Efficacy of modified vacuum-assisted dressing versus conventional betadine dressing in wound healing of open fractures.

Aim: This study aimed to compare the effectiveness of a customized vacuum-assisted dressing to traditional betadine dressings for wounds in open fractures. Materials and methods: In this prospective comparative study, 30 patients from two groups-group A receiving V.A.C. while group B receiving traditional dressing-were given data from sixty participants with open fractures. Wound was evaluated on days 0, 3, 7, 11, and 15 of the study. Results: In group A, there was a statistically significant decrease in the mean dimension of the wound overall (15.66 mm vs. 7.4 mm in group B), and it took an average of 9.83 days for healthy granulation tissue to emerge. In contrast to the 21 patients who had split skin grafting, five patients needed a flap as a final closure surgery. In group B, it took an average of 17 days for healthy granulation tissue to emerge. The authors used split skin grafting to close the wounds in 18 patients, and the wound was allowed to heal by secondary intention in 8 patients, while the flap was used in 4 patients. Conclusion: On comparing the modified Vacuum-assisted dressing to the standard dressing, there was considerable wound contraction and accelerated healing. Therefore, the authors observed that vacuum-assisted dressing treatment is superior to traditional betadine dressing in open fractures.

Pretreatment of mosquito larvae with ultraviolet-B and nitropolycyclic aromatic hydrocarbons induces increased sensitivity to permethrin toxicity.

Nitropolycyclic aromatic hydrocarbons (Nitro-PAH) are highly toxic PHA derivatives. Nitro-PHAs are emitted by carbonaceous materials and PHA post-emission transformation, which causes water and environmental pollution and also exists as carcinogenic and immunotoxic agents. UV light has been shown to cause DNA damage and improves the covalent binding of PAH to DNA significantly. Mosquito breeding grounds are pools of water that can be large open zones or encased ponds with varying levels of sunlight exposure. This research was performed to assess the combined effects of UV-B exposure and Nitro-PAH on the physiological function of Culex quinquefasciatus larvae. To assess the impact of UV-B irradiation and Nitro-PAH exposure on mosquito vectors, parameters were examined: (1) Nitro-PAH availability and its impact on cell fatalities; (2) the detoxifying abilities of cytochrome P450, glutathione-S-transferase, and esterase; (3) the reactions to Reactive Oxygen Species; and (4) The resistance of mosquito larvae to three synthetic pesticides (temephos, imidacloprid, and permethrin). UV-B and Nitro-PAH treatment caused cellular damage and increased major detoxification enzymes such as α & ß-esterase, cytoP450, CAT, GST, and POX. The levels of oxidative stress, ROS and protein carbonyl content, nitrite, ascorbic acid and thiobarbituric acid were decreased significantly. Toxicology bioassays revealed that UV-B + Nitro-PAH exposure significantly increased larval susceptibility. The current study concludes that prior exposure to Nitro-PAHs and UV-B may make mosquito larvae more vulnerable to chemical insecticides.

Isolation and identification of entomopathogenic fungus from Eastern Ghats of South Indian forest soil and their efficacy as biopesticide for mosquito control.

The repeated usage of chemical insecticides, responsible for insecticide resistance in mosquitoes and environmental toxicity. Currently effective and environmental-safe control strategies are needed for the control disease-vector mosquitoes. Entomopathogens can be an effective alternative to chemical insecticide. Herein we isolated and tested 46 soil-borne entomopathogenic fungi belonging to six genera, namely Beauveria sp., Metarhizium sp., Fusarium sp., Aspergillus sp., Trichoderma sp., and Verticillium sp., fungi conidia were tested on Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus larvae. Bioassays results show that M. anisopliae fungal isolate causes a 100%, 98.6% and 92% mortality within six days, on Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus, respectively. M. anisopliae treated three mosquito larvae have lower lifetime with LT50 values in A. stephensi, 2.931 days; A. aegypti, 2.676 days and C. quinquefasciatus, 3.254 days. 18 s rDNA sequence analysis confirmed that the isolated fungus are belonging to the genus of M. anisopliae-VKKH3, B. bassiana-VKBb03, and V. lecanii-VKPH1. Our results clearly show that M. anisopliae has good potential, as a low-cost, environmentally safe tool for the control of A. aegypti, A. stephensi, and C. quinquefasciatus mosquitoes.

