scholarly journals Molecular analysis of Blastocystis sp. and its subtypes from treated wastewater routinely used for irrigation of vegetable farmlands in Iran

2019 ◽  
Vol 17 (5) ◽  
pp. 837-844 ◽  
Author(s):  
Ehsan Javanmard ◽  
Hanieh Mohammad Rahimi ◽  
Maryam Niyyati ◽  
Hamid Asadzadeh Aghdaei ◽  
Meysam Sharifdini ◽  
...  
Keyword(s):  
Potential Role ◽  
Treated Wastewater ◽  
Rrna Gene ◽  
Pathogenic Microorganisms ◽  
Chain Reaction ◽  
Blastocystis Sp ◽  
Polymerase Chain ◽  
Wastewater Samples ◽  
Pcr Targeting

Abstract Treated wastewater samples were collected, filtered using sterile 47-mm cellulose nitrate membrane and DNA extracted from the filtered materials. The presence of Blastocystis sp. was confirmed via polymerase chain reaction (PCR) targeting the SSU rRNA gene of Blastocystis sp. in 5/12 of samples. Based on the subtype analysis after sequencing, 2, 2 and 1 of ST2, ST6 and ST8 were detected among the isolates, respectively. Furthermore, both ST6s were allele 139, alleles 11 and 138 were identified in ST2 and the only ST8 was allele 95. The phylogenetic tree showed that one of ST2 was clustered together with those ST2 that were already reported from humans and animals. The presence of Blastocystis sp. in treated wastewater can indicate the potential role of this type of water for irrigation in the transmission of pathogenic microorganisms to downstream farmlands.

A study of genes involved in adipocyte differentiation

2015 ◽  
Vol 28 (1-2) ◽  
Author(s):  
Shunming Zhu ◽  
Gong Cheng ◽  
Huolan Zhu ◽  
Gongchang Guan
Keyword(s):  
Potential Role ◽  
Adipocyte Differentiation ◽  
Late Phase ◽  
Redox Status ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Lipid Droplet Formation ◽  
Polymerase Chain ◽  
Microarray Technique

AbstractWith the use of the microarray technique, genes expressed in the late phase of adipocyte differentiation were investigated. These genes play an important role in stimulating adipocyte growth and lipid droplet formation. Therefore, they contribute a great deal to the onset of obesity.With the use of SW872 adipocytes and the microarray technique, genes related to adipocyte differentiation were tested and compared with undifferentiated preadipocytes 14 days after induction. Real-time reverse transcription polymerase chain reaction (RT-PCR) was used for confirmation.More than 21,329 transcriptors were expressed and determined, of which 1326 increased and 687 decreased undifferentiated adipocytes. Among them, 21 were highly expressed by more than 10-fold. With RT-PCR, 12 were confirmed, including apelin, CIDEC, PID1, LYRM1, ADD1, PPARγ2, ANGPTL4, ADIPOQ, ACOX1, FIP1L1, MAP3K2 and PEX14. Furthermore, genes involved in lipid metabolism, signal transduction, DNA replication, redox status and transcription factors were determined as well. Novel genes involved in adipogenesis (e.g., apelin) were detected.A variety of genes were discovered and validated with RT-PCR at the late phase of adipocyte differentiation. This may help us better understand the onset of obesity and the potential role of adipocytes in other organs.

scholarly journals Identification of latent neosporosis in sheep in Tehran, Iran by polymerase chain reaction using primers specific for the Nc-5 gene

2016 ◽  
Vol 83 (1) ◽  
Author(s):  
Mohsen Arbabi ◽  
Amir Abdoli ◽  
Abdolhossein Dalimi ◽  
Majid Pirestani
Keyword(s):  
Polymerase Chain Reaction ◽  
Latent Infection ◽  
Neospora Caninum ◽  
Clinical Signs ◽  
Ovis Aries ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Pcr Targeting

