Enrichment of laccase production by Phoma herbarum isolate KU4 under solid-state fermentation by optimizing RSM coefficients using genetic algorithm.

The process parameters were optimized to obtain enhanced enzyme activity from the fungus Phoma herbarum isolate KU4 using rice straw and saw dust as substrate under solid-state fermentation using Response surface methodology (RSM). Genetic algorithm was used to validate the RSM for maximum laccase production. Six variables, viz., pH of the media, initial moisture content, copper sulphate concentration, concentration of tannic acid, inoculum concentration and incubation time were found to be effective and optimized for enhanced production. Maximum laccase production was achieved by RSM at pH 5·0 and 86% of initial moisture content of the culture medium, 150 µmol l-1 of CuSO4 , 1·5% tannic acid and 0·128 g inoculum g-1 dry substrate inoculum size on the fourth day of fermentation. The highest laccase activity was observed as 79 008 U g-1 , which is approximately sixfold enhanced production compared to the unoptimized condition (12 085·26 U g-1 ).

Assessment of DNA Methylation in p15, p16 and E-Cadherin Genes as a Screening Tool for Early Carcinoma Cervix.

Cancer cervix is diagnosed late in women due to anatomical inaccessibility of the area. Hence, a robust screening strategy will help detect carcinoma cervix early which will significantly decrease the mortality and morbidity due to this disease. We evaluated DNA methylation of three tumour suppressor genes p15, p16 and E-Cadherin on cervical smears to assess DNA methylation as a screening tool for detection of early cervical cancer in comparison to PAP smears. DNA was extracted from cervical smears of 20 cases and 30 controls. The DNA was bisulphite modified. Methylation specific PCR (MSP) was performed to assess the methylation status of the promoter region of each of the genes. MSP results were compared with PAP smears to assess the utility of DNA methylation of these genes in screening for cervical cancer. DNA methylation was detected in 55% subjects in p15 gene, 45% in p16 gene and 40% in E-Cadherin gene. This was statistically significant when compared to the controls. DNA methylation of E-Cadherin, and p15 genes as a panel has a sensitivity and specificity of 80% and 90% respectively, which is better than the sensitivity of PAP smear for detection of early cancer cervix. Increased DNA methylation is seen in p15, p16 and E-Cadherin genes in early cancer cervix. p15 and E-Cadherin in combination can be used as a screening tool for detection of early cancer cervix.

PAI-1 Study in Thalassemia Major Patients Receiving Multiple Blood Transfusion.

Thalassemia is a congenital hemolytic disease which is treated by repeated blood transfusion. Chronic iron overload is currently considered to be the primary cause of mortality in ß-thalassemia, mainly due to the induction of left-sided cardiac failure. Iron overload results from a number of mechanisms associated with the disease itself. In addition to chronic iron overload thalassemic patients are more prone for procoagulant status which in turn lead to clinical thrombotic events. The hypercoagulable state in thalassemia is due to multiple elements, a combination of which is often the drive behind a clinical thromboembolic events. PAI-1 study was done in thalassemia major patients receiving multiple blood transfusion as a marker for procoagulant status. Total of 30 thalassemic patients on repeated blood transfusion was included in the study and total of 30 healthy age and sex matched controls were included in the study. It was also found that there was significant differences between cases and controls. The mean level of PAI 1 in controls was 3047 ± 414 pg/ml, the value in cases was 3683 ± 358 pg/ml. The level was significantly increased (p < 0.05) in the cases compared to controls. PAI-1 levels were also compared with the total number of blood transfusion which correlates well.

Comparative study of serum amylase and lipase in acute pancreatitis patients.

To establish utility of single enzymatic marker for the diagnosis of acute pancreatitis. This is a cohort study. Tertiary care centre proven cases of acute pancreatitis (n = 50) admitted in surgery ward between December 2011 and May 2013 were included in the study. Serum amylase and lipase were performed along with many analytes. All relevant data including serum lab values and imaging were collected. All 50 patients included in the study had raised serum lipase, 42 patients had both amylase and lipase raised, 8 patients had amylase normal but lipase raised. In smaller hospitals where limited lab and radiological facilities are available, estimation of serum lipase will be a better choice over serum amylase in diagnosis of acute pancreatitis.