Effect of entomopathogenic nematode of Heterorhabditis indica infection on immune and antioxidant system in lepidopteran pest Spodoptera litura (Lepidoptera: Noctuidae).

Entomopathogenic nematodes form excellent tools to study insect immunity in response to during infection. Insects activate as several defense mechanisms, namely Phenoloxidase, haemocytes, detoxification and antioxidant enzymes. However little mechanistic information is available about the sublethal effects of entomopathogenic nematodes infection on detoxification and immune mechanisms in lepidopteran insects. In the present study, the effects of infection on antioxidant, detoxification and immune systems of Spodoptera litura larvae were studied. Results show a significant reduction in Total Haemocyte Count observed after 3 h of infection. A significant increase Superoxide dismutase, Catalase, Glutathione S-transferase, Glutathione Peroxidase and Acid phosphatase were observed 6 h after infection and, progressive decrease in Peroxidase, Alkaline phosphatase and Lipid peroxidation was also observed. This study shows that increased detoxification enzyme levels in response to nematode infection are a protective mechanism in insects. Nematode infection suppresses insect immune response, which is evident from low haemocyte count and Phenoloxidase levels to ultimately cause larval mortality.

Spectral characterization and antibacterial activity of an isolated compound from Memecylon edule leaves.

Memecylon edule Roxb. (Melastamataceae family) is a small evergreen tree reported as having ethnobotanical and pharmacological properties. The present study was aimed to investigate the spectral characterization and antibacterial activity of isolated pure compound (3ß-hydroxyurs-12-en-28-oic acid (ursolic acid)) from Memecylon edule leaves by performing bioassay guided isolation method. The structure derivation of isolated compound was done by different spectral studies like UV, FT-IR, LC-MS, CHNS analysis, 1D (1H, 13C and DEPT-135) and 2D-NMR (HSQC and HMBC), respectively. About 99.29% purity of the compound was found in LC analysis. 1H NMR spectrum results of compound shown 48 protons appear at different shielded region and most of the protons were present in aliphatic region. Whereas, 13C NMR spectral data resulted seven methyl carbons (CH3), nine methylene carbons (CH2), seven methine carbons (CH) and six non-hydrogenated carbons (C) which are characteristic of pentacyclic triterpene. The isolated pure compound was tested for its antibacterial properties against targeted human pathogens by performing agar well diffusion, MIC and MBC assays and the result exhibits better growth inhibitory effects against S. epidermidis and S. pneumoniae, with the MIC values of 1.56 and 3.15µg/ml. The outcome of this study suggests that the bioactive compound is used for development of plant based drugs in pharmaceutical industry for combating microbial mediated diseases.

A novel amperometric catechol biosensor based on α-Fe2O3 nanocrystals-modified carbon paste electrode.

In this work, we designed an amperometric catechol biosensor based on α-Fe2O3 nanocrystals (NCs) incorporated carbon-paste electrode. Laccase enzyme is then assembled onto the modified electrode surface to form a nanobiocomposite enhancing the electron transfer reactions at the enzyme's active metal centers for catechol oxidation. The biosensor gave good sensitivity with a linear detection response in the range of 8-800 µM with limit of detection 4.28 µM. We successfully employed the sensor for real water sample analysis. The results illustrate that the metal oxide NCs have enormous potential in the construction of biosensors for sensitive determination of phenol derivatives.

Green synthesis of selenium nanoparticles conjugated Clausena dentata plant leaf extract and their insecticidal potential against mosquito vectors.

Mosquitoes are major vectors for the transmission of many diseases like chikungunya, malaria, dengue, zika, etc. worldwide. In the present study, selenium nanoparticles (SeNPs) were synthesized from Clausena dentata and were tested for their larvicidal efficacy against the fourth-instar larvae of Anopheles stephensi, Aedes Aegypti, and Culex quinquefasciatus. The synthesized nanoparticles were characterized using UV-Vis spectroscopy, Fourier Transform Infrared Radiation (FTIR) spectroscopy, EDaX, and SEM. The results recorded from UV-Vis spectroscopy show the peak absorption spectrum at 420 nm. In FTIR, the maximum peak value is 2922.25 cm-1 assigned to N-H group (amide group). In EDaX analysis shows peak around 72.64 which confirm the binding intensity of selenium. In SEM analysis, the synthesized SeNPs sizes were ranging from 46.32 nm to 78.88 nm. The synthesized SeNPs produced high mortality with very low concentration (LC50) were 240.714 mg/L; 104.13 mg/L, and 99.602 mg/L for A. stephensi, A. Aegypti, and C. quinquefasciatus, respectively. These results suggest that the C. dentata leaf extract-mediated biosynthesis of SeNPs has the potential to be used as an ideal ecofriendly approach toward the control of mosquito vectors at early stages.

