Detection of circulating miRNAs in the maternal serum during early pregnancy in the pig by qRT-PCR and digital RT-PCR

2014 ◽  
Author(s):  
Reliszko Zaneta P ◽  
Zdzislaw Gajewski ◽  
Kaczmarek Monika M
Keyword(s):  
Early Pregnancy ◽  
Maternal Serum ◽  
Circulating Mirnas ◽  
Rt Pcr ◽  
Qrt Pcr

scholarly journals Development of an RNA Extraction Protocol for a Norovirus from Raw Oysters and Detection by qRT-PCR and Droplet-Digital RT-PCR

Foods ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1804
Author(s):  
Daniel Plante ◽  
Julio Alexander Bran Barrera ◽  
Maude Lord ◽  
Irène Iugovaz ◽  
Neda Nasheri
Keyword(s):  
Hepatitis A Virus ◽  
Rna Extraction ◽  
Complex Nature ◽  
Murine Norovirus ◽  
Rt Pcr ◽  
Pcr Inhibitors ◽  
Qrt Pcr ◽  
Extraction Protocol ◽  
Viral Particles ◽  
Hands On

Foodborne viruses such as norovirus and hepatitis A virus cause frequent outbreaks associated with the consumption of raw or undercooked oysters. Viral particles are bioaccumulated in the oyster’s digestive glands, making RNA extraction and RT-PCR detection difficult due to the complex nature of the food matrix and the presence of RT-PCR inhibitors. Herein, we have developed a viral RNA extraction protocol from raw oysters using murine norovirus (MNV) as a surrogate for human noroviruses. The method combines lysis in Tri-Reagent reagent, followed by RNA extraction using Direct-Zol purification columns and lithium chloride precipitation. Viral load quantification was performed by both qRT-PCR and droplet-digital RT-PCR. We have demonstrated that this method can efficiently remove RT-PCR inhibitors, and is sensitive enough to reliably detect viral contamination at 25 PFU/0.2 g. We have also compared the efficiency of this method with the ISO 15216-1:2017 method and Method E developed by Quang and colleagues, and observed significantly higher efficiency compared with the ISO 15216-1 method and comparable efficiency with Method E, with less steps, and shorter hands-on time.

scholarly journals Cloning, Sequencing, and the Expression of the Elusive Sarcomeric TPM4α Isoform in Humans

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Dipak K. Dube ◽  
Syamalima Dube ◽  
Lynn Abbott ◽  
Ruham Alshiekh-Nasany ◽  
Charles Mitschow ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Western Blot ◽  
Human Heart ◽  
Amino Acid Sequences ◽  
Rt Pcr ◽  
Adult Heart ◽  
Qrt Pcr ◽  
Long Time ◽  
Alternate Promoters ◽  
Computational Analyses

In mammals, tropomyosin is encoded by four known TPM genes (TPM1, TPM2, TPM3, and TPM4) each of which can generate a number of TPM isoforms via alternative splicing and/or using alternate promoters. In humans, the sarcomeric isoform(s) of each of the TPM genes, except for the TPM4, have been known for a long time. Recently, on the basis of computational analyses of the human genome sequence, the predicted sequence of TPM4α has been posted in GenBank. We designed primer-pairs for RT-PCR and showed the expression of the transcripts of TPM4α and a novel isoform TPM4δ in human heart and skeletal muscle. qRT-PCR shows that the relative expression of TPM4α and TPM4δ is higher in human cardiac muscle. Western blot analyses using CH1 monoclonal antibodies show the absence of the expression of TPM4δ protein (~28 kDa) in human heart muscle. 2D western blot analyses with the same antibody show the expression of at least nine distinct tropomyosin molecules with a mass ~32 kD and above in adult heart. By Mass spectrometry, we determined the amino acid sequences of the extracted proteins from these spots. Spot “G” reveals the putative expression of TPM4α along with TPM1α protein in human adult heart.

A prospective study of early-pregnancy thyroid markers, lipid species, and risk of gestational diabetes mellitus

2021 ◽  
Author(s):  
Yi Wang ◽  
Fengjiang Sun ◽  
Ping Wu ◽  
Yichao Huang ◽  
Yi Ye ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Gestational Diabetes ◽  
Gestational Diabetes Mellitus ◽  
Early Pregnancy ◽  
Maternal Serum ◽  
Thyroid Peroxidase ◽  
Free Triiodothyronine ◽  
Composite Effect ◽  
Lipid Species ◽  
Increased Risk

