scholarly journals Metagenome Analysis of Buccal Mucosal Biofilms to Identify Bacterial Prevalence in Subjects with and without type II Diabetes

2020 ◽  
Vol 11 (2) ◽  
pp. 2018-2029
Author(s):  
Hari Priya A ◽  
Kannan A ◽  
Krithika C L
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Type Ii Diabetes ◽  
Bacterial Flora ◽  
Control Group ◽  
Type Ii ◽  
Metagenome Analysis ◽  
Blast Analysis ◽  
Copy Numbers ◽  
Age Range

To quantify the two most prevalent bacteria among Type II diabetic individuals and controls from the buccal mucosal biofilms using molecular methods. To compare the percent prevalence of Veillonella and Granulicetella bacteria in uncontrolled Type II diabetic individuals with a control group. Subjects selected randomly and categorized into two groups within the age range of 25 to 40 years diagnosed with and without Type II diabetes based on their HbA1c values. The samples of buccal mucosa biofilms are collected in sterile swabs and stored in bacterial lysis buffer, which was later subjected to quantification of DNA followed by 16S rRNA amplification and sequencing. The sequence obtained is then surveyed using BLAST Analysis to define the bacterial flora and two bacteria, namely Veillonella and Granulicatella are selected for further amplification and quantification by real-time PCR to express the bacteria in copy numbers. From the collected buccal mucosal biofilm samples (n=24), which was categorized into Type II diabetes (12) and non-diabetic (12). The sequence subjected to BLAST analysis gave a List of bacteria from which Veillonella sp. and Granulicatella sp. were selected and administered to real-time PCR for amplification and quantification, which revealed an increased bacterial prevalence in Type II diabetic subjects to non-diabetic subjects which was also proved statistically. Based on the results obtained, there is a significant prevalence of bacterial content in Type II diabetic subjects compared to non-diabetic subjects

Relationship between uric acid and creatinine in pre-diabetic and diabetic patients: Vidarbha region of Maharashtra

2020 ◽  
Vol 11 (3) ◽  
pp. 3412-3417
Author(s):  
Ranjit S. Ambad ◽  
Rakesh Kumar Jha ◽  
Lata Kanyal Butola ◽  
Nandkishor Bankar ◽  
Brij Raj Singh ◽  
...  
Keyword(s):  
Uric Acid ◽  
Serum Uric Acid ◽  
Type Ii Diabetes ◽  
Fasting Blood Glucose ◽  
General Medicine ◽  
Control Group ◽  
Diabetic Patients ◽  
Type Ii ◽  
Develop Type ◽  
Low Levels

Prediabetes is a glucose homeostasis condition characterized by decreased absorption to glucose or reduced fasting glucose. Both of these are reversible stages of intermediate hyperglycaemia providing an increased type II DM risk. Pre-diabetes can therefore be viewed as a significant reversible stage which could lead to type II DM, and early detection of prediabetes may contribute to type II DM prevention. Prediabetes patients are at high risk for potential type II diabetes, and 70 percent of them appear to develop Type II diabetes within 10 years. The present study includes total 200 subjects that include 100 Prediabetic patients, 50 T2DM patients and 50 healthy individual. Blood samples were collected from the subjects were obtained for FBS, PPBS, Uric acid and Creatinine estimation, from OPD and General Medicine Wards. Present study showed low levels of Serum Uric Acid in prediabetic and T2DM patients were decreased as compared to control group, while the level of creatinine in prediabetic and diabetic were elevated as compared to control group, were not statically significant. Serum Uric Acid was high in control group and low in prediabetic and diabetic patients. Serum creatinine was declined in control group and increased in prediabetic and diabetic patients with increasing Fasting blood glucose level.

scholarly journals The association of male infertility with telomere length: A case control study

2021 ◽  
Vol 8 (4) ◽  
pp. 325-332
Author(s):  
Kate Deepali Rajesh ◽  
Puranam Vatsalaswamy ◽  
Manvikar Purshotam Rao
Keyword(s):  
Male Infertility ◽  
Telomere Length ◽  
Real Time ◽  
Real Time Pcr ◽  
Case Control Study ◽  
Case Control ◽  
Control Group ◽  
Significant Difference ◽  
Control Study ◽  
Sperm Telomere Length