Correlation of Microalbuminuria with Estimated GFR (eGFR) by Cockcroft-Gault and MDRD Formula in Type 2 Diabetics and Hypertensives.

Increase in urine albumin excretion rate (AER) precede a fall in glomerular filtration rate in patients developing diabetic chronic kidney disease (CKD). Our results have shown that 7 (50 %) of diabetic and hypertensive individuals with decreased GFR do not have increased AER. In this cross-sectional study, we measured AER of 75 patients with type 2 diabetes and hypertension by immunoturbidimetric method. We correlated the results with eGFR values obtained by Cockcroft-Gault and MDRD method. The method used was not a compensated method. We measured serum creatinine by modified Jaffe's kinetic method in autoanalyzer XL-600. Analysis of data showed positive correlation between eGFR and microalbuminuria by both the methods with eGFR <60 mL/min/1.73 m(2). Pearson's correlation co-efficient (r) was 0.9 (p = 0.0001) by Cockcroft-Gault formula and 0.69 (p = 0.0063) by MDRD formula. Our results concluded that there was positive correlation between AER and eGFR <60 mL/min/1.73 m(2). We have recognized that these two parameters provide a complimentary benefit in management of cases with CKD.

A pilot study on the contribution of folate gene variants in the cognitive function of ADHD probands.

Genetic abnormalities in components important for the folate cycle confer risk for various disorders since adequate folate turnover is necessary for normal methylation, gene expression and chromosome structure. However, the system has rarely been studied in children diagnosed with attention deficit hyperactivity disorder (ADHD). We hypothesized that ADHD related cognitive deficit could be attributed to abnormalities in the folate cycle and explored functional single nucleotide polymorphisms in methylenetetrahydrofolate dehydrogenase (rs2236225), reduced folate carrier (rs1051266), and methylenetetrahydrofolate reductase (rs1801131 and rs1801133) in families with ADHD probands (N = 185) and ethnically matched controls (N = 216) recruited following the DSM-IV. After obtaining informed written consent for participation, peripheral blood was collected for genomic DNA isolation and PCR-based analysis of target sites. Data obtained was analyzed by UNPHASED. Interaction between sites was analyzed by the multi dimensionality reduction (MDR) program. Genotypic frequencies of the Indian population were strikingly different from other ethnic groups. rs1801133 "T" allele showed biased transmission in female probands (p < 0.05). Significant difference in genotypic frequencies for female probands was also noticed. rs1801131 and rs1801133 showed an association with low intelligence quotient (IQ). MDR analysis exhibited independent effects and contribution of these sites to IQ, thus indicating a role of these genes in ADHD related cognitive deficit.

Serum urea:albumin ratio as a prognostic marker in critical patients with non-chronic kidney disease.

Routine laboratory investigations play an important role in estimating the risk of mortality in intensive care unit (ICU) patients. The significance of urea:albumin ratio (UAR) in predicting the stay and mortality of ICU patients is not known. It is a retrospective study of patients admitted to ICU (n = 412) with non-chronic kidney disease (non-CKD). Receiver-operating characteristics (ROC) analysis for predicting mortality was carried out to find area under curve (AUC) and threshold levels. Analysis of survival probability was carried out by Kaplan-Meier method and Log-rank test. The AUC to predict mortality were 0.695, 0.767 and 0.791 for serum albumin, urea and UAR, respectively. The threshold levels for albumin, urea and UAR were 2.8 g/dL, 53 mg/dL, and 23.44 mg/g, respectively. The highest odds ratio (OR) of 9.75 to predict mortality at threshold level was observed for UAR, while OR were 7.0 and 3.62 for serum urea and albumin, respectively. The serum urea above and albumin below threshold level were associated with increase in ICU stay of >3 days but the highest OR of 4.73 to predict stay of >3 days was observed for UAR. Kaplan-Meier survival analysis shows significant (p < 0.001) difference at the threshold value of UAR. Serum urea and albumin are found to be an independent predictor for the mortality and stay; however an increased UAR value is the best parameter in predicting mortality and stay in ICU patients with non-CKD illness.

