Peripheral Blood Mesenchymal Stem Cells Isolated from Indonesia Long Tailed Monkey (Macaca fascicularis)

2018 ◽  
Vol 6 (2) ◽  
pp. 56-69
Author(s):  
Agus Harsoyo ◽  
Dondin Sajuthi ◽  
Arief Boediono ◽  
Yoga Yuniadi ◽  
Irma H Suparto
Keyword(s):  
Macaca Fascicularis ◽  
Block Design ◽  
Age Groups ◽  
Pcr Amplification ◽  
Rt Pcr ◽  
Messenger Ribonucleic Acid ◽  
Incomplete Block Design ◽  
Polymerase Chain ◽  
The Impact ◽  
Cd Markers

An experiment to compare age of Macaca fascicularis (Mf) as pheripheral blood (PB) mesenchymal stem cell (MSC) isolate sources and the impact of its concentration on the pheriperal blood mononucleous cells (PBMC) development has been conducted. Twelve male Mf were used in this experiment. Three different age groups (infant (A1), juvenil (A2) and adult (A3)) of the Mfs were compared as treatments. Isolate of pheriperal blood MSC were created by taking 1 ml, 5 ml or 10 ml the Mfs pheriperal blood, processed them into PBMC, counted, isolated, cultured, subcultured, pelleted, extracted for their messenger Ribonucleic Acid (mRNA). Reverse transcriptase - polymerase chain reaction (RT-PCR) were conducted to obtain complentary Deoxyribonucleic Acid (cDNA). PCR amplification were performed to look cluster differentiation (CD) of the MSC gene expression. Incomplete block design was used and the data were analysed using descriptive statistic and T-Test. The results showed that PBMC counted from infant, juvenil and adult were 6.78 – 7.28, 6.18 – 7.30, and 6.01 – 7.34 log cell, respectively. The subculture and pelleting cells were only obtained from A3 with positive 73, 90, 105 and negative 34, 45 CD markers. It is concluded that pheriperal blood of adult Mf can be utilized as MSC source.

Operation Principle and Simulation of Loop-Type Microfluidic Biochips

2006 ◽  
Vol 505-507 ◽  
pp. 649-654
Author(s):  
Yaw-Jen Chang ◽  
Yeon Pun Chang ◽  
Kai Yuan Cheng
Keyword(s):  
Pcr Amplification ◽  
Channel Length ◽  
Biochemical Reactions ◽  
Chip Design ◽  
Micro Channels ◽  
Loop Channel ◽  
Impact Area ◽  
Tangent Direction ◽  
Polymerase Chain ◽  
The Impact

Biochip is an emerging technology and has evoked great research interests in recent years. In this paper, a novel air-driven loop-type microfluidic biochip was investigated. Differing from conventional micro channels, this chip has a micro loop-channel and 3 sets of driving conduits with valveless design in their intersections so that the microfluid can be driven smoothly in unidirectional circular movements. The driving efficiency reaches the highest if the entry angle of driving conduits is in the tangent direction of the loop-channel. However, the smaller the included angle, the impact area the larger, leading to comparatively serious reflow phenomenon. Furthermore, the microfluid can be controlled to stop almost instantaneously in the loop segment. Therefore, this loop-type biochip is suitable for biochemical reactions under repeated multiple temperature operations such as polymerase chain reaction. A full circular movement completes a cycle of PCR amplification. Besides, this biochip has its merits including simpler chip design, shorter channel length, and flexible controllability for biochemical reactions.

scholarly journals Persistent Detection and Infectious Potential of SARS-CoV-2 Virus in Clinical Specimens from COVID-19 Patients

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1384
Author(s):  
Michael Zapor
Keyword(s):  
Pcr Amplification ◽  
World Health ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Distinctive Features ◽  
Clinical Specimens ◽  
Polymerase Chain ◽  
Health Organization ◽  
Infectious Potential ◽  
Infective Potential

The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) that emerged in December 2019 as the causative agent of Coronavirus 2019 (COVID-19) and was declared a pandemic by the World Health Organization in March 2020 has several distinctive features, including extensive multiorgan involvement with a robust systemic inflammatory response, significant associated morbidity and mortality, and prolonged persistence of viral RNA in the clinical specimens of infected individuals as detected by Reverse Transcription Polymerase Chain Reaction (RT-PCR) amplification. This review begins with an overview of SARS-CoV-2 morphology and replication and summarizes what is known to date about the detection of the virus in nasal, oropharyngeal, and fecal specimens of patients who have recovered from COVID-19, with a focus on the factors thought to contribute to prolonged detection. This review also provides a discussion on the infective potential of this material from asymptomatic, pre-symptomatic, and convalescing individuals, to include a discussion of the relative persistence and infectious potential of virus in clinical specimens recovered from pediatric COVID-19 patients.

