Cholestasis Alters miR-34c-5p Expression in the Testes of Male Wistar Rats

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Amir Hossein Hasani Fard ◽  
Hanieh Jalali ◽  
Homa Mohseni Kouchesfehani
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Target Genes ◽  
Serum Levels ◽  
Testicular Tissue ◽  
Adult Rats ◽  
Altered Expression ◽  
Rat Models ◽  
Duct Ligation ◽  
Male Wistar Rats

Background: Cholestasis is a pathophysiological condition, significantly reducing spermatozoa production. MiR-34c is highly expressed in adult male testicles and controls different stages of spermatogenesis. Objectives: Here, we aimed to investigate miR-34c expression in the testes of rat models of cholestasis. The expressions of THY-1, FGF-2, and CASP-3 genes, that are targeted by mirR-34c were also investigated. Methods: Cholestasis was induced in six adult rats via bile duct ligation. Four weeks after cholestasis induction, sera and testicular tissues were collected for further examinations. The levels of liver enzymes were measured using the ELISA. The structure of the testes was evaluated by histological examination. Total RNA was extracted from testes using a special kit and converted to cDNA. The expressions of miR-34c-5p, THY-1, FGF-2, and CASP-3 genes were determined by Real-Time PCR. Results: The serum levels of ALP, AST, and ALT were significantly elevated in the rat models of cholestasis (P < 0.001). Real-Time PCR revealed that the expressions of miR-34c-5p, THY-1, and FGF-2 genes decreased while CASP-3 gene was upregulated in the testes of cholestatic animals (all differences were significant at P < 0.05). Conclusions: Our study indicated that cholestasis was associated with reduced expression of miR-34c and altered expression of its target genes in the testis. Our results highlight the potential effects of cholestasis, a hepatobiliary disease, on testicular tissue function and male fertility.

Diagnosis and Causative Species of Visceral Leishmaniasis in Southwest Saudi Arabia

2021 ◽  
Author(s):  
Aymen Abdelhaleem ◽  
Nabil Dhayhi ◽  
Mohamed Salih Mahfouz ◽  
Ommer Daffalla ◽  
Mansour Mubarki ◽  
...  
Keyword(s):  
Saudi Arabia ◽  
Visceral Leishmaniasis ◽  
Real Time ◽  
Real Time Pcr ◽  
Leishmania Donovani ◽  
Target Genes ◽  
Sample Volume ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Robust Evidence

Visceral leishmaniasis (VL) is the most severe clinical form of the disease and has been reported in the Jazan region of southwest Saudi Arabia. This study aimed to diagnose VL by real-time polymerase chain reaction (PCR) and the direct agglutination test (DAT) and to identify the causative Leishmania species. A total of 80 participants, including 30 suspected VL patients, 30 healthy endemic control individuals, and 20 malaria disease controls, were enrolled in this study. Blood samples were collected and tested for Leishmania DNA by real-time PCR and for antibody by the DAT. Sequencing of some amplified PCR products was used to identify the causative Leishmania species. The diagnosis of VL was successfully achieved by both real-time PCR and by DAT with 100% sensitivity. Leishmania donovani and Leishmania infantum species were detected by sequencing both by the kDNA and ITS1 target genes, followed a BLASTn search. The detection of VL antibody by the DAT followed by the confirmatory detection of Leishmania DNA in patient blood by PCR could promote the adoption of the much less invasive and more sensitive methods for the routine diagnosis of VL. Further study with high sample volume to evaluate the PCR and the DAT are needed, to generate more robust evidence. Based on the sequencing results, emerging studies on VL should focus on the causative Leishmania species, reservoirs, and vectors that are important in the study area.

Validation of reference genes for quantitative real-time PCR in valproic acid rat models of autism

2016 ◽  
Vol 43 (8) ◽  
pp. 837-847 ◽  
Author(s):  
Jinlong Zhou ◽  
Xiaozheng Zhang ◽  
Junrong Ren ◽  
Ping Wang ◽  
Junfeng Zhang ◽  
...  
Keyword(s):  
Valproic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Reference Genes ◽  
Quantitative Real Time Pcr ◽  
Rat Models

scholarly journals Serum and Hepatic Autofluorescence as a Real-Time Diagnostic Tool for Early Cholestasis Assessment

