21 research outputs found
The Elk1 gene effect on prostate cancer cell line wound healing ability
In this study prostate cancer cell line model was used selecting three type of them (Pc3, DU145 & LNCaP) , gene transfecting was done by using the liposome into the cell culture cells then detecting the healing ability by the Wound Healing or (scratching) test that shows clear effect of ELK1 gene in this cells comparing with the control cell transfected with the knock down gene, using for each type of cells 6 repeats ,for each type there was two groups 1st for control ELK1 and the 2nd was knock down or (sh) ELK1, all works was done in Johns Hopkins University / School of Medicine / Pathology Department ( MD, USA
ELK1 Gene Transfection Effect in Prostate Cancer Cell Line Proliferation Activity
Three type of prostate cancer cell line was selected for this study (PC3, DU145 & LNCaP) as a model, transfected by liposome with the ELK1 gene ( control & knock down) , then detecting the proliferation ability of the cultured cell lines by the Mtt or (proliferation) assay that shows clear effect of ELK1 gene in this cells comparing with the control cell transfected with the knock down gene, using for each type of cells 6 repeats ,for each type there was two groups 1st for control ELK1 and the 2nd was knock down or (sh) ELK1, all works was done in Johns Hopkins University / School of Medicine / Pathology Department ( MD, USA
Human Blood Identification in Crime Scene as a Forensic Clue
مقدمة:
الدم هو أحد أكثر الأدلة شيوعًا التي يعثر عليها في مسرح الجريمة. يتم تشخيص الدم البشري وفقا للسياق الاتي أولاً موقع بقع الدم ، ثم إجراء الاختبارات الافتراضية ، ثم يتم عمل الاختبارات التاكيدية . تعتبر الاختبارات الجينية والاختبارات المصلية من الاختبارات المهمة في الطب الشرعي..
طرق العمل:
تم جمع ثلاثين عينة دم في أنابيب EDTA ، تم جمع 25 منها من البشر وخمس عينات من الأغنام. استمرت هذه الدراسة خمسة أشهر من 23/3 إلى 22/8/2022. أجريت هذه الدراسة في جامعة بابل - كلية العلوم - قسم الأحياء. كان الهدف من هذه الدراسة هو تقييم طقم Hightop للكشف عن الدم البشري. تم تعريض الدم لظروف جوية مختلفة . ثم فحص باستخدام هذه العدة لتحديد قدرة المستضد على تحمل الظروف المختلفة وحساسية القياس لهذه العدة.
النتائج:
أظهرت النتائج الإيجابية لبقع الدم التي تعرضت لظروف جوية مختلفة لمدة 30 يومًا باستثناء بقع الدم التي وضعت على سطح أملس اظهرت نتائج سلبية بعد 10 أيام من التعرض عكس التي كانت مدفونة بالتربة او البقع الدموية على الملابس التي تحملت الضروف الجوية مدة 30 يوما
الاستنتاجات:
من نتائج تبين عدة Hightop مناسبًا في مجال الطب الشرعي لتحديد دم الإنسان الذي يتعرض لظروف بيئية قاسية و كذلك لها القابلية لتميز الدم البشري عن الدم الحيواني و المستضد الهدف لها يتحمل الظروف الجوية.Background:
Blood is one of the most common clues found at crime scenes. Human blood is diagnosed according to the following context: first, the location of the blood spots, then virtual tests, then confirmatory tests. Genetic testing and serological testing are important tests in forensic medicine..
Materials and Methods:
Thirty blood samples were collected in EDTA tubes, of which 25 were collected from humans and five from sheep. This study lasted five months from 23/3 to 22/8/2022. This study was conducted at Babylon University - College of Science - Department of Biology. The aim of this study was to evaluate the Hightop kit for detection of human blood. The blood was exposed to different weather conditions. Then it was examined using this kit to determine the ability of the antigen to withstand different conditions and the sensitivity of the measurement for this kit.
