TLR-9 Expression in Human Bronchial Epithelial Cells induced with Lipopolysaccharide

The study conducted here was to investigate the gene and protein expression of Toll like receptor 9 when human bronchial epithelial cells were induced by lipopolysaccharide. TLRs are pattern recognition receptor (PRR) which plays a key role in innate immunity. They recognise the molecules that are shared by pathogens but distinguishes from the host which is referred to as pathogen associated molecular patterns (PAMPs). The HBECs were first cultured and once it becomes confluent they were induced with 10ng/ml of LPS. In order to find out whether the gene was expressed in the cells, the RNA was isolated and RT-PCR was carried out. Before carrying out the RT-PCR, RNA gel electrophoresis was carried out to show the 28S and 18S bands. Once the gene expression was analysed protein expression was carried out by extracting the protein using RIPA buffer and running the SDS-PAGE followed by staining. Silver staining and western blotting results revealed clear bands at 116kDa illustrating that the TLR9 protein had been expressed. RT-PCR results showed that a smear of bands for TLR9 and the in protein expression bands were seen in TLR9 protein region which indicates that TLR9 was expressed in the HBEC by LPS

Human Biological Monitoring of Diisononyl Phthalate and Diisodecyl Phthalate: A Review

High molecular-weight phthalates, such as diisononyl phthalate (DINP), and diisodecyl phthalate (DIDP), are widely used as plasticizers in the manufacturing of polymers and consumer products. Human biological monitoring studies have employed the metabolites of DINP and DIDP as biomarkers to assess human exposure. In this review, we summarize and analyze publicly available scientific data on chemistry, metabolism, and excretion kinetics, of DINP and DIDP, to identify specific and sensitive metabolites. Human biological monitoring data on DINP and DIDP are scrutinised to assess the suitability of these metabolites as biomarkers of exposure. Results from studies carried out in animals and humans indicate that phthalates are metabolised rapidly and do not bioaccmulate. During Phase-I metabolism, ester hydrolysis of DINP and DIDP leads to the formation of hydrolytic monoesters. These primary metabolites undergo further oxidation reactions to produce secondary metabolites. Hence, the levels of secondary metabolites of DINP and DIDP in urine are found to be always higher than the primary metabolites. Results from human biological monitoring studies have shown that the secondary metabolites of DINP and DIDP in urine were detected in almost all tested samples, while the primary metabolites were detected in only about 10% of the samples. This indicates that the secondary metabolites are very sensitive biomarkers of DINP/DIDP exposure while primary metabolites are not. The NHANES data indicate that the median concentrations of MCIOP and MCINP (secondary metabolites of DINP and DIDP, resp.) at a population level are about 5.1 μg/L and 2.7 μg/L, respectively. Moreover, the available biological monitoring data suggest that infants/children are exposed to higher levels of phthalates than adults

Fabrication of Chitosan/TPP Nano Particles as a Carrier Towards the Treatment of Cancer

Cancer one of the deadliest disease and resistance of cancer cells has been the major issue in both the modern molecular targeted therapeutics as well as in the conventional chemotherapeutics. As on date chitosan has been the ideal choice as a carrier for delivering anticancer drugs like doxorubicin and cisplatin for targeted delivery. The present study focusses on the fabrication of chitosan nanoparticles using TPP by ionic gelation method loaded with 5-FU, an anticancer drug. The morphology of the nanoparticles synthesized were analysed using SEM and the size, zeta potential using particle size analyser. The mechanism of drug release was studied by fitting the release kinetics with Peppas model. Cellular uptake of the drug was analysed and using MTT the cytotoxicity & metabolic activity of the normal and the cancerous cells were evaluated. Results of the study showed that the synthesized nanoparticles possessed a positive charge and the release of the drug was through diffusion and degradation mediated. Also the cellular uptake of the drug showed the amount of drug uptake reduced with the cancerous cell as the time increase which was further confirmed using the MTT assay which showed the metabolic activity of the cancerous cell got reduced. The results of the study showed that the fabricated nanoparticles could potentially be used as a carrier towards the treatment of cancer

The Phytochemical Composition of Medicinal Plants: Brazilian Semi-Arid Region (Caatinga)

Carnauba wax, the most important vegetable wax under the economic and extracted from the leaves of the carnauba (Copernicia prunifera (Miller) H. E. Moore), is extensively applied in food due to its physiochemical characteristics with a majority of esters. p-Methoxycinnamic acid diesters obtained from the ceriferous powder of carnauba wax (PCO-C) have been associated with biological actions. However, being a versatile product, many types of research have been carried out seeking to expand the possibilities of applications of this raw material. Furthermore, different experimental studies on the pharmacological activities have also been undertaken in recent years and have tested various biological activities, such as hypolipidemic, hypocholesterolemic and hypoglycemic effects in mice. Therefore, in this book chapter, it is reviewing the development of a process of extraction of 4-hydroxycinnamic acid diesters of carnauba wax powder and investigates their biological actions and physical and chemical characteristics

Benzimidazolium picrate single crystal: Synthesis, growth, characterization and their biological activity

