Veena Garg

Antihyperglycemic, antihyperlipidemic and antioxidative potential of Prosopis cineraria bark

Alloxan administration in male Swiss albino mice, induced diabetes by increasing blood glucose concentration and reducing hepatic glycogen content as compared to normal control group. Besides, serum lipid profile parameters such as total-cholesterol, triglyceride, low-density lipoprotein and very low-density lipoprotein-cholesterol were also elevated, whereas, the level of high-density lipoprotein-cholesterol was reduced significantly (P<0.05) in diabetic mice. Treatment of diabetic animals with crude ethanolic extract of bark of Prosopis cineraria (P. cineraria) for 45 days, significantly lowered blood glucose level, elevated hepatic glycogen content and maintained body weight and lipid-profile parameters towards near normal range. Declined activity of antioxidant enzymes and concentration of non-enzymatic antioxidants were also normalized by drug treatment, thereby reducing the oxidative damage in the tissues of diabetic animals and hence indicating the anti-diabetic and antioxidant efficacy of the extract.

Analysis of imidacloprid residues in fruits, vegetables, cereals, fruit juices, and baby foods, and daily intake estimation in and around Lucknow, India

A total of 250 samples-including fruits, fruit juices, and baby foods (50 samples each), vegetables (70 samples), and cereals (30 samples)-were collected from Lucknow, India, and analyzed for the presence of imidacloprid residues. The QuEChERS (quick, easy, cheap, effective, rugged, and safe) method of extraction coupled with high-performance liquid chromatographic analysis were carried out, and imidacloprid residues were qualitatively confirmed by liquid chromatography-mass spectrometry. Imidacloprid was not detected in samples of fruit juices and baby foods. It was, however, detected in 38 samples of fruits, vegetables, and cereals, which is about 15.20% of the total samples. Of samples of fruits, 22% showed the presence of imidacloprid, and 2% of samples showed residues above the maximal residue limit. Although imidacloprid was detected in 24% of vegetable samples, only 5.71% showed the presence of imidacloprid above the maximal residue limit. However, 33% of cereal samples showed the presence of imidacloprid, and about 3% of samples were above the maximal residue limit. The calculated estimated daily intake ranged between 0.004 and 0.131 µg/kg body weight, and the hazard indices ranged from 0.007 to 0.218 for these food commodities. It is therefore indicated that lifetime consumption of vegetables, fruits, fruit juices, baby foods, wheat, rice, and pulses may not pose a health hazard for the population of Lucknow because the hazard indices for imidacloprid residues were below one.

Disposition and acute toxicity of imidacloprid in female rats after single exposure.

Single dose of imidacloprid (IMI-20mg/kg bodyweight) was orally administered in female rats. Its disposition along with two metabolites 6-chloro nicotinic acid (6-CNA) and 6-hydroxy nicotinic acid (6-HNA) was monitored in organs (brain, liver, kidney, and ovary) and bodily fluids (blood, urine) at 6, 12, 24 and 48h and faeces at 24 and 48h. Maximum concentration (Cmax) of IMI and metabolites in each organ and bodily fluid occurred after 12h. Area under curve (AUC) of IMI ranged from 35 to 358μg/ml/h; 6-CNA: 27.12-1006.42μg/ml/h and 6-HNA: 14.98-302.74μg/ml/h in different organs and bodily fluids. Clearance rate of IMI was maximum in ovary followed by kidney, liver, brain, faeces, blood and urine. Percent inhibition of acetyl-cholinesterase (AChE) was comparable in brain and Red Blood Cells (RBC) at 6-48h which suggests the RBC-AChE as valid biomarker for assessing IMI exposure. It is evident that IMI was absorbed, metabolized, and excreted showing increased level of serum enzymes like Glutamic oxaloacetic transaminase (GOT), Glutamic pyruvic transaminase (GPT) and biochemical constituents like billirubin and Blood Urea Nitrogen (BUN) at 48h. These data suggest that IMI is widely distributed, metabolized and induced toxicology effects at 20mg/kg bodyweight to female rats.

Development of Polymeric Nanopaclitaxel and Comparison with Free Paclitaxel for Effects on Cell Proliferation of MCF-7 and B16F0 Carcinoma Cells