Results:
The positive results of blood stains that were exposed to different weather conditions for 30 days, except for blood stains that were placed on a smooth surface, showed negative results after 10 days of exposure, as opposed to those that were buried in the soil or blood stains on clothes that endured weather conditions for 30 days..
Conclusion:
The Hightop kit is suitable in the field of forensic medicine to determine human blood that is exposed to harsh environmental conditions, as well as the ability to distinguish human blood from animal blood and its target antigen bears the weather conditions
Effect of a Mixture of Plants Extracts on Genomic DNA, Insulin Receptor, and Insulin Receptor Substrate-1 Genes in Alloxan-Induced Diabetic Male Rats.
This study was designed to study the effects of methanol-watery extracts mixture of five selective medicinal plants on genomic DNA, insulin receptor, and insulin receptor substrate-1 genes in alloxan-induced diabetic male rats. The molecular assays used in this study was DNA fragmentation test and polymerase chain reaction of insulin receptor and insulin receptor substrate-1. Phytochemical assays (thin layer chromatography (TLC) and antioxidant activity by ?- carotene spray) were carried out for characterization of methanol-watery extracts of each plants used (Trigonella faenum-graecum (fenugreek) seeds, Nigella sativa (black cumin) seeds, Zingiber officinale (ginger) rhizomes, Olea europeae (olive) leaves, and Fraxinus ssp.(ash) seeds). The results indicate presence of number of antioxidant bands with different retardation factors (Rf). Molecular study of DNA showed fragmentation in both DNA extracted from WBC and renal cortex in diabetic group while treatment with mixture of plants extracts significantly reduced DNA fragmentation in both DNA extracted from WBC and renal cortex in all treatment groups except treatment for 45 days which still had lyses level 5000 bp of DNA extracted from renal cortex. On the other hand, insulin receptor gene of diabetic and diabetic-treated groups for different periods (45, 60, 75 days) showed precense of one band (200 bp) compared with normal control which had two bands while insulin receptor substrate-1 has normal band (320 bp) in normal and treated groups. Keywords: alloxan, insulin receptor, insulin receptor substrate -1, antioxidant, DNA fragmentatio
Genetic Diversity of Iraqi Date Palm (Phoenix dactylifera L.) by using RAPD Technique
في هذه الدراسة اثبتت جميع مؤشرات التفاعل الثضاعفي لسلسلة الدنا متعدد الاشكال نجاحا مع 65 مستزرع من نخيل التمر العراقي والتي جمعت من مدينة الحلة في العراق٬ وذلك لتحديد البصمة الوراثية والقيمة المتباينة الاشكال والعلاقة الوراثية بين تلك الانواع وكذلك مع الانواع نفسها المستعملة في الدراسة. وقد اظهر تحليل البيانات لعشرة من المؤشرات الجزيئية المستعملة في هذه الدراسة́٬ مجموع عدد القطع المتضخمة لقطع الدنا هي 592 حزمة ونسبة متعدد الاشكال لكل المؤشرات هي 64.2%٬ وان قيمة الفعالية لمؤشرات الدنا كانت 0.09%٬ كذلك اظهرت النسبة المئوية للتشابه الوراثي بين الانواع 67% الى 100% ما بين المستزرعات التي تعود الى نفس المستزرع. العلاقة الوراثية ل24 مستزرع من نخيل الثمر تقسم الى مجموعتين رئيستين٬ المجموعة الاولى تقع ضمن المدى الوراثي 0.74 الى 1.30 وتمثل (مدني واشرسي ورصاصي وسيسمي وسكري) والمجموعة الثانية تقع ضمن المدى الوراثي من 0.25 الى 0.60 وتقسم الى ثلاثة تحت مجموعات وهي: تحت المجموعة الاولى وتمثل (سلطاني وخستاوي وبريم وسبع ذراع وحمراوي وبربن وخضراوي) ٬ وتحت مجموعة الثانية تمثل (زهدي وتبرزل وبرحي وجبجاب وفوم الرمان) اما تحت مجموعة الثالثة تمثل (اسطة عمران ونيرسي ونجدي وكنطار وشويثي وغنامي احمر).In this study provided all molecular markers of Random amplified polymorphic (RAPD) successfully with the sixty five Iraqi date palm (Phoenix dactylifera L.) cultivars, which collected from Hilla city in Iraq, to determine fingerprinting, polymorphic value, and relationships among varieties of date palm cultivars, and also with the same type of cultivars. Data analysis of ten RAPD has been revealed. Number of amplified DNA fragments were (592) bands, polymorphism per all primers were (%64.2), primer efficiency was 0.1, and discriminatory value was (%0.09), which revealed a high percentage similarity about %67 to %100 between cultivars belong to the same variety. There are relationships with twenty four genotypes, divided in to two clusters, clusterΙ ranged distance from 0.74 to 1.30 represented(Maddany, Ashrasi, Greatli, Smeasmi and sukkary) and clusterII ranged distance from 0.25 to 0.60 which divided into three sub group, there are sub group I represented (Sultana, Khestawi, Breem, Sabb Drrah, Hamrawi, Brban, and Khadrawi), sub groupiesII represented (Zahdi, Tebarzal, Maktom, brahi, Chipchab and Fom Alrman), sub groupies III represented (Usta Umran, Nersi, Najdi, Guntar, Shwethi and Ghanami Ahmer)
