A mixed-apoptotic effect of Jurinea mesopotamica extract on prostate cancer cells: a promising source for natural chemotherapeutics.

This research investigates the potential use of Jurinea mesopotamica Hand.-Mazz. (Asteraceae) in cancer treatment. In this study, a plant extract was prepared using all parts of J. mesopotamica, and its effect on the proliferation of cancer and normal cells was tested using the MTT method. It was found to have a selective cytotoxic effect on prostate cancer cells, with the lowest IC50 (half-maximal inhibitory concentration) of 10µg/mL found in the butanol extract (JMBE). The extract suppressed the proliferation of prostate cancer cells (67 %), disrupted organelle integrity (49 %), increased reactive oxidative stress (66 %), and triggered cell death (51 %). In addition, apoptotic gene expressions and protein levels increased, and the profile of amino acids related to energy metabolism was elevated. Based on LC-MS/MS results, the plant contained higher levels of flavonoids, including isoquercitrin, cosmosiin, astragalin, nicotiflorin, luteolin, and apigenin. These results suggest that J. mesopotamica has a selective effect on prostate cancer due to its high flavonoid content and might be a promising natural alternative for cancer treatment.

Minor variant of AHSG gene 767C>G polymorphism may decrease the risk of gestational diabetes mellitus.

Insulin resistance plays a central role in the development of gestational diabetes mellitus (GDM). The fetuin A molecule, of which serum level increases during pregnancy, is an inhibitor of insulin receptor tyrosine kinase and it is associated with insulin resistance. The aim of this study is to research the relationship of -843A>T (rs2248690) and 767C>G (rs4918) polymorphisms in the alpha-2-Heremans Schmid glycoprotein (AHSG) gene which is responsible for the synthesis of fetuin A and its association with (GDM). In this study, 83 pregnant women with GDM who applied to the Obstetrics and Gynaecology Clinics and 100 normal pregnants enrolled as the control group. Genotyping of AHSG gene polymorphisms was performed by using the TaqMan allelic discrimination kit with real time PCR device. In our study, homozygous GG genotype which was polymorphic in the 767C>G polymorphism of AHSG gene was found significantly low in the patient group (p < .05). Genotype distribution of AHSG gene -843A>T polymorphism was not statistically significant between the patient and control groups (p > .05). Our results showed that homozygous GG variant of AHSG gene 767C>G polymorphism may have protective effect against the development of GDM.Impact statementWhat is already known on this subject? Insulin resistance has a central role in the development of gestational diabetes mellitus (GDM). The fetuin A molecule is an inhibitor of insulin receptor tyrosine kinase and it is associated with insulin resistance. The -843T>A and 767G>C polymorphisms of AHSG gene encoding fetuin A are affects serum fetuin A level. In a single study investigating the relationship between GDM and AHSG gene 767G>C polymorphism, there was no significant difference in genotype distribution but it was reported that the frequency of G allele increased in GDM group and this increase provided a weak risk or predisposition.What the results of this study add? The present study revealed that homozygous GG variant of AHSG gene 767C>G polymorphism may decrease the risk of GDM.What the implications are of these findings for clinical practice and/or further research? Protective effect of homozygous GG variant of AHSG gene 767C>G polymorphism, can be used as a molecular biomarker to predict the development of GDM. These results should be supported by further research in larger sample sizes.

miRNA-mediated apoptosis activation through TMEM 48 inhibition in A549 cell line.