RT-PCR analysis of Na+/H+ exchanger mRNAs in rat medullary thick ascending limb

1995 ◽  
Vol 268 (6) ◽  
pp. F1224-F1228 ◽  
Author(s):  
P. Borensztein ◽  
M. Froissart ◽  
K. Laghmani ◽  
M. Bichara ◽  
M. Paillard
Keyword(s):  
Rat Kidney ◽  
Pcr Amplification ◽  
Restriction Enzymes ◽  
Pcr Analysis ◽  
Thick Ascending Limb ◽  
Rt Pcr ◽  
Medullary Thick Ascending Limb ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Nhe Isoforms

The thick ascending limb (TAL) of rat kidney absorbs bicarbonate secondary to proton secretion, but displays both basolateral and luminal Na+/H+ exchange (NHE) activity. Several NHE genes, including NHE-1, NHE-2, NHE-3, and NHE-4, are expressed in the kidney. To identify the NHE isoforms expressed in the rat medullary TAL (MTAL), we used the reverse transcription-polymerase chain reaction (RT-PCR) to detect the mRNAs for NHE in microdissected MTAL. RT-PCR amplification from total RNA was performed between two specific primers for each NHE isoform. In rat kidney homogenate, the four NHE isoform mRNAs were detected, and the identity of the PCR products was demonstrated by the sizes of the fragments, digestion with restriction enzymes, and Southern blot analysis. In microdissected rat MTAL, NHE-3 was strongly expressed and NHE-1 mRNA was also detected, whereas NHE-2 and NHE-4 mRNAs were not detected. Therefore, NHE-3 could be the apical Na+/H+ exchanger, and NHE-1 could be the basolateral isoform in the MTAL.

scholarly journals Symptoms COVID 19 Positive Vapers Compared to COVID 19 Positive Non-vapers

2022 ◽  
Vol 13 ◽  
pp. 215013192110626
Author(s):  
David D. McFadden ◽  
Shari L. Bornstein ◽  
Robert Vassallo ◽  
Bradley R. Salonen ◽  
Mohammed Nadir Bhuiyan ◽  
...  
Keyword(s):  
Nasopharyngeal Swab ◽  
Symptom Experience ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Age And Gender ◽  
Robust Variance ◽  
Polymerase Chain ◽  
And Gender ◽  
Variance Estimates ◽  
The Impact

Objectives: The purpose of the present study was to assess and describe the severity of symptoms reported by Covid-19 positive patients who vaped (smoked e-cigarettes) when compared to those who did not vape or smoke at the time of the diagnosis of Covid-19. Methods: Patients from this study are from a well-characterized patient cohort collected at Mayo Clinic between March 1, 2020 and February 28, 2021; with confirmed COVID-19 diagnosis defined as a positive result on reverse-transcriptase–polymerase-chain-reaction (RT-PCR) assays from nasopharyngeal swab specimens. Among the 1734 eligible patients, 289 patients reported current vaping. The cohort of vapers (N = 289) was age and gender matched to 1445 covid-19 positive patients who did not vape. The data analyzed included: date of birth, gender, ethnicity, race, marital status, as well as lifestyle history such as vaping and smoking and reported covid-19 symptoms experienced. Results: A logistic regression analysis was performed separately for each symptom using generalized estimating equations (GEE) with robust variance estimates in order to account for the 1:5 age, sex, and race matched set study design. Patients who vaped and developed Covid-19 infection were more likely to have chest pain or tightness (16% vs 10%, vapers vs non vapers, P = .005), chills (25% vs 19%, vapers vs non vapers, P = .0016), myalgia (39% vs 32%, vapers vs non vapers, P = .004), headaches (49% vs 41% vapers vs non vapers, P = .026), anosmia/dysgeusia (37% vs 30%, vapers vs non vapers, P = .009), nausea/vomiting/abdominal pain (16% vs 10%, vapers vs non vapers, P = .003), diarrhea (16% vs 10%, vapers vs non vapers, P = .004), and non-severe light-headedness (16% vs 9%, vapers vs non vapers, P < .001). Conclusion: Vapers experience higher frequency of covid-19 related symptoms when compared with age and gender matched non-vapers. Further work should examine the impact vaping has on post-covid symptom experience.

