Correlation between major depressive disorder and circulating natural killer cells

 Background: Major depressive disorder is a mental disorder characterized by a pervasive and persistent low mood that is accompanied by low self-esteem and loss of interest or pleasure in normally enjoyable activities. Depression is associated with multiple immunological disorders. Aim of the present study was to determine correlation between percentage of circulating NK cells and major depressive disorder.Materials and Methods: Patients older than 18 years with the desire to participate were enrolled in this study. For depression evaluation, we used the Hamilton Depression Rating Scale and for determination of percentage of NK cells in peripheral blood, flow cytometry method was used. Results: Our results showed that in patients with major depressive disorder, numbers of circulating NK cells have significantly reduced. Conclusion: According to our findings, depression is associated with “immune suppression”. NK cells are important in early phase of immunological surveillance versus viral infections and tumors. Indeed, depressive patients are susceptible to cancers and infections

A study on drug delivery tracing with radiolabeled mesoporous hydroxyapatite nanoparticles conjugated with 2DG/DOX for breast tumor cells

Background: Mesoporous nanoparticles have a great potential in targeted therapy approaches due to their ideal properties for encapsulation of various drugs, proteins and also biologically active molecules. Material and methods: We used mesoporous hydroxyapatite (HA) nanoparticles as a drug carrier and developed radiolabeled mesoporous HA containing of 2-deoxy-D-glucose (2DG) and Doxorubicin (DOX) with technetium-99m (99mTc) for imaging in in vitro and in vivo studies. Results: 2DG and DOX in presence of mesoporous HA nanoparticles more reduced the fraction of viable cells in the MDA-MB-231, MCF-7 human and MC4-L2 Balb/c mice breast cancer cells. The radiochemical purity of the nano-2DG-DOX complex with 99mTc was calculated to 96.8%. The results of cellular uptake showed a 44.77% increase in uptake of the [99mTc]-nano-2DG-DOX compared to the complex without nanoparticles (p < 0.001). Conclusion: Radioisotopic imaging demonstrated a high biochemical stability for [99mTc]-nano-2DG-DOX complex. The results demonstrated that [99mTc]-nano-2DG-DOX, may be used as an attractive candidate in cancer imaging and treatment managing.BACKGROUND: Mesoporous nanoparticles have a great potential in targeted therapy approaches due to their ideal properties for encapsulation of various drugs, proteins and also biologically active molecules. MATERIAL AND METHODS: We used mesoporous hydroxyapatite (HA) nanoparticles as a drug carrier and developed ra­diolabeled mesoporous HA containing of 2-deoxy-D-glucose (2DG) and Doxorubicin (DOX) with technetium-99m (99mTc) for imaging in in vitro and in vivo studies. RESULTS: 2DG and DOX in presence of mesoporous HA nanoparticles more reduced the fraction of viable cells in the MDA-MB-231, MCF-7 human and MC4-L2 Balb/c mice breast cancer cells. The radiochemical purity of the nano-2DG-DOX complex with 99mTc was calculated to 96.8%. The results of cellular uptake showed a 44.77% increase in uptake of the [99mTc]- nano-2DG-DOX compared to the complex without nanoparticles (p &lt; 0.001). CONCLUSIONS: Radioisotopic imaging demonstrated a high biochemical stability for [99mTc]-nano-2DG-DOX complex. The results demonstrated that [99mTc]-nano-2DG-DOX, may be used as an attractive candidate in cancer imaging and treatment managing.

Recombinant expression and purification of L2 domain of human epidermal growth factor receptor

