Peripheral Lymphocyte Cultures Research Articles

Chromosomal aberrations in Ukrainian red-spotted dairy breed cows when using different breeding methods

Aim. The aim of this study was a comparative analysis of karyotypes of purebred, inbred, and crossbred cows of the Ukrainian red-spotted dairy breed in order to detect their degree of chromosomal aberrations. Methods. The cyto- genetic study involved 156 cows of the Ukrainian red-spotted dairy breed from two research farms of M.V. Zubets Institute of Animal Breeding and Genetics, the NAAS. Cultures of peripheral lymphocytes, stimulated with phytohemagglutinin (PHA, Difco, USA) were used according to the method of Moorhead et al. (1960). The chromosomal preparations were stained with 2 % Giemsa solution (Giemsa, Merk) and analyzed under an Axiostar plus (Carl Zeiss, Germany) light microscope at 1,000 times magnification. Metaphase slides were photographed with a digital camera Olympus D-460 ZOOM. The biometric processing of the study results was conducted by ANOVA using Microsoft Excel 2010. Results. Genomic mutations (aneuploidy and polyploidy) and structural aberrations of chromosomes were found in the karyotypes of the investigated purebred, inbred, and crossbred cows. The total share of aberrant cells varied from 11.4 ± 0.39 % in purebred, 11.8 ± 0.40 % in inbred to 17.30 ± 0.85 % in crossbred cows. In two animals the Robertson translocation, rob (1 : 29) was found. A statistically significant difference between purebred, inbred, and crossbred animals was found regarding cells with aneuploidy and structural chromosomal aberrations. Conclusions. The karyological analysis yielded data showing changes in genetic structure that can have an influence on fertility and production in different breeding programs.

Down’s Syndrome Presented with Transmission of Maternal Translocation of 2; 21 Chromosomes. A Case Report

Background: Down's syndrome is a genetic condition marked by distinctive physical characteristics and some degree of cognitive impairment. Down's syndrome is mostly caused by trisomy of chromosome 21, while chromosome translocations are also frequent. Objective: To evaluate a rare 2;21 translocation in the proband's family that was associated with Down syndrome. Methods: Chromosomal analysis was carried out using the G-banding technique and traditional peripheral lymphocyte culture. Case: The proband was a 9 months baby boy of non-consanguineous parents. The doctors clinically diagnosed him as having Down's syndrome with all typical features. The proband was found to have trisomy 21 associated with a 2;21 translocation inherited from his mother because his mother has the same type of translocation without any phenotypic features. Maternal age at the time of the study was 35 years and first pregnancy ended in stillbirth at 26th weeks of gestation, the proband was the second issue. His maternal aunt and cousin brother both had the same type of translocation. In chromosomal analysis, the proband's father and uncle had normal genotypic distribution. The current example was a Down's syndrome case with one normal 21 no chromosome and one Reciprocal translocation t (2;21). Conclusion: The present case of Down’s syndrome occurs due to reciprocal translocation (2;21) probably has arisen by familial transmission. Once an imbalanced translocation in the fetus/child has been found, the prenatal cytogenetic analysis is critical for the next pregnancies. Bangladesh Medical Res Counc Bull 2023; 49(2): 143-147

Candidate genes for infertility: an in-silico study based on cytogenetic analysis

BackgroundThe cause of infertility remains unclear in a significant proportion of reproductive-age couples who fail to conceive naturally. Chromosomal aberrations have been identified as one of the main genetic causes of male and female infertility. Structural chromosomal aberrations may disrupt the functioning of various genes, some of which may be important for fertility. The present study aims to identify candidate genes and putative functional interaction networks involved in male and female infertility using cytogenetic data from cultured peripheral blood lymphocytes of infertile patients.MethodsKaryotypic analyses was done in 201 infertile patients (100 males and 101 females) and 201 age and gender matched healthy controls (100 males and 101 females) after 72 h peripheral lymphocyte culturing and GTG banding, followed by bioinformatic analysis using Cytoscape v3.8.2 and Metascape.ResultsSeveral chromosomal regions with a significantly higher frequency of structural aberrations were identified in the infertile males (5q2, 10q2, and 17q2) and females (6q2, 16q2, and Xq2). Segregation of the patients based on type of infertility (primary v/s secondary infertility) led to the identification of chromosomal regions with a significantly higher frequency of structural aberrations exclusively within the infertile males (5q2, 17q2) and females (16q2) with primary infertility. Cytoscape identified two networks specific to these regions: a male specific network with 99 genes and a female specific network with 109 genes. The top enriched GO terms within the male and female infertility networks were “skeletal system morphogenesis” and “mRNA transport” respectively. PSME3, PSMD3, and CDC27 were the top 3 hub genes identified within the male infertility network. Similarly, UPF3B, IRF8, and PSMB1 were the top 3 hub genes identified with the female infertility network. Among the hub genes identified in the male- and female-specific networks, PSMB1, PSMD3, and PSME3 are functional components of the proteasome complex. These hub genes have a limited number of reports related to their respective roles in maintenance of fertility in mice model and humans and require validation in further studies.ConclusionThe candidate genes predicted in the present study can serve as targets for future research on infertility.