Shortened Telomere Length in White Blood Cells of Patients with Non-Insulin Dependent Diabetes Mellitus (NIDDM)
Type 2 diabetes(TIIDM) aging-related disorder, is caused by a combination of peripheral insulin resistance and ?-cell dysfunction .Recent evidence, however, suggests that TIIDM is additionally characterized by impaired ?-cell regeneration and reduced ?-cell mass .Shortened telomeres have been previously associated with diabetes in several small-scale studies(Zhu et al.,2011). Measurement of telomere length in diabetic patients in different age and different duration of disease and compared with control. Also included the comparison telomere length between male and female for both control and diabetic groups and between patients from genetic origin (mother origin or father origin) and patients from non genetic origin . Genomic DNA was prepared from whole blood extraction using genaid kit and was quantified by Nanodrop .Terminal restriction fragment (TRF) lengths were measured using the Southern-blotting technique .This study was conducted between November 2010- November 2012 and, it was carried out at the diabetic Centre / Merjan Teaching Hospital in Babel Province by taking 54 diabetic patients(Type II DM) with disease duration (0-5),(>5-10 )and (>10 )years ,with age average (35-65 year)and most of them were on oral hypoglycemic drugs. While the study included 18 people apparently healthy that included 9 male and 9 female with age average (35-65 year) .The study revealed that telomere length(TL) were differences between males and females of control group. Age- adjusted telomere length were shorter in males than in females of control subjects (13,200 bp ) vs. (11,600 bp), (14,200 bp) vs. (13,000 bp) and (15,100 bp) vs. (13,800 bp) ,and this differences in TL between males and females decreased as aging increased, while this gender differences in TL was not observed among the diabetic patients .In both the controls group and diabetic subjects, the telomere length were shorter in older subjects than younger for both males and females. The result show differences in TL between control and diabetic subjects. TL were shorter in the patients with Type 2diabetes compared with the control subjects ,and TL in first duration of male diabetic patients(2,100 bp) in male(55 years) and (1,900 bp) in male (65 years) ,and second duration of female diabetic patients (3,600 bp) in female (55 years) and (2,900 bp) in female (65 years) were shorter compared with other duration. The differences in TL in patients with family history of disease shown shorter than patients without family history of disease and this differences in TL in patients with family history of disease from mother origin shorter than patients with family history of disease from father origin. The result of telomere length (TL) show that TL were shorter in males than females of control group while this differences absent in diabetic patients .TL was shorter in first duration of disease in males diabetic patients and in second duration of females diabetic patients compared with other duration of disease. TL was shorter in older subjects than younger for both groups and for both gender ,also shorter in diabetic patients from genetic origin of disease than non-genetic origin. Positive correlation between TL and estrogen in female control group and between TL and adiponectin in diabetic patients was found. Keywords: Telomere, DNA, TR
Study the levels of adiponectin, FSH, LH and Sex hormones in Type 2 diabetes (NIDDM)