Epidermal growth factor receptor (EGFR) is one of the key molecules in cell growth and multiplication and plays an important role in some malignant processes. L2 domain of extra-cellular part of this receptor involved in ligand binding and its inhibition can prevent activation of related signaling pathways. The aim of the present study was cloning and expressing the fragment coding for L2 region of human EGFR for the production of recombinant L2 protein. The total RNA from A431 cells line was extracted and used for amplification of the sequence coding for L2 domain of EGFR by reverse transcriptase-polymerase chain reaction (RT-PCR) technique. The product was cloned into the PGEM-T vector and used for sequencing. In the next step, the insert was removed from the PGEM-T vector and subcloned into the PET22 expression vector. The expression construct was transformed into the Escherichia coli BL21 (DE3) and recombinant protein expression was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Analyzing the expression of produced recombinant protein showed that the recombinant L2 can be highly expressed by this expression system. This recombinant protein can be used for the production of specific mAb, screening for specific ligands and competitive inhibition of the EGFR.Keywords: Human epidermal growth factor receptor, domain L2, recombinant expressionAfrican Journal of Biotechnology Vol. 9(33), pp. 5292-5296, 16 August, 201

Immuno-biosensor for Detection of CD20-Positive Cells Using Surface Plasmon Resonance

Purpose: Surface plasmon resonance (SPR) sensing confers a real-time assessment of molecular interactions between biomolecules and their ligands. This approach is highly sensitive and reproducible and could be employed to confirm the successful binding of drugs to cell surface targets. The specific affinity of monoclonal antibodies (MAb) for their target antigens is being utilized for development of immuno-sensors and therapeutic agents. CD20 is a surface protein of B lymphocytes which has been widely employed for immuno-targeting of B-cell related disorders. In the present study, binding ability of an anti-CD20 MAb to surface antigens of intact target cells was investigated by SPR technique. Methods: Two distinct strategies were used for immobilization of the anti-CD20 MAb onto gold (Au) chips. MUA (11-mercaptoundecanoic acid) and Staphylococcus aureus protein A (SpA) were the two systems used for this purpose. A suspension of CD20-positive Raji cells was injected in the analyte phase and the resulting interactions were analyzed and compared to those of MOLT-4 cell line as CD20-negative control. Results: Efficient binding of anti-CD20 MAb to the surface antigens of Raji cell line was confirmed by both immobilizing methods, whereas this MAb had not a noticeable affinity to the MOLT-4 cells. Conclusion: According to the outcomes, the investigated MAb had acceptable affinity and specificity to the target antigens on the cell surface and could be utilized for immuno-detection of CD20-positive intact cells by SPR method

Recombinant expression and purification of L2 domain of human epidermal growth factor receptor

Epidermal growth factor receptor (EGFR) is one of the key molecules in cell growth and multiplication and plays an important role in some malignant processes. L2 domain of extra-cellular part of this receptor involved in ligand binding and its inhibition can prevent activation of related signaling pathways. The aim of the present study was cloning and expressing the fragment coding for L2 region of human EGFR for the production of recombinant L2 protein. The total RNA from A431 cells line was extracted and used for amplification of the sequence coding for L2 domain of EGFR by reverse transcriptase-polymerase chain reaction (RT-PCR) technique. The product was cloned into the PGEM-T vector and used for sequencing. In the next step, the insert was removed from the PGEM-T vector and subcloned into the PET22 expression vector. The expression construct was transformed into the Escherichia coli BL21 (DE3) and recombinant protein expression was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Analyzing the expression of produced recombinant protein showed that the recombinant L2 can be highly expressed by this expression system. This recombinant protein can be used for the production of specific mAb, screening for specific ligands and competitive inhibition of the EGFR

Targeted Co-Delivery of Docetaxel and cMET siRNA for Treatment of Mucin1 Overexpressing Breast Cancer Cells