Paclitaxel is hydrophobic in nature and is recognized as a highly toxic anticancer drug, showing adverse effects in normal body sites. In this study, we developed a polymeric nano drug carrier for safe delivery of the paclitaxel to the cancer that releases the drug in a sustained manner and reduces side effects. N-isopropylacrylamide/ vinyl pyrrolidone (NIPAAm/VP) nanoparticles were synthesized by radical polymerization. Physico- chemical characterization of the polymeric nanoparticles was conducted using dynamic light scattering, transmission electron microscopy, scanning electron microscopy and nuclear magnetic resonance, which confirmed polymerization of formulated nanoparticles. Drug release was assessed using a spectrophotometer and cell viability assays were carried out on the MCF-7 breast cancer and B16F0 skin cancer cell lines. NIPAAm/ VP nanoparticles demonstrated a size distribution in the 65-108 nm range and surface charge measured -15.4 mV. SEM showed the nanoparticles to be spherical in shape with a slow drug release of ~70% in PBS at 38° over 96 h. Drug loaded nanoparticles were associated with increased viability of MCF-7 and B16F0 cells in comparison to free paclitaxel. Nano loaded paclitaxel shows high therapeutic efficiency by sustained release action for the longer period of time, i increasing its efficacy and biocompatibility for human cancer therapy. Therefore, paclitaxel loaded (NIPAAm/VP) nanoparticles may provide opportunities to expand delivery of the drug for clinical selection.

Effect of extraction solvents on polyphenolic composition and antioxidant, antiproliferative activities of Himalyan bayberry (Myrica esculenta)

Himalyan bayberry (Myrica esculenta) fruit in Indian Himalayan region was evaluated for total polyphenol contents, antioxidant, and anticancer activities in 4 different solvent systems namely 80% each of methanol, acidicmethanol, acetone, and acidic-acetone to ensure the optimum recovery. Himalyan bayberry acidic-acetone (MeAA) extracts showed highest recovery of phenolics, flavonoids, freeradical scavenging, and ferric-reducing activities followed by acetone (MeA), acidic-methanol (MeAM), and methanol (MeM) extracts. Both MeA and MeAA extracts showed potent anticancer activities leading to 70–92% reduction in the viability of C33A, SiHa, and HeLa cells while exhibiting no cytotoxicity towards normal transformed cell lines. The RP-HPLC analysis revealed abundance of gallic acid [793.74 mg/100 g of fruit weight (f.w.) in MeA], myricetin (345.6 mg/100 g f.w. in MeAA), caffeic acid (246.6 mg/100 g f.w. in MeM), catechin (190.181 mg/100 g f.w. in MeAA) while traces of chlorogenic acid (17.67 mg/100 g f.w. in MeAA), transcinnamic acid (5.91 mg/100 g f.w. in MeA), p-coumaric acid (5.84 mg/100 g f.w. in MeA), and ellagic acid (2.21 mg/100 g f.w. in MeA) corroborating the high antioxidant and anticancer activities of Himalyan bayberry fruit extracts.

Consortium of Phosphate-solubilizing Bacteria and Fungi for Promotion of Growth and Yield of Chickpea (Cicer arietinum)

The combined inoculation of two or more microbial species, such as phosphate-solubilizing bacteria (PSB) and arbuscular mycorrhizal fungus (AMF) and PSB and nitrogen fixers, has often been reported to exert positive effect on growth and yield of various crops; however, reports on dual inoculation of PSB and free-living phosphate-solubilizing fungi (PSF) are hardly available, which makes this study relevant. P-solubilization efficiency (SE) was highest for Bacillus sp.; SE values for RM-2 were highest on the 2nd day of inoculation, whereas those for Aspergillus niger S-36 continuously increased up to the 5th day of incubation. We performed an experiment to evaluate the co-inoculation effect of two phosphate-solubilization-efficient strains of bacteria and fungi in a pot trial. RM-2 strain produced chitinase that might have conferred biocontrol properties on chickpea. The overall growth of plants via dual inoculation was significantly (P = 0.05) greater than that for control and single inoculations in a pot trial, indicating the positive synergistic effect of co-inoculation of PSB and PSF for crop improvement, which could be useful for farmers and sustainable agriculture.

Expression pattern of pluripotent markers in different embryonic developmental stages of buffalo (Bubalus bubalis) embryos and putative embryonic stem cells generated by parthenogenetic activation.

In this study, we describe the production of buffalo parthenogenetic blastocysts and subsequent isolation of parthenogenetic embryonic stem cell (PGESC)-like cells. PGESC colonies exhibited dome-shaped morphology and were clearly distinguishable from the feeder layer cells. Different stages of development of parthenogenetic embryos and derived embryonic stem cell (ESC)-like cells expressed key ESC-specific markers, including OCT-4, NANOG, SOX-2, FOXD3, REX-1, STAT-3, TELOMERASE, NUCLEOSTEMIN, and cMYC. Immunofluorescence-based studies revealed that the PGESCs were positive for surface-based pluripotent markers, viz., SSEA-3, SSEA-4, TRA 1-80, TRA 1-60, CD-9, and CD-90 and exhibited high alkaline phosphatase (ALP) activity. PGEC cell-like cells formed embryoid body (EB)-like structures in hanging drop cultures and when cultured for extended period of time spontaneously differentiated into derivatives of three embryonic germ layers as confirmed by RT-PCR for ectodermal (CYTOKERATIN8, NF-68), mesodermal (MSX1, BMP-4, ASA), and endodermal markers (AFP, HNF-4, GATA-4). Differentiation of PGESCs toward the neuronal lineage was successfully directed by supplementation of serum-containing media with retinoic acid. Our results indicate that the isolated ESC-like cells from parthenogenetic blastocyst hold properties of ESCs and express markers of pluripotency. The pluripotency markers were also expressed by early cleavage-stage of buffalo embryos.

