Bioprospecting of pods of Moringa oleifera Lam. as novel antibacterial agent- Supplementary Data

In the treatment of bacterial diseases, increasing resistance to traditional chemotherapeutics has drawn the necessity for substitute remedies. In this context, here, we evaluated the bactericidal activity of pods of Moringa oleifera Lam., an ethno medicinal plant, against eight pathogenic bacterial strains, both Gram positive (Bacillus licheniformis, B. mycoides, B. subtilis and Staphylococcus aureus) and Gram negative (Escherichia coli, Pseudomonas aeruginosa, P. fluorescens and P. putida). Different organic solvent extracts, like ethyl acetate, acetone and alcohol, of pods of M. oleifera were examined for bactericidal activity against test microorganisms. Minimum inhibitory concentration, chromatographic analyses along with infrared spectroscopy, gas chromatography-mass spectroscopy and nuclear magnetic resonance spectroscopy was carried out for chemical characterization of active ingredient responsible for antibacterial activity. Both the Gram positive and Gram negative organisms showed variable sensitivity to different solvent extracts of M. oleifera pods. Ethyl acetate extracts showed maximum antibacterial activity with MIC value ranging from 1.30 to 4.10 mg/mL. IR analysis provided preliminary information about the amines, amides, aromatics and sulphur containing compounds of the active ingredient. GC-MS and NMR analyses indicated the presence of principal bioactive antibacterial compound 2-(benzoylsulfanyl)-1,3- thiazol,4-yl, benzoate with molecular formula C17H11NO3S2 from ethyl acetate extract of M. oleifera pods. The study concludes that the compound 2-(benzoylsulfanyl)-1,3-thiazol,4-yl, benzoate from ethyl acetate extract of pods of M. oleifera possess the antibacterial activity against the tested strains.

Bioprospecting of pods of Moringa oleifera Lam. as novel antibacterial agent

In the treatment of bacterial diseases, increasing resistance to traditional chemotherapeutics has drawn the necessity for substitute remedies. In this context, here, we evaluated the bactericidal activity of pods of Moringa oleifera Lam., an ethno medicinal plant, against eight pathogenic bacterial strains, both Gram positive (Bacillus licheniformis, B. mycoides, B. subtilis and Staphylococcus aureus) and Gram negative (Escherichia coli, Pseudomonas aeruginosa, P. fluorescens and P. putida). Different organic solvent extracts, like ethyl acetate, acetone and alcohol, of pods of M. oleifera were examined for bactericidal activity against test microorganisms. Minimum inhibitory concentration, chromatographic analyses along with infrared spectroscopy, gas chromatography-mass spectroscopy and nuclear magnetic resonance spectroscopy was carried out for chemical characterization of active ingredient responsible for antibacterial activity. Both the Gram positive and Gram negative organisms showed variable sensitivity to different solvent extracts of M. oleifera pods. Ethyl acetate extracts showed maximum antibacterial activity with MIC value ranging from 1.30 to 4.10 mg/mL. IR analysis provided preliminary information about the amines, amides, aromatics and sulphur containing compounds of the active ingredient. GC-MS and NMR analyses indicated the presence of principal bioactive antibacterial compound 2-(benzoylsulfanyl)-1,3- thiazol,4-yl, benzoate with molecular formula C17H11NO3S2 from ethyl acetate extract of M. oleifera pods. The study concludes that the compound 2-(benzoylsulfanyl)-1,3-thiazol,4-yl, benzoate from ethyl acetate extract of pods of M. oleifera possess the antibacterial activity against the tested strains.