In vitro study of the effect of resveratrol purified from the skin of Iraqi black grape (Vitis vinifera) on lymphocyte cultures isolated from the blood of patients with lymphoma.

The natural stilbene compound resveratrol (RSV) was extracted and purified locally from the black grape skin (Vitis vinifera) cultivated in Iraq. Cultures of human peripheral lymphocytes were obtained from the blood samples of patients with and without lymphoma to be treated with RSV at different concentrations. Three RSV concentration levels were subjected to isolated lymphocytes from blood samples of Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL), and without lymphoma to estimate the change in TNF-α and IL-10. Resveratrol seemed to differently affect cytokines level in normal and lymphoma lymphocytes in relation to its concentration. The lowest resveratrol concentration (50 µg/ml) decreased TNF-α levels for patients without lymphoma and all NHL patients in contrast to the HL sample. Treating normal lymphocytes with a higher dose (1000 µg/ml) might elevate the levels of TNF-α in almost all samples. There was an inverse relationship between both cytokines in most treatments; with the increase in TNF-α level, there was a decrease in IL-10 level except in HL and normal lymphocytes treatment. The locally purified resveratrol could serve as a multi-target drug that modulates the immune system to improve body defense in patients suffering from lymphoma and in patients without lymphoma by altering cytokine levels in response to different resveratrol concentrations in a different manner.

Cytogenetic and Oxidative Status of Cultured Lymphocytes after Exposure to Extracts from Rhizopogon Mushrooms (Agaricomycetes) In Vitro.

In this study, acetone and water extracts obtained from edible mushrooms, Rhizopogon luteolus Fr. and Rh. roseolus (Corda) Th. Fr., containing important bioactive components were used. Total antioxidant capacity (TAC) and total oxidative stress (TOS) levels were examined on human peripheral lymphocyte culture treated with the respective extracts. Levels of genetic damage and cytotoxic effects of the respective extracts on lymphocytes were also tested. In general, when TAC levels of the extracts on cells were examined, a concentration-dependent increase was observed; a negative correlation was found between TOS data and concentration. Genotoxicity tests (chromosome aberration and micronucleus analysis) revealed that the concentrations of the extract applications did not significantly (P > 0.05) change genotoxicity on human peripheral lymphocyte culture compared to the negative control. Considering all of the results obtained, it was determined that applications of Rh. luteolus and Rh. roseolus extracts, especially at concentrations of 50 and 100 mg/L, increased the TAC value of lymphocytes, which play an important role in the human immune system, without causing genetic or oxidative stress.

Cytogenetic and oxidative effects of three lichen extracts on human peripheral lymphocytes.

In the present study, we investigated cytogenetic and oxidative [total antioxidant capacity (TAC), totaloxidant status (TOS)] effects of methanol and water extracts of Cladonia chlorophaea (Flörke ex Sommerf.) Sprengel, Dermatocarpon miniatum (L.) W.Mann and Parmelia saxatilis (L.) Ach. on cultured human lymphocytes. In addition, different phenolic compounds in the extracts were quantified by high performance liquid chromatography (HPLC) analysis. As a result of HPLC analysis, methanol extracts of all lichen species tested had higher phenolic compounds. Likewise, methanol extracts of eachlichen increased TAC levels in lymphocytes more thanwater extracts. The TOS levels of the cells treated with different concentrations (1-100mg/L) of the extracts decreased due to the increasing concentration of the extracts. Genotoxicity experiments revealed that the tested lichen extracts did not significantly increase (p>0.05) thelevel of genotoxicity on human peripheral lymphocyte culture compared to the negative control group. The results showed that C.chlorophaea, D.miniatum and P.saxatilis lichens, which were found to be a rich source of phenolic compounds, might be of interest in the pharmaceutical and food industries.