Expression of CK14 and vimentin in adenomatoid odontogenic tumor and dentigerous cyst.

BACKGROUND: Origin of adenomatoid odontogenic tumor (AOT) has long been a controversy, and the issue of it being a neoplasm or hamartoma was a subject of debate for a long time. Earlier it was grouped under a mixed group of odontogenic tumors considering the varying degrees of inductive changes. Recently, the WHO classification states that the presence of hard tissue within AOT was not due to induction but was rather a metaplastically produced mineralization and hence the tumor was reclassified under a group of tumors arising from odontogenic epithelium. This study is an attempt to identify if both epithelial (cytokeratin 14 [CK14]) and mesenchymal (vimentin) markers are expressed in the follicular and extrafollicular variants of AOT and to compare the expression with dentigerous cyst (DC) as this cyst is known to arise from reduced enamel epithelium which expressed CK14. This is done to possibly relate the origin of AOT with reduced enamel epithelium. AIMS AND OBJECTIVES: To study, analyze and correlate the expression of CK14 and vimentin in AOT and DC. MATERIALS AND METHODS: Retrospective study on paraffin embedded tissues. Sixteen cases of AOT and 15 cases of DC were retrieved from the departmental archives and subjected to CK14 and vimentin immunostaining. STATISTICAL METHODS: Measures of central tendency was used to analyze the results. RESULTS AND OBSERVATIONS: Ninety percent of cases of follicular AOT (FAOT) and 100% cases of extra-follicular AOTs (EAOTs) showed positivity for CK14 and all cases of DC showed positivity for CK14. Vimentin was positive in 44% and negative in 56% cases of both FAOT and EAOT taken together. CONCLUSION: The CK14 expression profile in AOT and DC supports its odontogenic epithelial specific nature. The possible role of reduced enamel epithelium and dental lamina in histogenesis of AOT and DC is strongly evident by their CK14 expression pattern.

Effect of lambda cyhalothrin and temephos on detoxification enzyme systems in Culex quinquefasciatus (Diptera: Culicidae).

Mosquitoes serve as vector for transmitting diseases. Among mosquitoes, Culex quinquefasciatus transmits lymphatic filariasis, yellow fever Japanese encephalitis etc. Application of chemical insecticides is still the best option for vector control programmes. Continuous use of these chemicals on mosquito reduces its effects. The present study determined the baseline susceptibility of Cx. quinquefasciatus in response to λ-cyhalothrin and temephos treatments. In addition, the biochemical mechanisms and zymogram analysis involved in insecticide detoxification among larval mosquitoes were studied. The larval bioassay indicated high LC50 value for λ-cyhalothrin (0.1484ppm) as compared to temephos (0.01092ppm). While AChE assay showed increased activity in temephos treatments, glutathione reductase (GR) and esterase levels were increased at both the treatments. Esterase quantitative analysis revealed the expression of three bands at 43kDa, 67kDa and 245kDa. The findings suggest that insensitivity of AChE, esterase and high GR activity may play an important role in developing resistance to synthetic pyrethroid and organophosphate insecticides in Cx. quinquefasciatus population.

Susceptibility status of Aedes aegypti (L.) (Diptera: Culicidae) to temephos from three districts of Tamil Nadu, India.

BACKGROUND & OBJECTIVES: Dengue is the most rapidly expanding arboviral disease in India. Aedes aegypti is the primary vector of dengue fever. Chemical insecticides have long been used in the vector control programmes along with other control measures. However, continuous use of insecticides targeting Ae. aegypti may lead to development of insecticide resistance. Though resistance in Ae. aegypti has been reported, the mutation in ace-1 gene associated with temephos resistance is not reported in natural populations. The present study aims to evaluate the susceptibility/resistance status of Ae. aegypti to temephos from three districts of Tamil Nadu. METHODS: Ae. aegypti larvae were sampled from different locations in three districts, viz., Dharmapuri, Salem and Namakkal. The standard WHO larval bioassay, biochemical assays and spotting of specific mutation (G119S) in the acetylcholinesterase gene, which is associated with organophosphate resistance, were carried out by PCR and sequencing. RESULTS: The results showed that larvae from Namakkal (NKL) population had an alteration in their susceptibility status (RR = 6.9 fold), while the other populations were moderately susceptible to insecticides. Biochemical assay showed increased activity for α- and ß-esterase in NKL, as well as evidence of acetylcholinesterase insensitivity. G119S mutation was detected in this population with high frequency of 0.24. INTERPRETATION & CONCLUSION: The high activity of esterase, mixed-function oxidase (MFO) and ace-1 mutation frequency were closely associated with temephos resistance. Early detection of resistance alleles in natural vector population could be useful for the successful implementation of insecticide resistance management strategies. The results of this study provide baseline data on temephos resistance in Ae. aegypti populations.