The hypothalamic/pituitary/gonadal (HPG)axis is central to the mammalian reproductive system . Pulsatile release of GnRH from neurons in the hypothalamus stimulates the secretion of LH and FSH from gonadotropes in the anterior pituitary. It has long been recognized that reproductive function is closely associated with energy balance, and metabolic dysregulation is linked with reproductive abnormalities (Lu et al.,2008). Compare the differences in levels of adiponectin, FSH, LH, testosterone and estradiol between the diabetic patients and control group and in diabetic patients according to the durations of disease for both males and females groups .Also study the relationship between adiponectin and hormones for both gender and for both diabetic groups and control also in diabetic patients according to the durations of disease. About five milliliters of venous blood were collected from each subject in the study. The blood was separated by centrifugation at (3000 rpm) for 15 min. The sera were stored frozen at (-20 ºC) until assayed. This study was conducted between November 2010- November 2012 and, it was carried out at the diabetic Centre / Merjan Teaching Hospital in Babel Province, by taking 120 diabetic patients non- insulin dependent diabetes mellitus (NIDDM) (60 male and 60 female)with disease durations(0-5),(>5-10 )and (>10 )years ,and with age average (35-65 year), most of them were on oral hypoglycemic drugs, and the study included 40 people apparently healthy that included 20 male and 20 female with age average (35-65 year) ,as control matched with disease groups. The statistical analysis of this study showed that patients with Type 2 diabetes of both males and females had significantly lower in adiponectin levels than control group (P<0.05),but this level of adiponectin significantly higher in females than males for both diabetic groups and control (p<0.05) ,while the levels of FSH and LH had significantly higher in diabetes group than control (p<0.05) ,also significantly higher in females than males for both diabetic groups and control (P< 0.05). The result of hormonal analysis show there were no significant differences in estradiol and testosterone between diabetic groups and control (p > 0.05). According to the durations of disease, the results show significant elevation in adiponectin level in third duration of disease for both males and females groups as compared with first and second duration (P <0.004), while the levels of FSH, LH ,estradiol and testosterone show no significant differences among duration(P>0.05). According to the gender ,levels of adiponectin ,FSH, LH, and estradiol were significantly higher in females than males(P<0.05) , whereas levels of testosterone was significantly higher in males than females (P<0.05) . Correlation analysis showed an inverse correlation between adiponectin and FSH in female diabetic patients. Also an inverse correlation between adiponectin and estradiol was found in females of control group, while the correlation of LH and testosterone with adiponectin appears no significant correlation for both groups and for both males and females . According to the durations of disease, in first duration (0-5year)the study revealed positive correlation between adiponectin and FSH in male diabetic patients . In third duration (>10 year) an inverse correlation between adiponectin and FSH in female diabetic patients. The result also appears no significant correlation among adiponectin and LH, estradiol and testosterone in both gender among duration . The results of the present study indicate that the patients with Type 2 diabetes of both males and females had significantly lower in adiponectin levels than control group . levels of adiponectin significantly higher in females than males for both diabetic groups and control. Adiponectin levels significantly increased with duration of disease also with age. Levels of FSH and LH had significantly higher in diabetic patients than control also significantly higher in females than males for both groups . Correlation analysis showed inverse correlation between adiponectin and both FSH and estradiol in female diabetic patients also inverse correlation between adiponectin and both FSH and estradiol in third duration of female diabetic patients
Study the relationship between adiponectin with thyroid hormones and cortisol in Type 2 diabetic patients (NIDDM)