Quantitative expression of pluripotency-related genes in parthenogenetically produced buffalo (Bubalus bubalis) embryos and in putative embryonic stem cells derived from them.

Parthenogenetically produced embryos and embryonic stem (ES) cells derived from them offer a unique model for investigating the role of transcription factors in embryonic genome activation (EGA), pluripotent lineage specification and in pluripotency and self-renewal of ES cells because of the unique nature of these embryos. There is little information on the quantitative expression of important genes in parthenogenetically produced embryos and in ES cells derived from them. The present study examined the quantitative expression of some important genes in parthenogenetically produced buffalo embryos and in putative parthenogenetic ES cells (pES) cells. The quantitative expression of OCT-4, SOX-2, NANOG, REX-1, FOXD-3 and NUCLEOSTEMIN, which is very low in immature and mature oocytes, and in embryos at 2-, 4- and 8- to 16-cell stage, increases significantly at morula and blastocyst stage. The expression level of TELOMERASE, c-MYC and STAT-3, which is high in immature oocytes decreases during embryonic development followed by either an increase at the morula stage (TELOMERASE) or a low expression level maintained throughout development till blastocyst stage (c-MYC and STAT-3). There is a progressive decline in the expression level of OCT-4, SOX-2, c-MYC, REX-1, NUCLEOSTEMIN, TELOMERASE and STAT-3 during long term culture of pES cells.

Comparative purification and characterization of two distinct extracellular monocrotophos hydrolases secreted by Penicillium aculeatum and Fusarium pallidoroseum isolated from agricultural fields.

The present study aimed at a comparative characterization of two distinct extracellular monocrotophos hydrolases, from Penicillium aculeatum ITCC 7980.10 (M3) and Fusarium pallidoroseum ITCC 7785.10 (M4), isolated from agricultural fields. The MCP hydrolases were purified by Sephadex G-100 column and DEAE-Sepharose CL-6B ion-exchange column followed by SDS–PAGE analysis, which showed the presence of two hydrolases, of 33 and 67 kDa respectively. Both enzymes were most active at alkaline pH and were stable over a wide range of temperatures (60–70 °C). Between the strains, the MCP hydrolases from M3 were 2-fold more active than that from M4. Enzyme kinetic studies showed lowest K m (33.52 mM) and highest V max (5.18 U/mg protein) for OPH67 of M3 in comparison to the K m and V max of the other hydrolases purified from M3 and M4, suggesting that M3 OPH67 was the most efficient MCP hydrolase. To the best of our knowledge, this is the first report of the purification of two distinct extracellular thermostable MCP hydrolases from fungal strains Penicillium aculeatum ITCC 7980.10 and Fusarium pallidoroseum ITCC 7785.10. Owing to its potential MCP hydrolyzing activity, M3 OPH67 can perhaps used directly or in the encapsulated form for remediation of MCP contaminated sites.

Proteomic Investigation of Photorhabdus Bacteria for Nematode-Host Specificity.

Majority of animals form symbiotic relationships with bacteria. Based on the number of bacterial species associating with an animal, these symbiotic associations can be mono-specific, relatively simple (2–25 bacterial species/animal) or highly complex (>102–103 bacterial species/animal). Photorhabdus (family-Enterobacteriaceae) forms a mono-specific symbiotic relationship with the entomopathogenic nematode Heterorhabditis. This system provides a tractable genetic model for animal-microbe symbiosis studies. Here, we investigated the bacterial factors that may be responsible for governing host specificity between nematode and their symbiont bacteria using proteomics approach. Total protein profiles of P. luminescens ssp. laumondii (host nematode- H. bacteriophora) and P. luminescens ssp. akhurstii (host nematode- H. indica) were compared using 2-D gel electrophoresis, followed by identification of differentially expressed proteins by MALDI-TOF MS. Thirty-nine unique protein spots were identified - 24 from P. luminescens ssp. laumondii and 15 from P. luminescens ssp. akhurstii. These included proteins that might be involved in determining host specificity directly (for e.g. pilin FimA, outer membrane protein A), indirectly through effect on bacterial secondary metabolism (for e.g. malate dehydrogenase Mdh, Pyruvate formate-lyase PflA, flavo protein WrbA), or in a yet unknown manner (for e.g. hypothetical proteins, transcription regulators). Further functional validation is needed to establish the role of these bacterial proteins in nematode-host specificity.