Promoter Hypermethylation of Tumor suppressor Genes correlates with Tumor grade and Invasiveness in patients with Urothelial Bladder Cancer

Purpose: To investigate the promoter methylation status at selected loci which encode for key proteins involved in apoptosis, DNA repair, cell cycle control and progression in urothelial cell carcinoma of bladder and compare the findings from tissue samples with that of plasma. Methods: Total genomic DNA was isolated from 43 non-muscle invasive (low grade) and 33 muscle invasive (high grade) urothelial bladder cancer samples along with 10 control cases of normal bladder mucosa. Promoter methylation status was investigated for RASSF1A, APC, MGMT, CDKN2A and CDKN2B genes using real-time methylation-specific PCR with SYBR® green. Plasma samples from 16 patients with muscle invasive high grade bladder cancer were also subjected to similar analyses. Results: Promoter hypermethylation was frequently observed in RASSF1A, APC and MGMT gene promoters (p-value \u3c 0.001). The methylation was more prominent in the muscle invasive high grade bladder cancer when compared to non-muscle invasive low grade group (p-value \u3c 0.001) and normal bladder mucosa (p-value \u3c 0.05). The RNA expression of RASSF1A, APC and MGMT was also found to be decreased in the muscle-invasive high grade bladder cancer when compared to the non muscle invasive low grade group (p-value \u3c 0.05). RASSF1A, MGMT and CDKN2A showed comparable results when data from 16 plasma samples was compared to the corresponding tissue samples. Conclusion: Our results suggest that epigenetic silencing of RASSF1A, APC and MGMT genes is strongly associated with invasive high grade urothelial bladder cancer. Thus, status of promoter methylation has the potential to serve as valuable tool for assessing aggressiveness of urothelial cell carcinoma of bladder

Are inert glasses really inert?

Abstract Objectives The aim of this study was to investigate the degradation of inert glass fillers which are commonly used in conventional resin-based composites to provide radiopacity, reduce the polymerization shrinkage and improve the mechanical properties. Methods 75 mg of five different glass powders (1 µm) was immersed separately into 50 mL of acetic acid (pH 4) and tris buffer (pH 7.4) for up to 4 weeks. At each time point the glass powder was filtered and dried for characterization using ATR-FTIR and XRD to assess the degradation behavior and crystallization. ICP-OES, ISE and pH measurements were performed on the supernatant solutions to monitor the pH and ion release. Results Although FTIR and XRD analysis showed no significant glass degradation or crystallization upon immersion, there was a substantial release of ions from the inert fillers, especially from BABFG and CDL. Barium release for these fillers were 270 and 165 ppm respectively. G018–373 glass presented the lowest ion release followed by GM27884 and BABG. The ion release was more pronounced in acidic conditions compared to neutral conditions apart from the fluoride release. Significance Inert glasses are not as inert as previously thought. This may result in leaching of ions, potentially causing toxicity, reduction in mechanical properties, increased wear and subsequent failure of the composite material. The ions released from the inert glass may interfere with other glass fillers such as bioactive glass fillers, inhibiting degradation of the bioactive glass, beneficial ion release from the bioactive glass, pH neutralization and apatite formation

The Serological Findings of Parvo virus B19 and Neopterin Detection among Sickle cell Disease Patients and Blood Doners in the Kingdom of Bahrain

Introduction. Parvovirus B19 (PV B19) is a small, non-enveloped, ss DNA virus with an icosahedral capsid having a size of 18–26 nm. PV B19 transmits through respiratory droplets, blood transfusion and nosocomial infections that have also been documented recently. The virus targets the actively dividing Erythroid Progenitor Cells (EPCs) that are found in the human bone marrow, fetal liver and human umbilical cord. Methods. The study was particularly conducted on Sickle Cell Disease (SCD) patients and focuses on the determination of parvovirus B19 among Bahraini population by relying on their clinical status. The serological study of PV B19 was performed using Enzyme Linked Immunosorbent Assay (ELISA) technique and includes 150 SCD patientsand 100 healthy blood donors in which both males and females were employed. The samples were taken from the emergency unit of Salmania Medical Complex (SMC) and Ibrahim Khalil Kano Center (IKKC). Results. Of the 150 SCD patients, 100 were with vaso-occlusive crisis(VOC) and 50 non-vaso-occlusive crisis (NVOC). The three groups showed significantly higher percentages of PV B19 IgG but the percentage in SCD was relatively high compared to the control group of age-matched healthy donors – 70% of the VOC patients, 76% of the NVOC cases and 57% of blood donors were found to be IgG sero-positive. Discussion and Conclusion. PV B19 is a pathogenic virus and sometimes considered as life-threatening specifically for those individuals who have SCD due to which a risk of transient aplastic crisis increases. This virus is only associated with those patients whohave some hematological disorders such as hemolytic anemia and erythro-cytopenia. An effective screening test must be performed in the future to reduce the risk of PV B19 infection. Keywords: Aplastic Crisis, Seroprevalence, Bahraini Population, Genotypes, Vasoocclusive crisis, Neopteri