2018 ◽  
Vol 19 (9) ◽  
pp. 2634 ◽  
Author(s):  
Anna Croce ◽  
Giovanni Bottiroli ◽  
Laura Di Pasqua ◽  
Clarissa Berardo ◽  
Veronica Siciliano ◽  
...  
Keyword(s):  
Bile Duct ◽  
Real Time ◽  
Diagnostic Tool ◽  
Bile Duct Ligation ◽  
Fiber Optic Probe ◽  
Duct Ligation ◽  
Male Wistar Rats ◽  
Optic Probe ◽  
Oxidative Products ◽  
Liver Changes

While it is well established that various factors can impair the production and flow of bile and lead to cholestatic disease in hepatic and extrahepatic sites, an enhanced assessment of the biomarkers of the underlying pathophysiological mechanisms is still needed to improve early diagnosis and therapeutic strategies. Hence, we investigated fluorescing endogenous biomolecules as possible intrinsic biomarkers of molecular and cellular changes in cholestasis. Spectroscopic autofluorescence (AF) analysis was performed using a fiber optic probe (366 nm excitation), under living conditions and in serum, on the livers of male Wistar rats submitted to bile duct ligation (BDL, 24, 48, and 72 h). Biomarkers of liver injury were assayed biochemically. In the serum, AF analysis distinctly detected increased bilirubin at 24 h BDL. A continuous, significant increase in red-fluorescing porphyrin derivatives indicated the subversion of heme metabolism, consistent with an almost twofold increase in the serum iron at 72 h BDL. In the liver, changes in the AF of NAD(P)H and flavins, as well as lipopigments, indicated the impairment of mitochondrial functionality, oxidative stress, and the accumulation of oxidative products. A serum/hepatic AF profile can be thus proposed as a supportive diagnostic tool for the in situ, real-time study of bio-metabolic alterations in bile duct ligation (BDL) in experimental hepatology, with the potential to eventually translate to clinical diagnosis.

scholarly journals Common Deletion (CD) in mitochondrial DNA of irradiated rat heart

2014 ◽  
Vol 86 (2) ◽  
pp. 685-694 ◽  
Author(s):  
RAQUEL G. SIQUEIRA ◽  
DAYSE A. DA SILVA ◽  
LUIZ D.B. DE MELO ◽  
ELIZEU F. DE CARVALHO ◽  
SAMARA C. FERREIRA-MACHADO ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Ionizing Radiation ◽  
Real Time ◽  
Real Time Pcr ◽  
Rat Heart ◽  
Wistar Rats ◽  
Post Irradiation ◽  
Male Wistar Rats ◽  
The Common ◽  
Common Deletion

The purpose of this study was to map the common deletion (CD) area in mtDNA and investigate the levels of this deletion in irradiated heart. The assays were developed in male Wistar rats that were irradiated with three different single doses (5, 10 or 15 Gy) delivered directly to the heart and the analyses were performed at various times post-irradiation (3, 15 or 120 days). The CDs area were sequenced and the CD quantified by real-time PCR. Our study demonstrated that the CD levels progressively decreased from the 3rduntil the 15th day after irradiation, and then increased thereafter. Additionally, it was observed that the levels of CD are modulated differently according to the different categories of doses (moderate and high). This study demonstrated an immediate response to ionizing radiation, measured by the presence of mutations in the CD area and a decrease in the CD levels.

scholarly journals Quantitative Detection of Listeria monocytogenes and Listeria innocua by Real-Time PCR: Assessment of hly, iap, and lin02483 Targets and AmpliFluor Technology

2004 ◽  
Vol 70 (3) ◽  
pp. 1366-1377 ◽  
Author(s):  
David Rodr�guez-L�zaro ◽  
Marta Hern�ndez ◽  
Mariela Scortti ◽  
Teresa Esteve ◽  
Jos� A. V�zquez-Boland ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Real Time ◽  
Real Time Pcr ◽  
Target Genes ◽  
Dynamic Range ◽  
Significant Proportion ◽  
Quantitative Detection ◽  
Target Sequence ◽  
Food Borne ◽  
Highly Sensitive

