A Novel Approach on Leukodepletion Filters: Investigation of Synergistic Anticancer Effect of Purified α-Defensins and Nisin.

Purpose: There are number of reports available regarding defensins activity against mammalian cells besides their antimicrobial and immune regulatory activities. This study aims to investigate anticancer and apoptosis activity of the purified defensins from leukodepletion filters alone or in synergism with bacterial peptide, nisin, on prostate and colorectal cancer. Methods: Leucoflex LCR-5 filters were backflushed by an optimized elution system. Isolated granulocytes were sonicated and the supernatant treated before further purification by high performance liquid chromatography (HPLC). SDS-PAGE and western blot testing verified the fraction. Cell culture on PC-3 (human prostate adenocarcinoma), and HCT-116 (human colorectal carcinoma) were conducted following by MTT assays in addition to annexin flow cytometry for sole and synergistic effects with peptide nisin. Results: Viable and active neutrophils could recover, and α-defensins were extracted and purified. Combinations of an optimal dose of α-defensins and nisin showed a remarkable synergistic effect on cancer cell lines (over 90% and 70% for PC-3 and HCT-116, respectively). Conclusion: It also observed that less than 40% of both cells could survive after co-treatment with optimal dose. Also, apoptosis was increased after treatment by these peptides together. Annexin Vpositive populations significantly increased in percentage in comparison with control.

Different Immune Responses in Dangerous O Blood Donors.

BACKGROUND & OBJECTIVE: Dangerous O is very important to transfusion medicine and there has been reports by Food and Drug Administration (FDA) regarding some death relating incidences. As high iso-antibody production is closely associated with different immune reactions, a survey on the different immune response of dangerous O donors can lead to understanding their immune response profile. Objectives were to assess different immune responses in dangerous O cases. METHODS: Two groups of donors were selected after performing titration as a high titer (>512) and non-high titer (<128). Then CBC, CD markers, total immunoglobulin, complement assay, anti-VZV, -CMV, -EBV, -HSV, -Rubella, -Toxoplasma gondii, -HBV, -ASO, total protein and albumin, protein electrophoresis, lymphocyte proliferation, and gene expression of INF-gamma, IL2/4/10 were evaluated on both study groups. RESULTS & CONCLUSION: Total IgG, IgM, and IgA was higher in high titer group. Moreover, after using PHA and LPS, gamma globulins and lymphocyte proliferation were significantly higher in high titer cases. Real-time PCR also showed higher IL-2 production in high titer group. Identification of high responder's characteristics can be efficient in many complications. Moreover, high titer donors are dangerous for transfusion medicine. This pilot study showed differences in immune responses between HR and LR O blood donors for the first time. So, other aspects of the immune system such as genetic differences can be surveyed.

The Effect of TAX-1 Gene of Human T-cell Leukemia Virus Type-1 on the Expression of CCR5 in K562 Cell Line.

BACKGROUND: Tax-1 protein of Human T-cell Leukemia Virus type 1(HTLV-1) serves as a key transcriptional regulatory gene product and has a crucial role in transactivating genes of infected cells by employing their transcriptional factors. This modulation includes induction of genes which encode CC-chemokines and their receptors. In this study, a recombinant vector containing Tax-1 gene was made and tested for its ability to induce CCR5 (CC chemokine receptor 5) expression in K562 cell line. METHODS: In order to perform this research, two blood samples of HTLV-1 positive were obtained from Urmia blood transfusion center. After DNA extraction, a complete sequence of Tax-1 gene was amplified by specific primers. Recombinant vectors carrying Tax-1 gene were synthesized and transformed into Escherichia coli (E. coli). After bacteria transformation, bacteria containing recombinant plasmid were selected and purified. Then, the recombinant shuttle vectors, pCDNA3.1-TAX, were transfected into the cell culture (K562 cell line). Expression of CCR5 was measured after 72 hr by Syber Green Real-Time PCR method compared to control cell culture. Normalization was done with GAPDH as a standard gene. RESULTS: Cloning of Tax-1 gene in the vector, pCDNA3.1 was confirmed by colony PCR, restriction digestion, and sequencing methods. Expression of Tax-1 and CCR5 genes were confirmed by real time PCR and also, expression of CCR5 gene showed an 8-fold increase in comparison to mock-treated controls (p<0.05). CONCLUSION: Our data suggested that recombinant Tax-1 may have the enhancing effect on CCR5 expression rate at mRNA levels in K562 cell line. Further studies are necessary to evaluate the effect of pCDNA3.1-TAX on cell surface CCR5 expression.