In silico pathway analysis based on chromosomal instability in breast cancer patients

BackgroundComplex genomic changes that arise in tumors are a consequence of chromosomal instability. In tumor cells genomic aberrations disrupt core signaling pathways involving various genes, thus delineating of signaling pathways can help understand the pathogenesis of cancer. The bioinformatics tools can further help in identifying networks of interactions between the genes to get a greater biological context of all genes affected by chromosomal instability.MethodsKaryotypic analyses was done in 150 clinically confirmed breast cancer patients and 150 age and gender matched healthy controls after 72 h Peripheral lymphocyte culturing and GTG-banding. Reactome database from Cytoscape software version 3.7.1 was used to perform in-silico analysis (functional interaction and gene enrichment).ResultsFrequency of chromosomal aberrations (structural and numerical) was found to be significantly higher in patients as compared to controls. The genes harbored by chromosomal regions showing increased aberration frequency in patients were further analyzed in-silico. Pathway analysis on a set of genes that were not linked together revealed that genes HDAC3, NCOA1, NLRC4, COL1A1, RARA, WWTR1, and BRCA1 were enriched in the RNA Polymerase II Transcription pathway which is involved in recruitment, initiation, elongation and dissociation during transcription.ConclusionThe current study employs the information inferred from chromosomal instability analysis in a non-target tissue for determining the genes and the pathways associated with breast cancer. These results can be further extrapolated by performing either mutation analysis in the genes/pathways deduced or expression analysis which can pinpoint the relevant functional impact of chromosomal instability.

Antigenotoxic and Antimutagenic Effects of Andrographis paniculata , a Traditional Medicinal Herb against Genotoxicity of Cyclophosphamide: An In Vitro Study on Human Peripheral Lymphocytes

Andrographis paniculata (family: Acanthaceae) is a medicinal herb—used in Indian system of medicine (Ayurveda, Siddha, and Unani), traditional and folk systems to treat various illnesses. This study examined the phytochemical constituents of ethanol extract from A. paniculata and its protective effect against genotoxicity caused by cyclophosphamide (CPA). Phytochemical screening and estimation of total phenolic content were analyzed using standard methods. The bioactive components from the ethanol extract of A. paniculata (EAP) were analyzed using gas chromatography-mass spectrometry. To investigate the protective effect of EAP against CPA-induced genotoxicity, human peripheral lymphocyte cultures were used. To test the antigenotoxic and antimutagenic effects of EAP, lymphocytes were treated with different concentrations of extract (50∼250 mg/mL) alone and co-treated along with CPA+EAP for 48 h. The cells were analyzed for structural chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) in control, CPA treated, and CPA+ EAP co-treated lymphocytes. Results of the study revealed that the lymphocyte cultures which had 48 h continuous exposure to EAP (50∼250 mg/mL) did not show any significant changes in CAs and SCE frequencies. These results substanti-ate the antimutagenic nature of the extract. Furthermore, the lymphocytes co-treated with CPA along with extract showed a significant reduction in CAs (reduced from 26.50±2.50% to 11.00±1.00%) and SCEs (reduced from 9.92±0.63 per cell to 4.56±0.18 per cell). These results suggest that A. paniculata is protective against CPA induced genotoxicity and put forward its possible use as a supplement with chemotherapeutic drugs.

Cytokinesis-Block Micronucleus Assay Using Human Lymphocytes as a Sensitive Tool for Cytotoxicity/Genotoxicity Evaluation of AgNPs

