Acceleration of cutaneous wound healing by Lucilia sericata maggots in diabetic Wistar rats.

The study was aimed to evaluate the effectiveness of maggot therapy in healing of cutaneous infected wound in streptozotocin (STZ) induced diabetic Wistar rat. For live maggots, the sterilized eggs of Lucilia sericata were obtained from colonies established in laboratory. Diabetes model was established in 48 male Wister rat by intra-peritoneal injection of STZ at the dose of 60 mg/kg body-weight. Cutaneous wounds exposed with mixed colonies of bacteria like Staphylococcus aureus, E. coli and Pseudomonas aeruginosa were prepared in all rat. The animals equally divided in 4 groups with 12 rats each being presented as treatment group of control, antibiotic, maggot and maggot with antibiotic in combination. All treatments were done once and hold for 24 hours. Wound kinetics and bacterial bio burden were measured at weekly interval to till complete healing. Significant reduction in wound area with maximum contraction was found (>95%) in maggot treated group when compared to antibiotic treated (79%) and control (72%). In maggot as well as maggot and antibiotic in combination group showed early elimination of bacterial bio-burden 7.88±0.03log CFU/ml to 1.12±0.65log CFU/ml and 7.86±0.04) log CFU/ml to 1.54±0.52log CFU/ml respectively in three weeks of time. Early healing indication was also experienced on histomorphological examination of wounded tissue of maggot treated groups by early and better epithelialization, collagenation and neovascularization with complete healing of wound in three weeks in comparison to antibiotic and control respectively. However, the present study did not show any difference in healing of wound with use of maggot alone or in antibiotic combination. Live maggot of Lucilia sericata effectively lower bacterial bioburden and and accelerate healing of infected cutaneous wound in diabetic conditions.

Acceleration of cutaneous wound healing by Lucilia sericata maggots in diabetic Wistar rats

@#The study was aimed to evaluate the effectiveness of maggot therapy in healing of cutaneous infected wound in streptozotocin (STZ) induced diabetic Wistar rat. For live maggots, the sterilized eggs of Lucilia sericata were obtained from colonies established in laboratory. Diabetes model was established in 48 male Wister rat by intra-peritoneal injection of STZ at the dose of 60 mg/kg body-weight. Cutaneous wounds exposed with mixed colonies of bacteria like Staphylococcus aureus, E. coli and Pseudomonas aeruginosa were prepared in all rat. The animals equally divided in 4 groups with 12 rats each being presented as treatment group of control, antibiotic, maggot and maggot with antibiotic in combination. All treatments were done once and hold for 24 hours. Wound kinetics and bacterial bio burden were measured at weekly interval to till complete healing. Significant reduction in wound area with maximum contraction was found (>95%) in maggot treated group when compared to antibiotic treated (79%) and control (72%). In maggot as well as maggot and antibiotic in combination group showed early elimination of bacterial bio-burden 7.88±0.03log CFU/ml to 1.12±0.65log CFU/ml and 7.86±0.04) log CFU/ml to 1.54±0.52log CFU/ml respectively in three weeks of time. Early healing indication was also experienced on histomorphological examination of wounded tissue of maggot treated groups by early and better epithelialization, collagenation and neovascularization with complete healing of wound in three weeks in comparison to antibiotic and control respectively. However, the present study did not show any difference in healing of wound with use of maggot alone or in antibiotic combination. Live maggot of Lucilia sericata effectively lower bacterial bioburden and and accelerate healing of infected cutaneous wound in diabetic conditions.

Enhanced Biosynthesis Synthesis of Copper Oxide Nanoparticles (CuO-NPs) for their Antifungal Activity Toxicity against Major Phyto-Pathogens of Apple Orchards.