A Study on the Association between CCRΔ32 Mutation and HCV Infection in Iranian Patients.

BACKGROUND: Mutations in the coding region of the Chemokine Receptor 5 (CCR5) genes reduce or eliminate CCR5 expression in immune cells and progression of HCV infection. This study aimed to investigate the role of this mutation in HCV infection in Iranian patients in comparison with healthy individuals. METHODS: 100 HCV infected patients and 100 healthy individuals were randomly selected. The CCR5Δ32 genotypes were determined using specific primers and PCR method. RESULTS: The agarose gel electrophoresis showed a189-bp fragment from wild type for both alleles of CCR5 gene. The CCR5-Δ32 allele was not found in any HCV infected and healthy subjects. CONCLUSION: The mutation in CCR5 gene was not detected in any of the two groups; therefore, the role of CCR5 gene expression in immune cells and progression of HCV infection needs to be studied in larger samples in our country.

Foods, Dietary Patterns and Occupational Class and Leukocyte Telomere Length in the Male Population.

Telomeres contain TTAGGG repetitive sequences and are located at the end of human chromosomes. Telomere dysfunction is associated with some age-related and chronic diseases, but its relationship with foods, dietary patterns, and occupational class in the young male population is not yet known. In this cross-sectional study, 300 healthy men, residents of Tehran, aged 25-40 years were enrolled from January to December 2016. We employed a cross-sectional study of 300 healthy people, residents of Tehran, aged 25-40 years. A food frequency questionnaire was used to obtain food intakes of all participants and converted into actual food intake (g/day). The principal components analysis was used to determine dietary patterns and other demographic characteristics. Leukocyte telomere length (TL) was measured by quantitative real-time polymerase chain reaction (PCR) to measure number of telomere repeat copy number (T) to the relative number of 36B4 copies (S) (T/S ratio). T/S in office-workers, waste recyclers, and other workers were 1.22 ± 0.4, 1.08 ± 0.3, and 1.094 ± 0.34, respectively. The results of multivariate linear regression adjusted for age, body mass index (BMI), and smoking were showed that whole grains (ß = 0.02; p = .05), refined grains, fruits and vegetables, fish and dairy products were associated with an increase in log-T/S, but consumption of nuts and seeds (ß = -0.00072; p = .06), meats (ß = -0.00043; p = .9), produced meats (ß = -0.00238; p = .03), oils and solid fats (ß = -0.00146; p = .03) had a negative relationship with log-T/S in all studied occupational classes. A positive relationship was reported between the healthy (ß = 0.017; p = .2) and traditional dietary pattern (ß = 0.012; p = .4) with log-T/S, but western pattern identified negative relationship (ß = -0.004; p = .7). Adherence to a healthy (with consumption whole grains, refined grains, dairy, and cereals) and then traditional pattern with increased consumption of fruits, vegetables and whole grains, fish and dairy products are necessary to prevent TL destruction in all studied occupational classes.

Sensing of Alzheimer's Disease and Multiple Sclerosis Using Nano-Bio Interfaces.

It is well understood that patients with different diseases may have a variety of specific proteins (e.g., type, amount, and configuration) in their plasmas. When nanoparticles (NPs) are exposed to these plasmas, the resulting coronas may incorporate some of the disease-specific proteins. Using gold (Au) NPs with different surface properties and corona composition, we have developed a technology for the discrimination and detection of two neurodegenerative diseases, Alzheimer's disease (AD) and multiple sclerosis (MS). Applying a variety of techniques, including UV-visible spectra, colorimetric response analyses and liquid chromatography-tandem mass spectrometry, we found the corona-NP complexes, obtained from different human serums, had distinct protein composition, including some specific proteins that are known as AD and MS biomarkers. The colorimetric responses, analyzed by chemometrics and statistical methods, demonstrate promising capabilities of the technology to unambiguously identify and discriminate AD and MS. The developed colorimetric technology might enable a simple, inexpensive and rapid detection/discrimination of neurodegenerative diseases.

Is Leukocyte Telomere Length Related with Lung Cancer Risk?: A Meta-Analysis.