Silver nanoparticles (AgNPs) are the most used nanomaterials worldwide due to their excellent antibacterial, antiviral, and antitumor activities, among others. However, there is scarce information regarding their genotoxic potential measured using human peripheral blood lymphocytes. In this work, we present the cytotoxic and genotoxic behavior of two commercially available poly(vinylpyrrolidone)-coated silver nanoparticle (PVP–AgNPs) formulations that can be identified as noncytotoxic and nongenotoxic by just evaluating micronuclei (MNi) induction and the mitotic index, but present enormous differences when other parameters such as cytostasis, apoptosis, necrosis, and nuclear damage (nuclear buds (NBUDs) and nucleoplasmic bridges (NPBs)) are analyzed. The results show that Argovit (35 nm PVP–AgNPs) and nanoComposix (50 nm PVP–AgNPs), at concentrations from 0.012 to 12 μg/mL, produce no changes in the nuclear division index (NDI) or micronuclei (MNi) frequency compared with the values found on control cultures of human blood peripheral lymphocytes from a healthy donor. Still, 50 nm PVP–AgNPs significantly decrease the replication index and significantly increase cytostasis, apoptosis, necrosis, and the frequencies of nuclear buds (NBUDs) and nucleoplasmic bridges (NPBs). These results provide evidence that the cytokinesis-block micronucleus (CBMN) assay using human lymphocytes and evaluating the eight parameters provided by the technique is a sensitive, fast, accurate, and inexpensive detection tool to support or discard AgNPs or other nanomaterials, which is worthwhile for continued testing of their effectiveness and toxicity for biomedical applications. In addition, it provides very important information about the role played by the [coating agent]/[metal] ratio in the design of nanomaterials that could reduce adverse effects as much as possible while retaining their therapeutic capabilities.

Identification and copy number profiling of sex chromosome specific gene fragments in crossbred pigs with numerically normal karyotype

We examined the copy number profile of sex chromosome specific genes in crossbred pigs with numerically normal karyotype. A total of 30 (15M + 15F) Landlly (Landrace crossbred) pigs from a research farm were cyto-screened by karyotyping to determine the number of chromosomes per diploid cell using short-term peripheral lymphocyte culture technique. All pigs had numerically normal karyotype with 38, XX (female) and 38, XY (male). Genomic DNA was extracted from cyto-screened pigs of both sexes. A set of sex chromosome (X and Y) specific single copy gene fragments along with an autosomal gene fragment were selected out of 12 pairs of primers on the basis of male specific PCR amplification, PCR and qPCR specificity. Quantitative real-time PCR was performed to study the relative copy number change of selected Phosphate repressible alkaline phosphatase X linked (PHOX) gene, using the Eukaryotic Translation Initiation Factor 1A Y-Linked (EIF1AY) gene as control and Breast Cancer Metastasis-Suppressor 1-Like (BRMS1 L- autosomal gene) as the reference gene. The relative copy number of the PHOX gene in females was found to be 1.873 times higher compared with the EIF1AY gene in males. The present study indicates that the real-time quantitative real-time PCR based copy number analysis can be helpful for the detection of sex chromosome ratio in pigs to aid as a preliminary screening for numerical sex chromosomal aneuploidies, adding to the rate of throughput of traditional cyto-screening.

The Determination of the Effects of Wild Thyme (Thymbra spicata L.) and Cumin (Cuminum cyminum L.) Extracts on SCE (Sister Chromatid Exchange) in Human Lymphocyte Chromosomes

The purpose of this work is to determine of T. spicata L. and C. cyminum L. plant extracts on sister chromatid exchange rate in human peripheral lymphocyte culture as in vitro . Human blood lymphocyte cells, T. spicata L. and C. cyminum L. plant extracts were allowed to interact with doses of 0.05 µL/mL, 0.10 µL/mL, 0.15 µL/mL and 0.20 µL/mL for 24 hours. When trial groups where plant extracts were applied were compared with negative control and mitomycin-C (MMC) which was used as positive control, it was determined that the extract doses applied led to an increase in sister chromatid exchange rate. Also, it was found that increasing concentrations of plant extracts caused cell replication index to decrease. Between C. cyminum L. doses and RI, a negative correlation (r = -0.95) was observed and between T. spicata L . doses and RI a negative correlation was observed (r = -0.94) respectively. As a result of this study; T. spicata L. and C. cyminum L. plant extracts were found to have genotoxic and clastogenic effects on human peripheral lymphocytes.