To study the relevance of sperm telomere length and infertility in men. : Our case-control study included twenty-five males in couple with sub-fertility/infertility (test group) and twenty five healthy males (control group) with proven paternity in the age group 25 to 35 years. The Absolute Sperm Telomere length (aSTL) was measured by real-time PCR. We investigated whether any significant difference in the aSTL value existed between the groups and analysed the relationship between aSTL and other sperm parameters.The mean (SE) aSTL recorded in the infertile cases was significantly shorter than for the control group being 140.60 (6.66) Kb/genome and 239.63 (12.32) Kb/genome respectively (p <0.001) A weak correlation was eminent between aSTL kb/genome and the total sperm count mil/ml (rho= 0.04, p - 0.86), progressive sperm motility (rho= - 0.02, p=0.934) and sperm viability (rho= - 0.07 p=0.741) in the infertile group. The measurement of aSTL by real-time PCR is a simple and rapid method that offers further paramount information with respective to the quality of sperm. It is befitted for epidemiological studies, hence opening new perspectives in the evaluation of male infertility. Limitations - Our study was confined to men aged between 25 and 35 years. Further comparative studies are needed to explore the significance of STL and infertility in older males. Additional studies will help illumine the significance of aSTL as a prognostic biomarker in assisted reproduction.

scholarly journals Astragaloside positively regulated osteogenic differentiation of pre-osteoblast MC3T3-E1 through PI3K/Akt signaling pathway

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Wei Bing Jing ◽  
Hongjuan Ji ◽  
Rui Jiang ◽  
Jinlong Wang
Keyword(s):  
Western Blot ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
Signaling Pathway ◽  
Real Time Pcr ◽  
Akt Signaling ◽  
Calcium Deposition ◽  
Control Group ◽  
Akt Signaling Pathway ◽  
Western Blot Assay

Abstract Background Osteoporosis is a widespread chronic disease characterized by low bone density. There is currently no gold standard treatment for osteoporosis. The aim of this study was to explore the role and mechanism of Astragaloside on osteogenic differentiation of MC3T3-E1 cells. Methods MC3T3-E1 cells were divided into control and different dose of Astragaloside (10, 20, 40, 50, and 60 μg/ml). Then, ALP and ARS staining were performed to identify the effects of Astragaloside for early and late osteogenic capacity of MC3T3-E1 cells, respectively. Real-time PCR and western blot were performed to assess the ALP, OCN, and OSX expression. PI3K/Akt signaling pathway molecules were then assessed by Western blot. Finally, PI3K inhibitor, LY294002, was implemented to assess the mechanism of Astragaloside in promoting osteogenic differentiation of MC3T3-E1 cells. Results Astragaloside significantly increased the cell viability than the control group. Moreover, Astragaloside enhanced the ALP activity and calcium deposition than the control groups. Compared with the control group, Astragaloside increased the ALP, OCN, and OSX expression in a dose-response manner. Western blot assay further confirmed the real-time PCR results. Astragaloside could significantly increase the p-PI3K and p-Akt expression than the control group. LY294002 partially reversed the promotion effects of Astragaloside on osteogenic differentiation of MC3T3-E1 cells. LY294002 partially reversed the promotion effects of Astragaloside on ALP, OCN, and OSX of MC3T3-E1 cells. Conclusion The present study suggested that Astragaloside promoted osteogenic differentiation of MC3T3-E1 cells through regulating PI3K/Akt signaling pathway.

scholarly journals Should Calibration Curve Retire From Real-Time PCR Assays?

2021 ◽  
Author(s):  
David An
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Fluorescent Dyes ◽  
Historical Data ◽  
Genetic Disorder ◽  
Calibration Curves ◽  
Copy Numbers ◽  
Biological Technique ◽  
Pcr Assays ◽  
Number Of Cycles

Abstract Real-time polymerase chain reaction (real-time PCR) is a biological technique that collects data of target nucleotides as PCR occurs by integrating fluorescent dyes as visual indicators into the amplification cycles. This enables the detection and quantification of the DNA segments in a sample through measurements of the fluorescent's intensity. The cycle threshold (Ct) is defined as the number of cycles required for the fluorescent signal to pass a specified threshold and is inverse to the copy number, the initial number of nucleotides in the sample. Calibration curves are commonly used to approximate the copy numbers of experimental samples using standards with known copy numbers. This study is a retrospective review of historical data to help evaluate the efficacy and accuracy of calibration curves in a real-time PCR assay which have been used for screening of a genetic disorder in laboratories. The hypothesis is that including calibration curves in real-time PCR assays may decrease the screening specificity and accuracy, resulting in more false positives and additional retests. Three different scenarios were designed to replay the historical data and evaluate the relative accuracy of assays without calibration curves. The outcomes of all the scenarios conclude that calibration curves are not helpful for detecting target DNA fragments with low copy numbers, suggesting a reconsideration of their implantation in real-time PCR assays.