BACKGROUND: Epidemiological studies have probed the correlation between telomere length and the risk of lung cancer, but their findings are inconsistent in this regard. The present meta-analysis study has been carried out to demonstrate the association between relative telomere length in peripheral blood leukocytes and the risk of lung cancer using an established Q-PCR technique. METHODS: A systematic search was carried out using PubMed, EMBASE, and ISI before 2015. A total of 2925 cases of lung cancer and 2931 controls from 9 studies were employed to probe the relationship between lung cancer and telomere length .ORs were used at 95% CI. Random-effects models were used to investigate this relationship based on the heterogeneity test. Heterogeneity among studies was analyzed employing subgroup analysis based on type studies and the year of publication. RESULTS: Random-effects meta-analysis revealed that patients with lung cancer were expected to have shorter telomere length than the control (1.13, 95% CI: 0.82-1.81, P=0.46). The summary of the pooled ORs of telomere length in adenocarcinoma lung cancer patients was 1 (95%CI=0.68-1.47, I2=93%) compared to patients with squamous cell lung cancer, which was 1.78 (95% CI=1.25-2.53, I2=3.9%). The meta-regression revealed that the effect of telomere length shortening, decreased and increased with the year of publication and the age of risks to lung cancer, was clearly related to short telomeres lengths. CONCLUSION: Lung cancer risks clearly related with short telomeres lengths. In patients with breathing problems, lung cancer risk can be predicted by telomere length adjustment with age, sex, and smoking.

Frequency of West Nile Virus Infection in Iranian Blood Donors.

West Nile virus (WNV) can be transmitted by blood transfusions and organ transplants. This study was a retrospective study which was performed in Blood Transfusion Center to evaluate the WNV infection in blood donors in Iran. A total of 540 blood samples were taken from volunteer healthy donors who referred for blood donation to Chabahar Blood Center. The presence of WNV was studied by detecting immunoglobulin G (IgG) WNV by enzyme linked immune sorbent assay (ELISA). Demonstration of elevated WNV IgG confirmed by immunoflouorescence assay (IFA) Euroimmun kit. Out of the 540 samples 17.96 % (97 cases) were seropositive by ELISA and 1.48 % (8 cases) was seropositive by IFA. This means that 8.24 % of ELISA seropositive samples were confirmed by IFA. Special attention should be paid to criteria of donor selection, albeit positive results may be due to a previous infection in these donors.

Quality evaluation of human serum albumin prepared by heat denaturation in Iran: an experience for developing countries.

Blood and plasma are unique resources and access to these products save life. In this study, albumin demand and surplus plasma makes it possible to use local experiences in plasma industry for preparation of albumin so plasma was heated after stabilization; afterward denatured proteins were precipitated and separated by continuous centrifuge system. The supernatant contained albumin was filtrated, diafiltrated, ultrafiltrated, formulated and pasteurized. Albumin preparation in pilot scale with heat denaturation was performed for the first time in Iran. This method using surplus plasma is recommended for all countries that have no access to plasma fractionation industry. Therefore with more attention it has potential for use in the production of safe plasma derived products and thereby it can be used as a safe product in clinic.

Frequency of dengue virus infection in blood donors in Sistan and Baluchestan province in Iran.

Risk of dengue virus in the blood supply has been demonstrated in recent studies. In this paper, Chabahar in Sistan and Baluchestan province, south east of Iran, was selected for studying dengue infection because of its climatic and geographical situation in the middle of the transit way between East Asia and other countries. The blood samples were taken from volunteer healthy donors who were referred to the Chabahar Blood Center for blood donation. The presence of dengue virus (DENV) was studied by detecting IgG to DENV by enzyme linked immune sorbent assay (ELISA). Reactive ELISA results were confirmed by an immune flouorescence assay (IFA). According to the results, some of the healthy donors were infected by DENV, which could not been recognized in donor selection. Therefore, special attention should be paid to the criteria of donor selection and additional screening tests are recommended.

Personalized protein coronas: a "key" factor at the nanobiointerface.

It is now well known that the primary interactions of biological entities (e.g., tissues and cells) with nanoparticles (NPs) are strongly influenced by the protein composition of the "corona" (i.e., the NP surface attached proteins). The composition of the corona strongly depends on the protein source (e.g., human plasma). Because the protein source determines the NP corona, it is reasonable to hypothesize that humans with specific disease(s) may have specific NP coronas. To test this hypothesis, we incubated two different hydrophobic/hydrophilic types of NPs (polystyrene and silica) with plasma from human subjects with different diseases and medical conditions (e.g., breast cancer, diabetes, hypercholesterolemia, rheumatism, fauvism, smoking, hemodialysis, thalassemia, hemophilia A and B, pregnancy, common cold and hypofibrinogenemia). Our results demonstrate that the type of disease has a crucial role in the protein composition of the NP corona. Based on these results, we introduce the concept of the "personalized protein corona" (PPC) as a determinant factor in nano-biomedical science. This study will help researchers rationally design experiments based on the "personalized protein corona" for clinical and biological applications.