ABSTRACT We developed and assessed real-time PCR (RTi-PCR) assays for the detection and quantification of the food-borne pathogen Listeria monocytogenes and the closely related nonpathogenic species L. innocua. The target genes were hly and iap for L. monocytogenes and lin02483 for L. innocua. The assays were 100% specific, as determined with 100 Listeria strains and 45 non-Listeria strains, and highly sensitive, with detection limits of one target molecule in 11 to 56% of the reactions with purified DNA and 3 CFU in 56 to 89% of the reactions with bacterial suspensions. Quantification was possible over a 5-log dynamic range, with a limit of 15 target molecules and R 2 values of >0.996. There was an excellent correspondence between the predicted and the actual numbers of CFU in the samples (deviations of <23%). The hly-based assay accurately quantified L. monocytogenes in all of the samples tested. The iap-based assay, in contrast, was unsuitable for quantification purposes, underestimating the bacterial counts by 3 to 4 log units in a significant proportion of the samples due to serovar-related target sequence variability. The combination of the two assays enabled us to classify L. monocytogenes isolates into one of the two major phylogenetic divisions of the species, I and II. We also assessed the new AmpliFluor technology for the quantitative detection of L. monocytogenes by RTi-PCR. The performance of this system was similar to that of the TaqMan system, although the former system was slightly less sensitive (detection limit of 15 molecules in 45% of the reactions) and had a higher quantification limit (60 molecules).

Evaluation of the effects of pulsed wave LLLT on tibial diaphysis in two rat models of experimental osteoporosis, as examined by stereological and real-time PCR gene expression analyses

2016 ◽  
Vol 31 (4) ◽  
pp. 721-732 ◽  
Author(s):  
Zhaleh Mohsenifar ◽  
Mohammadjavad Fridoni ◽  
Mahdi Ghatrehsamani ◽  
Mohammad-amin Abdollahifar ◽  
Hojjatallah Abbaszadeh ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Rat Models ◽  
Tibial Diaphysis ◽  
Experimental Osteoporosis ◽  
Gene Expression Analyses

scholarly journals Real-time PCR to assess the Leishmania load in Lutzomyia longipalpis sand flies: Screening of target genes and assessment of quantitative methods

2011 ◽  
Vol 129 (3) ◽  
pp. 234-239 ◽  
Author(s):  
Diana R. Bezerra-Vasconcelos ◽  
Luciana M. Melo ◽  
Érica S. Albuquerque ◽  
Maria C.S. Luciano ◽  
Claudia M.L. Bevilaqua
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Methods ◽  
Target Genes ◽  
Lutzomyia Longipalpis ◽  
Sand Flies

scholarly journals AN INVESTIGATION INTO THE EFFECT OF H9N2 INFLUENZA VIRUS ON APOPTOSIS OF TESTICULAR CELLS IN CHICKEN EMBRYO USING SYBR-GREEN REAL-TIME PCR

2020 ◽  
Vol 17 (34) ◽  
pp. 1003-1011
Author(s):  
Roshanak Bahrami NAZARABADI ◽  
Mohammad Javad MEHRABANPOUR ◽  
Mohammad Amin EDALATMANESH ◽  
Mehrdad SHARIATI
Keyword(s):  
Gene Expression ◽  
Cell Death ◽  
Influenza Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Testicular Tissue ◽  
H9n2 Influenza Virus ◽  
Testicular Cells ◽  
Molecular Studies ◽  
Apoptotic Genes

Influenza viruses can induce cell death in their host through apoptosis or necrosis.The H9N2 is a subtype of the Avian Influenza Virus (AIV) that can cause severe damage to reproductive organs of laying hens. The present study aimed to investigate the effect of the H9N2 influenza virus on apoptosis of testicular cells in chicken embryos. To this end, A/Chicken/Tehran/ZMT-101/99(H9N2) was inoculated to 210 embryonated fifteen-day-old eggs laid by SPF hens. Then, according to the experiment design, live embryos were dissected on the 19th and 21st days of fetal life and on the 25th day after birth for pathological and molecular studies and evaluation of gene expression in testicular tissue. Dissected tissues were stained with hematoxylin and eosin (HE) and studied under an optical microscope at 400x magnification. For molecular studies, viral RNA was extracted from testicular tissues, replicated by RT-PCR, and finally evaluated H9N2. Then, the genes expression involved in the testicular tissue cells apoptosis was evaluated through real-time PCR. Pathological studies indicated that H9N2 caused lesions in testicular tissue orchitis, seminiferous, and nephritis of the host. Molecular studies also showed that H9N2 replication in the host body increases BAX and Caspase 3 expression and reduces the expression of Bcl-2 and Bcl-XL. These changes in gene expression increased apoptosis and induced cell death in the host. In summary, the study findings suggested that H9N2 can increase the expression of pro-apoptotic genes and reduce the expression of anti-apoptotic genes, resulting in severe destruction of testicular tissue caused by cell apoptosis.

scholarly journals The Role of Exercise in Reducing Hyperlipidemia-Induced Neuronal Damage in Apolipoprotein E-Deficient Mice