Lung has critic function in gas exchange, supplying oxygen to all cells. Rapid metastasis and the high rate of mortality characterises lung cancer. There are two types of this disease, small cell and non-small cell, which differs from each other according to histopathologic features. To date, many therapeutic approaches have been developed to destroy this deadly type of cancer, which one of them is mRNA targeted therapies through miRNA. miRNAs are 19-25 base paired molecules be able to suppress and destruct mRNA and found to be involved in development and progression of lung cancer. Transmembrane Protein 48 (TMEM48) is localised on nuclear pore complex and plays critic roles in nuclear traffic. Known that TMEM48 gene overexpressed in non-small lung cancer cells. Growing TMEM48 suppressed therapeutic studies indicated that decreased TMEM48 level might reveal a therapeutic effect for non-small cell lung cancers. TMEM48 studies based on the same strategy of gene-silencing, however, to our knowledge, any report has been published evaluates TMEM48's regulation by miRNAs. We aimed to clarify if miR-421 might be therapeutic player for non-small cancer cell lines (A549), hereby we suppressed TMEM48 by miR-421 and performed advanced molecular tests. Consequently, we recorded that while miR-421 is significantly suppressing TMEM48 expression; it increased apoptotic and tumor suppressor players CASPASE 3, PTEN and TP53 in A549 line, which is consistent with Annexin V - PI results: 30,6% of A549 observed to be apoptotic - 68,5% of A549 was in GO/G1. Our study indicated that miR-421 can suppress TMEM48 so that leads the cells to apoptosis. But it is not entirely clear how miR-421 triggers apoptosis and whether it interacts with the other cellular death pathways in A549.

Monitoring of failure of chloroquine treatment for Plasmodium vivax using polymerase chain reaction in Sanliurfa province, Turkey.

Malaria is a complex disease that varies widely in epidemiology and clinical manifestation in the southeastern part of Turkey. In many regions of the world, chloroquine (CQ) has been the standard treatment for Plasmodium vivax. However, the resistance of the Plasmodium species to antimalarial drugs has become an increasing problem and a concern worldwide. Our target was to determine the Plasmodium species in the southeast region of Turkey and the therapeutic efficacy of CQ used in the treatment of malaria. Blood samples were collected from 180 patients infected with malaria before and after CQ treatment and were subjected to DNA isolation. The isolated DNA was amplified by a seminested multiplex polymerase chain reaction (SnM-PCR) including primers selected on Plasmodium small subunit ribosomal RNA (ssrRNA) genes for identification of the malaria species. The SnM-PCR results showed that only P. vivax exists in this province. It was also determined that there is a therapeutic failure to CQ in 9.5% of patients. These were the second report on identification of P. vivax and the third report on determination of the therapeutic failure in patients who used CQ to cure human malaria in the southeastern region of Turkey. Our results demonstrate that the SnM-PCR is a sensitive, specific, and a rapid tool for the differentiation of malaria species.

Single Nucleotide Polymorphism Analysis in HIV and Kaposi's Sarcoma Disease by Microarray Technique.

BACKGROUND: Emergence of Kaposi's Sarcoma in the cases other than HIV, following the use of immunosuppressant drugs, demonstrates that it is related to weak immunity. The fact that this malignancy does not occur in every HIV-positive patient suggests that genetic predisposition may also be effective. Replacement of one of the base pairs of adenine, guanine, cytosine, and thymine that constitute the DNA sequence in the human genome with another base pair can affect susceptibility to disease, response to treatment, and immunity. OBJECTIVE: The purpose of this study is to analyze the Single Nucleotide Polymorphism that could predispose to Kaposi's sarcoma of an HIV-infected patient and to identify which nucleotides such SNPs correspond to, using the microarray technology. MATERIALS AND METHODS: The blood samples of individuals, one of whom was diagnosed with Kaposi's Sarcoma HIV (+) visiting the outpatient clinic of infectious diseases polyclinic of Harran University Research and Practice Hospital and of a healthy individual with no Kaposi's Sarcoma, were used in the study. Following the DNA isolation of the blood samples taken from the respective individuals, a SNP analysis was conducted on the microarray device. 204,000 SNPs obtained were scanned later on in the databases in an attempt to identify the SNPs related to Kaposi's Sarcoma. RESULTS: In the 204,000 SNP screenings, we scrutinized the SNPs that differ in the case of Kaposi's Sarcoma [KS (+) and HIV (+)] on the basis of Control [KS(-) and HIV(-)] and HIV+ [KS(-)], and two SNPs of the ENDRA gene, three SNPs of the ADRA1A gene, six SNPs of the STIM1 gene, four SNPs of the EFNB2 gene, and one SNP of the CD209 gene were found to be different. However, when it comes to all SNPs (all the 204.000 SNPs) screened in terms of allele, it was observed that the AA and BB alleles were lower in the patient with Kaposi's Sarcoma [KS (+) and HIV (+)] compared to other groups and AB alleles were found to be higher than others in the patient with Kaposi's sarcoma [KS] (+) and HIV (+)]. CONCLUSION: In the microarray study we have conducted, 204,000 SNPs were screened for Control (HIV-) HIV (+) and HIV (+) patient with Kaposi's Sarcoma. It was found that 32,362 of those SNPs had different alleles in the Kaposi's Sarcoma [KS + HIV (+)] patient, while they had the same ones in the control [KS (-) and HIV (-)] and HIV + [KS (-)] group. 16 of the 32,362 SNPs took place among the genes related to Kaposi's Sarcoma. In the cases of Kaposi's Sarcoma with suspected diagnosis, it can be used as a beneficial laboratory test.

