Inactivation of Mad2B Enhances Apoptosis in Human Cervical Cancer Cell Line upon Cisplatin-Induced DNA Damage

Mad2B (Mad2L2), the human homolog of the yeast Rev7 protein, is a regulatory subunit of DNA polymerase ζ that shares sequence similarity with the mitotic checkpoint protein Mad2A. Previous studies on Mad2B have concluded that it is a mitotic checkpoint protein that functions by inhibiting the anaphase-promoting complex/cyclosome (APC/C). Here, we demonstrate that Mad2B is activated in response to cisplatin-induced DNA damage. Mad2B co-localizes at nuclear foci with DNA damage markers, such as proliferating cell nuclear antigen and gamma histone H2AX (γ-H2AX), following cisplatin-induced DNA damage. However, unlike Mad2A, the binding of Mad2B to Cdc20 does not inhibit the activity of APC/C in vitro. In contrast to Mad2A, Mad2B does not localize to kinetochores or binds to Cdc20 in spindle assembly checkpoint-activated cells. Loss of the Mad2B protein leads to damaged nuclei following cisplatin-induced DNA damage. Mad2B/Rev7 depletion causes the accumulation of damaged nuclei, thereby accelerating apoptosis in human cancer cells in response to cisplatin-induced DNA damage. Therefore, our results sug-gest that Mad2B may be a critical modulator of DNA damage response.

Salivary AsHPX12 influences pre-blood meal associated behavioral properties in Anopheles stephensi

Background & objectives: A successful blood meal acquisition process by an adult female mosquito is accomplished through salivary glands, which releases a cocktail of proteins to counteract the vertebrate host’s immune homeostasis. Here, we characterize a salivary-specific Heme peroxidase family member HPX12, originally identified from Plasmodium vivax infected salivary RNAseq data of the mosquito Anopheles stephensi. Methods: To demonstrate we utilized a comprehensive in silico and functional genomics approach. Results: Our dsRNA-mediated silencing experiments demonstrate that salivary AsHPX12 may regulate pre-blood meal-associated behavioral properties such as probing time, probing propensity, and host attraction. Altered expression of the salivary secretory and antennal proteins expression may have accounted for salivary homeostasis disruption resulting in the unusual fast release of salivary cocktail proteins and delayed acquisition of blood meal in the AsHPX12 knockdown mosquitoes. We also observed a significant parallel transcriptional modulation in response to blood feeding and P. vivax infection. Interpretation & conclusion: With this work, we establish a possible functional correlation of AsHPX12 role in the maintenance of salivary physiological-homeostasis, and Plasmodium sporozoites survival/ transmission, though the mechanism is yet to unravel.

Autoimmune Sclerosing Cholangitis in Children: A Prospective Case-Control Study / 대한소아소화기영양학회지

Purpose@#In children overlap of autoimmune hepatitis (AIH) and primary sclerosing cholangitis is labelled as autoimmune sclerosing cholangitis (ASC). The only prospective pediatric study showed a high prevalence of ASC by using endoscopic retrograde cholangiopancreatography. Aims of our study were to find the prevalence of ASC by using magnetic resonance cholangiography (MRC) in AIH and in non-AIH cirrhosis and to compare clinical presentation and outcome of AIH and ASC. @*Methods@#Prospectively we did MRC in 38 children with AIH (cases) and 19 disease controls (Wilson disease). Multiple biliary strictures with proximal dilatation on MRC were taken as definitive changes of ASC. Detail clinical, laboratory parameters, liver histopathology and treatment outcome were recorded. @*Results@#The median age of cases was 11.5 (3–18) years, 22 (57.9%) were girls and 28 (73.7%) were diagnosed as type 1 AIH. MRC was done in 11 children (28.9%) at the time of diagnosis and in 27 (71.1%) after a median follow-up of 2.5 (0.3–10) years. Abnormal MRC changes were seen in 14/38 (36.8%) of AIH and 8/19 (42.1%) of controls. However, definite changes of ASC were present in four (10.5%) children in AIH and none in controls. None of the clinical, laboratory, histological parameters and treatment response were significantly different between ASC and AIH groups. @*Conclusion@#The prevalence of ASC in children with AIH was just 10.5%. We suggest MRC in select group with cholestatic features, inflammatory bowel disease and in those who showed poor response to immunosuppression instead of all children with AIH.