Knowledge on charge-transfer complexes of drugs is important to understand the drug-receptor binding mechanism. Here, we have been synthesized the organic charge transfer complex of Benzimidazolium picrate (BP) and confirmed its molecular structure and carbon skeleton by 1H and 13C NMR spectral analysis. Asymmetric unit of BP consists of one benzimidazolium cation and a monovalent anion of picric acid. Decomposition pattern and thermal stability BP have been investigated using TG/DTA analysis. The interaction of the compound with calf thymus DNA is investigated by electronic absorption spectroscopy which indicates that the compound could interact with DNA through intercalation. The CT complex demonstrated significant free radical scavenging with 2,2-diphenyl-2-picryl-hydrazyl. The BP complex has been also screened for its antimicrobial activity

Benzimidazolium picrate single crystal: Synthesis, growth, characterization and their biological activity

1305-1312Knowledge on charge-transfer complexes of drugs is important to understand the drug-receptor binding mechanism. Here, we have been synthesized the organic charge transfer complex of Benzimidazolium picrate (BP) and confirmed its molecular structure and carbon skeleton by 1H and 13C NMR spectral analysis. Asymmetric unit of BP consists of one benzimidazolium cation and a monovalent anion of picric acid. Decomposition pattern and thermal stability BP have been investigated using TG/DTA analysis. The interaction of the compound with calf thymus DNA is investigated by electronic absorption spectroscopy which indicates that the compound could interact with DNA through intercalation. The CT complex demonstrated significant free radical scavenging with 2,2-diphenyl-2-picryl-hydrazyl. The BP complex has been also screened for its antimicrobial activity

Sample preparation procedure for the determination of polycyclic aromatic hydrocarbons in petroleum vacuum residue and bitumen

This paper describes a novel method of sample preparation for the determination of trace concentrations of polycyclic aromatic hydrocarbons (PAHs) in high-boiling petroleum products. Limits of quantitation of the investigated PAHs in materials of this type range from tens of nanograms per kilogram to <20 μg/kg. The studies revealed that in order to separate most of interferences from the analytes without a significant loss of PAHs, it is necessary to use size exclusion chromatography as the first step of sample preparation, followed by adsorption using normal-phase liquid chromatography. The use of orthogonal separation procedure described in the paper allows the isolation of only a group of unsubstituted and substituted aromatic hydrocarbons with a specific range of molar mass. The lower the required limit of quantitation of PAHs, the larger is the scale of preparative liquid chromatography in both steps of sample preparation needed. The use of internal standard allows quantitative results to be corrected for the degree of recovery of PAHs during the sample preparation step. Final determination can be carried out using HPLC-FLD, GC-MS, or HPLC-UV–VIS/DAD. The last technique provides a degree of identification through the acquired UV–VIS spectra

Distinguishing Type 2 Diabetes from Type 1 Diabetes in African American and Hispanic American Pediatric Patients

To test the hypothesis that clinical observations made at patient presentation can distinguish type 2 diabetes (T2D) from type 1 diabetes (T1D) in pediatric patients aged 2 to 18.Medical records of 227 African American and 112 Hispanic American pediatric patients diagnosed as T1D or T2D were examined to compare parameters in the two diseases. Age at presentation, BMI z-score, and gender were the variables used in logistic regression analysis to create models for T2D prediction.The regression-based model created from African American data had a sensitivity of 92% and a specificity of 89%; testing of a replication cohort showed 91% sensitivity and 93% specificity. A model based on the Hispanic American data showed 92% sensitivity and 90% specificity. Similarities between African American and Hispanic American patients include: (1) age at onset for both T1D and T2D decreased from the 1980s to the 2000s; (2) risk of T2D increased markedly with obesity. Racial/ethnic-specific observations included: (1) in African American patients, the proportion of females was significantly higher than that of males for T2D compared to T1D (p<0.0001); (2) in Hispanic Americans, the level of glycated hemoglobin (HbA1c) was significantly higher in T1D than in T2D (p<0.002) at presentation; (3) the strongest contributor to T2D risk was female gender in African Americans, while the strongest contributor to T2D risk was BMI z-score in Hispanic Americans.Distinction of T2D from T1D at patient presentation was possible with good sensitivity and specificity using only three easily-assessed variables: age, gender, and BMI z-score. In African American pediatric diabetes patients, gender was the strongest predictor of T2D, while in Hispanic patients, BMI z-score was the strongest predictor. This suggests that race/ethnic specific models may be useful to optimize distinction of T1D from T2D at presentation

DNA2 drives processing and restart of reversed replication forks in human cells

Accurate processing of stalled or damaged DNA replication forks is paramount to genomic integrity and recent work points to replication fork reversal and restart as a central mechanism to ensuring high-fidelity DNA replication. Here, we identify a novel DNA2- and WRN-dependent mechanism of reversed replication fork processing and restart after prolonged genotoxic stress. The human DNA2 nuclease and WRN ATPase activities functionally interact to degrade reversed replication forks with a 5'-to-3' polarity and promote replication restart, thus preventing aberrant processing of unresolved replication intermediates. Unexpectedly, EXO1, MRE11, and CtIP are not involved in the same mechanism of reversed fork processing, whereas human RECQ1 limits DNA2 activity by preventing extensive nascent strand degradation. RAD51 depletion antagonizes this mechanism, presumably by preventing reversed fork formation. These studies define a new mechanism for maintaining genome integrity tightly controlled by specific nucleolytic activities and central homologous recombination factors