Investigation of IVS14+ 1G > A polymorphism of DPYD gene in a group of Turkish patients with colorectal cancer.

BACKGROUND: Dihydropyrimidine dehydrogenase (DPD) is a critical enzyme in the catabolism of 5-fluorouracil (5-FU), a drug frequently used in cancer therapy. One of the possible causes of severe 5-FU toxicity is genetic polymorphisms in the DPYD gene, such as IVS14+1G > A. In this study we aimed to investigate the frequency of the IVS14+1G > A mutation in the DPYD gene in Turkish patients with colorectal cancer (CRC) and healthy controls. MATERIALS AND METHODS: Blood samples were collected from 218 individuals (56 patients with CRC and 162 healthy individuals), and the DNA was isolated. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect the frequency of the IVS14+1G > A mutation in our population. RESULTS: The IVS14+1G > A mutation (heterozygous) in the DPYD gene was identified in two healthy subjects in this Turkish population. CONCLUSION: The apparently high prevalence (allele frequency of 0.6%) of the IVS14+1G > A mutation warrants genetic screening for this mutation in cancer patients before the administration of 5-FU.

Patterns of active and passive smoking, and associated factors, in the South-east Anatolian Project (SEAP) region in Turkey.

BACKGROUND: Smoking is an important health threat in Turkey. This study aimed to determine the frequency of and main factors associated with smoking in persons of 15 years and over, and the frequency of passive smoking in homes in the South-east Anatolian Project (SEAP) Region in Turkey. METHODS: A cross sectional design was employed. The sample was chosen by the State Institute of Statistics using a stratified cluster probability sampling method. 1126 houses representing the SEAP Region were visited. Questionnaires about tobacco smoking and related factors were applied to 2166 women and 1906 men (of 15 years old and above) in their homes. Face-to-face interview methods were employed. Participants were classified as current, ex, and non-smokers. The presence of a regular daily smoker in a house was used as an indication of passive smoking. The chi-square and logistic regression analysis methods were used for the statistical analysis. RESULTS: The prevalence of smoking, in those of 15 years and over, was 11.8% in women and 49.7% in men. The prevalence of current smokers was higher in urban (34.5%) than in rural (22.8%) regions. The mean of total cigarette consumption was 6.5 packs/year in women and 17.9 packs/year in men. There was at least one current smoker in 70.1% of the houses. CONCLUSION: Smoking is a serious problem in the South-eastern Anatolian Region. Male gender, middle age, a high level of education and urban residency were most strongly associated with smoking.

Attitudes towards family planning in the Southeast Anatolian Project (SEAP) region of Turkey.

OBJECTIVE: To determine attitudes towards family size and last pregnancies in order to improve family planning services in the Southeast Anatolian Project (SEAP) region. METHODS: A questionnaire survey in the nine SEAP regional provinces was carried out under the auspices of the 'SEAP Public Health Project' from 2001 to 2002. The participants comprised 1756 women and 661 men from 1126 households. RESULTS: For men and women aged 15 years and over the median ideal number of children was three. The rate of unintended last pregnancies (43.1%) in the present study was very high compared to the national average of 18.8%. Some 30.1% of the last pregnancies were unwanted by either partner. CONCLUSIONS: The number of pregnancies and children in this region is approximately twice as high as the ideal number. Families in the region are having more children than they want. Basic education must be given to women, particularly non-Turkish speakers, to improve their knowledge and use of family planning. Family planning education for men in rural areas also needs special attention.

