CD20-targeting in B-cell malignancies: novel prospects for antibodies and combination therapies

Expression of CD20 antigen by the most of transformed B cells is believed to be the driving force for targeting this molecule by using anti-CD20 monoclonal antibodies. While it is true that most lymphoma/leukemia patients can be cured, these regimens are limited by the emergence of treatment resistance. Based on these observations, development of anti-CD20 monoclonal antibodies and combination therapies have been recently proposed, in particular with the aim to optimize the cytotoxic activity. Here we outline a range of new experimental agents concerning the CD20 positive B-cell tumors which provide high benefit from conventional therapy. © 2016, Springer Science+Business Media New York

In vitro assessment of cytotoxicity of giomer on human gingival fibroblasts

Root coverage on restored root surfaces has been considered as a challenging issue. The evaluation of cytotoxic effects of restorative materials is a fundamental requirement for sustaining the cell attachment and the clinical success of root coverage. The aim of the present study was to compare the human gingival fibroblast cytotoxicity of the recently introduced giomer composite (GC) with resin ionomer (RI) restorative material. Discs (6x2 mm) of GC and RI restorative materials were prepared using sterile Teflon mold. Extracts from the materials were incubated to cell culture medium for 24, 48 and 72 h. Human gingival fibroblasts (HGF) were exposed to the extracts of the materials while the un-incubated media served as the control group. The cytotoxicity of the materials were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. In order to compare the mean values of the measured parameters a Kruskal-Walis test was carried out. MTT assay indicated that human gingival fibroblasts proliferated well in the presence of GC extract. The proliferation rate was higher in cells incubated with GC compared to RI extracts but the differences were not statistically significant (p= 0.09). This in vitro study indicated that GC is a non-toxic material for HGF. However, further studies are needed to assess the other biologic and clinical behavior of this material prior to it being considered as a potentially suitable restorative material to restore the carious root lesions candidated to root coverage procedures

Co-solute assistance in refolding of recombinant proteins

Prokaryotic expression system is the most widely used host for the production of recombinant proteins but inclusion body formation is a major bottleneck in the production of recombinant proteins in prokaryotic cells, especially in Escherichia coli. In vitro refolding of inclusion body into the the proteins with native conformations is a solution for this problem but there is a need for optimization of condition for each protein specifically. Several approaches have been described for in vitro refolding; most of them involve the use of additives for assisting correct refolding. Co-solutes play a major role in refolding process and can be classified according to their function as, aggregation suppressors and folding enhancers. This study presents a review of additives that are used in refolding process of insoluble recombinant proteins in small scale and industrial process.Key words: Refolding, protein aggregation, low-molecular-weight additives, arginine

Neuroplasticity in the mammalian clock : the effect of aging and seasons

Many organisms have developed an internal clock to cope with the daily and seasonal cycles in the environment. In mammals, suprachiasmatic nuclei (SCN) of the hypothalamus control circadian rhythms in behavior and physiology. Evidence links the proper function of circadian clock to mental and physical health. Aging disturbs the accurate function of the SCN and impairs many rhythms such as sleep-wake cycle. Hence improvement of clock function can aid healthy aging. In chapters 3 and 4 I show the ensemble output of the SCN neuronal network is more robust than individual cells__ output suggesting a compensatory role of the network in aging. Seasonal changes affect the physiology and reproduction success of many organisms. The SCN encodes for day-length by adjusting the pattern of its electrical activity rhythm.. In chapters 5 and 6 I reveal that plasticity in interneuronal and cell-intrinsic functions in the SCN helps the organism to adjust to yearly natural changes in photoperiod. These results imply that extensive artificial light in modern society may alter neurotransmitters action in the SCN. A better understanding of SCN network function and cellular properties facilitate alleviation of modern life-related diseases caused by circadian disturbances and aging.UBL - phd migration 201

Determination of cephalosporin acylase activity by biological and colorimetric method in bacteria

The effective production of 7-aminocephalosporanic acid (7-ACA) is a matter of concern in the pharmaceutical industry because it is a starting material for the synthesis of semi synthetic cephalosporin. Therefore screening for new source of cephalosporin acylase positive bacteria is veryimportant. The cephalosporin acylase can be found in several Pseudomonas sp. and other bacteria. To facilitate the attempts of obtaining the microorganisms with higher cephalosporin acylase activity from natural environments, development of new and specific methods for screening environmental microorganisms with cephalosporin acylase activity is very important. In this study, a biological and colorimetric method was evaluated for determination of cephalosporin acylase product in bacteria. Samples were cultured in general and selective media, and the routine biochemical laboratory tests were used for diagnosis of Pseudomonas sp. All of the isolated strains were tested for cephalosporinacylase by a biological and colorimetric method. A total of 180 Pseudomonas sp. out of 350 samples were isolated. Two strains capable of producing cephalosporin acylase were identified from 180 candidates. The Pseudomonas bacteria isolated in this study is a source for cloning and cephalosporin acylase enzyme production

Early age onset familial Mediterranean fever associated with compound heterozygote M680I /M694V mutation

Familial Mediterranean fever (FMF) is an autosomal recessive genetic disorder characterized by acute episodes of fever accompanied by severe abdominal pain, pleurisy, arthritis, and skin rash. The clinicalvariability of the disease has been mainly attributed to MEFV gene allelic heterogeneity and partly to the influence of additional genetic and/or environmental factors. We present a 6-month-old boy who suffered from recurrent fever accompanied by abdominal pain and skin rashes. Molecular screening by polymerase chain reaction (PCR) and sequencing for common mutations causing FMF revealed presence of a 694V/680I compound heterozygote mutation in exon 10 of the related gene. This is thefirst report of early onset and severe phenotype FMF case associated with a 694V/680I compound heterozygote mutation