Little is known about latent infection and molecular characterisation of Neospora caninum in sheep (Ovis aries). In this study, 330 sheep samples (180 hearts and 150 brains) were analysed for N. caninum DNA by nested polymerase chain reaction (PCR) targeting the Nc-5 gene. Neospora caninum DNA was detected in 3.9% (13/330) of sheep samples. The parasite’s DNA was detected in 6.7% of heart samples (12/180) and 0.7% (1/150) of brain samples. No clinical signs were recorded from infected or uninfected animals. Sequencing of the genomic DNA revealed 96% – 99% similarity with each other and 95.15% – 100% similarity with N. caninum sequences deposited in GenBank. To our knowledge, this is the first report on the use of PCR to identify latent neosporosis in sheep in Iran. The results of this study have the potential to contribute to our understanding of the role of N. caninum-infected sheep in the epidemiology of neosporosis.

Activated Leukocyte Cell Adhesion Molecule Expression Is Up-Regulated in the Development of Endometrioid Carcinoma

2011 ◽  
Vol 21 (3) ◽  
pp. 523-528 ◽  
Author(s):  
Shumei Liang ◽  
Cuiping Huang ◽  
Shuangzheng Jia ◽  
Bo Wang
Keyword(s):  
Polymerase Chain Reaction ◽  
Cell Adhesion Molecule ◽  
Potential Role ◽  
Myometrial Invasion ◽  
Atypical Hyperplasia ◽  
Chain Reaction ◽  
Normal Endometrium ◽  
Strong Staining ◽  
Polymerase Chain

BackgroundActivated leukocyte cell adhesion molecule (ALCAM/CD166) is a member of the immunoglobulin superfamily that contributes to cell migration. The present study investigated the potential role of ALCAM in the transition from normal endometrium to endometrioid adenocarcinoma (EEC).MethodsTo clarify the role of ALCAM in endometrial tumorigenesis, we determined the levels of protein and messenger RNA expression of ALCAM in human endometrial tissue (proliferative phase [n = 20], secretory phase [n = 20], simple hyperplasia [n = 15], complex hyperplasia [n = 12], atypical hyperplasia [AH, n = 14], EEC [n = 42]) using immunohistochemistry, Western blot, and semiquantitative reverse transcription-polymerase chain reaction, respectively.ResultsExpression of ALCAM detected by immunohistochemistry showed a gradual increase from normal endometrium to atypical hyperplasia in a membranous pattern; in addition, cytoplasmic staining emerged in a few cases of simple hyperplasia and complex hyperplasia, which also showed an increasing tendency. Most cases of EEC showed a homogenously strong staining in all parts of the tumor; other cases showed either membranous or cytoplasmic strong staining; heterogeneous loss of membranous staining was also found in some cases. Similar results of ALCAM expression were detected by reverse transcription-polymerase chain reaction and Western blot. In EEC, ALCAM expression was significantly increased in high-grade tumors and cases with myometrial invasion; however, no correlation was found between ALCAM expression and surgical pathological stages.ConclusionsThe up-regulation of ALCAM expression during endometrial carcinogenesis and the correlations of ALCAM expression with grade and myometrial invasion suggest its potential role as a diagnostic and prognostic biomarker.

scholarly journals Detection of putative virulence genes in Aeromonas isolates from humans and animals

2014 ◽  
Vol 8 (11) ◽  
pp. 1398-1406 ◽  
Author(s):  
Hanifi Körkoca ◽  
Yusuf Alan ◽  
Sedat Bozari ◽  
Mustafa Berktas ◽  
Yaşar Goz
Keyword(s):  
Virulence Genes ◽  
Potential Role ◽  
Virulence Gene ◽  
Potential Importance ◽  
Human Pathogens ◽  
Chain Reaction ◽  
Pcr Screening ◽  
Polymerase Chain ◽  
Animal Isolates

