Assessment the Possible Association Between Neuromyelitis Optica and Cytomegalovirus as a Provocative Factor

Background: Neuromyelitis Optica (NMO) is an autoimmune inflammation of the central nervous system in which autoantibodies are released against Aquaporin-4 (AQP-4), astrocytic water channels. The disease is characterized by transverse myelitis and optic neuritis. Viruses could be inflammatory agents in the brain. Due to such inflammatory reactions, autoantibodies would cross the blood brain barrier. Therefore, recognizing the responsible viral agent may help us prevent or treat NMO. Objectives: To investigate the probable association between Cytomegalovirus infection (CMV) and Neuromyelitis Optica. Materials & Methods: This descriptive study was performed on 25 patients with NMO, 30 patients with Multiple Sclerosis (MS) referring to an academic MS Clinic and 30 healthy individuals in Isfahan City, Iran in 2016. In order to detect DNA of CMV in the sera of the studied groups, real-time PCR technique was used with hydrolyzing probes such as TaqMan. Beacon Designer 7 was used to design a primer and probe. The Chi-square test was used for statistical analysis in SPSS 16. Results: Three study groups had no significant difference in terms of age (P=0.33) and gender (P=0.599). All of the samples were negative for CMV DNA. There was no significant difference between three groups of study (P=0.33). Conclusion: Regarding the negative real-time PCR results of all samples, and especially using higher specificity of primers and probes in detecting this virus, it can be inferred that no significant correlation exists between CMV infection and NMO disease.

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The Influence of Age and Gender on Mandibular Indices among the Libyan Population

Journal of Oral & Dental Health ◽

10.33140/jodh.03.02.2 ◽

2019 ◽

Vol 3 (2) ◽

Keyword(s):

Negative Correlation ◽

Bone Mineralization ◽

Age Groups ◽

T Test ◽

Chi Square ◽

Age And Gender ◽

Significant Difference ◽

Chi Square Test ◽

And Gender ◽

Libyan Population

Radiographic Mandibular Indices serve as easy and relatively cheap tools for evaluating bone mineralization. Objectives: To examine the effect of age and gender on three mandibular indices: the panoramic mandibular index (PMI), the mandibular ratio (MR) and the mandibular cortical index (MCI), among Libyan population. Methods: The three indices were measured on 317 digital (OPGs) of adult humans (155 males, 162 females). The sample was divided into six age groups (from 18-25 years through 56-65 years). The measurements were analyzed for interactions with age and sex, using SPSS (Statistical Package for Social Studies) software version no. 22. The tests employed were two way ANOVA, the unpaired T-test and chi-square test. Results: The mean PMI fluctuated between 0.37 s.d. 0.012 and 0.38 s.d. 0.012. among the sixth age groups. One-way ANOVA statistical test revealed no significant of age on PMI. On the other hand gender variation has effect on PMI, since independent sample t-test disclosed that the difference between the male and female PMI means statistically significant. ANOVA test showed that the means of MR among age groups showed a negative correlation i.e. MR mean declined from 3.01 in 18-25 age groups to 2.7 in 55-65 age groups. In contrary, the gender showed no effect on MR according two sample t-test at p> 0.05. In regards with MCI, statistical analysis showed that it affected by age that is C1 was decreasing by age while C2 and C3 were increased by age. Using chi square test the result indicated that there is a significant difference among the different age group and the two genders in MCI readings. Conclusion: PMI was influenced significantly by age but minimally by the gender. MR is not affected by gender but has a negative correlation with age. MCI is affected by both age and gender

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Success Rate of Obturation of Root Canals by Different Techniques in Primary Molars: A Comparative Clinical Study

Current Dentistry ◽

10.2174/2542579x02999200503032916 ◽

2020 ◽

Vol 2 (1) ◽

pp. 36-44

Author(s):

Satyawan G. Damle ◽

Ritika Bansal ◽

Dhanashree D. Sakhare

Keyword(s):