Purpose: Targeted treatment of breast cancer through combination of chemotherapeutic agents and siRNA had been drawing much attention in recent researches. This study was carried out to evaluate mucin1 aptamer-conjugated chitosan nanoparticles containing docetaxel and cMET siRNA on SKBR3 cells. Methods: Nano-drugs were characterized by transmission electron microscope, Zetasizer and loading efficiency calculation. siRNA entrapment onto nanoparticles, stability of siRNA-loaded nanoparticles and conjugation of mucin1 aptamer to nanoparticles were evaluated via separate electrophoresis. Cellular uptake of the targeted nanoparticles was evaluated through GFP-plasmid expression in mucin1+ SKBR3 vs. mucin1- CHO cells. Protein expression, cell viability and gene expression were assessed by Western Blotting, MTT assay, and Quantitative Real Time-PCR, respectively. Results: Characterization of nano-drugs represented the ideal size (110.5± 3.9 nm), zeta potential (11.6± 0.8 mV), and loading efficiency of 90.7% and 88.3% for siRNA and docetaxel, respectively. Different gel electrophoresis affirmed the conjugation of aptamers to nanoparticles and entrapment of siRNA onto nanoparticles. Increased cellular uptake of aptamer-conjugated nanoparticles was confirmed by GFP expression. cMET gene silencing was confirmed by Western Blotting. The significant (p ≤0.0001) impact of combination targeted therapy vs. control on cell viability was shown. Results of Quantitative Real Time-PCR represented a remarkably decreased (p ≤0.0001) expression of the studied genes involving in tumorigenicity, metastasis, invasion, and angiogenesis (STAT3, IL8, MMP2, MMP9, and VEGF) by targeted combination treatment vs. control. Conclusion: The mucin1 aptamer-conjugated chitosan nanoparticles, containing docetaxel and cMET siRNA, is suggested for treatment of mucin1+ metastatic breast cancer cells. However, further studies should be conducted on animal models

The Study of HLA-G Gene and Protein Expression in Patients withRecurrent Miscarriage

Purpose: Although it has been frequently confirmed that HLA-G plays an important role in thereproduction and pregnancy, the pattern of HLA-G gene and its protein expression are rarelyaddressed in studies. Therefore we conducted this study in regard to evaluate the HLA-G geneand its protein expression in the women’s placenta with recurrent miscarriage.Methods: Placental samples were obtained from the women who were admitted for deliveryor abortion in Al Zahra and Taleghani hospitals, Tabriz, Iran. HLA-G gene expression wasdetermined by real-time polymerase chain reaction (PCR) and HLA-G protein expression wasassessed by western blotting and immunofluorescence staining in the tissue samples.Results: The results showed a significant decrease in the expression of gene and proteins ofHLA-G in the women with recurrent miscarriage compared to the control placental tissues.Conclusion: According to the obtained results, it was concluded that the decrement of HLA-Ggene and protein expressions are associated with recurrent miscarriage. Since there areconflicting results from other studies, it is suggested to conduct a more comprehensive similarstudy with greater sample size

False positive seroreactivity to brucellosis in tuberculosis patients: a prevalence study

Mojtaba Varshochi1,2, Jafar Majidi2, Marjan Amini1, Kamyar Ghabili3, Mohammadali M Shoja31Department of Infectious Disease, Tabriz University of Medical Sciences, Tabriz, Iran; 2Infectious Disease and Tropical Medicine Research Center, Tabriz University of Medical Sciences, Tabriz, Iran; 3Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, IranBackground: The rising worldwide incidence of tuberculosis (TB) increases the demand for knowledge about its potential seroreactivity with other microbial agents. A few reports and the authors&amp;rsquo; experiences indicate that tuberculosis may result in a false-positive brucellosis serology. This may cause a diagnostic challenge because of the close clinical resemblance of these two infections.Objective: The aim of the present prevalence study was to elucidate brucellosis seroreactivity in patients with active TB.Methods: Ninety-eight patients with newly diagnosed and active TB were studied using an enzyme-linked immunosorbent assay (ELISA) and Wright&amp;rsquo;s and Coombs&amp;ndash;Wright&amp;rsquo;s tests. Seventy-five healthy individuals were used as controls. The patients showed signs of recovery after starting a standard anti-TB regimen and had no clinical evidence of brucellosis at a subsequent 6-month follow-up. The data were analyzed statistically by Fisher&amp;rsquo;s exact test using SPSS 11.0.Results: We found that 9.2% of TB patients versus 1.3% of healthy controls had positive results on the anti-Brucella IgG ELISA (P = 0.04). Five TB patients were found to have agglutination on Wright&amp;rsquo;s tests, while none of the controls showed agglutination.Conclusion: Active TB patients may have some seroreactivity with Brucella antigens, and Brucella IgG ELISA may give a false positive in these patients. Clinicians should consider false positive brucellosis seroreactivity in patients with active TB.Keywords: false positive serology, ELISA, diagnosi