Ribonucleic Acid Expression Research Articles

Apoptotic Molecules In Chronic Leukemia

Background: Chronic myeloid leukemia (CML) is characterized by the spread of malignant cells that exhibit resistance to caspase-mediated cell death (apoptosis) and this mechanism is proposed to play an important role in myeloid cell growth. However, the extent to which caspase-mediated cell death plays a crucial role in the regulation of myelopoiesis remains controversial.Objectives & Proceedure: The objectives of this study were to examine whether or not caspase-mediated cell death-related proteins take part in the development of CML and to also to detect the relationship between Fas, p53 and caspase-mediated cell death protease activating factor (Apaf-1) in 5 patients with CML using the real-time quantitative polymerase chain reaction. We demonstratedthatp53 and Apaf-1 messenger ribonucleic acid (mRNA) expression was moderately elevated (up to 5 fold, p<0.05) in 4 out of 5 CML patients. One patient with a p53 point mutation, exhibited a far greater elevation of p53 mRNA expression throughout their blast crisis, but in contrast, displayed a significant reduction in levels of Apaf-1 mRNA and Fas mRNA.Conclusion: Our results show in-vivo linkages between Fas, p53 and Apaf-1 transcription parameters suggesting that the key genes involved in the caspase-mediated cell death might contribute to CML disease development.Bangladesh Journal of Medical Science Vol.15(4) 2016 p.651-654

Targeted myocardial gene expression in failing hearts by RNA sequencing

BackgroundMyocardial recovery with left ventricular assist device (LVAD) therapy is highly variable and difficult to predict. Next generation ribonucleic acid (RNA) sequencing is an innovative, rapid, and quantitative approach to gene expression profiling in small amounts of tissue. Our primary goal was to identify baseline transcriptional profiles in non-ischemic cardiomyopathies that predict myocardial recovery in response to LVAD therapy. We also sought to verify transcriptional differences between failing and non-failing human hearts.MethodsRNA was isolated from failing (n = 16) and non-failing (n = 8) human hearts. RNA from each patient was reverse transcribed and quantitatively sequenced on the personal genome machine (PGM) sequencer (Ion torrent) for 95 heart failure candidate genes. Coverage analysis as well as mapping the reads and alignment was done using the Ion Torrent Browser Suite™. Differential expression analyses were conducted by empirical analysis of digital gene expression data in R (edgeR) to identify differential expressed genes between failing and non-failing groups, and between responder and non-responder groups respectively. Targeted cardiac gene messenger RNA (mRNA) expression was analyzed in proportion to the total number of reads. Gene expression profiles from the PGM sequencer were validated by performing RNA sequencing (RNAseq) with the Illumina Hiseq2500 sequencing system.ResultsThe failing sample population was 75% male with an average age of 50 and a left ventricular ejection fraction (LVEF) of 16%. Myosin light chain kinase (MYLK) and interleukin (IL)-6 genes expression were significantly higher in LVAD responders compared to non-responders. Thirty-six cardiac genes were expressed differentially between failing and non-failing hearts (23 decreased, 13 elevated). MYLK, Beta-1 adrenergic receptor (ADRB1) and myosin heavy chain (MYH)-6 expression were among those significantly decreased in failing hearts compared to non-failing hearts. Natriuretic peptide B (NPPB) and IL-6 were significantly elevated. Targeted gene expression profiles obtained from the Ion torrent PGM sequencer were consistent with those obtained from Illumina HiSeq2500 sequencing system.ConclusionsHeart failure is associated with a network of transcriptional changes involving contractile proteins, metabolism, adrenergic receptors, protein phosphorylation, and signaling factors. Myocardial MYLK and IL-6 expression are positively correlated with ejection fraction (EF) response to LVAD placement. Targeted RNA sequencing of myocardial gene expression can be utilized to predict responders to LVAD therapy and to better characterize transcriptional changes in human heart failure.