Success Rate ◽

Study Groups ◽

Primary Molars ◽

Control Group ◽

Second Primary ◽

Chi Square ◽

Exact Test ◽

Root Canals ◽

Significant Difference ◽

Comparative Clinical Study

Objective: To compare the success rate of different obturation procedures in primary mandibular second molars clinically and also by digital radiovisiography. Methods: A total of 40 children aged between 4-8 years with deeply carious mandibular second primary molars indicated for single session pulpectomy were selected. Canals were obturated with Metapex. The 3 study groups (Endodontic plugger, Handheld lentulospiral, Navi Tip syringe) were compared with the control group (reamer) both clinically and radiovisiographically. The data collected were statistically analyzed using Pearson’s Chi-square and Fisher’s exact test. Results: The use of Navi tip syringe led to the least number of voids followed by Endodontic plugger and Reamer and the highest number of voids was reported with Lentulospiral. Navitip presented maximum number of optimally filled cases followed by Endodontic plugger and Lentulospiral and least number of optimally filled cases with reamer. However, there was no statistically significant difference (p>0.05) in any of the groups with clinical (pain and tenderness to percussion) and radiographic parameters (presence or absence of voids and length of obturation). Conclusion: Within the limitations of the present study, though the clinical outcome was statistically insignificant, Navitip syringe exhibited encouraging results and is a promising option for obturation in primary teeth.

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Genome-wide screening of m6A-modified transcript in the Wilms’ tumor

10.21203/rs.3.rs-197376/v1 ◽

2021 ◽

Author(s):

Zhuo Liu ◽

Feng He ◽

Jing Liu ◽

Shengrong OuYang ◽

Zexi Li ◽

...

Keyword(s):

Gene Ontology ◽

Real Time ◽

Real Time Pcr ◽

Wilms Tumor ◽

Expression Patterns ◽

Mrna Levels ◽

Gene Ontology Analysis ◽

Quantitative Real Time Pcr ◽

Related Factors ◽

Significant Difference

Abstract Background Wilms’ tumor, also called nephroblastoma, is the most common pediatric renal malignancy. The pathogenesis of Wilms’ tumor has been attributed to several genetic and epigenetic factors. However, the most pervasive internal mRNA modification that affects almost every process of RNA metabolism, RNA N6-Methyladenosine (m6A) methylation, has not been characterized in Wilms’ tumor. Methods Wilms’ tumor (WT) and adjacent non-cancerous (NC) tissue samples were obtained from 23 children with nephroblastoma, and the global m6A levels were measured by mass spectrometry. Analyses by m6A-mRNA epitranscriptomic microarray and mRNA microarray were performed, and m6A-related mRNAs were validated by quantitative real-time PCR for input and m6A-immunoprecipitated RNA samples from WT and NC tissues. Gene ontology analysis and KEGG pathway analysis were performed for differentially expressed genes, and expression of RNA methylation-related factors was measured by quantitative real-time PCR. Results The total m6A methylation levels in total RNA of WT samples and NC samples were (0.21 ± 0.01)% and (0.22 ± 0.01)%, respectively, with no statistically significant difference. Fifty-nine transcripts were differentially m6A-methylated between the WT and NC groups, which showed distinct m6A modification patterns. Gene ontology analysis indicated that m6A-modified genes were enriched in cancer-associated pathways, including the mTOR pathway, and conjoint analysis of the unique methylation and gene expression patterns in WT samples suggested an association with metabolic pathways.The mRNA levels of the m6A-related “reader” genes, YTHDF1, YTHDF2 and IGF2BP3, were statistically higher in WT samples than in NC samples. Conclusion This is the first study to determine the m6A modification profiles in Wilms’ tumor. Our data provide novel information regarding patterns of m6A modification that correlate with carcinogenesis in Wilms’ tumor.

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DETEKSI CLOSTRIDIUM DIFFICILE TOKSIGENIK MENGGUNAKAN UJI CEPAT TOKSIN DAN REAL TIME POLYMERASE CHAIN REACTION (Toxigenic Clostridium Difficile Detection Using Toxin Rapid Test and Real Time Polymerase Chain Reaction)

INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY ◽

10.24293/ijcpml.v22i1.1217 ◽

2018 ◽

Vol 22 (1) ◽

pp. 22

Author(s):

Ika Yasma Yanti ◽

Dalima Ari Wahono Astrawinata

Keyword(s):