Antimicrobial, hemolytic and thrombolytic activities of some new N-substituted-2-({5-[(1E,3E)F-4-(1,3-benzodioxol- 5-yl)-1,3-butadienyl]-1,3,4-oxadiazol-2-yl}sulfanyl) propanamides

Purpose: To synthesize and evaluate the bioactivity of some N-substituted-2-({5-[(1E,3E)-4-(1,3- benzodioxol-5-yl)-1,3-butadienyl]-1,3,4-oxadiazol-2-yl}sulfanyl)propanamides (9a-o) of the alkaloid piperine (1) extracted from P. nigrum (black pepper).Methods: Extract 1 was subjected to basic hydrolysis to obtain piperic acid (2). The heterocyclic 1,3,4- oxadiazole ring was synthesized from 2 through the formation of an ester and carbohydrazide. A series of electrophiles, 8a-o, were synthesized in the presence of 10 % Na2CO3. The final compounds, 9a-o, were synthesized by stirring 5-[(1E,3E)-4-(1,3-benzodioxol-5-yl)-1,3-butadienyl]-1,3,4-oxadiazol-2-thiol (5) and 8a-o in LiH/DMF. Spectral analysis was performed using infrared (FTIR), proton nuclear magnetic resonance (1H-NMR) and electron impact mass spectrometry (EI-MS) to determine the structures of 9a-o. Antimicrobial activity was evaluated as zone of inhibition by disc diffusion method. Hemolytic and thrombolytic activities were determined by measuring absorbance before and after incubation of blood cells with test compound.Results: Compound 9d strongly inhibited Bacillus subtilis and Escherichia coli with zone of inhibition values of 16 mm for each. The reference drug, rifampicin, showed zone of inhibition of 21 and 23 mm against B. subtilis and E. coli, respectively. Compound 9a strongly inhibited Aspergillus niger with a zone of inhibition of 18 mm compared to the reference drug, fluconazole, with a zone of inhibition of 19mm.Conclusion: The newly synthesized compounds are more active antimicrobial agents than piperine. Compounds 9a and 9d are the most active.Keywords: Piper nigrum, Piperine, Propanamide, Hemolytic, Thrombolytic, Black peppe

Epidermal Growth Factor Receptor (EGFR) as a Prognostic Marker: an Immunohistochemical Study on 315 Consecutive Breast Carcinoma Patients

Objective: To assess the independent and interdependent prognostic value of epidermal growth factor receptor (EGFR) in carcinoma of breast in female population. The Type I family of growth factor receptors includes epidermal growth factor receptor (EGFR also known as EGFRI). Methods: The expression of EGFR protein was analysed immunohistochemically on 315 tumour specimens of infiltrating ductal carcinoma of breast. These patients also had axillary lymph nodes sampling. Results: Overexpression and/or amplification of EGFR was observed in 70 (22.00%) tumours. Eleven (16%) were grade I, 43 (61%) grade II and 16 (23%) grade Ill tumours. Axillary lymph node metastasis had significant correlation with intensified positivity of EGFR (p\u3c 0.05). Significant number of EGFR positive patients developed local recurrence and distant metastases to brain, liver and bone (p\u3c 0.05). EGFR positivity showed significant correlation with the disease free and overall survival (p\u3c 0.05). At a median follow-up of48 (4 years) months in EGFR positive patients, the overall survival was 3.39 years and disease free survival was 2.86 years. EGFR negative tumour patients showed a better survival. In this group the overall survival was 4.62 years and the disease free survival was 4 years. Conclusion: EGFR analysis can be a useful indicator for the selection of patients who are at the high risk, for hormonal therapy decisions and can be useful as a target for new treatment modalities

Calidad y seguridad microbiológica de aceites extraídos de semillas de almendras (Prunus dulcis Mill.) sometidas a radiación gamma

The physicochemical attributes and microbial decontamination of oils extracted from gamma-irradiated almond (Mission and Price varieties) seeds, to the absorbed doses of 2-10 kGy, have been evaluated. Gamma irradiation exerted no considerable effect on the proximate seed composition. The physicochemical properties such as density and refractive index of the oils, extracted from gammairradieted seeds, were almost unaffected; the iodine value decreased while saponification value, unsaponifiable matter and free fatty acids increased. The oxidative status and tocopherol content of almond oils were negatively affected while the fatty acid profile slightly changed due to irradiation stress. Interestingly, these effects on the oil quality attributes were more pronounced at higher irradiation doses (> 6 kGy). Besides, microbial contamination was completely eliminated in the oils irradiated to an absorbed dose of 6.0 kGy. It could be concluded from the present findings that irradiation has a considerably positive or negative effect on some attributes of the almond oil. Therefore, an appropriate magnitude of gamma irradiation should be exercised to treat almond seeds in order to retain maximum nutritive benefits.Se ha evaluado las caracteristicas fisico-químicas y la descontaminación microbiana de aceites extraídos de semillas de almendras (variedades Misión y Price) gamma-irradiadas a dosis absorbidas de 2-10 kGy. La radiación gamma no ejerce ningún efecto considerable en la composición próximal de las semillas. Las características fisico-químicas tales como la densidad y el índice de refracción de los aceites, extraídos a partir de semillas gamma-irradiadas, permanecieron casi sin afectar; el índice de yodo disminuye mientras que el valor de saponificación, la materia insaponificable y los ácidos grasos libres aumentan. El estado oxidativo y el contenido de tocoferoles de los aceites de almendra se vieron afectados negativamente, mientras que el perfil de ácidos grasos se modifica ligeramente debido al estrés de la radiación. Curiosamente, los efectos sobre los atributos de calidad del aceite fueron más pronunciados a dosis de radiación más altas (> 6 kGy). Además, la contaminación microbiana se eliminó por completo en los aceites irradiados a una dosis absorbida de 6,0 kGy. Se puede concluir a partir de los presentes hallazgos que la radiación tiene un efecto positivo o negativo considerable en algunos atributos de la aceite de almendras. Por lo tanto, se debe aplicar una magnitud apropiada de radiación gamma para el tratamiento de semillas de almendra con el fin de retener los máximos beneficios nutritivos