Abstract Context While the associations between thyroid markers and gestational diabetes mellitus (GDM) have been extensively studied, the results are inconclusive and the mechanisms remain unclear. Objective We aimed to investigate the prospective associations of thyroid markers in early gestation with GDM risk, and examine the mediating effects through lipid species. Methods This study included 6068 pregnant women from the Tongji-Shuangliu Birth Cohort. Maternal serum thyroid markers (free triiodothyronine (fT3), free thyroxine (fT4), thyroid-stimulating hormone, thyroid peroxidase antibody, and thyroglobulin antibody) were measured before 15 weeks. Deiodinase activity was assessed by fT3/fT4 ratio. Plasma lipidome were quantified in a subset of 883 participants. Results Mean age of the participants was 26.6 ± 3.7 years, and mean gestational age was 10.3 ± 2.0 weeks. Higher levels of fT4 were associated with a decreased risk of GDM (OR=0.73 comparing the extreme quartiles; 95% CI 0.54, 0.98, Ptrend =0.043), while higher fT3/fT4 ratio was associated with an increased risk of GDM (OR=1.43 comparing the extreme quartiles; 95% CI 1.06, 1.93, Ptrend =0.010) after adjusting for potential confounders. Multiple linear regression suggested that fT3/fT4 ratio was positively associated with alkylphosphatidylcholine 36:1, phosphatidylethanolamine plasmalogen 38:6, diacylglyceride 18:0/18:1, sphingomyelin 34:1, and phosphatidylcholine 40:7 (false discovery rate adjusted P<0.05). Mediation analysis indicated 67.9% of the association between fT3/fT4 ratio and GDM might be mediated through the composite effect of these lipids. Conclusions Lower concentration of serum fT4 or higher fT3/fT4 ratio in early pregnancy was associated with an increased risk of GDM. The association of fT3/fT4 ratio with GDM was largely mediated by specific lipid species.

scholarly journals Evaluation of the performance of the COVID-19 rapid antigen tests in a tertiary level hospital in Bangladesh.

2021 ◽  
Author(s):  
Mohammad Jahidur Rahman Khan ◽  
Md. Shahadat Hossain ◽  
Samshad Jahan Shumu ◽  
Md. Selim Reza ◽  
Farzana Mim ◽  
...  
Keyword(s):  
Viral Load ◽  
Real Time ◽  
Sensitivity And Specificity ◽  
Detection Rate ◽  
High Sensitivity ◽  
Rt Pcr ◽  
College Hospital ◽  
Medical College ◽  
Qrt Pcr ◽  
Antigen Tests

Abstract Background: While the COVID-19 pandemic is a worldwide crisis, tests with high sensitivity and specificity are essential for identifying and managing COVID-19 patients. Globally, several rapid antigen tests RATs for COVID-19 have been developed, but their clinical efficacy has not been well established. This study aimed to evaluate the performance of several rapid antigen tests (RATs) to diagnose SARS-CoV-2 infection.Methods: This prospective observational study was conducted at Shaheed Suhrawardy Medical College hospital from February 2021 to April 2021 in Dhaka, Bangladesh. This study included the patients admitted in this hospital at the COVID-19 isolation unit or referred from the triage facility of the outdoor department of this hospital suspected as COVID-19 case. Two nasopharyngeal samples were collected simultaneously. one sample was used on the spot for the RAT. The other was sent to the adjacent Shaheed Suhrawardy Medical College COVID-19 RT-PCR laboratory for real-time reverse transcription-polymerase chain reaction (qRT-PCR). The performance of the RAT was evaluated using the results of qRT-PCR as a reference.Results: A total of 223 patients were included in this study, and the real-time RT-PCR detected SARS-CoV-2 in 84 (37.7%) patients. Of these 84 patients, 9 (10.7%) were asymptomatic. The overall sensitivity and specificity of RATs were 78.6% and 99.3%, respectively. The sensitivity was 81.3% in symptomatic cases and 55.6% in asymptomatic cases. False-negatives were observed in 18 patients, 3 of whom were asymptomatic and had a low viral load (cycle threshold (Ct) > 30). The detection rate of RATs was 100% when the Ct value was up to 24. The detection rate was 42.3% when the Ct was >29. The detection rate of RATs was 92.3% when the onset of symptoms was within three days. The detection rate was 33.3% when the onset of symptoms was >7 days.Conclusions: RATs for COVID-19 used in this study delivered an acceptable performance in patients with high viral load and within the first week of the onset of symptoms. They can be used as a supplementary method to RT-PCR for the diagnosis of COVID-19 patients.

Maternal serum thyroid antibodies in early pregnancy and fetal Down's syndrome

1988 ◽  
Vol 8 (6) ◽  
pp. 439-445 ◽  
Author(s):  
Howard Cuckle ◽  
Nicholas Wald ◽  
Rossana Stone ◽  
James Densem ◽  
James Haddow ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Down's Syndrome ◽  
Down’S Syndrome ◽  
Maternal Serum ◽  
Thyroid Antibodies

scholarly journals Absolute measurement of androgen receptor mRNA in peripheral blood mononuclear, preputial skin and urethral mucosa cells of control individuals with phimosis using qRT-PCR