A real-world experience of SARS-CoV-2 infection in a tertiary referral centre of Montréal: Unexpected low prevalence and low mortality

2021 ◽  
pp. e20210022
Author(s):  
Isaac Ruiz ◽  
Geneviève Huard ◽  
Claire Fournier ◽  
Julien Bissonnette ◽  
Hélène Castel ◽  
...  
Keyword(s):  
Real World ◽  
Rt Pcr ◽  
Referral Centre ◽  
Tertiary Referral ◽  
Tertiary Referral Centre ◽  
Health Care Centre ◽  
Liver Unit ◽  
Polymerase Chain ◽  
Low Prevalence ◽  
The Impact

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in patients with chronic liver disease (CLD) and liver transplant (LT) recipients remains a concern. The aim of this study was to report the impact of coronavirus disease 2019 (COVID-19) infection among patients at the tertiary health care centre Centre hospitalier de l’Université de Montréal (CHUM) during the first wave of the SARS-CoV-2 pandemic. Methods: This real-world, retrospective cohort included all patients admitted to our liver unit and/or seen as an outpatient with CLD with or without cirrhosis and/or LT recipients who tested positive to SARS-CoV-2 infection. Cases were considered positive as defined by the detection of SARS-CoV-2 by reverse-transcription polymerase chain reaction (RT-PCR) on nasopharyngeal swabs. Results: Between April 1 and July 31, 2020, 5,637 were admitted to our liver unit and/or seen as outpatient. Among them, 42 were positive for SARS-CoV-2. Twenty-two patients had CLD without cirrhosis while 16 patients had cirrhosis at the time of the infection (13, 2, and 1 with Child–Pugh A, B, and C scores, respectively). Four were LT recipients. Overall, 15/42 patients (35.7%) were hospitalized; among them, 7/42 (16.7%) required respiratory support and 4/42 (9.5%) were transferred to the intensive care unit (ICU). Only 4/42 (9.5%) patients died: 2 with CLD without cirrhosis and 2 with CLD with cirrhosis. Overall survival was 90.5%. Conclusion: This real-world study demonstrates an unexpectedly low prevalence and low mortality in the context of SARS-CoV-2 infection among patients with CLD with or without cirrhosis and LT recipients.

Fatal Hantavirus Pulmonary Syndrome in an Adolescent

PEDIATRICS ◽  
1995 ◽  
Vol 95 (2) ◽  
pp. 276-280 ◽  
Author(s):  
Ali S. Khan ◽  
Thomas G. Ksiazek ◽  
Sherif R. Zaki ◽  
Stuart T. Nichol ◽  
Pierre E. Rollin ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Immunoglobulin M ◽  
Hantavirus Pulmonary Syndrome ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sin Nombre Virus ◽  
Rt Pcr ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Polymerase Chain ◽  
Formalin Fixed

Hantavirus pulmonary syndrome (HPS) is a recently recognized viral zoonosis characterized by a febrile prodrome progressing to severe noncardiogenic pulmonary edema.1-4 This syndrome is caused by at least three newly described hantaviruses: the first, Sin Nombre virus, is the Southwestern hantavirus that caused an outbreak of respiratory failure during the summer of 1993; the second, Black Creek Canal virus, caused a case of HPS in Florida; and the third hantavirus was identified in lung tissue from a patient in Louisiana. Diagnosis is by enzyme-linked immunosorbent assay (ELISA) serology with elevated immunoglobulin M (IgM) titers against heterologous and homologous hantaviral antigens, positive immunohistochemistry on formalin-fixed tissue, or reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of hantaviral nucleotide sequence from frozen tissue.

Regeneration of vgb-transgenic poplar (Populus alba×P. glandulosa) and the primary observation of growth

2006 ◽  
Vol 3 (1) ◽  
pp. 59-64 ◽  
Author(s):  
Zhang Bing-Yu ◽  
Su Xiao-Hua ◽  
Li Yi-Liang ◽  
Huang Qin-Jun ◽  
Zhang Xiang-Hua ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Blot Hybridization ◽  
Pcr Amplification ◽  
First Year ◽  
Populus Alba ◽  
Rt Pcr ◽  
Dot Blot ◽  
Transgenic Lines ◽  
Polymerase Chain ◽  
Morphological Differences