Thyroid hormones has profound effects of lipid metabolism and carbohydrate homeostasis . Abnormalities in serum lipids and lipoproteins are frequent findings in thyroid dysfunction, mainly in hypothyroidism. Impaired glucose tolerance and insulin resistance have been documented in patients with thyroid dysfunction. In addition, thyroid hormones are remarkable regulators of energy metabolism, being the adipose tissue the largest fuel storage compartment . Furthermore, thyroid hormones share some physiological actions with adiponectin, such as reduction of body fat by increasing thermogenesis and lipid oxidation .However, a few number of studies have found low adiponectin levels in hypothyroid subjects (Díez and Iglesias, 2009). Cortisol excess in man is characterized by abdominal obesity, hypertension, glucose intolerance or diabetes and dyslipidemia. All these features share a state of insulin resistance , and contribute to high cardiovascular risk typical of this condition. Glucocorticoids negatively regulate adiponectin mRNA in human visceral adipose tissue. Cortisol counteracts the action of insulin at multiple sites, and increases hepatic gluconeogenetic efficiency (Fallo et al.,2004). Study the relationship between adiponectin with thyroid hormones and cortisol for both gender and for both control and diabetic groups also in diabetic patients according to duration of disease. This study was conducted between November 2010- November 2012 and, it was carried out at the diabetic Centre / Merjan Teaching Hospital in Babel Province by taking 120 diabetic patients(Type II DM) (60 male and 60 female) with disease duration (0-5),(>5-10 )and (>10 )years ,with age average (30-65 year)and most of them were on oral hypoglycemic drugs. While the study included 40 people apparently healthy that included 20 male and 20 female with age average (30-65 year) .About five milliliters of venous blood were collected from each subject in the study. The blood was separated by centrifugation at (3000 rpm) for 15 min. The sera were stored frozen at (-20 ºC) until hormonal assayed. Correlation analysis showed an inverse correlation between adiponectin and cortisol in female diabetic patients, while in male diabetic patients there was positive correlation between adiponectin and T4 was found. The correlation of TSH and T3 with adiponectin appears no significant correlation for both groups and for both males and females. The study revealed positive correlation between adiponectin and T4 in second and third duration in male diabetic patients , while an inverse correlation between adiponectin and TSH in male diabetic patients in first duration. In female diabetic patients an inverse correlation between adiponectin and T3 in second duration,also found between adiponectin with cortisol in third duration of female diabetic patients. Correlation analysis showed an inverse correlation between adiponectin and cortisol in female diabetic patients, while positive correlation between adiponectin and T4 was found in male diabetic patients. This study show that Type 2 diabetic patients in first duration(0-5 year) an inverse correlation between adiponectin and TSH in male diabetic patients. In second duration(>5-10 year)an inverse correlation between adiponectin and T3 in female diabetic patients. In third duration(>10 year) an inverse correlation between adiponectin with cortisol of female diabetic patients, while positive correlation between adiponectin and T4 in second and third duration of male diabetic patients
Protective Effect of Plants Extracts Mixture on Sperm Abnormalities, Testicular and Epididymal Tissues in Diabetic Male Rats
Background: Diabetes has been associated with reproductive impairment in both men and women. About 90% of diabetic patients have disturbances in sexual function, including a decrease in libido, impotence and infertility. Attention has been paid to the search of effective drugs in the field of traditional medicine. Objective: The aim of the present study was to investigate the effect of diabetes mellitus type 2(T2DM) on the sperm abnormalities, testicular and epididymal tissues of male rats. Also to evaluate the efficiency of the mixture of methanol-watery extracts composed of five medicinal plants (Trigonella faenum-graecum seeds, Nigella sativa seeds, Zingiber officinale rhizomes, Olea europeae leaves and Fraxinus ssp.seeds) in reducing harmful effect of DM on the percentage of sperm abnormalities, testicular and epididymal tissues. Methods: Twenty-four mature male rats were used in six groups (n=4), the first group kept as a control. The second group received alloxan (120 mg/ kg body weight i.p.) for 3 days to induction of T2DM while the third, fourth, and fifth groups were diabetic rats received the plants extracts mixture orally for 