Anticancer Effect of dl-Glyceraldehyde and 2-Deoxyglucose in Ehrlich Ascites Carcinoma Bearing Mice and Their Effect on Liver, Kidney and Haematological Parameters.

Cancer cells generally exhibit increased glycolysis for ATP generation (the Warburg effect). Compounds that inhibit glycolysis have potential applications in cancer treatment. dl-glyceraldehyde (DLG) and 2-Deoxyglucose (2-DG) have been proven effective in the inhibition of glucose metabolism. Ehrlich ascites carcinoma (EAC) cells were injected intraperitoneally (i.p) in 10-12 weeks old Swiss albino mice, weighing between 20 and 30 g. The anticancer activity of DLG and 2-DG were determined by tumor volume, tumor weight, viable and nonviable tumor cell count, average survival time, percentage increase in life span and tumor inhibition ratio. The blood samples were obtained for biochemical analysis after 9 days of treatment to study the effect on liver, kidney and haematological parameters. Histopathological examination of liver and kidney was also performed. One-way ANOVA test and Dunnett's test were used for comparisons of parameters in study groups. Both DLG and 2-DG individually decreased the tumor weight, tumor volume, viable tumor cell count and significantly increased the life span of treated mice, however the combination was found to be better. The biochemical parameters of liver and kidney functions and haematological parameters were restored close to control group as compared with the EAC bearing mice. Histopathological examination of liver and kidney in EAC control group showed large areas of necrosis, congestion and mononuclear cell infiltration but such changes were not observed in liver and kidney sections observed after i.p injection of DLG and 2-DG for 9 days. Improvement was much better in the group where combination of these two drugs were used.

Usefulness of Diagnostic Ultrasound for Detection of Common Shoulder Abnormalities Prior to MRI.

Shoulder pain is a common musculoskeletal pain in the general population and results in significant disability, quality of life impairment and financial burden to the health care system. This cross-sectional study was carried out among purposively selected 61 adult patients with shoulder pain and or limited range of motion in the Department of Radiology and Imaging, Mymensingh Medical College Hospital, Mymensingh, Bangladesh from September 2018 to August 2020 to determine the usefulness of USG in detection of common shoulder abnormalities, as an initial imaging modality using MRI as reference standard. The majority of the patients 25(40.98%) were in age group of 51-60 years with mean age of 52.98±10.85 years. In the dectection of rotator cuff pathology, the overall sensitivity, specificity and accuracy of USG for any complete tear were 100.0% each, for any partial tear were 79.71%, 96.57% and 91.80%, for any rotator cuff tear were 83.33%, 96.25% and 91.80%, and for any tendinosis were 90.48%, 99.37% and 96.31% respectively. The sensitivity, specificity and accuracy of USG were 88.23%, 92.59% & 90.16% for long head of biceps tendon sheath effusion, 100.0% each for long head of biceps tendon dislocation, 71.11%, 87.50% and 75.41% for glenohumeral joint effusion, and 58.06%, 96.67% and 77.05% for bursal effusion respectively. From this study, it is concluded that high resolution USG showed high sensitivity, specificity and accuracy compared to MRI in detecting common shoulder abnormalities, and could be considered as the first line imaging modality in the evaluation of shoulder pain.

National TB Control Program of Bangladesh: System Failure and Loss of Effectiveness.

The National Tuberculosis Control Program (NTP) is one of the largest programs run by the state with many successes. However, underreporting is considered a real flaw of the current program. To report the characteristics and outcomes of TB patients registered in an upazila health complex in Kazipur, Sirajganj district, Bangladesh between September 2018 and February 2019 under the control of NTP was the objective of the study. This retrospective cohort study was conducted using routinely collected program data from the aforementioned site. Data retrieved from the hospital record form. Formal permission was obtained from the local authority. Consent statements and ethical aspects were waived due to the retrospective nature of the study. Analysis was performed using SPSS 20.0. A total of 207 tuberculosis cases were included with an average age of presentation of 43 years. Approximately 82.0% had pulmonary tuberculosis (PTB) and the rest had extrapulmonary tuberculosis (EPTB). PTB was more common in males, whereas EPTB was common in females (p=0.01). Of all cases, 84.0% were diagnosed by a positive sputum smear and 16.0% were diagnosed clinically with a negative sputum smear. Attendance at follow-up was 82.12%, 70.04% and 68.59% at months 2, 5 and 6 following index admission respectively. Overall, the cure rate was higher in PTB than EPTB [146 (85.9%) vs. 5 (13.5%), p<0.001). The rate of treatment completion was 25.1% (n=52) and the death count was 1.4% (n=3) [PTB-1.2 (n=2) vs. EPTB-2.7% (n=1)]. A gradual decline in reporting or completion of treatment was observed in this setting. However, a nationwide study is warranted to explore this issue in detail.