Autoimmune Sclerosing Cholangitis in Children: A Prospective Case-Control Study / 대한소아소화기영양학회지

Purpose@#In children overlap of autoimmune hepatitis (AIH) and primary sclerosing cholangitis is labelled as autoimmune sclerosing cholangitis (ASC). The only prospective pediatric study showed a high prevalence of ASC by using endoscopic retrograde cholangiopancreatography. Aims of our study were to find the prevalence of ASC by using magnetic resonance cholangiography (MRC) in AIH and in non-AIH cirrhosis and to compare clinical presentation and outcome of AIH and ASC. @*Methods@#Prospectively we did MRC in 38 children with AIH (cases) and 19 disease controls (Wilson disease). Multiple biliary strictures with proximal dilatation on MRC were taken as definitive changes of ASC. Detail clinical, laboratory parameters, liver histopathology and treatment outcome were recorded. @*Results@#The median age of cases was 11.5 (3–18) years, 22 (57.9%) were girls and 28 (73.7%) were diagnosed as type 1 AIH. MRC was done in 11 children (28.9%) at the time of diagnosis and in 27 (71.1%) after a median follow-up of 2.5 (0.3–10) years. Abnormal MRC changes were seen in 14/38 (36.8%) of AIH and 8/19 (42.1%) of controls. However, definite changes of ASC were present in four (10.5%) children in AIH and none in controls. None of the clinical, laboratory, histological parameters and treatment response were significantly different between ASC and AIH groups. @*Conclusion@#The prevalence of ASC in children with AIH was just 10.5%. We suggest MRC in select group with cholestatic features, inflammatory bowel disease and in those who showed poor response to immunosuppression instead of all children with AIH.

Prevalence of headache among patients with psychotic disorder: a cross sectional cohort study

Prevalence of headache among patients with psychotic disorder: a cross sectional cohort study. Aims-We plan thisstudy to explore the prevalence of headache among psychotic disorders. Methods : It was a cross sectional observational study. All subjects were assessed for inclusion – exclusion criteria, and on qualification they were requested to fill up Socio-demographic data sheet or asked verbally and filled up by investigators. Results-A total of 245 subjects with mean age of 35.38 years (± 5.17 years) with mean duration of psychotic illness of 2.32 ± 0.54 Years, 53.87% males and mostly belonging to hindu religion 80.40% and 64.08 %were unemployed.There was headache among 189 (77.14 %) of the sample, Migraine consisted 22.85%,Tension headache consisted 43.26 %, Cervicogenic headache 7.75% and remaining was others or unspecified types of headache 3.26%. Conclusions-This study finds a prevalence of 77.14 % of headache among psychotic patients, that includes migraine among 22.85%; tension headache 43.26%; cervicogenic 7.75 % amd others or unspecified 3.26%.

Synergistic Approach Of Ayurvedic Principles And Life Style Modifications To Combat Psychosomatic Disorders

The modern world has come to a challenge where most of the untreated conditions are resulting into complications and are burdened over society. To fetch all these, physicians are trying to limit such conditions by applying various scientific theories and methods to explain the basic underlying mechanism of health and diseases. Understanding of physical conditions through psychological approach is the mainstream of such conditions and is termed as Somatoform disorders or psychosomatic illnesses as explained by World Health Organization in international Statistical Classification of Diseases and Related Health Problems. Stress is the contributing factor that doesn't allow the individual to remain in harmony with the surroundings and threatens the body's ability to maintain homeostasis. Dietary changes, mind-body therapies including meditation, stress relaxation techniques and Yoga can bring harmony back to individual suffering from such disorders.