Reduced SIRT1 expression correlates with enhanced oxidative stress in compensated and decompensated heart failure.

Sirtuin-1 (SIRT1) is a longevity factor in mammals initiating the cell survival mechanisms, and preventing ischemic injury in heart. In the etiopathogenesis of heart failure (HF), impairment in cardiomyocyte survival is a notable factor. Oxidative stress comprises a critical impact on cardiomyocyte lifespan in HF. The aim of the present study was to investigate SIRT1 expression in patients with compensated (cHF) and decompensated HF (dHF), and its correlation with oxidative stress. SIRT1 expression in peripheral leukocytes was examined using quantitative RT-PCR in 163 HF patients and 84 controls. Serum total oxidant status (TOS) and total antioxidant status (TAS) were measured via colorimetric assays, and oxidative stress index (OSI) was calculated. Lipid parameters were also determined by routine laboratory methods. SIRT1 mRNA expression was significantly downregulated in HF with more robust decrease in dHF (p=0.002, control vs cHF; p<0.001, control vs dHF). Markedly increased oxidative stress defined as elevated TOS, OSI and low TAS levels were detected in HF patients comparing with the controls (TAS; p=0.010, control vs cHF, p=0.045 control vs dHF, TOS; p=0.004 control vs cHF; p<0.001 control vs dHF, OSI; p<0.001 for both comparisons, respectively). With SIRT1 expression levels, TAS, TOS, OSI, and high density lipoprotein levels in cHF and dHF were determined correlated. SIRT1 expression were significantly reduced in both HF subtypes, particularly in dHF. SIRT1 expression was correlated with the oxidant levels and antioxidant capacity. Data suggest that SIRT1 may have a significant contribution in regulation of oxidant/antioxidant balance in HF etiology and compensation status.

Induced abortion and effecting factors of ever married women in the Southeast Anatolian Project Region, Turkey: a cross sectional study.

BACKGROUND: Nearly 10% of the population of Turkey lives in the Southeast Anatolian Project (SEAP) region. The population growth rate and the rate of unintended pregnancies are high and family planning services are insufficient in this region. Lifetime induced abortion rate is also high in this region. Public health problems of the SEAP region were investigated in the "SEAP Public Health Project" in 2001 and 2002. As it is one of the most important health problems of the women living in this region; induced abortion was also investigated in this project. METHODS: An optimumsample size representing the rural and urban area of the region (n = 1150) was chosen by the State Institute of Statistics by a sampling method proportional to size. 1126 of the area's 1150 houses have been visited and data about induced abortions have been obtained by applying a questionnaire to 1491 ever married women who live in the region. RESULTS: It has been found that 9.0% of these women who had at least one pregnancy in their life had at least one induced abortion. The lifetime induced abortion per 100 pregnancies was found to be 2.45. The primary reason given for induced abortions was "wanting no more children" (64.6%). Lifetime induced abortions were 5.3 times greater with women using a family planning method than women not using family planning methods. Lifetime induced abortions were 4.1 times greater with unemployed women than working women. Most of the women have used private doctors in order to have an induced abortion. Although 32.29% have not yet begun to use a contraceptive method after their last induced abortion, 43.75% of the women have since started to use an effective contraceptive method. 23.96% of them have begun to use an ineffective contraceptive method. CONCLUSIONS: Induced abortion is still an important problem at the SEAP region. The results of the study remind us that unemployed women and women who have more than four children is our target group in the campaign against induced abortions. Most of the women use private doctors in order to have an induced abortion. Thus, priority must be given to educate private gynecologists with respect to induced abortion. After induced abortions, a qualified family planning consultant can be given to women and they can be secured to use a suitable contraceptive method.

Detection of VDR gene ApaI and TaqI polymorphisms in patients with type 2 diabetes mellitus using PCR-RFLP method in a Turkish population.