PURPOSE: The present study made an attempt to develop copper nanoparticles (Cu-NP) with antifungal property using green synthesis method. Copper oxide nanoparticles (CuO-NPs) botanically synthesized using Neem leaf extract (Azadirachta indica A. Juss) were characterized by using different techniques like; UV-visible spectrophotometry, FTIR, XRD, SEM and TEM. METHODS: Materials were chosen the disease free and fresh Azadirachta indica A. Juss were collected and identified at Center of Biodiversity and Taxonomy. The plant samples were vigorously washed with distilled water then shade dried followed by sterilization with 0.1% mercuric chloride for 20 s and again it was washed with distilled water. 15 g powder form of plant material was added to 200 ml double distilled, CO2 free and deionized water and kept in shaker at 80°C and 1500 rpm for six hours. After agitation, the extract was separated by regular centrifugation at 10,000 rpm followed by filtration by using whatmann filter paper. The final volume of 100 ml of supernatant was collected as pure extract and stored in cool place for further use. RESULTS: The final results confirm a significant inhibition of CuO-NPs for the test fungi. Additionally, CuO-NPs demonstrated an enhanced effect when combined with Neem leaf extract. A total of 20-30% improvement in activity was noticed after combination, which correlates with commonly used synthetic fungicides. The toxicity results reveal that A. indica extract and their combined fractions with CuO-NP were less toxic to the test seeds of experimental plant while as bulk Cu followed by biosynthesized CuO-NPs influenced the germination rate as compared to control pots. CONCLUSIONS: The study drops a concern of research and offers a promising route of developing Copper based green fungicides that can help to combat with modern issues of synthetic fungicides. An average size of 80 ± 15 nm monoclinic cupric oxide (CuO) and cubic cuprous oxides (Cu2O) nanocrystals that existed in mixed form were successfully developed.

Hookah smoking, nass chewing, and oesophageal squamous cell carcinoma in Kashmir, India.

BACKGROUND: Although cigarette smoking is an established risk factor for oesophageal squamous cell carcinoma (ESCC), there is little information about the association between other smoking and smokeless tobacco products, including hookah and nass, and ESCC risk. We conducted a case-control study in Kashmir Valley, India, where hookah smoking, nass chewing, and ESCC are common, to investigate the association of hookah smoking, nass use, and several other habits with ESCC. METHODS: We recruited 702 histologically confirmed ESCC cases and 1663 hospital-based controls, individually matched to the cases for age, sex, and district of residence from September 2008 to January 2012. Conditional logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (95% CIs). RESULTS: Ever-hookah smoking (OR=1.85; 95% CI, 1.41-2.44) and nass chewing (OR=2.88; 95% CI, 2.06-4.04) were associated with ESCC risk. These associations were consistent across different measures of use, including intensity, duration, and cumulative amount of use, and after excluding ever users of the other product and cigarette smokers. Our results also suggest an increased risk of ESCC associated with ever-gutka chewing and -bidi smoking. However, the latter associations were based on small number of participants. CONCLUSION: This study shows that hookah and nass use are associated with ESCC risk. As prevalence of hookah use seems to be increasing among young people worldwide, these results may have relevance not only for the regions in which hookah use has been a traditional habit, but also for other regions, including western countries.

Isolation of an Isometric Virus Causing Sunflower Necrosis Disease in India.