Comparative Study Of Different Anand-bhairav Rasa And Its Standarization (w.s.r. To Bhaishajya Ratnavali)

Introduction: Standardization of Ayurvedic drug is an important criteria for selection, processing, efficacy & safety wise, to meet the WHO guidelines for the world wide acceptability of Ayurvedic formulations. Therefore different preparations of Anandbhairav rasa from classic Bhaishajya Ratnavali has been selected and studied for standariztion. Materials & Methods: Three samples of Anand Bhairav Rasa were prepared according to Jwaratisar prakaran of Bhaishajya Ratnavaliand three samples according to Atisar Prakaran of Bhaishajya Ratnavali were made and subjected to various physico-chemical analyses so that their physical as well as chemical changes can be analyzed. Conclusion: Both formulations showed the difference in pharmaceutically, organoleptic examination as well as in chemical analysis. However, the results obtained from physico chemical analysis of all the three samples are very close together and within fixed physico-chemical norms as described in pharmacopeial standards for Ayurvedic formulations.

Rationality Behind Fast Acting Drugs In Ayurveda

Ayurveda like any other life sciences, has its integrity and potential to understand and determining the cure of rather potential serious and emergency conditions. There are numerous examples of such clinical success stories mentioned in our ancient texts ranging from various surgical and Para-surgical operations done by Acharya Sushruta during emergency, to the treatment of the patients who are nearby death due to intake of certain poisons incidentally or accidentally, or due to any other chronic medical illness such as fever, Rakta-pitta, Kshataj-kasa, Grahni, Ura-Kshtaja etc., due to excessive blood loss or to any Marmabhighata (injury to vital organs). All these require a highly skilled health care system which is beneficial not only for its preventing aspects but a curative one too and need of fast acting substances or procedures. Most of people who have certain experiences or knowledge of Ayurveda commonly say that Ayurvedic medicines or treatments are slower in action. Is it true ? if yes, why they are so? If not, then what is truth?

Level of PAX5 in differential diagnosis of non-Hodgkin’s lymphoma.

Background & objectives: The PAX5, a paired box transcription factor and B-cell activator protein (BSAP), activates B-cell commitment genes and represses non-B-cell lineage genes. About 14 transcript variants of PAX5 have been observed in human. Any alteration in its expression pattern leads to lymphogenesis or associated diseases and carcinogenesis in non-lymphoid tissues. Its mechanisms of function in pathophysiology of non-Hodgkin’s lymphoma (NHL) are unclear. This study was intended to explore influence of PAX5 in cascade of NHL pathogenesis and diagnosis. Methods: Samples of 65 patients were evaluated by immunohistochemical staining for cellular localization of PAX5, CD19, CD3, cABL, p53, Ras and Raf and by TUNEL assay, RNA-isolation and reverse transcriptase (RT)-PCR, Western blot analysis, and lactate dehydrogenase (LDH) specific staining. Results: B-cell type NHL patients were positive for PAX5, p53, Ras, CD19, Raf and CD3. All of them showed TUNEL-positive cells. The differential expression pattern of PAX5, CD19, p53, CD3, ZAP70, HIF1α, Ras, Raf and MAPK (mitogen-activated protein kinase) at the levels of transcripts and proteins was observed. The LDH assay showed modulation of LDH4 and LDH5 isoforms in the lymph nodes of NHL patients. Interpretation & conclusions: The histological observations suggested that the patients represent diverse cases of NHL like mature B-cell type, mature T-cell type and high grade diffuse B-cell type NHL. The findings indicate that patients with NHL may also be analyzed for status of PAX5, CD19 and ZAP70, and their transcriptional and post-translational variants for the differential diagnosis of NHL and therapy.

Transcriptome analysis of Anopheles stephensi embryo using expressed sequence tags.