Drumstick Mucilage Microspheres For Controlled Release Of Lamivudine: Design, Optimization And In Vitro Evaluation

Objective: The objective of this study was to design and evaluate controlled release mucoadhesive microspheres of lamivudine using mucoadhesive polymers and mucilage. Methods: Mucoadhesive microspheres of lamivudine were formulated by ionic gelation method. The response surface methodology was adapted for optimization of formulation using central composite design (CCD) for two factors at three levels each was employed to study the effect of independent variables, Sodium alginate-drumstick mucilage (X1) and calcium chloride (CaCl2) concentration (X2) on dependent variables, namely drug encapsulation efficiency (DEE) and particle size (PS). Optimized drumstick mucilage mucoadhesive microspheres of lamivudine were obtained by using numerical optimization of desirability approach. The observed microspheres were coincided well with the predicted values by the experimental design. Results: The microspheres formed were spherical in shape, and Particle size (PS) ranged between 681.63-941.57μm. Drug encapsulation efficiency (DEE) was ranged between 69.63-94.56 %. The drug release for an optimized formulation was 96.58 %. The mechanism of drug release from microspheres followed Korsemeyer’s-Peppas and exponential ‘n’ value was greater than 0.45, indicating the drug release was non-fickian i.e., swelling followed by erosion mechanism. Conclusion: This work suggests that mucoadhesive microspheres, an effective drug delivery system for lamivudine, can be prepared using drumstick mucilage in improving the bioavailability of the drug.

Gender Determination Using Peripheral Blood Smear

Background: Sex chromatin is a chromatin mass of 1 micron size usually seen at the periphery of nucleus infemales. The term sex chromatin comprises of two superficially dissimilar structures known as Barr body inepithelial cells, other tissue cells and as Drumstick appearance in polymorphonuclear leucocytes. Aim of thestudy is gender determination by drumstick appearance of neutrophils using Leishman’s stain and Field stain.Materials and Methods: The present study included 60 individuals ranging from 20-50 age group of both thesexes to observe the morphology of sex chromatin in neutrophil by using Leishman’s and Field stain for thegender determination and comparison between the 2 staining procedure, using research microscope.Results: Presence of drumsticks, sessile nodule and non specific appendages like racket, tag, hook, minor lobe,small club, were found in neutrophils. In males 37% of drumstick, 7% of sessile nodule, 20% of racket structure,3% small club, 13% minor lobe, 7% hook, 3% tag, was seen and in females 10% of drumstick, 60% of sessilenodule, 17% of racket structure, 7% small club, 1% minor lobe, 0% hook, 3% tag, was seen and field’s stainshowed better results in appreciating the morphology of neutrophil.Conclusion: Observation on morphology and comparison between 2 different stain presented a moderate agreementin gender determination.

Moringa oleifera Lam. seed extract prevents fat diet induced oxidative stress in mice and protects liver cell-nuclei from hydroxyl radical mediated damage.

High fat diet (HFD) prompts metabolic pattern inducing reactive oxygen species (ROS) production in mitochondria thereby triggering multitude of chronic disorders in human. Antioxidants from plant sources may be an imperative remedy against this disorder. However, it requires scientific validation. In this study, we explored if (i) Moringa oleifera seed extract (MoSE) can neutralize ROS generated in HFD fed mice; (ii) protect cell-nuclei damage developed by Fenton reaction in vitro. Swiss mice were fed with HFD to develop oxidative stress model (HFD group). Other groups were control, seed extract alone treated, and MoSE simultaneously (HS) treated. Treatment period was of 15 days. Antioxidant enzymes with tissue nitrite content (TNC) and lipid peroxidation (LPO) were estimated from liver homogenate. HS group showed significantly higher (P <0.05) superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH) activity, and ferric reducing antioxidant power (FRAP) compared to only HFD fed group. Further, TNC and LPO decreased significantly (P <0.05) in HS group compared to HFD fed group. MoSE also protected hepatocytes nuclei from the hydroxyl radicals generated by Fenton reaction. MoSE was found to be polyphenol rich with potent reducing power, free radicals and hydroxyl radicals scavenging activity. Thus, MoSE exhibited robust antioxidant prospective to neutralize ROS developed in HFD fed mice and also protected the nuclei damage from hydroxyl radicals. Hence, it can be used as herbal medication against HFD induced ROS mediated disorders.