AbstractIncreasing the growth rate is especially important for low-quality wood applications, so this has become an important goal in poplar breeding. The present study describes the transfer of Vitreoscilla haemoglobin (VHb) gene (vgb) driven by constitutive promoters, by Agrobacterium tumefaciens into poplar (Populus alba×P. glandulosa). From about 450 leaf discs used for transformation, 60 Kan-resistant plants were obtained, and 52 proved to be true transgenic plants. The transgenic nature of these plants was confirmed by polymerase chain reaction (PCR) amplification and Southern dot blot hybridization. The expression of vgb gene in transgenic plants was confirmed by reverse transcriptase-PCR (RT-PCR). The performance of the transgenic lines was evaluated during the first year of growth in a greenhouse. These plants showed no significant stable morphological differences from the untransformed plants. Among them, three vgb-transgenic lines exhibited noticeably higher growth rates in terms of height and diameter.

scholarly journals Evaluation of the Sensitivity of Reverse-Transcription Polymerase Chain Reaction to Detect Porcine Reproductive and Respiratory Syndrome Virus on Individual and Pooled Samples from Boars

2007 ◽  
Vol 19 (5) ◽  
pp. 502-509 ◽  
Author(s):  
Albert Rovira ◽  
Travis Clement ◽  
Jane Christopher-Hennings ◽  
Bob Thompson ◽  
Mark Engle ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Biological Samples ◽  
Acute Infection ◽  
Respiratory Syndrome Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
The Impact ◽  
Pooled Samples

Boar studs are continuously monitored for the presence of porcine reproductive and respiratory syndrome virus (PRRSV) by testing different biological samples by reverse-transcription polymerase chain reaction (RT-PCR). In most cases, samples are run in pools, even though the impact of pooling on the sensitivity of RT-PCR is unknown. The objective of this study was to evaluate the feasibility of using PCR on pooled samples through the estimation of the sensitivity of RT-PCR on different biological samples run individually, in pools of 3 and in pools of 5. Twenty-nine boars were inoculated with a low virulent PRRSV isolate. Serum, blood swab, and semen samples were obtained from each boar every 2 to 3 days for 2 weeks. Each sample was tested by RT-PCR undiluted or diluted 1:3 and 1:5 with negative samples. Eleven of the 29 boars did not appear to get infected from the inoculum, as evidenced by no seroconversion 15 days after inoculation. Data from the other 18 boars showed that serum was the best sample to detect PRRSV during acute infection, with the blood swab sample performing almost as well. Semen samples failed to detect PRRSV infection in most of the cases. Pooling samples at pool sizes of 3 and 5 resulted in a decrease in the sensitivity of RT-PCR. Sensitivity was reduced by 6% and 8%, respectively, when serum or blood swab samples were run in pools of 5. The impact of pooling on the sensitivity of PCR was higher in samples taken during the beginning of the viremic period.

scholarly journals Rare spondylodiscitis due to Mycobacterium mucogenicum

2020 ◽  
Vol 11 ◽  
pp. 289
Author(s):  
Massimo Furnari ◽  
Gianluca Scalia ◽  
Giuseppe Emmanuele Umana ◽  
Massimiliano Giuffrida ◽  
Giancarlo Ponzo ◽  
...  
Keyword(s):  
Drug Abuse ◽  
Pcr Amplification ◽  
Rt Pcr ◽  
Ct Guided ◽  
Decompressive Laminectomy ◽  
Ct Guided Biopsy ◽  
Underlying Causes ◽  
Granulomatous Tissue ◽  
Polymerase Chain ◽  
Mycobacterium Mucogenicum

Background: Nontuberculous mycobacteria (NTM) represents an important cause of infection, particularly in immunocompromised patients. Spondylodiscitis is unusual and may be associated with underlying causes such as drug abuse. Timely diagnosis and treatment are critical, as without this, patients will demonstrate progressive neurological deterioration. Here, we present a rare case of Mycobacterium mucogenicum spondylodiscitis in a 36-year-old male, along with a focused literature review. Case Description: A 36-year-old female with previous drug abuse presented with 3-years of progressive thoracolumbar pain. The MRI of the spine revealed paravertebral abscesses from Th10–L1 with vertebral lesions involving Th11–Th12 levels (e.g., vertebral body collapse/deformity and destruction of the posterior vertebral walls). After a needle CT-guided biopsy of the paravertebral tissues, real time-polymerase chain reaction (RT-PCR) amplification documented NTM; the final identification was M. mucogenicum. The patient then underwent a Th11–Th12 decompressive laminectomy, facetectomy, granulomatous tissue debridement, and posterior pedicle screw fusion from Th8–Th10, and L1–L3. Postoperatively, the patient’s pain resolved, and she was left with residual lower extremities dysesthesias; 6-months later, she could walk without assistance. Conclusion: Spondylodiscitis caused by M. mucogenicum is rare, and the medical and surgical treatment is comparable to that for other NTM groups.

Sign in / Sign up

Export Citation Format

Share Document