Psychological Assessment of Health-Care Workers Working during COVID-19 Pandemic Condition in Bangladesh.

The study was aimed to assess the psychological aspects and relevant factors of the health-care workers (HCWs) working in COVID 19 pandemic condition in Bangladesh. This online cross-sectional survey was conducted from different tertiary, secondary and primary hospitals in Bangladesh. Eligible 638 HCWs who were directly involved in the caring of confirmed or suspected COVID-19 patients were recruited in this study. The mental health was assessed by the Patient Health Questionnare-9 (PHQ-9), Generalized Anxiety Disorder-7 (GAD-7) and Athens Insomnia Scale (AIS). High frequency of depression 536(84.0%), anxiety 386(60.5%) and insomnia 302(47.3%) was found among the HCWs, which were significantly higher in physicians (p<0.001) than nurses. Moderate to severe depression was significantly higher in female, whereas minimal to mild depression was significant in male HCWs (p=0.014). Symptoms of depression (p<0.001), anxiety (p<0.001) and insomnia (p=0.004) were significantly higher among the HCWs of primary and secondary compared to the tertiary level. The HCWs developed psychological trauma due to family health (45.3%) and contagious disease property (66.6%). After adjusting confounders, multivariable logistic regression analysis showed that physicians and HCWs of secondary hospital had significant symptoms of severe depression (OR=2.95, 95% CI=0.50-17.24; p<0.001), anxiety (OR=2.64, 95% CI=0.80-8.72; p<0.001) and insomnia (OR=2.67, 95% CI=1.23-5.84; p=0.018); whereas female HCWs had more risk of developing symptoms of severe insomnia (OR= 1.84; 95% CI=1.23-2.75; p=0.003). High rate of depression, anxiety and insomnia was found among HCWs working in the COVID-19 pandemic condition in this survey.

Sociodemographic and psychopathologic predictors of first incidence of DSM-IV substance use, mood and anxiety disorders: results from the Wave 2 National Epidemiologic Survey on Alcohol and Related Conditions.

The objective of this study was to present nationally representative findings on sociodemographic and psychopathologic predictors of first incidence of Diagnostic and Statistical Manual of Mental Disorders, 4th edn (DSM-IV) substance, mood and anxiety disorders using the Wave 2 National Epidemiologic Survey on Alcohol and Related Conditions. One-year incidence rates of DSM-IV substance, mood and anxiety disorders were highest for alcohol abuse (1.02), alcohol dependence (1.70), major depressive disorder (MDD; 1.51) and generalized anxiety disorder (GAD; 1.12). Incidence rates were significantly greater (P<0.01) among men for substance use disorders and greater among women for mood and anxiety disorders except bipolar disorders and social phobia. Age was inversely related to all disorders. Black individuals were at decreased risk of incident alcohol abuse and Hispanic individuals were at decreased risk of GAD. Anxiety disorders at baseline more often predicted incidence of other anxiety disorders than mood disorders. Reciprocal temporal relationships were found between alcohol abuse and dependence, MDD and GAD, and GAD and panic disorder. Borderline and schizotypal personality disorders predicted most incident disorders. Incidence rates of substance, mood and anxiety disorders were comparable to or greater than rates of lung cancer, stroke and cardiovascular disease. The greater incidence of all disorders in the youngest cohort underscores the need for increased vigilance in identifying and treating these disorders among young adults. Strong common factors and unique factors appear to underlie associations between alcohol abuse and dependence, MDD and GAD, and GAD and panic disorder. The major results of this study are discussed with regard to prevention and treatment implications.

