Pharmacophore modelling, atom-based 3D-QSAR generation and virtual screening of molecules projected for mPGES-1 inhibitory activity.

COX-2 inhibitors exhibit anticancer effects in various cancer models but due to the adverse side effects associated with these inhibitors, targeting molecules downstream of COX-2 (such as mPGES-1) has been suggested. Even after calls for mPGES-1 inhibitor design, to date there are only a few published inhibitors targeting the enzyme and displaying anticancer activity. In the present study, we have deployed both ligand and structure-based drug design approaches to hunt novel drug-like candidates as mPGES-1 inhibitors. Fifty-four compounds with tested mPGES-1 inhibitory value were used to develop a model with four pharmacophoric features. 3D-QSAR studies were undertaken to check the robustness of the model. Statistical parameters such as r2 = 0.9924, q2 = 0.5761 and F test = 1139.7 indicated significant predictive ability of the proposed model. Our QSAR model exhibits sites where a hydrogen bond donor, hydrophobic group and the aromatic ring can be substituted so as to enhance the efficacy of the inhibitor. Furthermore, we used our validated pharmacophore model as a three-dimensional query to screen the FDA-approved Lopac database. Finally, five compounds were selected as potent mPGES-1 inhibitors on the basis of their docking energy and pharmacokinetic properties such as ADME and Lipinski rule of five.

Egg production performance, egg yolk antioxidant profile and excreta concentration of corticosterone in golden pheasants (Chrysolophus pictus) fed diets containing different levels of green vegetables.

In order to study the effect of feeding graded levels of green vegetables on egg production performance and egg yolk antioxidant status, 27 female golden pheasants (GP) were randomly distributed into three groups of nine each in an experiment based on completely randomized design. The diets of the birds in groups T1 , T2 and T3 contained 1.4%, 2.7% and 5.0% of green vegetables respectively. Feeding of experimental diets started on 12th February (day 1) and was continued till 30 June 2012. Average number of eggs laid and egg mass produced by the hens in group T3 was higher (p < 0.004) than those of T1 and T2 . Hen day egg production was lowest (p < 0.001) in group T1 . Roche yolk colour score was highest (p < 0.01) in T3 followed by T2 and was lowest in T1 . Other external and internal egg quality parameters were similar among the groups. Total antioxidant capacity (TAC) of egg yolk was higher (p < 0.04) in group T3 as compared to other two groups. Egg yolk concentration of total carotenoids, ß-carotene, total sulfhydryl, protein-bound sulfhydryl and non-protein-bound sulfhydryl was higher (p < 0.05), whereas concentration of malondialdehyde was lower (p < 0.024) in group T3 as compared to other two groups. Excreta concentration of corticosterone was highest (p < 0.012) in T1 , followed by T2 , and was lowest in T3 . Egg yolk concentration of TAC and TSH was positively and that of malondialdehyde was negatively correlated with carotenoids intake. Excreta concentration of corticosterone was negatively correlated with carotenoids intake. It was concluded that incorporation of green vegetables in the diet at 5% on dry matter basis would improve egg production, egg yolk antioxidant status and the ability of the captive GP to combat stress.

A case of medulloblastoma in adult patient affected by anaplastic oligoastrocytoma.

Medulloblastomas and high-grade gliomas (HGG) are two distinct brain tumor, with different peculiarities in terms of age of onset, localizations and prognosis. The coexistence of the two neoplasms in the same adult patient is an extremely rare event. We present the case of a woman treated with radio-chemotherapy for an HGG, who developed a cerebellar medulloblastoma 7 years later. Considering the poor prognosis of these tumors, the lack of knowledge about the mechanisms of onset as well as effective therapies, it is necessary to determine the exact role of irradiation and the presence of any potential molecular genetic abnormalities in the developing of the two tumors.

Radiation therapy outcomes in muscle invasive urinary bladder cancer: A single institution experience.