Biological relevance of CuFeO2 nanoparticles: Antibacterial and anti-inflammatory activity, genotoxicity, DNA and protein interactions

Heterometal oxide nanoparticles of bioessential metals are shedding new light to nanoparticle-inspired bioapplications. Pairing bioreactive elements like copper and iron can affect the redox dynamic and biological profile of the nanomaterial. Given the complexity of physicochemical properties, biological activity and toxicity concerns, extensive exploration is demanded, especially when active and less active oxidation states participate as in case of cuprous-ferric delafossite CuFeO2 (copper(I)-iron(III)), a less widespread nanomaterial. In that vein, CuFeO2 nanoparticles were synthesized and biological profile was evaluated in comparison with cuprous oxide (Cu2O NPs) counterpart, an already established antimicrobial agent. Interactions with bacteria, proteins and DNA were examined. Cu2O NPs exhibited stronger antibacterial activity (IC50 < 25 μg/ml) than CuFeO2 NPs (IC50 > 100 μg/ml). In vitro exposure of nanoparticles on plasmid DNA unveiled toxicity in the form of DNA damage for Cu2O and enhanced biocompatibility for CuFeO2 NPs. Genotoxicity estimated by the frequency of sister chromatid exchanges, cytostaticity based on the proliferating rate indices and cytotoxicity based on the mitotic indices at human peripheral lymphocyte cultures were all significantly lower in the case of CuFeO2 NPs. Furthermore, through in vitro albumin denaturation assay, CuFeO2 NPs showed better performance in protein denaturation protection, correlating in superior anti-inflammatory activity than Cu2O and similar to acetylsalicylic acid. Synergy of copper(I)-iron(III) in nanoscale is apparent and gives rise to fruitful bioapplications and perspectives.

Resveratrol Attenuates LPS-induced Apoptosis via Inhibiting NF- κ B Activity in Chicken Peripheral Lymphocyte Cultures

Resveratrol Attenuates LPS-induced Apoptosis via Inhibiting NF- κ B Activity in Chicken Peripheral Lymphocyte Cultures

Cytogenetic screening in couples with Habitual Abortions

ObjectiveHabitual abortion (HA) is defined at least three consecutive pregnancy losses. One of the etiologic causes is parental chromosomal anomalies. In this study, we aimed to that investigate the effect of parental chromosomal abnormalities on HA. MethodsThe cytogenetic results of patients with at least three abortions referred to our university hospital between January 2010 - March 2017 were evaluated. A total of 1154 couples with HA were analysed. Peripheral lymphocyte cultures incubated for 72 h were used for karyotype analysis via the Giemsa banding technique. ResultsOf a total 1154 couples (2308 patients) 37 female (3.2%) and 17 male (1.47%) had abnormal karyotypes. Reciprocal translocation carriage (n = 26; 1.12%) was the most commonly detected structural anomaly, followed by X chromosome mosaicism (n = 16; 0.69%),Robertsoniantranslocation (n = 9; 0.38%), Chromosomal inversion (n = 6; 0.26%). Chromosomal polymorphisms, which are considered minor chromosomal changes, were detected in 221 (9.57%) individuals. ConclusionOur study exhibits that chromosomal analysis in patient with HA is an appropriate approach to elucidate the aetiology of HA. Data from cytogenetic screening can be used in guiding couples planning future pregnancies and in prenatal diagnosis of chromosomal anomalies in the foetus.

An Analysis of Cytogenetic and Clinical Phenotype of Klinefelter Syndrome Over 17 Years

Background: Clinical phenotype in Klinefelter syndrome (KS) shows utmost contrariety according to the genetic presentation. The karyotype 47, XXY is one of the commonest types of sex chromosomal abnormality in males presenting with infertility, hypogonadism, small penis, gynaecomastia and tall stature. Cytogenetic study is the only way to differentiate between chromosomal abnormality and other androgen deficiency disorders. The aim of this study was to investigate cytogenetic and phenotypic profile of Klinefelter syndrome in a group of referred patients with suspected genetic disorders.Methods: This observational study was carried out at the Cytogenetic Laboratory of the Department of Immunology BIRDEM General Hospital for a period of seventeen years from 2000 to 2016. A total of 9,216 patients suspected for different chromosomal abnormalities (e.g. numerical chromosomal disorders, primary amenorrhoea, ambiguous genitalia etc.) were included in this study referred by physicians of various discipline from different areas of Bangladesh. From the patients referred for cytogenetic study, detailed family history and physical findings were noted. Complete genetic examination and pedigree construction was done to exclude non-chromosomal causes of anomalies. For cytogenetic analysis, peripheral lymphocyte culture by the standard method using the G-banding technique was employed.Results: In this study 1.67% (154) of referred patients were diagnosed as Klinefelter syndrome in cytogenetic study and most of them were diagnosed in their adulthood between 20-29 years of age. Classical cytogenetic form of KS-47, XXY (87%) were most common followed by other mosaic and supernumerary X chromosome aneuploidy. Most of the patients presented with tall stature (61.7%) followed by other features such as gynaecomastia (45.5%), eunuchoid skeleton (29.8%), sexual dysfunction (34.41%), small penis (22.7%), and delayed development of secondary sex characteristics (22.7%).Conclusion: Diagnosis of Klinefelter syndrome in early age before puberty is needed to be differentiated from other related disorders and thereby improving the quality of life by providing appropriate and timely treatment. Therefore, we have to focus to improve our overall capacity to diagnose genetic disorders for proper interventionBirdem Med J 2018; 8(2): 126-131