Sunflower necrosis disease (SND) is becoming a potential threat to sunflower (Helianthus annuus L.) cultivation in the Indian subcontinent. The disease was first recorded in parts of Karnataka state in 1997. Since then the disease has become increasingly important in Andhra Pradesh, Karnataka, Maharashtra, and Tamil Nadu, the four major sunflower-growing states of India, and is a limiting factor in sunflower production; up to 80% of the plants of some open pollinated and hybrids were affected during the 1999 survey in sunflower growing areas. Field symptoms of the disease include extensive necrosis of leaf lamina, petiole, stem and floral calyx and severe stunting with malformation of flowering head when plants are infected early. The association of a tospovirus, antigenically related to groundnut bud necrosis (GBNV) and watermelon silver mottle (WSMV) viruses, with the disease has been reported (1). However, the etiology of the disease remains unaddressed. In this study a sap-transmissible isometric virus was transferred to cowpea (cvs. Pusa Komal and C152) inciting localized chlorotic and necrotic lesions and systemic veinal necrosis. Electron-microscopic studies of leaf-dip preparations from field samples revealed two types of particles (isometric measuring 25 to 28 nm in diameter and flexuous rods with a length of about 600 nm). The sap-inoculated cowpea and sunflower contained only the isometric particles. Some preparations also showed the presence of tubules containing virus particles. The presence of flexuous particles in field samples could be due to mixed infection as the mosaic disease, known to be caused by a flexuous virus, was common in the sunflower fields surveyed in the present investigations. Extracts from the field collected samples or sap-inoculated plants did not react with antisera to cucumber mosaic (CMV) or potato Y (PVY) viruses in direct antigen-coated (DAC)-ELISA and immunosorbent electron microscopy tests. The isometric virus isolated from sunflower was purified from sap-inoculated cowpea plants by differential and sucrose density-gradient centrifugations. The virus was sap transmitted back to sunflower (cv. Morden), which developed symptoms identical to those observed under field conditions. Disease symptoms were also reproduced on sunflower upon mechanical inoculation with the purified virus. Polyclonal antiserum raised in rabbits using purified virus preparations, detected the virus from field and glasshouse collected sunflower plants in DAC-ELISA tests. This will help in epidemiological studies and breeding for disease resistance. The particle size and structure and the presence of tubule containing virus particles in plant extracts suggest that the virus belongs to ILAR group. An ILAR virus is reported to infect sunflower (2), but details of its natural occurrence are not known. This is the first report on the etiology of the sunflower necrosis disease in India. Further studies are in progress. References: (1) Anon. 2000. Annual Report (1999-2000), Indian Agricultural Research Institute, New Delhi, India. (2) A. A. Brunt et al. CAB International, Wallingford, UK, 1210, 1996.

Effects of phytoestrogens and synthetic combinatorial libraries on aromatase, estrogen biosynthesis, and metabolism.

Approximately 60% of breast cancer patients have hormone-dependent breast cancer containing estrogen receptors and requiring estrogen for tumor growth. The extent of estrogen biosynthesis and metabolism in the breast cancer tissue microenvironment influences breast-tumor development and growth, and endogenous and exogenous agents may alter the levels of hormonally active estrogens and their metabolites. Isoflavonoid phytoestrogens such as genistein exhibit numerous biochemical activities; however, their effects on estrogen biosynthesis and metabolism in breast cancer cells have not been fully examined. MCF-7 cells (hormone-dependent) and MBA-MB-231 cells (hormone-independent) were treated with genistein (100 nM) for five days and then incubated with radiolabeled estradiol (100 nM, 2.5 microCi) for 0 to 48 h. Media were extracted with ethyl acetate, and the organic residues analyzed by reverse-phase HPLC with a radioactivity flow detector. The major metabolite formed in all cases is estrone, although differences were observed between the cell lines and the various drug treatments. The formation of estrone in untreated MCF-7 cells (approximately 9.3% of radioactivity at 24 h) is relatively limited, in contrast to untreated MDA-MB-231 cells (approximately 32.0% of radioactivity at 24 h). Treatment of MCF-7 cells with 100 nM genistein increased the conversion of estradiol to estrone up to 19.5% in 24 h. The effect of genistein on estrone formation in MDA-MB-231 cells resulted in 37.7% of the radioactivity being estrone. Thus, genistein treatment of breast cancer cells resulted in increased 17-betahydroxysteroid dehydrogenase activity and elevated formation of estrone. Increased levels of oxidative 17-betahydroxysteroid dehydrogenase activity (Type II) were confirmed by Western blots. Therefore, exposure of breast cancer cells to genistein results in elevated conversion of estradiol to estrogenically weaker or inactive metabolites. The regulation of breast-tissue aromatase by exogenous agents such as drugs and environmental agents is being investigated. The benzopyranone-ring system is a molecular scaffold of considerable interest, and this scaffold is found in flavonoid natural products that have weak aromatase inhibitory activity. Medicinal chemistry efforts focus on diversifying the benzopyranone scaffold and utilizing combinatorial chemistry approaches to construct small benzopyranone libraries as potential aro- matase inhibitors. Several compounds in the initial libraries have demonstrated moderate aromatase inhibitory activity in screening assays.