INTRODUCTION: To audit the survival outcomes and loco-regional control in muscle invasive urinary bladder cancer patients treated with external beam radiation therapy (RT). MATERIALS AND METHODS: From November 2008 through December 2011, 50 consecutively diagnosed muscle invasive urinary bladder carcinoma (T2-4a N0-2, M0) patients were included in this retrospective study. All these patients received external beam RT to a median dose of 60 Gy (range 30-66 Gy), and were not suitable for radical surgery due to patients' preference or medical comorbidities. A stepwise procedure using proportional hazard regression was used to identify prognostic factors with respect to survival. RESULTS: Completion trans-urethral resection of bladder tumor was done in 38 (76%) patients of the cohort and 47 (94%) had transitional cell carcinoma on histopathology. Clinical stage T2 was diagnosed in 40 (80%) patients. The median follow-up for the entire cohort was 14 ± 8.9 months (range 1-36 months). In conclusion, 24 patients (48%) were free of disease, 5 patients (10%) had residual disease, and 13 patients (26%) had died of disease. Two-year and 3 year overall survival of intact bladder for the entire cohort was 58% and 43.6%, respectively. Cox regression modeling strongly suggested clinical stage (P = 0.01) and RT dose (P = 0.001) as being predictors for overall survival. CONCLUSION: RT shows reliable outcomes and excellent compliance in this advanced disease. Prescribing a higher RT dose could potentially correlate to better intact bladder control rates while maintaining good quality of life in selected patients.

Influence of dietary protein on serum metabolites and antioxidant status: A study in Chrysolophus amherstiae.

This experiment was conducted to study the effect of feeding graded levels of dietary crude protein (CP) on serum biochemical profile of Lady Amherst's pheasants (LAP). Eighteen male LAP were randomly distributed into three groups of six each in an experiment based on completely randomized design. The CP content of the diets was 13.4%, 16.5%, and 19.1%, in groups I, II, and III, respectively. Serum concentrations of uric acid was lowest (P < 0.05) in group I. Relationship between serum concentration of uric acid and nitrogen intake was linear (R(2) = 0.39, P < 0.01). Concentrations of other serum metabolites and enzymes were similar among the groups. Serum concentration of triiodothyronine (T3 ) was highest (P < 0.05) in group I, followed by groups II and III. Total antioxidant capacity (TAC) of serum was lower (P < 0.007), whereas serum concentration of malondialdehyde (MDA) was higher (P < 0.001) in group I as compared to groups II and III. Regression of serum concentration of TAC (R(2) = 0.74, P < 0.01) and MDA (R(2) = 0.39, P < 0.05) was polynomial. Heterophil to lymphocyte ratio was higher (P < 0.007) in group I as compared to groups II and III. Relationship between H/L ratio and nitrogen intake was polynomial (R(2) = 0.69, P < 0.05). Cell mediated immune response measured as foot web index was similar among the groups. Based upon the results, it was concluded that a diet containing 16.5% crude protein would be optimum for improving antioxidant defense and the ability of Lady Amherst's pheasant to combat stress. Zoo Biol. 35:346-354, 2016. © 2016 Wiley Periodicals, Inc.

Transverse myelitis due to Staphylococcus aureus may occur without contiguous spread.

STUDY DESIGN: A case report of staphylococcal transverse myelitis. OBJECTIVES: To illustrate the clinical presentation of acute transverse myelitis due to Staphylococcus aureus, without a contiguous source of infection. SETTING: National Neuroscience Institute. CASE REPORT: A 79-year-old female was diagnosed with acute transverse myelitis. Clues to an infectious etiology included fever, raised inflammatory markers and cerebrospinal fluid neutrophilic pleocytosis. Staphylococcal etiology was established based on cerebrospinal fluid and blood cultures. Despite extensive investigations, no contiguous or systemic source of infection could be identified. She was treated with appropriate antibiotics; however, neurological recovery was poor. CONCLUSIONS: Bacterial myelitis may occur in isolation and the diagnosis should not be discounted when evaluation shows an absence of a contiguous or systemic source of infection.

Superior vena cava obstruction (SVCO) in patients with advanced non small cell lung cancer (NSCLC).