45, 60, 75 days, respectively after induction of T2DM. The sixth group received the plants extracts mixture for 60 days. By the end of the experiment, animals were sacrificed and the testis and epididymis were excised and sperm was collected for morphological abnormalities of the sperm shape. Also histopathological examination of the testes and epididymis were examined. Results: The sperm shape abnormalities significantly increased (p<0.05) in alloxan – induced diabetic rats compared with normal control group. Histopathological examination showed degenerative changes of both testes and epididymis in diabetic rats which include significant decrease (p<0.05) in both diameter of seminiferous tubules and their germinal epithelium heights and non-significant decrease (p>0.05) in both diameters of cauda epididymal tubules and height of epithelium of cauda epididymal tubules. On the other hand, atrophied seminiferous tubules, atrophied spermatogenic cells and the lumen of semineferous tubules free from sperm was detected. Furthermore, expansion of interstitial space between seminiferous tubules were seen . In contrast , oral administration of the plants extracts mixture to the diabetic rats at three period of study 45, 60, 75 days, respectively statistically decreased (p<0.05) most sperms abnormalities compared with diabetic group and caused non-significant differences compared with control group besides enhancing the histological changes of both testes and epididymis. Conclusion: DM has negative effect in male testis and epidydimis of alloxan-induced rats and may contribute in reduction of fertility while the plants extracts mixture used in this study can reduce most degerenative changes occurred in testes and epidydimis in diabetic groups treated with the mixture of plants extracts. Therefore, plants extracts mixture can be effectively used by DM patients for therapeutic purpose. Key words: Alloxan- induced diabetes, Sperm Abnormalities, Histopathological Changes of testis and epididymis, rats
Seminal fluid; the Natural Guard of Seminal DNA
The ability of foreign DNA to be introduced into the sperm cells faces many problems. These problems have been emerged from the existence of some inhibitory factors available in the seminal fluid. Add to that, other several factors are involved in this internalization. The purpose of this review is to evaluate the protective or interactive roles of these seminal fluid proteins in the process of foreign DNA internalization into the head of the sperm. The ability of foreign DNA to be internalized inside sperm cell is become obvious before more than four decades (Brackett et al., 1971). But, the mechanism by which this foreign DNA has the ability to do so is still under debate. However, several papers described certain factors involved in this process. Thus, in order to describe the mechanism of sperm transfection by exogenous DNA, it is necessary to understand the following natural factors that are playing main roles in this scene and they include; inhibitory factor I (IF1), seminal fluid DNase, DNA binding proteins (DBPs), CD4, major histoincompatibility complex class II (MHCII), topoisomerase type II (TOPO II) and reverse transcriptase (RT). They can be further classified into three main groups. The First group is represented by two factors found in seminal plasma; IF1 and DNase, this group is responsible about inhibiting the internalization process of exogenous DNA. The fact which refers to the existence of one or more factors in seminal plasma that able to block sperm permeability must be taken into account. This means, extensive washing steps of ejaculate to remove seminal plasma is necessary and should be made before incubating sperm with exogenous DNA. Lauria and Gandolfi reviewed that seminal fluid inhibitors have two ways of inhibition to exogenous DNA, either directly or indirectly (Lauria and Gandolfi, 1993). These seminal plasma inhibitory factors may prevent transfection of intact sperm by foreign DNA (Camaioni et al., 1992). Gandolfi showed that there is a consensus on the experiments made on seminal fluid of the ejaculated spermatozoa of mammals in the impermeability of sperm cell to the aggression of foreign DNA as long as seminal plasma is not removed (Gandolfi, 2000). Thus, seminal fluid prevents any foreign DNA from binding with its receptor on the sperm cell