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Yumeng Bai ◽  
Yali Feng ◽  
Bo Jiang ◽  
Yan Yang ◽  
Zuowei Pei ◽  
...  
Keyword(s):  
Exercise Training ◽  
Apolipoprotein E ◽  
Real Time ◽  
Real Time Pcr ◽  
High Fat Diet ◽  
Neuronal Injury ◽  
Serum Levels ◽  
Normal Diet ◽  
High Fat ◽  
Expression Levels

Hyperlipidemia causes nervous system-related diseases. Exercise training has developed into an established evidence-based treatment strategy that is beneficial for neuronal injury. This study investigated the effect of exercise on hyperlipidemia-induced neuronal injury in apolipoprotein E-deficient (ApoE-/-) mice. Male ApoE-/- mice (age: 8 weeks) were randomly divided into four groups as follows: mice fed a normal diet (ND), normal diet+swimming training (ND+S), high-fat diet (HD), and high-fat diet+swimming (HD+S). Exercise training consisted of swimming for 40 min/day, 5 days/week for 12 weeks. After 12 weeks, we measured serum levels of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-c). We also evaluated glial fibrillary acidic protein (GFAP) expression levels using immunohistochemistry, real-time PCR, and immunoblotting. In addition, NLR family pyrin domain-containing 3 (NLRP3), interleukin- (IL-) 18, caspase-1, Bax, Bcl-2, and phosphorylated extracellular signal-regulated kinase (p-ERK) expression levels were measured using immunoblotting. Serum levels of TG, TC, and LDL-c were lower in ApoE-/- HD+S mice than in ApoE-/- HD mice. Immunohistochemistry, real-time PCR, and immunoblotting showed increased levels of GFAP in the ApoE-/- HD group. Immunoblotting revealed increased levels of NLRP3, IL-18, caspase-1, Bax, Bcl-2, and p-ERK in the ApoE-/- HD group; however, they were significantly suppressed in the ApoE-/- HD+S group. Therefore, exercise has protective effects against neuronal injury caused by hyperlipidemia.

scholarly journals D-limonene in diabetic rats

2021 ◽  
Vol 10 (3) ◽  
pp. e24-e24
Author(s):  
Shahrokh Bagheri ◽  
Mostafa Moradi Sarabi ◽  
Mohammadreza Gholami ◽  
Vahideh Assadollahi ◽  
Reza Mohammadrezaei Khorramabadi ◽  
...  
Keyword(s):  
Real Time ◽  
Kidney Tissue ◽  
Serum Levels ◽  
Diabetic Control ◽  
Serum Glucose ◽  
Diabetic Rats ◽  
Tissue Homogenate ◽  
Control Group ◽  
Mrna Levels ◽  
Male Wistar Rats

Introduction: Diabetes mellitus (DM) is a multi-factorial condition associated with oxidative stress. Limonene, as a plant-derived antioxidant, can be used for treating DM. Objectives: An investigation on antioxidant effects in diabetic rats exposed to D-limonene. Materials and Methods: Sixty male Wistar rats were categorized into six groups as follows: control (healthy rats), diabetic control (untreated diabetic rats), sham glibenclamide, diabetic glibenclamide, sham limonene, and finally diabetic limonene. Alloxan (100 mg/dL) was infused intraperitoneally to induce type 1 diabetes in rats. Rats in certain groups were given limonene (100 mg/dL) and glibenclamide (10 mg/dL) orally for 8 weeks. Subsequently, animals were killed, and their kidneys were removed. Serum levels of biochemical factors (including serum creatinine, urea, and glucose) were determined, and factors such as nitric oxide (NO), malondialdehyde (MDA), glutathione (GSH), and myeloperoxidase (MPO) were measured in kidney tissue homogenate. The gene expression and enzymatic activity of glutathione peroxidase (GPx), catalase (CAT), and superoxide dismutase (SOD) in the kidney were measured by real-time polymerase chain reaction (real-time PCR) and spectrophotometry, respectively. Results: Limonene treatment significantly decreased serum glucose, creatinine, and urea. Additionally, MDA, MPO, and NO significantly decreased while GSH increased after treatment with limonene. Real-time RT-PCR showed significant elevation (P<0.05) in mRNA levels of GPx, CAT, and SOD in the limonene-treated compared with the diabetic control group. Conclusion: Our results demonstrated that limonene as an herbal antioxidant had better effects on antioxidant markers compared to glibenclamide in rat models of diabetes.

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