INTRODUCTION: To report on prognostic and treatment factors influencing the response of SVCO and related survival outcomes in advanced non small cell lung cancer. MATERIAL AND METHODS: From November 2008 through December 2011, 18 consecutively diagnosed NSCLC patients with SVCO were included in this study. The patient, tumor and treatment related factors were analyzed. Median overall survival (OS), Kaplan -Meier survival plots, T-test, Cox Proportional Hazards models were generated by multiple covariates (MVA) and analyzed on SPSS software (version 19.0; SPSS, Inc., Chicago, IL). RESULTS: Thirteen patients (72%) had presented with SVCO before the pathological diagnosis of underlying lung malignancy, while 5 (28%) progressed to SVCO after initiating treatment with chemotherapy. Twelve (68%) patients achieved subjective relief from the obstruction at the completion of palliative radiation therapy. Treating oncologists preferred 4 Gy per fraction in 11 (62%), while the median biologically equivalent dose delivered was 28 Gy. Six (33%) patients received chemotherapy during the course of treatment. Median OS of the entire cohort was 3±1.85mths and 1-year survival rate of 7%. Univariate analysis confirmed that SVCO patients with good performance score (p=0.02), and partial response to chemotherapy (p= 0.001) have superior OS. However, Cox regression modeling for MVA demonstrated only good performance SVCO patients (p = 0.05) have a better OS. CONCLUSION: RT effectively relieves SVCO but overall poor survival associated in our clinical scenario needs to be improved with multimodality approach. Adjuvant chemotherapy is to be considered after initial radiation therapy in good performance patients. KEYWORDS: superior vena cava obstruction (SVCO), radiation therapy, chemotherapy, non small cell lung cancer (NSCLC), BED (biologically equivalent dose).

Characterisation and in silico analysis of interleukin-4 cDNA of nilgai (Boselaphus tragocamelus) and Indian buffalo (Bubalus bubalis).

Interleukin-4 (IL-4) produced from Th2 cells modulates both innate and adaptive immune responses. It is a common belief that wild animals possess better immunity against diseases than domestic and laboratory animals; however, the immune system of wild animals is not fully explored yet. Therefore, a comparative study was designed to explore the wildlife immunity through characterisation of IL-4 cDNA of nilgai, a wild ruminant, and Indian buffalo, a domestic ruminant. Total RNA was extracted from peripheral blood mononuclear cells of nilgai and Indian buffalo and reverse transcribed into cDNA. Respective cDNA was further cloned and sequenced. Sequences were analysed in silico and compared with their homologues available at GenBank. The deduced 135 amino acid protein of nilgai IL-4 is 95.6% similar to that of Indian buffalo. N-linked glycosylation sequence, leader sequence, Cysteine residues in the signal peptide region, and 3' UTR of IL-4 were found to be conserved across species. Six nonsynonymous nucleotide substitutions were found in Indian buffalo compared to nilgai amino acid sequence. Tertiary structure of this protein in both species was modeled, and it was found that this protein falls under 4-helical cytokines superfamily and short chain cytokine family. Phylogenetic analysis revealed a single cluster of ruminants including both nilgai and Indian buffalo that was placed distinct from other nonruminant mammals.

Production of nuclear transfer embryos by using somatic cells isolated from milk in buffalo (Bubalus bubalis).

Somatic cells in milk are a potential source of nuclei for nuclear transfer to produce genetically identical animals; this is especially important in animals that are susceptible to risks of bacterial infection on biopsy collection. In this study, a minimum of 10 milk samples were collected from each of the three buffaloes representing Murrah breed. All the samples were processed immediately and cell colonies were obtained. Cell colonies from one buffalo (MU-442) survived beyond 10 passages and were evaluated by fluorescence microscopy and used in nuclear transfer experiments. In culture, these cells expressed vimentin, indicating they were of fibroblast origin similar to ear cells. We compared the effectiveness of cloning using those milk-derived fibroblast (MDF) cells and fibroblast cells derived from the ear derived fibroblast (EDF). Fusion and cleavage rates of MDF-NT and EDF-NT embryos were found to be similar (92.43 ± 1.28% vs 94.98 ± 1.24%, and 80.27 ± 1.75% vs 84.56 ± 3.73%, respectively; p > 0.01); however, development to blastocyst stage and total cell number was higher for EDF-NT embryos (50.24 ± 2.54%, 227.14 ± 13.04, respectively, p < 0.01), than for MDF-NT embryos (16.44 ± 0.75%, 170.57 ± 4.50 respectively). We conclude that somatic cells from milk can be cultured effectively and used as nucleus donor to produce cloned blastocyst-stage embryos.