scholarly journals EFFECTIVENESS OF AMLA JUICE IN REDUCING BLOOD GLUCOSE LEVEL AMONG PATIENTS WITH TYPE II DIABETES IN A SELECTED AREA OF KANYAKUMARI DISTRICT

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Mrs. Vanitha. S s ◽  
Dr. Pramjit kaur
Keyword(s):  
Blood Glucose ◽  
Standard Deviation ◽  
Blood Glucose Level ◽  
Glucose Level ◽  
Type Ii Diabetes ◽  
Control Group ◽  
Type Ii ◽  
Glucose Levels ◽  
Blood Glucose Levels ◽  
Experimental Group

Challenges in lifestyle, such as increasesin energy intake and decreasesin physical activity are causing overweight and obesity leading to epidemic increases in type II Diabetes Mellitus. The research approach used for this study was evaluative approach and the research design was true experimental design. 60 patients with type II diabetes, 30 in experimental group and 30 in control group were selected for this study by using purposive sampling technique. Data was collected with the help of self-structured interview schedule. Descriptive statistics (frequency, percentage, mean and standard deviation) and inferential statistics (chi-square, paired ‘t’ test) were used to analyse the data and to test the hypotheses. In the experimental group,the pre-test mean score was 2.966, mean percentage was 59% and standard deviation was 1.129 and in post-testmean score was 2.533, mean percentage was 50.66% and standard deviation was 1.074 with effectiveness of 8.34% and paired‘t’ test value of t=3.971,which was statistically significant (p<0.05) which is an evidence ofthe effectiveness of Amla juice in reducing blood glucose level. Comparison of blood glucose levels in experimental and control groups, shows that the value is statistically highly significant, as was observed from the unpaired ‘t’ test value of 13.39 with P value of <0.05, which is an evidence indicatingthe effect of Amla juice in reducing postprandial blood glucose levels. The resultsfound that the administration of Amla juice did have aneffect in reducing blood glucose level in the experimental group. By comparing the findings of pre-test and post test between the experimental group and the control group,the effect was identified (assessed). The study concluded that the Amlajuice is effective in reducing blood glucose level.

scholarly journals CHANGES IN GENE EXPRESSION ASSOCIATED WITH NON-CANCER EFFECTS OF THE CHORNOBYL CLEAN-UP WORKERS IN THE REMOTE PERIOD AFTER EXPOSURE

2020 ◽  
Vol 25 ◽  
pp. 456-477
Author(s):  
I. Ilienko ◽  
◽  
D. Bazyka ◽  
N. Golyarnyk ◽  
L. Zvarych ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Main Group ◽  
Control Group ◽  
Chronic Obstructive ◽  
Relative Level ◽  
Gstm1 Gene ◽  
Expression Of Genes ◽  
Immune Inflammation

