were estimated during the different growing seasons and developmental age of shoots in seven grape genotypes. Both, the total phenol content and catecholase activity were found significantly affected by the growing season, developmental age of shoot and the genotype. But, season as well as age of shoot could not influence cresolase activity. Spring as season; 30 days of sprouting as age; Gold and Anab-e-Shahi as genotypes were found better in comparison to summer as season and 15 days of sprouting as age in respect to level of phenols and PPO activity.     

Cytogenetic effects of methyl isocyanate exposure in Bhopal

Summary Among human survivors following the methyl isocyanate (MIC) gas tragedy the major complaints have been related to deep-seated suffocation, terrible pain in breathing, and severe ocular irritations. In order to assess the possible genetic effects we have used lymphocyte cultures and screened chromosomes by two techniques; one by looking for chromosomal aberrations and the other by estimating sister-chromatid exchange (SCE) frequencies. Both these paramaters are good indicators of genetic damage in chromosomal DNA. SCE frequencies in lymphocytes have been increased more than three times in MIC-exposed persons. The results were compared to two groups of controls (one group comprising persons present in the same house; the second group of persons were chosen from distant places, 20–50 km away from the incident). Chromosomal breaks have been observed in 10 out of 14 MIC-affected people (71.4%) studied while only 6 out of 28 (21.4%) controls had chromosomal breaks. Some MIC-exposed persons had chromatin bodies in addition to the normal 46 chromosomes. These observations suggest that chromosomal DNA has been damaged.