2011 ◽  
Vol 55 (8) ◽  
pp. 665-668 ◽  
Author(s):  
Tatiane Sousa e Silva ◽  
Flavio Richetti ◽  
Daniela Patricia Palmeira Santos Cunha ◽  
Antonio Carlos Moreira Amarante ◽  
Jovelino Quintino de Souza Leão ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Peripheral Blood ◽  
External Genitalia ◽  
Absolute Measurement ◽  
Rt Pcr ◽  
Peripheral Blood Mononuclear ◽  
Skin Cells ◽  
Qrt Pcr ◽  
Specific Manner ◽  
The Mean

INTRODUCTION: Androgen actions are exerted upon the androgen receptor (AR), and complete genital virilization of normal 46,XY individuals depends on adequate function and expression of the AR gene in a tissue-specific manner. OBJECTIVE: Standardization of normal ARmRNA in androgen-sensitive tissues. MATERIALS AND METHODS: In this study, we determined the quantitative amounts of ARmRNA in peripheral blood mononuclear, urethral mucosa and preputial skin cells of control subjects with phimosis by using RT-PCR. RESULTS: The mean (SD) values of AR expression in blood, urethra and prepuce were: 0.01 (0.01); 0.43 (0.32); 0.31 (0.36), respectively. CONCLUSION: The AR expression is low in blood and equivalent in urethral mucosa and preputial skin, which may be useful in the diagnosis of individuals with abnormal external genitalia.

scholarly journals Feeding of 1-Kestose Induces Glutathione-S-Transferase Expression in Mouse Liver

Foods ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 69 ◽  
Author(s):  
Takumi Tochio ◽  
Yuki Ueno ◽  
Yasuyuki Kitaura ◽  
Mikako Shinohara ◽  
Yoshihiro Kadota ◽  
...  
Keyword(s):  
Gene Expression ◽  
Functional Food ◽  
Mouse Liver ◽  
Antioxidative Activity ◽  
Epididymal Adipose Tissue ◽  
Activity Levels ◽  
Rt Pcr ◽  
Glutathione S Transferase ◽  
Food Ingredients ◽  
Qrt Pcr

Functional food ingredients, including prebiotics, have been increasingly developed for human health. The improvement of the human intestinal environment is one of their main targets. Fructooligosaccarides (FOS) are oligosaccharide fructans that are well studied and commercialized prebiotics. 1-Kestose, one of the components of FOS, is considered to be a key prebiotic component in FOS. However, to our knowledge, no studies have been reported on the physiological efficacy of 1-Kestose regarding its anti-oxidative activity. In the present study, we examined the effects of dietary 1-Kestose on gene expression of antioxidative enzymes in the liver, kidney and epididymal adipose tissue of mice by quantitative RT-PCR (qRT-PCR). We demonstrated that a 1-Kestose-rich diet increased mRNA and enzymatic activity levels of glutathione-S-transferase (GST) in mouse liver. These results suggest the possibility that dietary 1-Kestose as a prebiotic may enhance antioxidative activity in mice.

scholarly journals M20. CYTOKINE LEVELS THROUGHOUT PREGNANCY AND RISK FOR PSYCHOSIS IN ADULT OFFSPRING: EARLY PREGNANCY MATTERS

2020 ◽  
Vol 46 (Supplement_1) ◽  
pp. S141-S141
Author(s):  
Dana Allswede ◽  
Robert Yolken ◽  
Stephen Buka ◽  
Tyrone Cannon
Keyword(s):  
Psychotic Disorder ◽  
Early Pregnancy ◽  
Serum Levels ◽  
First Trimester ◽  
Underlying Mechanism ◽  
Maternal Serum ◽  
Serum Samples ◽  
Birth Complications ◽  
Cytokine Levels ◽  
Pregnancy And Birth

Abstract Background Schizophrenia has been associated with pregnancy and birth complications, and fetal exposure to inflammation is thought to be a common underlying mechanism. However, it is unclear whether the risk associated with inflammation is specific to particular phases of pregnancy, as no prior studies have examined maternal serum samples across multiple assessments from the first trimester onward. This study examined differences in longitudinal patterns of maternal serum levels of TNFa, IL-1b, IL-5, IL-6, IL-8, IL-10, and IL-17a across pregnancy for offspring who were later ascertained as having a psychotic disorder diagnosis, non-psychotic siblings of these cases, and unrelated, non-psychotic individuals who served as controls. Methods Participants included 90 offspring, 79 siblings, and 273 matched controls from the Philadelphia cohort of the National Collaborative Perinatal Project. Psychotic disorder diagnoses in adulthood were assessed with review of medical records and were confirmed with a validation study. Cytokine levels were assessed using a multiplex bead assay in archived maternal serum samples collected across prenatal visits and birth. Results Levels of pro-inflammatory TNFa, IL-1b, and IL-6 were significantly higher in maternal serum of offspring who later developed psychosis relative to maternal serum of non-psychotic siblings and matched controls. These differences were maximal in first half of pregnancy (7–20 weeks), tapering to non-significant during the second half of pregnancy. Discussion These findings elucidate the importance of exposure to elevated maternal pro-inflammatory cytokine levels in early pregnancy to the etiology of psychosis.

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