SAG2 locus genotyping of Toxoplasma gondii in meat products of East Azerbaijan Province, North West of Iran During 2010-2011

Toxoplasmosis is an infection caused by Toxoplasma gondii, an intracellular obligate parasite. Its transmission is usually attributed to ingestion of undercooked or raw meat. The aim of this study was the detection and genotyping of T. gondii in meat products using the molecular method in East Azerbaijan. DNA was extracted from 164 meat products sample obtained from 15 commercial establishments. Nested polymerase chain reaction with specific primers for Toxoplasma gondii SAG2 locus was used for detection of the parasite in samples. Genotyping was carried out by digestion of PCR products with restriction enzymes Cfo1 and Sau3AI. T. gondii DNA was detected in 16.6, 19.1, 15 and 56.6%, in salami, sausages, and hamburger and kebab samples, respectively. Genotyping by restriction fragment length polymorphism (RFLP) analysis of SAG2 locus revealed that all of the samples belonged to genotype I. The detection of the parasite in uncooked meat and commercial meat products, and the high ratio of seropositive slaughtered animals, emphasis that the risk still exists for food -born toxoplasmosis.Key words: Toxoplasma gondii, SAG2, genotyping, meat products

Evaluation of methylation pattern in promoter region of E-cadherin gene and its relation to tumor grade and stage in breast cancer

The epithelial cadherin gene (CDH1) has been identified as a tumor suppressor gene located within the 16q22.1 region. The CDH1 gene encodes a transmembrane glycoprotein involved in cell to cell adhesion and loss of CDH1 expression contributes to increased proliferation, invasion and metastasis in breast carcinoma. No mutation in CDH1 have been identified in invasive ductal carcinoma (IDC), suggesting that, other inactivation mechanisms are responsible for IDC oncogenesis. In order to analyze the role of promoter methylation in CDH1 gene inactivation in breast cancer, the CpG methylation status of Ecadherin promoter region by bisulfite sequencing PCR (BSP) was investigated. 10 CpG sites [nucleotide (nt) 863, 865, 873, 879, 887, 892, 901, 918, 920 and 940] in the promoter region were screened for methylation. The CDH1 methylation was detected in 94% (47 to 50) of breast tumors which was associated with higher tumor grade (p = 0.035), tumor stage (p = 0.000) and tumor metastasis (p = 0.000). There was also a significant correlation between tumor stage, grade and metastatic status with sites of methylation (p = 0.000). The data indicate that CDH1 promoter methylation might be a potential mechanism for epigenetic silencing of CDH1 in primary breast cancer suggesting a valuable molecular marker for detection of breast cancer progression.Key words: Breast cancer, E-cadherin, methylation pattern, tumor stage, tumor grade

Cloning and over-expression of Penicillin G acylase in Escherichia coli BL21

Penicillin G acylase (PGA) is one of the most important enzymes in the pharmaceutical industry. It is utilized in the process for production of semi-synthetic penicillins. Several different penicillin acylases with various characteristic have been isolated from different bacteria and identification of bacterial isolates harboring PGA enzyme with higher industrial compatibilities is of high interest. The aim of this study is to screen for PGA producing Escherichia coli isolates as well as the cloning and recombinant expression of PGA for high level enzyme production. Bacteria isolated from environmental and clinical samples were identified by standard microbiological tests and then E. coli isolates were subjected toDNA extraction and PCR screening using primers designed on conserved region of PGA genes. The PCR product from a positive isolate were cloned and subjected to sequencing. The gene encoding for full length PGA was expressed in E. coli under the T7 promoter. PCR screening identified several PGA positive E. coli. One of the positive isolates was cloned in pGEM -T easy vector. Sequencing of the cloned gene revealed that the gene encoding Penicillin G acylase from this wild E. coli isolate containsan open reading frame of 2538 nucleotide encoding 846 amino acids. Analysis of the sequencing results showed that the PGA gene is highly conserved among E. coli strains. Recombinant expression of PGA from wild isolate in E. coli BL 21 resulted in a high level expression of recombinant PGA which appeared as a dense band in SDS-PAGE analysis of induced culture

In vitro assessment of cytotoxicity of giomer on human gingival fibroblasts

Root coverage on restored root surfaces has been considered as a challenging issue. The evaluation of cytotoxic effects of restorative materials is a fundamental requirement for sustaining the cellattachment and the clinical success of root coverage. The aim of the present study was to compare the human gingival fibroblast cytotoxicity of the recently introduced giomer composite (GC) with resinionomer (RI) restorative material. Discs (6×2 mm) of GC and RI restorative materials were prepared using sterile Teflon mold. Extracts from the materials were incubated to cell culture medium for 24, 48and 72 h. Human gingival fibroblasts (HGF) were exposed to the extracts of the materials while the unincubated media served as the control group. The cytotoxicity of the materials were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. In order to compare the mean values of the measured parameters a Kruskal-Walis test was carried out. MTT assay indicated that human gingival fibroblasts proliferated well in the presence of GC extract. The proliferation rate washigher in cells incubated with GC compared to RI extracts but the differences were not statistically significant (p= 0.09). This in vitro study indicated that GC is a non-toxic material for HGF. However, further studies are needed to assess the other biologic and clinical behavior of this material prior to it being considered as a potentially suitable restorative material to restore the carious root lesions candidated to root coverage procedures