Antineoplastic potential of medicinal plants.

Cancer is one of the most dreaded diseases worldwide and the incidence is on the rise in both developing and developed countries. It is treated by chemotherapy, radiotherapy and surgery. In spite of advances in treatment strategies, cancer still remains a major cause of death. Research is on for development of better drugs which are more effective and simultaneously cause fewer side effects. Plants have been used for the treatment of various ailments of man and animals since ages. They are being screened extensively to explore the possibility of development of economically viable anticancer drugs. Natural products of plant origin currently constitute a considerable proportion of commercially available antineoplastic drugs. This review gives an insight into commercially available anticancer drugs of plant origin and also patents granted to plant derived components, extracts and polyherbal formulations possessing anticancer activity. The exhaustive work reviewed here on antineoplastic activity of various plants both in vitro and experimental models throughout the world will help design further research in this field.

Anti-tumor activity of Aloe vera against DMBA/croton oil-induced skin papillomagenesis in Swiss albino mice.

Human populations are increasingly exposed to various carcinogens such as chemicals, radiation, and viruses in the environment. Chemopreventive drugs of plant origin are a promising strategy for cancer control because they are generally nontoxic or less toxic than synthetic che-mopreventive agents, and can be effective at different stages of carcinogenesis. The present investigation was undertaken to explore the antitumor activity of topical treatment with aloe vera (Aloe vera) gel, oral treatment with aloe vera extract, and topical and oral treatment with both gel and extract in stage-2 skin carcinogenesis in Swiss albino mice induced by 7,12-dim ethylbenz(a)anthracene (DMBA) and promoted croton (Croton tiglium) oil. The animals were randomly divided into 4 groups and treated as follows: Group I, DMBA + croton oil only (controls); Group II, DMBA + croton oil + topical aloe vera gel; Group III, DMBA + croton oil + oral aloe vera extract; Group I V, DMBA + croton oil + topical aloe vera gel + oral aloe vera extract. Results showed that body weight was significantly increased from 78.6% in the control group (Group I) to 92.5%, 87.5%, and 90.0% in Groups II, III, and I V, respectively. A 100% incidence of tumor development was noted in Group I, which was decreased to 50%, 60%, and 40% in Groups II, III, and I V, respectively. Also in Groups II, III, and IV, the cumulative number of papillomas was reduced significantly from 36 to 12, 15, and 11; tumor yield from 3.6 to 1.2, 1.5, and 1.1; and tumor burden from 3.6 to 2.4, 2.50, and 2.75, respectively, after treatment with aloe vera. Conversely, the average latent period increased significantly from 4.9 (Group I) to 5.23, 5.0, and 6.01 weeks in Groups II, III, and I V, respectively. We conclude that aloe vera protects mice against DMBA/croton oil-induced skin papillomagenesis, likely due to the chemopreventive activity of high concentrations of antioxidants such as vitamins A, C, and E; glutathione peroxidase; several isozymes of superoxide dismutase; the minerals selenium and zinc; and polysaccharides in aloe vera.

Evaluation of chemopreventive response of two cycloxygenase-2 inhibitors, etoricoxib and diclofenac in rat colon cancer using FTIR and NMR spectroscopic techniques.

Non steroidal anti inflammatory drugs (NSAIDs) are efficacious in chemoprevention of colorectal cancer. Therefore, the potential ability of Etoricoxib, a selective cycloxygenase-2(COX-2) inhibitor and Diclofenac, a preferential COX-2 inhibitor are considered in the chemoprevention of 1, 2-dimethylhydrazine (DMH) induced colon carcinogenesis in rat model. DMH was injected s.c. for six weeks while Etoricoxib and Diclofenac were fed daily orally alone and also in combination with an weekly subcutaneous injection of 1,2-dimethylhydrazine dihydrochloride (DMH) to the rats. After the treatment period of 6 weeks the animals were sacrificed by an overdose of ether anesthesia and the colonic tissues were removed and studied by the FTIR and NMR Spectroscopic techniques to evaluate the changes occurring in the lipid bilayer of colonic membrane lipids. The alterations in wave number of FTIR spectra as well as the chemical shifts of NMR spectra were recorded which signify the modulation of membrane lipids during colon carcinogenesis and possible cancer prevention by the oral administration of NSAIDs in an experimental model of chemical induced colon carcinogenesis.