Yield-related salinity tolerance traits identified in a nested association mapping (NAM) population of wild barley

Producing sufficient food for nine billion people by 2050 will be constrained by soil salinity, especially in irrigated systems. To improve crop yield, greater understanding of the genetic control of traits contributing to salinity tolerance in the field is needed. Here, we exploit natural variation in exotic germplasm by taking a genome-wide association approach to a new nested association mapping population of barley called HEB-25. The large population (1,336 genotypes) allowed cross-validation of loci, which, along with two years of phenotypic data collected from plants irrigated with fresh and saline water, improved statistical power. We dissect the genetic architecture of flowering time under high salinity and we present genes putatively affecting this trait and salinity tolerance. In addition, we identify a locus on chromosome 2H where, under saline conditions, lines homozygous for the wild allele yielded 30% more than did lines homozygous for the Barke allele. Introgressing this wild allele into elite cultivars could markedly improve yield under saline conditions.Stephanie Saade, Andreas Maurer, Mohammed Shahid, Helena Oakey, Sandra M. Schmöckel, Sónia Negrão, Klaus Pillen and Mark Teste

Management of Fusarium Wilt using mycolytic enzymes produced by Trichoderma harzianum (Th. Azad)

The main aim of this study was to isolate the best chitinase and glucanase enzyme producing Trichoderma strain to manage the Fusarium wilt disease of Cicer aritenum under in vitro conditions. We also studied the effect of Trichoderma strains on the growth and development of C. aritenum plants. Seven strains of Trichoderma were screened against the Fusarium pathogen to isolate the best biocontrol agent causing maximum inhibition of Fusarium growth. Trichoderma harzianum (Th. Azad) was found to be the best strains among all the tested strains. Trichoderma treated plant exhibited the least disease incidence as compared to control plants. Trichoderma treated plant showed a significant stimulatory effect on all the tested eight parameters as compared to control.Key words: Trichoderma, antagonistic activity, chitinase, glucanase, Biocontrol agent, phytopathogenic fungi

COVID-19 Vaccine Hesitancy among Pregnant Women Attending Antenatal Clinics in Pakistan: A Multicentric, Prospective, Survey-Based Study

This study aimed to assess the vaccination status and factors contributing to vaccine hesitancy among pregnant women in the largest province of Pakistan. A multicentric, prospective, survey-based study using an interviewer-administered tool was conducted among pregnant women attending antenatal clinics between 1 December 2021 through 30 January 2022 across seven hospitals in Pakistan. The healthcare professionals providing care at the participating hospitals administered the survey. Four hundred and five pregnant women fully consented and completed the study. The majority of the study participants (70.6%, n = 286) were aged between 25 and 34 and had a previous successful pregnancy history. More than half of the study participants (56.0%, n = 227) did not receive COVID-19 vaccination at the time of data collection despite their family members (93.9%, n = 372) had already received at least one dose of COVID-19 vaccine. Among those who received COVID-19 vaccination (n = 173), vaccine efficacy, protection for the foetus, and risk of COVID-19-associated hospitalisation were the main driving factors for vaccine hesitancy. The majority of the unvaccinated women (77.8%, n = 182) had no intention of receiving the vaccine. However, more than two-thirds (85.7%, n = 342) consulted the doctor about COVID-19 vaccines, and most were recommended to receive COVID-19 vaccines by the doctors (80.7%, n = 280). Women were significantly more likely to be vaccinated if they had employment (odds ratio [OR] 4.47, 95% confidence interval [CI]: 2.31–8.64) compared with their counterparts who were homemakers, consulted their doctors (OR 0.12, 95% CI: 0.04–0.35), and if they did not have pregnancy-related issues (OR 6.02, 95% CI: 2.36–15.33). In this study, vaccine hesitancy was prevalent, and vaccine uptake was low among pregnant women. Education and employment did impact COVID vaccination uptake, emphasising the need for more targeted efforts to enhance the trust in vaccines