Influences of Pulsed Electric Fields on the Gene Expression of Pathogenic Bacteria

SUMMARYIn the biological and medical fields, sterilization using pulsed electric fields or discharges and electroporation have been studied actively in recent years. It is clear that a pulsed electric field has some sort of effect on an organism. At present, however, it is not sufficiently clear what effect occurs and how the mechanism of a pulsed electric field acts on the organism. The effects of pulsed electric fields on ribonucleic acid (RNA) expression in biological cells were studied with the objective of elucidating the bioelectric mechanism. A pulsed electric field was applied to Vibrio parahaemolyticus, a pathogenic bacteria, and the expression of RNAs, specifically thermostable direct hemolysin (TDH), VP1680, and VP1699, which represent pathogenic agents, were analyzed using the polymerase chain reaction (PCR) method. The level of expression of VP1680 increased when a pulsed electric field was applied, and the effect was at a peak when the electric field strength was 14 kV/cm. The electric field strength barely affected the expression of TDH and VP1699. It was found that the effects of the electric field were different among the RNAs. The expression level varied with time when expression was analyzed. These results suggest the possibility of selective control of gene expression by using pulsed electric fields and possible new applications such as the development of functional foods.

Analysis of Altered Micro RNA Expression Profiles in Focal Cortical Dysplasia IIB.

Focal cortical dysplasia type IIB is a commonly encountered subtype of developmental malformation of the cerebral cortex and is often associated with pharmacoresistant epilepsy. In this study, to investigate the molecular etiology of focal cortical dysplasia type IIB, the authors performed micro ribonucleic acid (RNA) microarray on surgical specimens from 5 children (2 female and 3 male, mean age was 73.4 months, range 50-112 months) diagnosed of focal cortical dysplasia type IIB and matched normal tissue adjacent to the lesion. In all, 24 micro RNAs were differentially expressed in focal cortical dysplasia type IIB, and the microarray results were validated using quantitative real-time polymerase chain reaction (PCR). Then the putative target genes of the differentially expressed micro RNAs were identified by bioinformatics analysis. Moreover, biological significance of the target genes was evaluated by investigating the pathways in which the genes were enriched, and the Hippo signaling pathway was proposed to be highly related with the pathogenesis of focal cortical dysplasia type IIB.

Biomarkers and genetics of brain injury risk in diabetic ketoacidosis: A pilot study.

Diabetic ketoacidosis (DKA) is the primary cause of death for children with diabetes, especially when complicated by cerebral edema. Central nervous system (CNS) involvement is common, however the mechanism of, and predictors of CNS dysfunction/injury are largely unknown. In this observational pilot study, blood was collected from pediatric DKA patients at three time points (consent, 12 hr and 24 hr after beginning treatment), to test genetic markers, ribonucleic acid expression and plasma biomarkers reflecting inflammation (tumor necrosis factor-alpha [TNF-α], interleukin-6 [IL-6]) and cerebral dysfunction and/or possible injury (S100β, glial fibrillary acidic protein [GFAP]). Thirty patients were enrolled in the study. The average age was 11.3 yr, 73% were new onset diabetes and 53% were female. Forty percent exhibited abnormal mentation (Glasgow Coma Scale <15), consistent with CNS dysfunction. IL-6 and TNF-α were elevated in plasma, suggesting systemic inflammation. GFAP was measurable in 45% of patients and correlated positively with GCS. Only two patients had detectable levels of S100β. In conclusion, children with DKA often present with evidence of acute neurologic dysfunction or injury. We have demonstrated the feasibility of exploring genetic and biochemical markers of potential importance in the pathophysiology of CNS dysfunction and/or possible injury in DKA. We have identified IL-6, TNF-α and GFAP as potentially important markers for further exploration. A larger, follow-up study will help to better understand the extent and type of CNS injury in DKA as well as the mechanism underlying this dysfunction/injury.

Nanowire Analysis of Cancer-Testis Antigens as Biomarkers of Aggressive Prostate Cancer