Bioactivity guided fractionation of Moringa oleifera Lam. flower targeting Leishmania donovani.

Leishmaniases is a group of diseases caused by the protozoan parasite belonging to the genus Leishmania. At least 20 species of Leishmania are known to infect humans transmitted by female sandflies, Phlebotomus spp. Leishmania donovani causes visceral leishmaniasis, considered most lethal among the common three forms of leishmaniasis. Lack of appropriate vaccines, emergence of drug resistance and side effects of currently used drugs stress the need for better alternative drugs, particularly from natural sources. Here, we conducted in vitro and in vivo experiments to study the efficacy of different parts of Moringa oleifera Lam. against Leishmania donovani promastigotes. The flower extract of M. oliefera (MoF) was found to be the most potent antileishmanial agent when compared to other parts of the plant like leaf, root, bark and stem. It imparted significant reduction in parasite number in infected macrophages. The bioactivity guided fractionation of MoF showed ethyl acetate fraction (MoE) as the most active and gave significant parasite reduction in the infected macrophages. Further, growth kinetics studies revealed loss of L. donovani promastigotes viability in the presence of MoE in both time and dose dependent manner. In vivo experiment in Balb/c mouse model of leishmaniasis supported the in vitro findings with a remarkable reduction of the parasite burden in both liver and spleen.

Identification and characterization of a potent anticancer fraction from the leaf extracts of Moringa oleifera L.https://docs.google.com/forms/d/1RkQLeYfiMvE7SKcdOXv9Acm5eXRjmvleic5FYjjX1TE/viewform?fbzx=6173639259593088622

Anticancer potential of Moringa oleifera L. extracts have been well established. However, there are no reports on the isolated molecules/fractions from these extracts which are responsible for the anticancer/cytotoxic activity. Thus, in the present study, we explored the same. The n-hexane, chloroform, ethyl acetate, methanol extracts of the M. oleifera leaves and 15 fractions (F1 to F15) of ethyl acetate extract were evaluated for their in vitro and in vivo anticancer activity using Hep-2 cell lines and Dalton’s lymphoma ascites model in mice, respectively. Among the tested samples, the F1 fraction showed potential cytotoxic effect in Hep-2 cell lines with a CTC50 value of 12.5 ± 0.5 µg/ml. In vivo studies with the doses 5 and 10 mg/kg, p.o. demonstrated significant reduction in body weight and increased the mean survival time compared to the control group. These results were also comparable to the standard, 5-Fluorouracil, treated animals. We have also successfully isolated and characterized the anticancer fraction, F1 from the leaves of M. oleifera L.

Sexual Dimorphism in Morphology and Morphometry of Neutrophil Drumsticks.

Introduction: Sex chromatin is a chromatin mass of 1 micron size usually seen at the periphery of nucleus in females. In the literature majority reported its absence inmales while few reported its low incidence inmales. The term ‘sex chromatin’ comprises two superficially dissimilar structures the “Barr body” present in epithelial and other tissue cells and the “Drumstick” of the polymorphonuclear leucocytes. Materials and methods: The present study was conducted to observe the morphology, morphometry and percentage incidence of Drumsticks in the blood neutrophils of 110 individuals ranging from 17-30 age group and both sexes using a calibrated ocular/eye piece micrometer. Results: The percentage incidence of drumsticks including non-specific appendages as well as the total number of true drumsticks in females exceeds that in males. Four different types of nonspecific appendages-sessile nodules, racket structures, minor lobes and small clubs were found in the blood neutrophils along with the drumsticks. A higher percentage of non-specific appendages i.e.minor lobes (46.2%), racket structures (42.3%), and small clubs (11.5%) were observed in males and sessile nodules were found only in females. Conclusion: Observations on morphology, morphometry and percentage incidence of polymorphonuclear drumsticks presented a valuable data on sex differences.