Germ band retraction (GBR) stage is one of the important stages during insect development. It is associated with an extensive epithelial morphogenesis and may also be pivotal in generation of morphological diversity in insects. Despite its importance, only a handful of studies report the transcriptome repertoire of this stage in insects. Here, we report generation, annotation and analysis of ESTs from the embryonic stage (16–22 h post fertilization) of laboratoryreared Anopheles stephensi mosquitoes. A total of 1002 contigs were obtained upon clustering of 1140 high-quality ESTs, which demonstrates an astonishingly low transcript redundancy (12.1%). Putative functions were assigned only to 213 contigs (21%), comprising mainly of transcripts encoding protein synthesis machinery. Approximately 78% of the transcripts remain uncharacterized, illustrating a lack of sequence information about the genes expressed in the embryonic stages of mosquitoes. This study highlights several novel transcripts, which apart from insect development, may significantly contribute to the essential biological complexity underlying insect viability in adverse environments. Nonetheless, the generated sequence information from this work provides a comprehensive resource for genome annotation, microarray development, phylogenetic analysis and other molecular biology applications in entomology.

Biological activity predictions, crystallographic comparison and hydrogen bonding analysis of cholane derivatives.

A total of eighteen molecules of cholane derivatives (I-XVIII) (a series of steroids) have been included to predict their pharmacological effects, specific mechanisms of action, known toxicities, drug-likeness, etc, by using the statistics of multilevel neighbourhoods of atoms (MNA) descriptors for active and inactive fragments. The biological activity spectra for substances have been correlated on SAR base (structure-activity relationships data and knowledge base), which provides the different P(a) (possibility of activity) and P(i) (possibility of inactivity). Most of the probable activities have been characterized by P(a) and P(i) values, which depict that all the molecules have high value of teratogen activity. The Lipinski's thumb rule predicts that all the cholane derivatives have stronger preponderance for "cancer-like-drug" molecules and some of their related analogous have entered in the ANCI (American National Cancer Institute) database. Some selected bond distances and bond angles of interest have been taken into account and deviation of bond distances/bond angles, vis-a-vis the substitutional group and X-H...A intra/intermolecular hydrogen bonds has been discussed in detail. X-H...A intra and intermolecular hydrogen bonds in the molecules have been described with the standard distance and angle cut-off criteria. D-theta and d-theta. scatter plots for intra- and intermolecular interactions are presented for better understanding of packing interactions existing among these derivatives. There exists only one C-H...O intramolecular bifurcated hydrogen bond. while high tendency of intermolecular bifurcated hydrogen bonds based on a defined O-H...O has been observed, in which O atom acts as a prototype donor as well as acceptor. The frequency of occurrence of C-H...O hydrogen bonds is predominant (i.e. 85.7%) in intramolecular interactions, whereas in intermolecular interactions, frequency of occurrence for O-H...O interactions is 62.9%. Solvent-solute/solute-solvent interactions have also been investigated to understand more complicated processes that occur for biomolecules in aqueous solutions. The number of hydrogen donors in each derivative is less than 5, except for molecule XVIII and which has 91.3% of drug-likeness, instead of observed range of 96.5-99.39%.

Identification and characterization of a new putative c-type lysozyme from malaria vector Anopheles stephensi.

Lysozyme (E.C. 3.2.1.17) activity is reported from the malaria vector Anopheles stephensi. The activity was detected in the salivary gland and midgut using bacteriolytic radial diffusion assay. Spectrophotometric analysis indicated that higher level of lysozyme activity was maintained in both midgut and salivary gland tissues. The activity reached the highest level in 4-8 days old mosquitoes. Genomic PCR amplification revealed the presence of at least two putative lysozyme genes in the mosquito genome. Preliminary analysis of one of the 413 bp genomic fragments showed 56% identity to the lysozyme of mosquito A. gambiae. However, the nature and origin of the putative cloned lysozyme gene remains elusive.