Introduction: Aeromonas are food- and water-borne bacteria that are considered to be zoonotic human pathogens. This study aimed to investigate the presence of genes associated with virulence in human and animal Aeromonas isolates and the potential role of animal isolates with regards to human Aeromonas infections. Methodology: The presence of aerA, hlyA, alt, ast, laf, ascF-G, stx1 and stx2 putative virulence genes in 40 human and animal Aeromonas isolates (16 human and 24 animal isolates) were examined by polymerase chain reaction (PCR). DNA fragments of expected sizes were purified and sequenced. BLAST in the NCBI was used to verify any amplified products. Results: PCR screening showed that hlyA, alt, and laf genes were determined at ratios of 6.25%, 50%, and 6.25%, respectively, in human isolates. The ratios of hlyA, alt, ascF-G, laf, stx2, and stx1 genes in animal isolates were 58.3%, 20.83%, 33.3%, 20.83%, 8.33%, and 4.17%, respectively. Neither aerA nor ast genes were detected in any isolates. Any one of eight putative virulence genes was not detected in seven human and eight animal isolates in the study. Conclusions: The current study is the first to investigate the presence of the virulence gene in gull Aeromonas isolates. The manifestation of the presence of the virulence gene and gene combinations was considerable, especially in fish and gull isolates when compared with clinical human isolates. The current study demonstrates the potential importance of fish and gulls in terms of human Aeromonas infections.

scholarly journals Phytoplasma occurrence in apple trees in the Czech Republic

2011 ◽  
Vol 39 (No. 1) ◽  
pp. 7-12 ◽  
Author(s):  
R. Fialová ◽  
M. Navrátil ◽  
P. Válová
Keyword(s):  
Fragment Length Polymorphism ◽  
Rrna Gene ◽  
Polymorphism Analysis ◽  
Apple Proliferation ◽  
Chain Reaction ◽  
Apple Trees ◽  
The Czech Republic ◽  
High Incidence ◽  
Phytoplasma Infection ◽  
Polymerase Chain

The presence of phytoplasmas in apple trees with proliferation symptoms, rubbery wood symptoms and no symp­toms was determined by using polymerase chain reaction assays with primers amplifying phytoplasma 16S rRNA gene. Phytoplasmas were detected in all trees with proliferation symptoms. Positive tests for phytoplasma in the group of trees with rubbery wood symptoms and of those without symptoms revealed a relatively high incidence of latent phytoplasma infection. Using restriction fragment length polymorphism analysis, phytoplasma of the same identity – apple proliferation phytoplasma (subgroup 16SrX-A) – was recorded in all positively tested trees.  

scholarly journals Chest CT texture-based radiomics analysis in differentiating COVID-19 from other interstitial pneumonia

2021 ◽  
Author(s):  
Damiano Caruso ◽  
Francesco Pucciarelli ◽  
Marta Zerunian ◽  
Balaji Ganeshan ◽  
Domenico De Santis ◽  
...  
Keyword(s):  
Interstitial Pneumonia ◽  
Potential Role ◽  
Texture Features ◽  
Roc Curves ◽  
Chest Ct ◽  
Preliminary Evaluation ◽  
Rt Pcr ◽  
Mean Intensity ◽  
Polymerase Chain

Abstract Purpose To evaluate the potential role of texture-based radiomics analysis in differentiating Coronavirus Disease-19 (COVID-19) pneumonia from pneumonia of other etiology on Chest CT. Materials and methods One hundred and twenty consecutive patients admitted to Emergency Department, from March 8, 2020, to April 25, 2020, with suspicious of COVID-19 that underwent Chest CT, were retrospectively analyzed. All patients presented CT findings indicative for interstitial pneumonia. Sixty patients with positive COVID-19 real-time reverse transcription polymerase chain reaction (RT-PCR) and 60 patients with negative COVID-19 RT-PCR were enrolled. CT texture analysis (CTTA) was manually performed using dedicated software by two radiologists in consensus and textural features on filtered and unfiltered images were extracted as follows: mean intensity, standard deviation (SD), entropy, mean of positive pixels (MPP), skewness, and kurtosis. Nonparametric Mann–Whitney test assessed CTTA ability to differentiate positive from negative COVID-19 patients. Diagnostic criteria were obtained from receiver operating characteristic (ROC) curves. Results Unfiltered CTTA showed lower values of mean intensity, MPP, and kurtosis in COVID-19 positive patients compared to negative patients (p = 0.041, 0.004, and 0.002, respectively). On filtered images, fine and medium texture scales were significant differentiators; fine texture scale being most significant where COVID-19 positive patients had lower SD (p = 0.004) and MPP (p = 0.004) compared to COVID-19 negative patients. A combination of the significant texture features could identify the patients with positive COVID-19 from negative COVID-19 with a sensitivity of 60% and specificity of 80% (p = 0.001). Conclusions Preliminary evaluation suggests potential role of CTTA in distinguishing COVID-19 pneumonia from other interstitial pneumonia on Chest CT.