An urban, water-borne outbreak of diarrhoea and shigellosis in a district town in eastern India.

BACKGROUND: In September 2007, the Gayeshpur municipality reported a cluster of cases with diarrhoea. We aimed to identify the causative agent and the source of the disease. METHODS: We defined a case as the occurrence of diarrhoea (> 3 loose stools/day) with fever or bloody stools in a resident of Gayeshpur in September-October 2007. We asked healthcare facilities to report cases, collected stool specimens from patients, constructed an epidemic curve, drew a map and calculated the incidence by age and sex. We also conducted a matched case-control study (58 in each group), calculated matched odds ratio (MOR) and population attributable fraction (PAF), as well as assessed the environment. RESULTS: We identified 461 cases (attack rate: 46/1000 population) and isolated Shigella flexneri (serotype 2a and 3a) from 3 of 4 stool specimens. The attack rate was higher among females (52/1000) and those in the age group of 45-59 years (71/1000). The outbreak started on 22 September, peaked multiple times and subsided on 12 October 2007. Cases were clustered distal to a leaking pipeline that crossed an open drain to intermittently supply non-chlorinated water to taps. The 58 cases and 58 controls were matched for age and sex. Drinking tap water (MOR: 10; 95% CI: 3-32; PAF: 89%), washing utensils in tap water (MOR: 3.7; 95% CI: 1.2-11.3) and bathing in tap water (MOR: 3.5; 95% CI: 1.1-11) were associated with the illness. CONCLUSION: This outbreak of diarrhoea and Shigella flexneri dysentery was caused by contamination of tap water and subsided following repair of the pipeline. We recommended regular chlorination of the water and maintenance of pipelines.

Conversion from mycophenolate mofetil to enteric-coated mycophenolate sodium in renal transplant recipients with gastrointestinal tract disorders.

OBJECTIVE: The objective of this prospective study was to evaluate the effect of conversion from mycophenolate mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) in renal transplant recipients with gastrointestinal tract (GI) reverse effects using patient-reported outcomes instrument. METHODS: A multicenter, open-label, prospective study was undertaken in renal transplant recipients treated with MMF. In patients experiencing GI tract symptoms, treatment was changed to equimolar EC-MPS (myfortic). At baseline and visit 2 (4-6 weeks after baseline), patients completed the Gastrointestinal Symptom Rating Scale (GSRS) and Gastrointestinal Quality of Life Index (GIQLI), and physicians completed the Overall Treatment Effect (OTE) scale at visit 2. A difference of 0.5 or greater in the OTE score is indicative of clinical relevance. RESULTS: Of 154 patients screened, 118 fulfilled the inclusion or exclusion criteria. Eighty-five men and 33 women with a mean age of 41.6 years participated in this study. Median time since transplantation was 12 months. Mean (SD) dose of MMF reported at baseline was 1209.4 (422.89) mg/d. More than 50% of patients reported MMF-associated nausea, dyspepsia, and abdominal pain. After conversion to an equimolar dose of EC-MPS, patients showed improvement in GI symptoms. This benefit was predominantly observed in patients with moderate to severe symptoms at baseline. On the GSRS, patients reported a significant (P < .05) reduction in symptom burden across all parameters (reflux, 36%; diarrhea, 38%; indigestion, 36%; constipation, 28%; and abdominal pain, 40%). On the GIQLI also, significant (P < .05) improvement was reported (symptoms, 18%; emotional status, 22%; physical functioning, 21%, and use of medical treatment, 18%). On the OTE scale, 84.7% of patients reported improvement in GI symptoms. CONCLUSION: In patients with moderate to severe GI symptoms, changing treatment from MMF to EC-MPS significantly reduces GI-related symptom burden and improves GI-specific quality of life.

Assessment of the impact of pesticide residues on microbiological and biochemical parameters of tea garden soils in India.