Objective. to establish the connection of radiation-induced changes in gene expression with the realized pathology of the broncho-pulmonary and cardiovascular systems in Chornobyl clean-up workers. Materials and methods. We examined 314 male Chornobyl clean-up workers (main group; age (58.94 ± 6.82) years (M ± SD); min 33, max 79 years; radiation dose (411.82 ± 625.41) mSv (M ± SD); min 1.74, max 3600 mSv) with various nosological forms of cardiovascular and broncho-pulmonary pathology (BPP) and 50 subjects of the control group: age (50.50 ± 5.73) years (M ± SD); min 41, max 67 years. The relative level of BCL2, CDKN2A, CLSTN2, GSTM1, IFNG, IL1B, MCF2L, SERPINB9, STAT3, TERF1, TERF2, TERT, TNF, TP53, CCND1, CSF2, VEGFA genes expression was determined in peripheral blood leukocytes by real-time PCR (7900 HT Fast Real-Time PCR System (Applied Biosystems, USA)). The «gene-disease» association was determined on statistical models stratified separately for each disease and gene. Logistic regression was used to calculate the odds ratio. Results. Increased GSTM1 gene expression and no changes in angiogenesis-related VEGFA gene expression were found in the main group of patients with coronary heart disease (CHD). It was established overexpression of TP53, VEGF and IFNG genes in the group of patients with arterial hypertension (AH). At combination of these diseases an increase of expression of СSF2, TERF1, TERF2 genes was established. The detected changes demonstrate an activation of the antioxidative defense system in patients with CHD, while AH is associated with the expression of genes of angiogenesis and immune inflammation. It was shown an increase in the expression of genes associated with apoptosis and kinase activity (BCL2, CLSTN2, CDKN2), immune inflammation (CSF2, IL1B, TNF) in Chornobyl clean-up workers with BPP. Expression of TP53 and GSTM1 (gene, associated with the glutathione system) was significantly upregulated in the group of individuals with chronic bronchitis, whereas in patients with chronic obstructive pulmonary disease, no increase was detected; the expression of SERPINB9 and MCF2L genes was downregulated. Conclusions. Changes in the expression of genes, associated with the development of somatic pathology in the remote period after irradiation, in particular the genes of the immune response and inflammatory reactions CSF2, IFNG, IL1B, TNF; expression of genes that regulate cell proliferation, aging and apoptosis TP53, BCL2, MCF2L, CDKN2A, SERPINB9, TERF1, TERF2, TERT; genes that regulate cell adhesion and angiogenesis CLSTN2, VEGF. Key words: gene expression, somatic pathology, radiation, Chornobyl.

Relationship between Netrin-1 Levels and Diabetes Mellitus Type 2

2021 ◽  
Vol 19 (10) ◽  
pp. 29-33
Author(s):  
Dr. Mohammed Abdulrazzaq Assi ◽  
Dr. Hanan Jasim Hammood ◽  
Zainab Mahdi Attia
Keyword(s):  
Diabetes Mellitus ◽  
Type Ii Diabetes ◽  
Diabetes Mellitus Type 2 ◽  
Fasting Blood Glucose ◽  
The Body ◽  
Blood Cholesterol ◽  
Control Group ◽  
Type Ii ◽  
Significant Drop

Objects: The study's purpose was to see if there was a link between netrin.1 and type II diabetes patients. Methodology: The present study carried out on (45) patients affected with diabetes mellitus from (120) persons were examined in the education of Diwaniyah hospital from 6th September to the 4th December 2020. The study includes measurement of the body mass index, fasting blood glucose level, HbA1c, lipid profile and level of netrin-1. Results: Our findings were revealed the netrin-1 level was significantly lower in the diabetes (1205.36±753.09) compared to control group (1477.79±700.26; P < 0.01). Our observations were appeared Significantly higher levels of blood cholesterol, TG, LDL, and VLDL-C and In diabetes individuals, there was a significant drop There was a 0.05 difference in HDL-C levels when compared to healthy controls. Conclusion: These findings indicate that low netrin1 concentration in serum are strongly linked to the occurrence of type II diabetes.

scholarly journals Co-infection of bovine papillomavirus type-1 and -10 in teat warts of dairy cattle

2013 ◽  
Vol 58 (No. 12) ◽  
pp. 605-608
Author(s):  
P. Kumar ◽  
BL Jangir ◽  
G. Saikumar ◽  
R. Somvanshi
Keyword(s):  
Dairy Cattle ◽  
Real Time ◽  
Real Time Pcr ◽  
Rice Grain ◽  
Bovine Papillomavirus ◽  
Viral Dna ◽  
Specific Primers ◽  
Copy Numbers ◽  
Pcr Techniques

The present study was carried out to investigate the involvement of different bovine papillomaviruses in the teat warts of cattle. A total of 11 teat wart samples showing rice grain-like and small, sessile elevated greyish or flesh-like growths were collected from dairy cattle. DNA was extracted from these teat wart samples and PCR and real time PCR techniques were applied using specific primers for BPV-1 and -10 to detect the presence of viral nucleic acid. PCR revealed the presence of viral DNA of BPV-1 and -10 in three and seven samples, respectively. Quantification using real time PCR revealed that the copy numbers of the viral DNA of BPV-1 and -10 DNA varied from 1.12E + 04 to 2.99E + 04 and 3.56E + 02 to 5.23E + 06, respectively. From the present study it can be concluded that BPV-1 and -10 are involved in production of rice grain-like and sessile elevated growths on the teats of cattle.

scholarly journals Validation of a real-time PCR assay for high-throughput detection of Avibacterium paragallinarum in chicken respiratory sites