Lymphoid tissue inducer cells: bridges between the ancient innate and the modern adaptive immune systems.

Phylogeny indicates that adaptive immunity evolved first in diffusely distributed lymphoid tissues found in the lamina propria (LP) of the gut. B follicular structures appeared later, probably initially in isolated lymphoid follicles in the LP and then in organized lymphoid tissues such as lymph nodes and Peyer's patches. The development of these new lymphoid structures was enabled by gene duplication and evolution of new tumor necrosis family members. Here, we argue that lymphoid tissue inducer cells (LTis) had a pivotal role, not only in the development of organized lymphoid structures, but also in the subsequent genesis of the CD4-dependent class-switched memory antibody responses. In this review, we concentrate on the latter function: the sustenance by LTis of CD4 T-cell responses for protective immunity.

Nuclear PTHrP targeting regulates PTHrP secretion and enhances LoVo cell growth and survival.

Parathyroid hormone-related protein (PTHrP) is expressed by human colon cancer tissue and cell lines; expression correlates with colon carcinoma severity. PTHrP is synthesized as a prepro isoform and contains two targeting sequences - a signal sequence and a nuclear localization signal (NLS). The signal peptide (SP) directs PTHrP to the secretory pathway, where it exerts autocrine/paracrine effects. The NLS directs PTHrP to the nucleus/nucleolus, where it exerts intracrine effects. In this study, we used the human colon cancer cell line LoVo as a model system to study the effects of autocrine/paracrine and intracrine PTHrP action on cell growth and survival, hallmarks of malignant tumor cells. We report that PTHrP increases cell growth and survival, protects cells from serum-starvation-induced apoptosis, and promotes anchorage-independent cell growth via an intracrine pathway. Conversely, autocrine/paracrine PTHrP action decreases cell growth and survival. We also show an inverse relationship between secreted and nuclear PTHrP levels, in that cells overexpressing NLS-deleted PTHrP secrete higher PTHrP levels than those overexpressing the wild-type isoform. Conversely, SP deletion results in higher nuclear PTHrP levels. These observations provide evidence of a link between intracrine PTHrP action and cell growth and survival. Targeting PTHrP production in colon cancer may thus prove therapeutically beneficial.

Sindbis virus replicon-based DNA vaccine encoding Rabies virus glycoprotein elicits specific humoral and cellular immune response in dogs.

A Sindbis virus (SINV) replicon-based DNA vaccine encoding Rabies virus (RABV) glycoprotein G developed previously (Saxena et al., Vaccine 26, 6592, 2008) was used for immunization of dogs against rabies. The intradermal injection of DNA vaccine into external ear generated protective level of virus neutralizing antibodies. The cellular immune response was specific to RABV, in particular by an increase in CD3+CD4+ and CD3+CD8+ lymphocytes. This study has demonstrated that the SINV replicon-based DNA vaccine encoding RABV G is capable of inducing the protective level of specific immune response in dogs.

Chemopreventive response of diclofenac, a non-steroidal anti-inflammatory drug in experimental carcinogenesis.

The chemopreventive response was evaluated of nonsteroidal anti-inflammatory drug, Diclofenac, a preferential cyclooxygenase-2 (COX-2) inhibitor in 1,2-dimethyhydrazine (DMH)-induced colon cancer in rat model. The signs of neoplasm were evident in the animals receiving 30mg of DMH per kg body weight in a weekly s.c injection for six weeks. The putative biomarker of carcinogenesis was visible in the form of multiple plaque lesions in DMH treatment and then regression seen in those animals which also received an oral dose of Diclofenac, 8 mg/kg body weight whereas no such macroscopic neoplastic lesions were seen in the animals receiving Diclofenac only or the control animals receiving the vehicle of the drug. Histopathological results showed the presence of early aberrant changes in the form of severe dysplasia and also numerous crypt fissions in the apical surface of the colonic mucosa. A very high expression of COX-2 was seen in the colonic epithelium of DMH-treated rats, as analyzed by immunohistochemistry. Also, the apoptotic events were assessed by terminal deoxynucleotidyl dUTP nick end labeling (TUNEL) assay, where the DMH group shows few number of TUNEL positive cells which dramatically increased in the Diclofenac treatment. The results suggest that Diclofenac could be an effective chemopreventive agent in colon cancer, where perhaps apoptosis plays a very dominant end effect in cancer cell killings.