Safety and persistence of WT1-specific T-cell receptor gene−transduced lymphocytes in patients with AML and MDS

AbstractWilms' tumor 1 (WT1) is constantly expressed in leukemic cells of acute leukemia and myelodysplastic syndrome (MDS). A T-cell receptor (TCR) that specifically reacts with WT1 peptide in the context of HLA-A*24:02 has been identified. We conducted a first-in-human trial of TCR–gene transduced T-cell (TCR–T-cell) transfer in patients with refractory acute myeloblastic leukemia (AML) and high-risk MDS to investigate the safety and cell kinetics of the T cells. The WT1-specific TCR-gene was transduced to T cells using a retroviral vector encoding small interfering RNAs for endogenous TCR genes. The T cells were transferred twice with a 4-week interval in a dose-escalating design. After the second transfer, sequential WT1 peptide vaccines were given. Eight patients, divided into 2 dose cohorts, received cell transfer. No adverse events of normal tissue were seen. The TCR-T cells were detected in peripheral blood for 8 weeks at levels proportional to the dose administered, and in 5 patients, they persisted throughout the study period. The persisting cells maintained ex vivo peptide-specific immune reactivity. Two patients showed transient decreases in blast counts in bone marrow, which was associated with recovery of hematopoiesis. Four of 5 patients who had persistent T cells at the end of the study survived more than 12 months. These results suggest WT1-specific TCR-T cells manipulated by ex vivo culture of polyclonal peripheral lymphocytes survived in vivo and retained the capacity to mount an immune reaction to WT1. This trial was registered at www.umin.ac.jp as #UMIN000011519.

Anticarcinogenic and antimutagenic activity of Alstonia scholaris on the albino mice bone marrow cells and peripheral human lymphocyte culture against methyl methane sulfonate induced genotoxicity.

Background:The use of medicinal plants in modern medicine for the prevention and treatment of cancer is an important aspect. For this reason, it is important to identify antitumor promoting agents present in medicinal plants commonly used by the human population.Materials and Methods:We used in vivo and in vitro methods using chromosomal aberrations (CAs), sister chromatid exchange (SCE) and replication index (RI) as markers, exposed by methyl methanesulfonate (MMS) as well as alcoholic extract of Alstonia scholaris in five increasing concentrations (200, 250, 300, 350 and 400 mg/kg body weight for in vivo and 150, 200, 250 and 300 μg/ml of culture) and of three different durations of 24, 48 and 72 h in the presence as well absence of S9 mix.Results:Extracts of Alstonia reduces the total aberrant cells ranges from 10.0% to 41.84% and frequencies of aberration in the aberrant cells ranges from 220 to 124 against 290 aberrations causes due to MMS in vivo. Similarly in the in vitro, it reduces CAs (39.62%, 32.83%, and 38.48%) and (45.31%, 44.46%, and 38.34%) at 24, 48, and 72 h of exposure respectively; in the absence as well as presence of liver S9 fraction. It also reduces SCE from 7.70 to 4.20 per cell and enhances RI from 1.45 to 1.64.Conclusion:Extracts of Alstonia significantly reduces the number of aberrant cells and frequency of aberration per cell at each concentration and duration of exposure in vivo; and CAs and SCE in vitro and enhances RI.