Diagnostic Accuracy of a Direct Panfungal Polymerase Chain Reaction Assay Performed on Stained Cytology Slides

2021 ◽  
pp. 030098582199156
Author(s):  
Alexandra N. Myers ◽  
Unity Jeffery ◽  
Zachary G. Seyler ◽  
Sara D. Lawhon ◽  
Aline Rodrigues Hoffmann
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Accuracy ◽  
Molecular Techniques ◽  
Fungal Culture ◽  
Chain Reaction ◽  
Identification Of Fungi ◽  
Fungal Dna ◽  
Polymerase Chain ◽  
Panfungal Pcr ◽  
Pcr Targeting

Molecular techniques are increasingly being applied to stained cytology slides for the diagnosis of neoplastic and infectious diseases. Such techniques for the identification of fungi from stained cytology slides have not yet been evaluated. This study aimed to assess the diagnostic accuracy of direct (without nucleic acid isolation) panfungal polymerase chain reaction (PCR) followed by sequencing for identification of fungi and oomycetes on stained cytology slides from dogs, cats, horses, and other species. Thirty-six cases were identified with cytologically identifiable fungi/oomycetes and concurrent identification via fungal culture or immunoassay. Twenty-nine controls were identified with no cytologically or histologically visible organisms and a concurrent negative fungal culture. Direct PCR targeting the internal transcribed spacer region followed by sequencing was performed on one cytology slide from each case and control, and the sensitivity and specificity of the assay were calculated. The sensitivity of the panfungal PCR assay performed on stained cytology slides was 67% overall, 73% excluding cases with oomycetes, and 86% when considering only slides with abundant fungi. The specificity was 62%, which was attributed to amplification of fungal DNA from control slides with no visible fungus and negative culture results. Direct panfungal PCR is capable of providing genus- or species-level identification of fungi from stained cytology slides. Given the potential of panfungal PCR to amplify contaminant fungal DNA, this assay should be performed on slides with visible fungi and interpreted in conjunction with morphologic assessment by a clinical pathologist.

scholarly journals Decreased Expression of GPER1 Gene and Protein in Goiter

2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Raquel Weber ◽  
Ana Paula Santin Bertoni ◽  
Laura Walter Bessestil ◽  
Ilma Simoni Brum ◽  
Tania Weber Furlanetto
Keyword(s):  
Estrogen Receptor ◽  
Quantitative Polymerase Chain Reaction ◽  
Thyroid Tissue ◽  
Chain Reaction ◽  
Normal Thyroid ◽  
Protein Levels ◽  
Protein Expressions ◽  
Polymerase Chain ◽  
G Protein Coupled

Goiter is more common in women, suggesting that estrogen could be involved in its physiopathology. The presence of classical estrogen receptors (ERαand ERβ) has been described in thyroid tissue, suggesting a direct effect of estrogen on the gland. A nonclassic estrogen receptor, the G-protein-coupled estrogen receptor (GPER1), has been described recently in several tissues. However, in goiter, the presence of this receptor has not been studied yet. We investigated GPER1 gene and protein expressions in normal thyroid and goiter using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot, respectively. In normal thyroid (n=16) and goiter (n=19), GPER1 gene was expressed in all samples, while GPER1 protein was expressed in all samples of normal thyroid (n=15) but in only 72% of goiter samples (n=13). When comparing GPER1 gene and protein levels in both conditions, gene expression and protein levels were higher in normal thyroid than in goiter, suggesting a role of this receptor in this condition. Further studies are needed to elucidate the role of GPER1 in normal thyroid and goiter.

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