Polymerase Chain Reaction ◽

Clostridium Difficile ◽

Antibiotic Therapy ◽

Real Time ◽

Real Time Pcr ◽

Rapid Test ◽

Chain Reaction ◽

Chi Square ◽

Polymerase Chain ◽

Clostridium Difficile Associated Diarrhea

Toxigenic Clostridium difficile infection, causing a Pseudo Membrane Colitis (PMC) and Clostridium Difficile Associated Diarrhea(CDAD) has increased sharply. The largest risk factor is the use of antibiotics. The purpose of this study was to know how to determinethe prevalence and characteristics of subjects with Toxigenic Clostridium difficile and to assess the ability of the toxin rapid test comparedto real-time PCR. Ninety adult subjects with antibiotic therapy more than two (2) weeks were enrolled in this study. The results of toxinrapid test and real-time PCR were presented in a 2x2 table, statistical test used was Chi square. The prevalence of Toxigenic Clostridiumdifficile based on the toxin rapid test and by real-time PCR was 27.3% and 37.5%, respectively. There were significant differences betweenstool consistency and number of antibiotics used with the detection of Toxigenic Clostridium difficile. There was a relationship betweenthe duration of antibiotic therapy with the detection of Toxigenic Clostridium difficile using real-time PCR (p=0.010, RR=2.116). Thesensitivity, specificity, PPV, NPV, PLR and NLR rapid test against real-time PCR were 69.7%; 98.2%; 95.8%; 84.4%; 39.2 and 0.31,respectively. This study concluded that the prevalence of Clostridium difficile in RSCM was higher compared to that in Malaysia, Thailandand India; the subjects with antibiotic therapy for more than four (4) weeks had a double risk to have Toxigenic Clostridium difficilethan subjects with antibiotic therapy for less than that time (4 weeks). Thus, in this study, toxin rapid test could be used as a tool todetect Toxigenic Clostridium difficile.

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

Indian Journal of Clinical Anatomy and Physiology ◽

10.18231/j.ijcap.2021.070 ◽

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.

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TLR, RAGE, and HMGB1 in the Inflammatory Response in Ph(-) Myeloproliferative Neoplasm

Blood ◽

10.1182/blood-2020-134988 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 31-32

Author(s):

Guanfang Shi ◽

Kiron Nair ◽

Preethi Ramachandran ◽

Chi Chen ◽

Ching Wong ◽

...

Keyword(s):

Inflammatory Response ◽

Real Time ◽

Inflammatory Cytokines ◽

Internal Control ◽

Real Time Pcr ◽

Mononuclear Cells ◽

Conflicts Of Interest ◽

Philadelphia Chromosome ◽

Myeloproliferative Neoplasm ◽

Significant Difference

Recent evidence of increased constitutional symptoms and inflammatory cytokines in Philadelphia chromosome negative (Ph (-)) MPN suggests that an inflammatory response is important in the pathogenesis of Ph (-) MPN. Toll-like receptors (TLR), Receptor for Advanced Glycation End products (RAGE) and High mobility group protein B1 (HMGB1) are the important pathways for the inflammatory response. All these three important pathway proteins were studied in MPN diseases in the current studies. Materials and Methods: TLR assay. TLR 2,3, 4, 7, 9 quantification was performed by immuno-staining of 1×106 mononuclear cells (peripheral blood) which were incubated with fluorescence-conjugated anti-TLR-2,3, 4, 7, 9 antibodies and assayed by flow cytometry. HMGB1assay:HMGB1 ELISA kit from Immuno-Biological Laboratories, Inc. (IBL-America) were used. The plasma samples were diluted four times with the provided sample dilution buffer, and assayed in duplicate according to the manufacturer's suggestion. RAGE (RT-PCR) Assay: Total RNA was extracted from normal control or patient mononuclear cells. Predesigned primers for RAGE, and internal control genes were ordered from Qiagen (Germantown, MD). Real-time PCR was performed using SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA) on Bio-Rad iQ5 Multicolor Real-Time PCR Detection System. At least three house-keeping genes (ribosomal protein L4, TATA box binding protein, and tubulin-α 1b) were used as normalization controls. The expression of RAGE were compared with each internal control. Average of three was used to calculate the ratio of final patient to normal Results: Total of 97 patients with MPN were studied 1) TLR: TLR 3,7,9 was not significantly different from controls. But TLR 2 was significantly increased in both PV, as well as in the MPN group when PV, ET and MF were grouped together as MPN (Fig A). TLR 4 was not significantly increased in PV, ET, MF individually but was found to be significantly increased than the controls, when they are grouped together as MPN (Fig B). 2) RAGE: No significant difference was found between ET, PV, MF individually or when they were grouped together as MPN than the controls (Fig C). 3) HMGB1: No significant difference was seen between ET, PV, MF or when they were grouped as MPN (Fig D). Conclusion: Current study suggests that TLR pathway especially TLR2, and to a lesser extent TLR4 are the important pathways for inflammatory response with increased inflammatory cytokines in MPN, while HMGB1 and RAGE pathways were not different from controls. Figure Disclosures No relevant conflicts of interest to declare.