Involvement of metabolic resistance and F1534C kdr mutation in the pyrethroid resistance mechanisms of Aedes aegypti in India.

Pesticide resistance poses a serious problem for worldwide mosquito control programs. Resistance to insecticides can be caused by an increased metabolic detoxification of the insecticide and/or by target site insensitivity. In the present study, we estimated the tolerance of Indian Aedes aegypti populations using adult bioassays that revealed high resistance levels of the field populations to permethrin (RR-6, 5.8 and 5.1 folds) compared to our susceptible population. Enzymatic assays revealed increased activities of glutathione S-transferase and carboxylesterase enzymes in the field populations comparatively to the susceptible population. PBO synergist assays did not confirm that cytochrome P450 monooxygenase metabolic detoxification acted as a major cause of resistance. Hence the role of target site resistance was therefore investigated. A single substitution Phe1534Cys in the voltage gated sodium channel was found in domain III, segment 6 (III-S6) of the resistance populations (allele frequency=0.59, 0.51 and 0.47) suggesting its potential role in permethrin resistance in A. aegypti.

Resistance selection and molecular mechanisms of cypermethrin resistance in red hairy caterpillar (Amsacta albistriga walker).

Amsacta albistriga is one of the important pests of oilseed crops in India. This pest has developed high resistance to organophosphate (OP) insecticide in field. Therefore, cypermethrin insecticide was used as an alternative for this pest. After 20 generations of selection with cypermethrin, the LD50 value for A. albistriga was increased by 21.5-folds. The synergism ratio of piperonyl butoxide (PBO) and triphenyl phosphate (TPP) was increased by 10- and 9.6-fold in resistant strains and comparatively, 3.9 and 4.2-fold in susceptible strains. Detoxification enzyme analysis and native PAGE electrophoresis of esterase isoenzyme further revealed that esterase and mixed function oxidase may be involved in cypermethrin resistance in CypRes strain. In addition to enzyme analysis overexpression of CYP4M44, CYP9A77 and CYP6B47 (ortholog) can confer metabolic resistance in the CypRes strain. These data provide a foundation for further study of cypermethrin resistance mechanism observed in A. albistriga.

Phyllanthus wightianus Müll. Arg.: a potential source for natural antimicrobial agents.

Phyllanthus wightianus belongs to Euphorbiaceae family having ethnobotanical importance. The present study deals with validating the antimicrobial potential of solvent leaf extracts of P. wightianus. 11 human bacterial pathogens (Bacillus subtilis, Streptococcus pneumoniae, Staphylococcus epidermidis, Proteus vulgaris, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella typhimurium, Escherichia coli, Shigella flexneri, Proteus vulgaris, and Serratia marcescens) and 4 fungal pathogens (Candida albicans, Cryptococcus neoformans, Mucor racemosus, and Aspergillus niger) were also challenged with solvent leaf extracts usingagar well and disc diffusion methods. Further, identification of the active component present in the bioactive extract was done using GC-MS analysis. Results show that all extracts exhibited broad spectrum (6-29 mm) of antibacterial activity on most of the tested organisms. The results highlight the fact that the well in agar method was more effective than disc diffusion method. Significant antimicrobial activity was detected in methanol extract against S. pneumoniae (29 mm) with MIC and MBC values of 15.62 µg/mL. GC-MS analysis revealed that 29 bioactive constituents were present in methanolic extract of P. wightianus, of which 9,12-octadecaenioic acid (peak area 22.82%; RT-23.97) and N-hexadecanoic acid (peak area 21.55% RT-21.796) are the major compounds. The findings of this study show that P. wightianus extracts may be used as an anti-infective agent in folklore medicine.