Type 2 diabetes mellitus (T2DM) is by far the most common type of diabetes and is characterized by insulin resistance and altered insulin secretion. Some genes, such as the vitamin D receptor gene (VDR, NM_001017535; GI: 7421), involved in its metabolic pathway have been regarded as good candidates for T2DM. In this study, we investigated whether there was an association of VDR: g.59979G>T or c.1025-49G>T (ApaIG>T) and g.60058T>C or c.1056T>C (TaqIT>C) polymorphisms in the 3' untranslated region of VDR with T2DM in a Turkish population. We collected blood samples from 241 individuals (72 patients with T2DM and 169 healthy individuals), and their DNA was isolated. Polymorphisms of the VDR were analyzed by DNA amplification with polymerase chain reaction and endonuclease digestion with ApaI and TaqI. Body mass index was higher in T2DM patients than in control individuals. However, the frequency of g.59979TT genotype in T2DM patients was not significantly increased compared to healthy subjects (37.5% vs. 36.1%, respectively). Although the VDR g.60058CC genotype in T2DM patients (19.4%) was higher than that in healthy individuals (11.2%), there was no significant difference. In the same way, there was no difference between the groups in allele frequencies. In conclusion, our study did not provide evidence for the association of two examined VDR polymorphisms with T2DM in a Turkish population.

Identification of Leishmania parasites in clinical samples obtained from cutaneous leishmaniasis patients using PCR-RFLP technique in endemic region, Sanliurfa province, in Turkey.

Antroponotic cutaneous leishmaniasis (ACL) is an endemic disease and one of the major health problems in Sanliurfa province located in the southeastern region of Turkey. Leishmania tropica is confirmed as the causative agent of ACL in this region. In Sanliurfa city alone, the recorded total cases of ACL were 6,817 between 2001 and 2006. We aimed to determine the effectiveness of a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identification and differentiation of the Leishmania parasite in comparison to direct microscopic examination of clinical samples. The lesion exudates were collected from 51 ACL suspected patients and used for smear-slide preparations and DNA isolation. The isolated DNA was amplified by PCR, including primers selected on repetitive DNA for identification of a Leishmania subgenus, and the amplified DNA was restricted by HaeIII restriction endonuclease. The PCR-RFLP results showed that only L. tropica exists in this province. It is also determined that the positivity rate with PCR was higher (96%) than by microscopic examination (64%) in the diagnosis of ACL. Our results indicate that the PCR-RFLP method is more sensitive and specific for the detection and differentiation of agents of ACL in this area.

The distribution of the intestinal parasitic diseases in the Southeast Anatolian (GAP=SEAP) region of Turkey.

OBJECTIVES: The physical alterations put in place by the Southeastern Anatolia Project will undoubtedly provide a remarkable economical growth and a social development in the area. In addition, the influence that formation of dam ponds, enlargement of irrigation areas, change of product and the way of cultivation, urbanization and industrialization will have an impact on the environment. To minimize the adverse effects of this process on human beings, a Community Health Project was completed by the teams participated by Ege, Dicle, Gaziantep and Harran Universities under the Directorate of Turkish Parasitology Association and by Southeastern Anatolia Project Regional Development Administration between 2001 and 2003. RESULTS: To identify individuals with parasite, feces samples were taken from a total of 4,470 individuals. Parasites were found in feces of 41.8% of men, 44.3% of women and 32.2% of children, 0-59 months old, who were included in the research and gave feces samples for parasites tests. These prevalence values indicate how widespread parasitic diseases are in the region. The high prevalence of parasitic diseases in this area is one of the causes of malnutrition in 40% of children. Parasites were detected in 44.2% of feces samples taken from rural areas and in 39.5% taken from urban areas. When the distribution of parasites detected in feces samples was studied, the most common parasites were Giardia intestinalis (18.1%), Entamoeba coli (11.8%), Ascaris lumbricoides (4.8%), Trichuris trichiura (4.5%) and Hymenolepis nana (3.9%). Distribution of parasites according to cities varied widely. The most frequently seen parasites were T. trichiura in Gaziantep; G. intestinalis in Batman, Mardin, Diyarbakir, Sirnak and Sanliurfa; and E. coli in Siirt, Kilis and Adiyaman. CONCLUSIONS: This study is the first investigation of intestinal parasite prevalence in a large region, specifically, in this GAP region and in Turkey, in general. There is no direct relationship between irrigating the cultivation areas and diffusion of parasitic diseases because the existence of intestinal parasites mentioned above is not related to the range of irrigation of cultivation areas, but is related to factors already discussed.