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Diversity of KIR genes and their HLA-C ligands in Ugandan populations with historically varied malaria transmission intensity

10.21203/rs.3.rs-45487/v2 ◽

2021 ◽

Author(s):

Stephen Tukwasibwe ◽

James A. Traherne ◽

Olympe Chazara ◽

Jyothi Jayaraman ◽

John Trowsdale ◽

...

Keyword(s):

Infectious Diseases ◽

Malaria Transmission ◽

Real Time ◽

High Throughput ◽

Real Time Pcr ◽

Association Studies ◽

Transmission Intensity ◽

Malaria Transmission Intensity ◽

Significant Difference ◽

Pcr Method

Abstract Background: Malaria is one of the most serious infectious diseases in the world. The malaria burden is greatly affected by human immunity, and immune responses vary between populations. Genetic diversity in KIR and HLA-C genes, which are important in immunity to infectious diseases, is likely to play a role in this heterogeneity. Several studies have shown that KIR and HLA-C genes influence the immune response to viral infections, but few studies have examined the role of KIR and HLA-C in malaria infection, and these have used low-resolution genotyping. The aim of this study was to determine whether genetic variation in KIR and their HLA-C ligands differ in Ugandan populations with historically varied malaria transmission intensity using more comprehensive genotyping approaches.Methods: High throughput multiplex quantitative real-time PCR method was used to genotype KIR genetic variants and copy number variation and a high-throughput real-time PCR method was developed to genotype HLA-C1 and C2 allotypes for 1,344 participants, aged 6 months to 10 years, enrolled from Ugandan populations with historically high (Tororo District), medium (Jinja District) and low (Kanungu District) malaria transmission intensity. Results: The prevalence of KIR3DS1, KIR2DL5, KIR2DS5 and KIR2DS1 genes was significantly lower in populations from Kanungu compared to Tororo (7.6% vs. 13.2%: p=0.006, 57.2% vs. 66.4%: p=0.005, 33.2% vs. 46.6%: p<0.001 and 19.7% vs. 26.7%: p=0.014 respectively) or Jinja (7.6% vs.18.1%: p<0.001, 57.2% vs. 63.8%: p=0.048, 33.2% vs. 43.5%: p=0.002 and 19.7% vs. 30.4%: p<0.001 respectively). The prevalence of homozygous HLA-C2 was significantly higher in populations from Kanungu (31.6%) compared to Jinja (21.4%), p=0.043, with no significant difference between Kanungu and Tororo (26.7%), p=0.296. Conclusions: The KIR3DS1, KIR2DL5, KIR2DS5 and KIR2DS1 genes may partly explain differences in transmission intensity of malaria since these genes have been positively selected for in places with historically high malaria transmission intensity. The high-throughput multiplex real-time HLA-C genotyping PCR method developed will be useful in disease association studies involving large cohorts.

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Major clinical risk factors for seizures in febrile children aged 6 to 60 months.

The Professional Medical Journal ◽

10.29309/tpmj/2020.27.05.3397 ◽

2020 ◽

Vol 27 (05) ◽

pp. 891-894

Author(s):

Shahid Ishaq ◽

Ejaz Mazari ◽

Fazal ur Rehman

Keyword(s):

Risk Factors ◽

Family History ◽

Febrile Seizures ◽

Study Groups ◽

Clinical Risk Factors ◽

P Value ◽

Chi Square ◽

Clinical Risk ◽

Significant Difference ◽

History Of

Objectives: Febrile seizures (FS) are the most common type of seizures and typically transpire in children with ages from 6 to 60 months. This study was planned to find out major clinical risk factors for seizures in febrile children who were aged 6 to 60 months. A total of 100 febrile children aged 6 to 60. Study Design: Analytical Study. Setting: Department of Neurology, Children’s Hospital and the Institute of Child Health, Multan. Period: From 1st April 2018 to 31st December 2018. Material & Methods: Group A had 40 children with febrile seizures while group B had 60 febrile children but without seizures. Demographic features along with family history of (H/O) epilepsy as well as family history of febrile seizure, types of seizure and infection diseases were noted and analyzed using SPSS version 20. Odds ratio was calculated for various risk factors. Chi square test was applied and P value < 0.05 was considered as significant. Results: Out of a total of 100 children, there were 54 (54.0%) male and 46 (46.0%) female. There was no statistical difference in terms of gender between the two groups (p value = 0.566). Overall, mean age of the children was 26.02 months with standard deviation of 13.4 months. There were 28 (70.0%) children who reported with simple seizures while complex seizures were found in 12 (30.0%) cases. Statistically significant difference (p value = 0.001) was seen in terms of types of infections between the two study groups. When risk of seizures for various risk factors was calculated, family H/O FS, family H/O epilepsy, and upper RTI were as 14, 7 and 3 times respectively and turned out to be the major risk factors for seizures in febrile children. Conclusions: Family H/O FS, family H/O epilepsy and upper RTIs are the major risk factors related with seizures in febrile children. Measures to prevent these risk factors can decrease the burden of FS in our population.