The main aim of this study was to assess the impact of pesticidal residues on soil microbial and biochemical parameters of the tea garden soils. The microbial biomass carbon (MBC), basal (BSR) and substrate induced respirations (SIR), beta-glucosidase activity and fluorescein diacetate hydrolyzing activity (FDHA) of six tea garden soils, along with two adjacent forest soils (control) in West Bengal, India were measured. The biomass and its activities and biochemical parameters were generally lower in the tea garden soils than the control soils. The MBC of the soils ranged from 295.5 to 767.5 micro g g(- 1). The BSR and SIR ranged from 1.65 to 3.08 mu g CO2-C g(- 1) soil h(- 1) and 3.08 to 10.76 micro g CO2-C g(- 1)h(- 1) respectively. The beta-glucosidase and FDHA of the soils varied from 33.3 and 76.3 micro g para-nitrophenol g(- 1) soil h(- 1) and 60.5 to 173.5 micro g fluorescein g(- 1)h(- 1)respectively. The tea garden soils contained variable residues of organophosphorus and organochlorine pesticides, which negatively affected the MBC, BSR, SIR, FDHA and beta -glucosidase activity. Ethion and chlorpyriphos pesticide residues in all the tea garden soils varied from 5.00 to 527.8 ppb and 17.6 to 478.1 ppb respectively. The alpha endosulfan, beta endosulfan and endosulfan sulfate pesticide residues in the tea garden soils ranged from 7.40 to 81.40 ppb, 8.50 to 256.1 ppb and 55 to 95.9 ppb respectively. Canonical correlation analysis shows that 93% of the total variation was associated with the negative impact of chlorpyriphos, beta and alpha endosulfan and endosulfan sulfate on MBC, BSR and FDHA. At the same time ethion had negative impact on SIR and beta-glucosidase. Data demonstrated that the pesticide residues had a strong impact on the microbial and biochemical components of soil quality.

Risk Factors for Coronary Artery Disease in Indians.

BACKGROUND: A case control study was carried out to study the emerging risk factors for coronary artery disease in Indians. METHODS: The diagnosis of coronary artery disease was based on correlation of clinical, biochemical, electrocardiography, echocardiography, treadmill testing and coronary arteriography findings. The study comprised 100 cases of coronary artery disease (acute coronary syndrome and chronic coronary artery disease) and 100 controls in two tertiary care service hospitals. The subjects were evaluated for total plasma homocysteine, insulin, C-reactive protein (CRP), lipoprotein fibrinogen and anti-chlamydial anti-bodies. RESULT: Male to female ratio was 10:1 in study group with similar predominance of males in controls. Mean age of the cases was 47 years (range 25-59 years) and that of controls was 43 years (range 23-56 years). 64% cases had acute coronary syndrome and 34% had chronic coronary artery disease. In the coronary artery disease population, 76% cases had hyperhomocyteinemia, 9% hyperinsulinaemia, 11% abnormal CRP values, 23% abnormal lipoprotein (a) levels, 40% IgG anti-chlamydial anti-bodies and only 11% had Ig M anti-chlamydial antibodies. In the control population, 72% had hyperhomocystinaemia and 6% hyperinsulinaemia while 23% and 9% controls had IgG and IgM anti chlamydial antibodies respectively. In control group 19% cases had abnormal lipoprotein(a) levels and only 2% had abnormal C reactive protein values. Significant correlation of CAD was seen with CRP values and Ig G anti-chlamydial antibodies. Both the study group and controls had higher homocysteine levels than that observed in some Indian and Western studies. CONCLUSION: High C reactive protein levels and Ig G anti-chlamydial antibodies are associated with coronary artery disease in Indians. Insulin, lipoprotein A, fibrinogen, lgM anti-chlamydial antibodies and higher levels of total plasma homocysteine have no significant association with coronary artery disease.

RING finger-dependent ubiquitination by PRAJA is dependent on TGF-beta and potentially defines the functional status of the tumor suppressor ELF.