A DNA vaccine that encodes rabies virus glycoprotein lacking transmembrane domain enhances antibody response but not protection.

Rabies virus (RV) glycoprotein (gp) consists of three domains: cytoplasmic, transmembrane and ectodomain. It occurs in a complete, membrane-bound form within the infected cell, but it is released from them in a deleted, secreted form lacking the transmembrane domain. This study was performed to test the importance of the transmembrane domain for the capability of the RV gp gene, introduced into mice via a recombinant plasmid (DNA vaccine), to induce immune response and protection against challenge. Although the antibody response to the secreted form of gp was higher than that to complete gp, the protective efficacy of the respective DNA vaccine against challenge was not better than that of the DNA vaccine inducing complete gp. This indicates that the transmembrane domain of RV gp is important for generating protection against rabies and should be present in RV DNA vaccines.

Fasciola gigantica cathepsin-L cysteine proteinase in the detection of early experimental fasciolosis in ruminants.

Cathepsin-L cysteine proteinase was purified from Fasciola gigantica regurgitant by two-step alcoholic fractionation, followed by ion-exchange chromatography. The purification strategy was evolved to eliminate other contaminating proteins co-precipitating with the purified proteinase during alcoholic fractionation. The enzyme was stable on long-term storage at -20 degrees C rendering it more suitable for field diagnostic use. The purified cathepsin-L cysteine proteinase was assayed for detection of F. gigantica experimental infection in sheep and buffaloes and could detect infection, as early as 4 weeks post-infection by ELISA, Western blotting and Dipstick ELISA. The 28-kDa cathepsin-L cysteine proteinase seems a promising antigen for the diagnosis of tropical fasciolosis in domestic animals.

Antioxidative effect of Rajgira leaf extract in liver of Swiss albino mice after exposure to different doses of gamma radiation.

The radioprotective effect of Rajgira leaf extract (800 mg/kg b.wt.) was studied in the liver of Swiss albino mice at various post-irradiation intervals between day 1 and 30 after its oral administration for 15 consecutive days prior to whole body gamma irradiation with 6, 8 and 10 Gy of gamma rays. In this study, abnormal and binucleated hepatocytes were counted in both the control and experimental sets because these hepatocytes are good indicators of radiation-induced damage. In the experimental (RLE + irradiation) sets, the percentage of abnormal and binucleated hepatocytes was lower compared with their respective control (irradiation alone) sets at each autopsy interval with all three radiation doses studied. The increase in the percentage of these hepatocytes was also found to be dose-dependent in the control as well as in the RLE treated (experimental) sets. Thus, Rajgira leaf extract (RLE) treatment given before irradiation protects mouse liver against radiation-induced lesions by increasing the GSH content and decreasing the LPO level.

Short-term zinc deficiency in diet induces increased oxidative stress in testes and epididymis of rats.

In order to determine the effects of Zinc deficient diet on oxidative stress in testis and epididymis, various parameters viz: total proteins, lipid peroxidation, hydroperoxides, antioxidant capacity and enzymatic activities are evaluated in rats fed on zinc deficient diet for 2, 4 and 6 weeks. Total proteins, water and lipid solouble antioxidant capacity decreased while lipid peroxidation (TBARS) and hydroperoxides concentration increased in testes, caput and cauda epididymis except in 2ZD (testes) where hydroperoxides revealed a significant decrease. GSH decreased in testes and caput and cauda epididymis. GPx and gamma-GT activities increased in testes and caput and cauda epididymis of zinc deficient rats. Further, GST increased in testes but exhibited decreases after 2 and 4 weeks and an increase after 6 weeks in caput and cauda epididymis of zinc deficient rats. GR activities decreased in testes but it increased in caput and cauda epididymis of zinc deficient rats. Thus, zinc deprivation results in increased sensitivity to oxidative stress. All these may have been as a consequence of increased ROS generation and/or decreased zinc dependent antioxidant processes.