Laboratory Use of Lectin Mitogens for Mitotic Stimulation of Human Lymphocytes

Lectins are believed to act as modulations of cell substratum interactions and to be essential for the normal differentiation and growth of all multicellular humans and animals. Although several lectins have been reported from microfungi, many more genera remain unexplored and their physiological role is also uncertain. The aim of this laboratory work was to make a comparison between self-made lectins (Indigenous) and commercial ones, following High Resolution Cell Synchronization technique (HRCS). Cytogenetic studies were performed in 175 normal healthy blood donor individuals of both genders and statistical analysis was performed. Our results indicated that the preparation of fresh phytohemagglutinin at the time of cell division and cell culture procedure reveals a satisfactory score. The overall frequency of mitotic index in our study was higher when compared with commercial imported Lectins (p < 0.05). The significant differences in the results may be due to fresh preparation. However, the cost effective, easy and nearest approach of this indigenous product, as well as the high demand for this product, among health care services can be considered.Highlights:Lectins act as essential factor of the normal differentiation and growth of all humans and animals.Phytohemagglutinin (PHA) is a lectin (mucoprotein) from Phaseolus vulgaris.Crude extract of PHA could be used in human leucocyte cultures as mitotic stimulator.The indigenous PHA have used to identify chromosome preparation in normal conditions and malignancies

Investigation of metabonomics technique by analyze of NMR data, which method is better? Mean center or auto scale?

The factors such as disease can disrupt homeostasis, resulting in perturbations of endogenous biochemicals that are involved in key metabolic profiles. Metabonomics is useful technique to quantitative description of endogenous metabolites present in a biological sample such as urine, plasma and tissue. High resolution 1H nuclear magnetic resonance (NMR)-based metabonomics is a technique used to analyze and interpret multivariate metabolic data that correlate with changes of physiological conditions. Before any explanation for metabolite data, preprocessing the spectroscopic data is essential. In this paper, we show scaling effects in metabonomics investigation of patients diagnosed with Crohn's and Celiac disease. two techniques of scaling were applied as follows: mean centering and auto scaling. Results reveal that the mean centering is more useful to segregate patients from healthy subjects in the data set of Crohn's and Celiac disease

Natural Killer Cells from the Subcutaneous Adipose Tissue Underexpress the NKp30 and NKp44 in Obese Persons and Are Less Active against Major Histocompatibility Complex Class I Non-Expressing Neoplastic Cells

There are many types of leukocytes reside in subcutaneous adipose tissue (SAT), and among them, natural killer cells (NKs) comprise a major part. We show that the NKs that reside in the SAT (adipose tissue-derived NK cells; ADNKs) of the abdominal region found with phenotypic differences from the NKs circulating in the peripheral blood derived NK cells (PBNKs). In this survey, flow cytometry phenotyping was used to study the differences between the natural cytotoxicity receptor expression on ADNKs and PBNKs of both obese and lean persons. Also, their cytotoxicity and cytokine production patterns were evaluated. The activation experiments on isolated and expanded NKs with IL-2, IL-15, and IL-21 cytokines revealed the main population of the CD56dim within the total ADNKs of obese persons has an under-expression of NKp30 and NKp44 despite the unchanged levels of NKG2D. The data suggest the suppressive condition of the adipose tissue niche on the NKs response against sensitive major histocompatibility complex class I non-expressing neoplastic cells. As the NKs are the first line of the body’s defense vs tumor formation, this change may lead to the development of transformed cells into the tumors

Downregulation of E-cadherin expression in breast cancer by promoter hypermethylation and its relation with progression and prognosis of tumor

Breast cancer is the most common cancer in women around the world, and novel prognosis strategies is needed to control more accurate and effective of this malignant disease. Among the latest prognostic markers is E-cadherin, which mediates cell-cell adhesion by associating with catenins. Loss of E-cadherin gene (CDH1) function by genetic or epigenetic alteration leads to tumorigenesis. The aim of our study was to investigate E-cadherin gene promoter methylation in breast cancer, and its correlation with E-cadherin protein expression. Fifty primary breast cancers tissue with ductal type and 50 normal breast sample from the same patients that was located adjacent to tumor region as controls were provided by Imam Reza-based referral and teaching hospital affiliated to Tabriz University of Medical Sciences, Tabriz, Iran. CDH1 promoter region CpG sites methylation and E-cadherin protein expression were determined by bisulfitespecific polymerase chain reaction and Western blot analysis, and the resulting products were sequenced on an ABI automated sequencer for firm conclusion. CDH1 hypermethylation in breast tumor specimen (ductal type) was observed in 94 % (47 of 50) comparing with normal samples methylation, and the significant difference was (p = 0.000). Protein expression in tumor samples tends to diminish with the CDH1 promoter region methylation. In the group of 50 ductal carcinomas cases, most of the cases showing CDH1 hypermethylation correlated inversely with the reduced levels of expression of E-cadherin proteins (95 % of full-methylated tumor samples had no protein expression, and 4.5 % of them had weak expression levels). Possible association was observed between CDH1 methylation and its protein expression (p = 0.000). The results of methylation analysis in promoter region in ten CpG sites (863, 865, 873, 879, 887, 892, 901, 918, 920, and 940) suggested that abnormal CDH1 methylation occurs in high frequencies in ductal breast tumors probably sounds the process of carcinogenesis progression

Introducing Transthyretin as a Differentially Expressed Protein in Washing Subtype of Obsessive-Compulsive Disorder

Introduction: Obsessive-Compulsive Disorder (OCD) as one of the important mental problems is valuable topic for proteomic research studies to better understand the underlying mechanisms of this disorder. Methods: In this paper, gel-based proteomic was used to investigate the proteome profile of 16 female patients with OCD, washing subtype before and after treatment with fluoxetine and comparing them with 20 healthy female controls. Results: One of the abnormally expressed protein spots in this study was introduced and examined for protein-protein interaction network analysis via Cytoscape and its plug-ins. Transthyretin (TTR) protein showed significant expression changes (fold change=1.7, P<0.05). While the expression level of TTR is significantly decreased in OCD patients before any treatments, the trend is partially normalized after treatment with fluoxetine in positive responders. Furthermore, TTR interaction profile shows that the proteins interacting with this protein may get affected as this protein expression trend changes in OCD patients. Conclusion: TTR can be considered for further studies to be validated as a potential biomarker for OCD

FITC immunostaining of apoptin expression in breast carcinoma cell lines.

<p>Immunostaining of apoptin protein showed that in can express in breast carcinoma cell lines after 12λ ZAP-CMV vector have not any apoptosis after 36 h as same as untreated cells.</p

Recovery of NBPs from nude mice tissues.

<p>There were no significant differences in recombinant NBPs titer that was recovered from the different tissues of the treated mice. The tumor site was the only part of the mice body to support the more NBPs accumulation.</p

Confirmation of apoptosis by flowcytometry.

<p>BT-474, SKBR-3 and ZR-75 cells were examined after 36 h of transfection by flowcytometry. All the cell lines were susceptible to NBPs apoptin-induced apoptosis. We have not apoptosis in vector treated group and untreated group.</p

Cytotoxicity evaluation of plasmid and vehicle.

<p>BT-474 breast carcinoma cell line transfected with λ ZAP-CMV-apoptin, λ ZAP-CMV vector and λ phage (vehicle) construct stained with FITC immunostaining and then visualized by fluorescence microscopy. There was no sign of cell necrosis after the treatments. There is only apoptotic morphology of cells after treatment with λ ZAP-CMV-apoptin.</p