In gastrointestinal cells, biological signals for transforming growth factor-beta (TGF-beta) are transduced through transmembrane serine/threonine kinase receptors that signal to Smad proteins. Smad4, a tumor suppressor, is often mutated in human gastrointestinal cancers. The mechanism of Smad4 inactivation, however, remains uncertain and could be through E3-mediated ubiquitination of Smad4/adaptor protein complexes. Disruption of ELF (embryonic liver fodrin), a Smad4 adaptor protein, modulates TGF-beta signaling. We have found that PRAJA, a RING-H2 protein, interacts with ELF in a TGF-beta-dependent manner, with a fivefold increase of PRAJA expression and a subsequent decrease in ELF and Smad4 expression, in gastrointestinal cancer cell lines (P < 0.05). Strikingly, PRAJA manifests substantial E3-dependent ubiquitination of ELF and Smad3, but not Smad4. Delta-PRAJA, which has a deleted RING finger domain at the C terminus, abolishes ubiquitination of ELF. A stable cell line that overexpresses PRAJA exhibits low levels of ELF in comparison to a Delta-PRAJA stable cell line, where ELF expression is high compared to normal controls. The alteration of ELF and/or Smad4 expression and/or function in the TGF-beta signaling pathway may be induced by enhancement of ELF degradation, which is mediated by a high-level expression of PRAJA in gastrointestinal cancers. In hepatocytes, half-life (t(1/2)) and rate constant for degradation (k(D)) of ELF is 1.91 h and 21.72 min(-1) when coupled with ectopic expression of PRAJA in cells stimulated by TGF-beta, compared to PRAJA-transfected unstimulated cells (t(1/2) = 4.33 h and k(D) = 9.6 min(-1)). These studies reveal a mechanism for tumorigenesis whereby defects in adaptor proteins for Smads, such as ELF, can undergo degradation by PRAJA, through the ubiquitin-mediated pathway.

Molecular evolution and functional characterisation of haplotypes of an important rubber biosynthesis gene in Hevea brasiliensis.

Hydroxy-methylglutaryl coenzyme-A synthase (HMGS) is a rate-limiting enzyme in the cytoplasmic isoprenoid biosynthesis pathway leading to natural rubber production in Hevea brasiliensis (rubber). Analysis of the structural variants of this gene is imperative to understand their functional significance in rubber biosynthesis so that they can be properly utilised for ongoing crop improvement programmes in Hevea. We report here allele richness and diversity of the HMGS gene in selected popular rubber clones. Haplotypes consisting of single nucleotide polymorphisms (SNPs) from the coding and non-coding regions with a high degree of heterozygosity were identified. Segregation and linkage disequilibrium analysis confirmed that recombination is the major contributor to the generation of allelic diversity, rather than point mutations. The evolutionarily conserved nature of some SNPs was identified by comparative DNA sequence analysis of HMGS orthologues from diverse taxa, demonstrating the molecular evolution of rubber biosynthesis genes in general. In silico three-dimensional structural studies highlighting the structural positioning of non-synonymous SNPs from different HMGS haplotypes revealed that the ligand-binding site on the enzyme remains impervious to the reported sequence variations. In contrast, gene expression results indicated the possibility of association between specific haplotypes and HMGS expression in Hevea clones, which may have a downstream impact up to the level of rubber production. Moreover, haplotype diversity of the HMGS gene and its putative association with gene expression can be the basis for further genetic association studies in rubber. Furthermore, the data also show the role of SNPs in the evolution of candidate genes coding for functional traits in plants.

Additional synthesis on thiophene-containing trisubstituted methanes (TRSMs) as inhibitors of M. tuberculosis and 3D-QSAR studies.

We earlier reported thiophene-containing trisubstituted methanes (TRSMs) as novel cores carrying anti-tubercular activity, and identified S006-830 as the phenotypic lead with potent bactericidal activity against single- and multi-drug resistant clinical isolates of Mycobacterium tuberculosis (M. tb). In this work, we carried out additional synthesis of several TRSMs. The reaction scheme essentially followed the Grignard reaction and Friedel-Crafts alkylation, followed by insertion of a dialkylaminoethyl chain. We also performed microbiological evaluations including in vitro screening against the virulent strain M. tb H37Rv, cytotoxicity assessment in the Vero C-1008 cell line, and 3D-QSAR studies with comparative molecular field analysis (CoMFA) and comparative molecular similarity index analysis (CoMSIA). CoMFA and CoMSIA models yielded good statistical results in terms of q2 and r2 values, suggesting the validity of the models. It was concluded that a para-substituted benzene ring with bulkier electron-donating groups and aminoalkyl chains are required for higher inhibitory capacity against M. tuberculosis. We believe that these insights will rationally guide the design of newer, optimal, TRSMs.