2019 ◽  
Vol 31 (5) ◽  
pp. 714-718 ◽  
Author(s):  
Kristin A. Clothier ◽  
Simone Stoute ◽  
Andrea Torain ◽  
Beate Crossley
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Limit Of Detection ◽  
Bacterial Flora ◽  
Clinical Samples ◽  
Single Tube ◽  
Avibacterium Paragallinarum ◽  
Normal Bacterial Flora ◽  
Close Relatives

Avibacterium paragallinarum is the causative agent of infectious coryza, a highly contagious respiratory disease in chickens. Given its fastidious nature, this bacterium is difficult to recover and identify, particularly from locations colonized by normal bacterial flora. Standard PCR methods have been utilized for detection but are labor-intensive and not feasible for high-throughput testing. We evaluated a real-time PCR (rtPCR) method targeting the HPG-2 region of A. paragallinarum, and validated a high-throughput extraction for this assay. Using single-tube extraction, the rtPCR detected 4 A. paragallinarum (ATCC 29545T and 3 clinical) isolates with a limit of detection (LOD) of 10 cfu/mL and a PCR efficiency of 89–111%. Cross-reaction was not detected with 33 non– A. paragallinarum, all close relatives from the family Pasteurellaceae. Real-time PCR testing on extracts of 66 clinical samples (choana, sinus, or trachea) yielded 98.2% (35 of 36 on positives, 30 of 30 on negatives) agreement with conventional PCR. Duplicate samples tested in a 96-well format extraction in parallel with the single-tube method produced equivalent LOD on all A. paragallinarum isolates, and 96.8% agreement on 93 additional clinical samples extracted with both procedures. This A. paragallinarum rtPCR can be utilized for outbreak investigations and routine monitoring of susceptible flocks.

scholarly journals Implementation and Validation of the Roche Light Cycler 480 96-Well Plate Platform as a Real-Time PCR Assay for the Quantitative Detection of Cytomegalovirus (CMV) in Clinical Specimens Using the Luminex MultiCode ASRs System

2020 ◽  
Vol 8 (1) ◽  
pp. 14
Author(s):  
Shengwen Calvin Li ◽  
Kara J. Sparks ◽  
Leonard S. Sender
Keyword(s):  
Stem Cell ◽  
Real Time ◽  
Real Time Pcr ◽  
Reference Range ◽  
Measurement Range ◽  
Quantitative Detection ◽  
Clinical Settings ◽  
Stem Cell Therapies ◽  
Cell Therapies ◽  
Copy Numbers

Allogenic stem-cell therapies benefit patients in the treatment of multiple diseases; however, the side effects of stem-cell therapies (SCT) derived from the concomitant use of immune suppression agents often include triggering infection diseases. Thus, analysis is required to improve the detection of pathogen infections in SCT. We develop a polymerase chain reaction (PCR)-based methodology for the qualitative real-time DNA detection of cytomegalovirus (CMV), with reference to herpes simplex virus types 1 (HSVI), Epstein–Barr virus (EBV), and varicella-zoster virus (VZV) in blood, urine, solid tissues, and cerebrospinal fluid. This real-time PCR of 96-well plate format provides a rapid framework as required by the Food and Drug Administration (FDA) for clinical settings, including the processing of specimens, reagent handling, special safety precautions, quality control criteria and analytical accuracy, precisely reportable range (analyst measurement range), reference range, limit of detection (LOD), analytical specificity established by interference study, and analyte stability. Specifically, we determined the reportable range (analyst measurement range) with the following criteria: CMV copies ≥200 copies/mL; report copy/mL value; CMV copies ≤199 copies/mL; report detected but below quantitative range; CMV copies = 0 with report <200 copies/mL. That is, with reference range, copy numbers (CN) per milliliter (mL) of the LOD were determined by standard curves that correlated Ct value and calibrated standard DNA panels. The three repeats determined that the measuring range was 1E2~1E6 copies/mL. The standard curves show the slopes were within the range −2.99 to −3.65 with R2 ≥ 0.98. High copy (HC) controls were within 0.17–0.18 log differences of DNA copy numbers; (2) low copy (LC) controls were within 0.17–0.18 log differences; (3) LOD was within 0.14–0.15 log differences. As such, we set up a fast, simple, inexpensive, sensitive, and reliable molecular approach for the qualitative detection of CMV pathogens. Conclusion: This real-time PCR of the 96-well plate format provides a rapid framework as required by the FDA for clinical settings.

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