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Investigation of the effect of Prunus Amygdalus Amara on the expression of some genes of apoptosis and immortality in breast cancer cells (MCF-7)

Current Drug Research Reviews ◽

10.2174/2589977513666211202094433 ◽

2021 ◽

Vol 13 ◽

Author(s):

Maryam Abdolahi-Majd ◽

Gholamhossein Hassanshahi ◽

Mahboubeh Vatanparast ◽

Mojgan Noroozi Karimabad

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Pcr Analysis ◽

Mrna Levels ◽

Prunus Amygdalus ◽

Mcf7 Cells ◽

Significant Difference ◽

Anti Cancer ◽

Bitter Almond ◽

Mcf 7

Background: Anti-cancer effects of almond nuts or oil have been approved, but there are a few pieces of research that have evaluated, in detail, almond and other seeds' effects on cancer. Therefore, in the present project, the aim was to explore the regulatory effect of the bitter almond extract (Prunus amygdalus Batsch) on the apoptotic and anti-cancer potency of MCF-7 cells. Objectives: In the current experimental research, the Almond effect on MCF7 cells was evaluated by investigating the expression and the balance between Bcl-2, Bax genes to unmark the potential molecular mechanism. Methods: For 24 and 48h, the MCF7 cells were treated with the bitter almond extract (187.5-3000 µg/mL). MTT assay was used to assess the viability, and Real-time-PCR was applied to determine the expression of Bax and Bcl-2, facing β-actin. Results: Our results revealed a significant difference between different extract concentrations on the viability of MCF7 cell lines in 24 and 48 h; cell viability decreased time-dependently (P < 0.05). After 24 and 48h of extract facing MCF7 cells, the evaluated IC50 value was 3000 and 1500 µg/mL, respectively. Based on Real Time-PCR analysis, after 24 and 48 h, the mRNA levels of BCL-2 decreased by the extract, whereas BAX was in the MCF-7 cell line. Conclusion: From the results, it can be concluded that bitter almond extract has anti-cancer properties that may influence the apoptotic pathways by regulating relative gene expression.

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Validation of iQ-Check E. coli O157:H7 Real-Time PCR Test Kit for Detection of Escherichia coli O157:H7 in Selected Foods

Journal of AOAC International ◽

10.1093/jaoac/92.4.1095 ◽

2009 ◽

Vol 92 (4) ◽

pp. 1095-1104 ◽

Cited By ~ 1

Author(s):

Wendy F Lauer ◽

Sylvie Tymciu ◽

Caroline D Sidi ◽

Pierre Sonigo

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Ground Beef ◽

Reference Method ◽

Target Sequence ◽

Apple Cider ◽

E Coli ◽

Test Kit ◽

Significant Difference ◽

The U.S

Abstract iQ-Check E. coli O157:H7 (Bio-Rad Laboratories, Hercules, CA) is a real-time PCR kit for detection of E. coli O157:H7 from selected foods. Specific fluorescent oligonucleotide probes are used to detect target DNA during the amplification, by hybridizing to the amplicons. These fluorescent probes are linked to a fluorophore which fluoresces only when hybridized to the target sequence. Three foods (ground beef, apple cider, fresh spinach) were selected to compare the performance of iQ-Check E. coli O157:H7 to the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook (MLG) reference method for ground beef and the U.S. Food and Drug Administration/Bacteriological Analytical Manual reference method for apple cider and fresh spinach. Three protocols were tested in this study: a shortened 8 h primary enrichment in buffered peptone water (BPW), a 24 h enrichment in BPW, and an enrichment in appropriate reference method enrichment broth. The iQ-Check E. coli O157:H7 method was able to identify more true/confirmed positive samples than the reference method. Inclusivity and exclusivity rates of the method were 100. iQ-Check E. coli O157:H7 performed as expected when minor procedural variations were introduced, validating the ruggedness of the method. There was no significant difference observed in performance over the shelf life of the kit.

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