Nutrition, HIV, and drug abuse: the molecular basis of a unique role for selenium.

HIV-infected injection drug users (IDUs) often suffer from serious nutritional deficiencies. This is a concern because plasma levels of micronutrients such as vitamin B12, zinc, and selenium have been correlated with mortality risk in HIV-positive populations. Injection drug use also increases lipid peroxidation and other indicators of oxidative stress, which, combined with antioxidant deficiencies, can stimulate HIV-1 replication through activation of NF-kappaB transcription factors, while weakening immune defenses. As detailed herein, these prooxidant stimuli can also increase the pathogenic effects of HIV-1 by another mechanism, involving viral selenoproteins. Overlapping the envelope coding region, HIV-1 encodes a truncated glutathione peroxidase (GPx) gene (see #6 in reference list). Sequence analysis and molecular modeling show that this viral GPx (vGPx) module has highly significant structural similarity to known mammalian GPx, with conservation of the catalytic triad of selenocysteine (Sec), glutamine, and tryptophan. In addition to other functions, HIV-1 vGPx may serve as a negative regulator of proviral transcription, by acting as an NF-kappaB inhibitor (a known property of cellular GPx). Another potential selenoprotein coding function of HIV-1 is associated with the 3' end of the nef gene, which terminates in a conserved UGA (potential Sec) codon in the context of a sequence (Cys-Sec) identical to the C-terminal redox center of thioredoxin reductase, another cellular regulator of NF-kappaB. Thus, in combination with known cellular mechanisms involving Se, viral selenoproteins may represent a unique mechanism by which HIV-1 monitors and exploits an essential micronutrient to optimize its replication relative to the host.

Selenium-dependent glutathione peroxidase modules encoded by RNA viruses.

Glutathione peroxidase (GPx) is the prototypical eukaryotic selenoprotein, with the rare amino acid selenocysteine (Sec) at the enzyme active site, encoded by the UGA codon in RNA. A DNA virus, Molluscum contagiosum, has now been shown to encode a functional selenium-dependent GPx enzyme. Using modifications of conventional sequence database searching techniques to locate potential viral GPx modules, combined with structurally guided comparative sequence analysis, we provide compelling evidence that Se-dependent GPx modules are encoded in a number of RNA viruses, including potentially serious human pathogens like HIV-1 and hepatitis C virus, coxsackievirus B3, HIV-2, and measles virus. Analysis of the sequences of multiple viral isolates reveals conservation of the putative GPx-related features, at least within viral subtypes or genotypes, supporting the hypothesis that these are functional GPx modules.

Frequency and duration of silent myocardial ischemia in patients with unstable angina before and after intensive medical therapy.

Frequency and duration of silent myocardial ischemia (SMI) on Holter recording were determined in 20 patients with unstable angina before and after 4 weeks of intensive triple drug therapy with oral nitrates (20-80 mg daily), betablocker (metoprolol 100-200 mg/day) and calcium channel blocker (nifedipine 40-80 mg/day). The number of ischemic episodes decreased from 445 (409 silent) to 149 (140 silent) (p less than 0.001), and mean duration of silent and symptomatic ischemia per patient decreased from 5.9 +/- 3.3 minutes to 2.4 +/- 2.6 minutes (p less than 0.001) and 4.1 +/- 5.5 minutes to 1.4 +/- 2.8 minutes (p less than 0.001) respectively. Intensive medical therapy is effective in ameliorating SMI in patients with unstable angina.