Khawaja Raees Ahmad

RNA-Seq analysis uncovers transcriptomic variations between morphologically similar in vivo- and in vitro-derived bovine blastocysts

BackgroundA valuable tool for both research and industry, in vitro fertilization (IVF) has applications range from gamete selection and preservation of traits to cloning. Although IVF has achieved worldwide use, with approximately 339,685 bovine embryos transferred in 2010 alone, there are still continuing difficulties with efficiency. It is rare to have more than 40% of fertilized in vitro cattle oocytes reach blastocyst stage by day 8 of culture, and pregnancy rates are reported as less than 45% for in vitro produced embryos. To investigate potential influences in-vitro fertilization (IVF) has on embryonic development, this study compares in vivo- and in vitro-derived bovine blastocysts at a similar stage and quality grade (expanded, excellent quality) to determine the degree of transcriptomic variation beyond morphology using RNA-Seq.ResultsA total of 26,906,451 and 38,184,547 fragments were sequenced for in vitro and in vivo embryo pools, respectively. We detected expression for a total of 17,634 genes, with 793 genes showing differential expression between the two embryo populations with false discovery rate (FDR) < 0.05. There were also 395 novel transcribed units found, of which 45 were differentially expressed (FDR < 0.05). In addition, 4,800 genes showed evidence of alternative splicing, with 873 genes displaying differential alternative splicing between the two pools (FDR < 0.05). Using GO enrichment analysis, multiple biological pathways were found to be significantly enriched for differentially expressed genes (FDR < 0.01), including cholesterol and sterol synthesis, system development, and cell differentiation.ConclusionsThus, our results support that IVF may influence at the transcriptomic level and that morphology is limited in full characterization of bovine preimplantation embryos.

Irrational use of antimalarial drugs in rural areas of eastern Pakistan: a random field study

BackgroundPrescription of antimalarial drugs in the absence of malarial disease is a common practice in countries where malaria is endemic. However, unwarranted use of such drugs can cause side effects in some people and is a financial drain on local economies. In this study, we surveyed the prevalence of malaria parasites in humans, and the prevalence of the malaria transmitting mosquito vectors in the study area. We also investigated the use of antimalarial drugs in the local people. We focused on randomly selected rural areas of eastern Pakistan where no malaria cases had been reported since May 2004.MethodsMass blood surveys, active case detection, passive case detection, and vector density surveys were carried out in selected areas of Sargodha district from September 2008 to August 2009. Data pertaining to the quantities and types of antimalarial drugs used in these areas were collected from health centers, pharmacies, and the district CDC program of the Health Department of the Government of the Punjab.ResultsSeven hundred and forty four blood samples were examined, resulting in a Blood Examination Rate (BER) of 3.18; microscopic analysis of blood smears showed that none of the samples were positive for malaria parasites. Investigation of the mosquito vector density in 43 living rooms (bedrooms or rooms used for sleeping), 23 stores, and 32 animal sheds, revealed no vectors capable of transmitting malaria in these locations. In contrast, the density of Culex mosquitoes was high. Substantial consumption of a variety of antimalarial tablets, syrups, capsules and injections costing around 1000 US

Fetal neurohistopathology of chloropyrifos in mice

, was documented for the region.ConclusionUse of antimalarial drugs in the absence of malarial infection or the vectors that transmit the disease was common in the study area. Continuous use of such drugs, not only in Pakistan, but in other parts of the world, may lead to drug-induced side effects amongst users. Better training of health care professionals is needed to ensure accurate diagnoses of malaria and appropriate prescription of antimalarial drugs delivered to communities.

Histological and micrometric effects of diazinon exposure on adrenal medulla and cortex in mice

Chlorpyrifos (CPF) was tested for feto-neuro-histopathological manifestations on fetal central nervous system (CNS) in mice at 3 maternally sub-toxic oral doses - 0, 9 and 18 mg/kg. Each dose group was further categorized as: single (gestation day (GD) 6) and triple exposures (9, and 12 respectively). Fetuses were exteriorized on GD18. No obvious signs of toxicity were seen in the dams at these exposures. Mean fetal weight showed a dose (9 and 18 mg/kg) and exposure (single and triple) dependent decrease compared to that of the 0 mg/kg group while the litter size remained unaffected. The neurohistopathological abnormalities include vacuolations of the medullary region along with cortical lesions in CNS in 9 and 18 mg/kg groups on triple exposure only. These neurohistopathological manifestations were considered as the indicatives of neuroglial cells necrosis apoptosis. Our findings suggest that gestational exposure of CPF at motherly safe dose levels in mice induce neuroglial cells apoptosis in fetal CNS.

Effects of diazinon on the ovarian micro-anatomical and micrometric parameters of pregnant mice

Histopathological effects of Diazinon (DZ) were studied on adrenal gland in virgin young female mice divided into five groups (10 animals each): (1) Control (untreated); (2) and (3) 9 and 18 mg kg−1 single exposure groups (given distilled water on day 1 and 2 and DZ on day 3); (4) and (5) 9 and 18 mg kg−1 multiple exposure groups (given respective DZ doses on days 1–3). All doses were applied by gavage. Animals were euthanized by cervical dislocation on day 6. Adrenals were removed surgically for histological and micrometric assessments of medulla and cortex. Histological examination of adrenal medulla revealed apoptotic changes and granular depletion in chromaffin cells while cortex gave impression of increased thickness of Zona Fasciculata following multiple DZ exposures. Mean cell sizes in cortical and medullary regions showed dose and exposure dependent significant decline. In conclusion, DZ exposure caused granular depletion in chromaffin cells indicating a rapid release of adrenaline and increased fascicular thickness indicating an anticipated shift from adrenal sex steroids to glucocorticoids biosynthesis and release. Apoptotic changes and significant decline in mean cell sizes of cortical and medullary regions were considered as the outcomes of oxidative and biosynthetic stresses of DZ exposure.

Species Composition and Population Dynamics of Spider Fauna of Trifolium and Brassica Field

Pregnant mice (10 in 4 groups each) were exposed to diazinon (DZN) at 10 and 20 mg/kg on gestation day 6 (GD 6), 9 and 12. A group of 10 pregnant animals was maintained as untreated control. The ovaries were exteriorized on GD 18 by euthanizing the animals. The tissues were processed for histopathological and micrometric analysis. Oocytic nuclear derangements in terms of size increase followed by leakage of nuclear contents with increased duration of exposure were seen. Follicular and luteal cells also showed necrosis. The morphometric data showed significant increase in oocyte and para-follicular size (P_0.0001) along with similar increase in nuclear sizes (P_0.001). These findings indicate that DZN is toxic to bring about histopathological and micrometric derangements in the ovaries of the treated pregnant females. Key words : Pesticides, diazinon, toxic, gestation, ovaries, micro-anatomical, micrometric, derangements.