Cytogenetic Analysis and Thrombophilia-Associated Gene Mutations of Couples with Recurrent Miscarriage

Introduction: Three or more pregnancy losses before 20 weeks of gestation are usually defined as recurrent miscarriage. Parental chromosomal translocations, thrombophilic gene polymorphisms, autoimmune factors, uterine, endocrine factors associated with recurrent miscarrige (RM). Factor V Leiden, prothrombin gene mutation G20210A, protein S/Protein C/antithrombin deficiency and MTHFR mutations responsible for hereditary thrombophilia and has been included to keep a very common practice of RM pathogenesis. This study aimed to determine the incidence of these factors believed to be the effects of RM. Materials and methods: All patients were took a full genetic analysis; full genetic examination and pedigree drawing was done to exclude known nonchromosomal causes of the anomaly. Cytogenetic analysis was done for 635 patients. The study included peripheral lymphocyte culture by a standard method using Leishmann-banding technique, centromerebanding (C-banding), and nucleolar organizing region staining was done when needed. Pyrosequencing was used to genotype the 392 individual. Results: Male/female in patients with an inherited thrombophilia tested four types of changes, including; MTHFR C677T/1298, FV Leiden G1691A and prothrombin G20210A. Among them, 152 men and woman did not carry any mutation.

Simple and convenient preparation of some bicyclic alcohols and epoxide derivatives: promising antibiotic activities of (rac)-dimethyl 3,8-dioxatricyclo[3.2.1.02,4]oct-6-ene-6,7-dicarboxylate

The biological activity of alcohol and epoxide groups containing bicyclic skeletal structures was investigated in terms of antibiotic and toxic effects at cellular level. The present protocol, which utilizes readily available starting materials and mild reaction conditions, provides an attractive approach to a series of functionalized bicyclic alcohol and epoxide derivatives with promising biological applications. In this protocol, while some of the bicyclic compounds were synthesized as in the literature, the other bicyclic compounds were synthesized by new synthetic methods. The antibiotic and toxic properties of the synthesized bicyclic compounds were investigated, and some of their biological activities were tested in prokaryotic and eukaryotic systems. (rac)-Dimethyl 3,8-dioxatricyclo[3.2.1.02,4]oct-6-ene-6,7-dicarboxylate, one of the synthesized molecules that showed antibiotic activity, was tested in two different living systems (prokaryotic and eukaryotic) in terms of biological and toxic effects at cellular level using a peripheral lymphocyte culture assay. Mitotic index for this compound was calculated in lymphocyte cultures.

Computer assisted human karyotyping – an analysis of 131 cases

Introduction: Karyotyping is the process of pairing and ordering all the chromosomes of an organism, thus providing a genome-wide snapshot of an individual’s chromosome. G-banded karyograms are routinely used to diagnose a wide range of chromosomal abnormalities in individuals. Although the resolution of chromosomal changes detectable by G-banded karyotyping is typically a few megabases, this can be sufficient to diagnose certain categories of abnormalities. Objectives: The primary aim of this study was to investigate the different types of chromosomal aberrations and their relative frequencies in a group of referred patients with suspected genetic disorders. Methods: This observational study was carried out at Armed Forces Institute of Pathology (AFIP) for a period of two years from January 2011 to December 2012. A total of 131 patients were included in this study. These patients were referred to AFIP from different Combined Military Hospitals (CMH) of Bangladesh Army and also from civil medical installations. All the patients were subjected to full genetic study; complete genetic examination and pedigree construction was done to exclude nonchromosomal causes of anomaly. Detailed history and physical findings were also noted in a prescribed format. The study included peripheral lymphocyte culture by a standard method using the G–banding technique. 32 JAFMC Bangladesh. Vol 9, No 2 (December) 2013 Results: Out of 131 patients, 54 (42.2%) were male and 77 (57.8%) were female with male to female ratio 0.7:1. The age limit of the patient ranges from 04 days to 70 years. Most of the patients (32.1%) were in the age group of 0–10 years followed by 21–30 year age group (30.5%). Consanguineous marriage was found in 15 (11.5%) cases in which 3 (2.3%) cases had chromosomal aberrations. Recurrent abortion was the main clinical indication (18.3%) followed by infertility (15.3%). Chromosomal aberrations were detected in 26 patients (19.8%); of these, 20 (15.2%) involved autosomes, while only 6 (4.6%) involved gonosomes. Trisomy 21 was detected in 12 (9.1%) patients and Philadelphia chromosome was found in 8 (6.1%) patients. Turner syndrome was detected in 5 (3.8%) patients and Klinefelter syndrome was found in 1 (0.8%) patient. Conclusion: The precise delineation of different types of chromosomal aberrations is only possible using clinical examination and advanced cytogenetic tools by experienced cytogeneticists. DOI: http://dx.doi.org/10.3329/jafmc.v9i2.21823 Journal of Armed Forces Medical College Bangladesh Vol.9(2) 2013