To demonstrate the ability of the nCounter Analysis System, a nanowire technology, to sensitively and accurately detect cancer-testis antigens (CTAs) in men with prostate cancer and correlate them with disease parameters. The clinical implementation of novel biomarkers is necessary to provide for individual disease treatment planning for men with prostate cancer. The CTAs, as cancer-associated biomarkers that may correlate with aggressive disease, have the potential to play an important role. Formalin-fixed, paraffin embedded samples were used from men undergoing radical prostatectomy for prostate cancer. The expression of CTAs along with control genes was measured from formalin-fixed, paraffin-embedded prostate cancer tissues using real-time polymerase chain reaction and the nCounter assay. Using a nanowire-based assay, ribonucleic acid (RNA) expression levels of the CTAs CSAG2 and NOL4 were found to be significantly higher in men with Gleason score (GS) 8-10 disease than those with GS≤4+3 disease. On the contrary, the RNA expression level of PAGE4 was lower in men with GS 8-10 disease than those with GS≤6 group. This study demonstrates that CTAs can be detected with a nanostring assay that is translatable and that a set of CTAs correlates with the clinical characteristics of the disease. CTAs represent unique, cancer-associated biomarkers with potential utility in the clinic. The nCounter nanowire technology provides an opportunity to evaluate this panel of CTAs associated with aggressive prostate cancer in a multi-institutional fashion.

Long noncoding ribonucleic acids maternally expressed gene 3 inhibits lung cancer tumor progression through downregulation of MYC.

Long noncoding ribonucleic acids (RNAs) nowadays emerge as important biomarkers or potential therapeutic targets discussed in human cancers. Among them, maternally expressed gene 3 (MEG3) is known to be decreased in a variety of malignancies. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to detect the expression of MEG3 in forty pairs of lung cancer (LC) tissues. Overexpression of MEG3 was carried out, and we determined its effect on cell proliferation, apoptosis, and migration evaluated by cell counting kit-8, flow cytometric, and transwell analysis. Messenger RNA and protein expression of MYC were determined by qRT-PCR and western blot, respectively. The expression of MEG3 was downregulated in LC tissues. Forced expression of MEG3 led to reduced abilities of cell proliferation and elevated apoptosis rate. It also slightly inhibited cell migration capacity in vitro. In addition, MYC was inhibited by MEG3 overexpression at both transcriptional and translational levels. Our findings revealed MEG3 could regulate LC progression and serve as an important target for LC treatment.

Effect of small intestinal submucosa sponges on the attachment and proliferation behavior of Schwann cells

The aim of the study was to investigate the effects of small intestinal submucosa (SIS) sponges, a natural biodegradable polymer scaffold, on the adhesion behavior and the proliferation of Schwann cells (SCs). SIS sponges were prepared by varying the contents of SIS powders (1%, 2%, and 3% SIS concentration) and SCs were seeded on the sponges and incubated to investigate the adhesion and proliferation of SCs. A series of analytical process was conducted to observe the water absorption rate of sponges and confirmed the proliferation and morphology of SCs by scanning electron microscopy (SEM), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, reverse transcription polymerase chain reaction (RT-PCR), and histological evaluation. The best adhesion behavior and the proliferation of SCs were observed on the sponge with 2% powder content. Also, analysis of messenger ribonucleic acid (mRNA) expression using the neuron-specific enolase (NSE) and neurofilament protein (NF) nerve markers showed that SC gene expression was excellent in the 2% SIS sponge. In vivo hematoxylin and eosin (H&E) staining and immunofluorescence results showed that cells and tissue formed well in the pores of sponges with 2% and 3% SIS powder contents, compared with formation in the 1% SIS sponge. The SIS sponges provided an unsuitable habitat for the adhesion and proliferation of cells in that the pore size was small. In this study, the adhesion behavior and the proliferation could be changed depending on the pore size and SIS powder content in the sponges. Thus, an appropriate powder content of SIS should be used to prepare SIS sponge scaffolds.

Involvement of anandamide transporter in calcitonin gene-related peptide expression stimulated by nitroglycerin and influence of ALDH2 Glu504Lys polymorphism.

The aim of this study was to investigate whether N-arachidonic acid ethanolamine (anandamide, AEA) transporter contributed to calcitonin gene-related peptide (CGRP) expression mediated by nitroglycerin (GTN) in peripheral blood mononuclear cells (PBMCs) of healthy volunteers and its association with the mitochondrial aldehyde dehydrogenase-2 (ALDH2) Glu504Lys (ALDH2*2) polymorphism. In 10 ALDH2*2-genotyped Chinese volunteers, we assessed the activity of AEA transporter and expression of CGRP messenger ribonucleic acid (mRNA) in cultured PBMCs treated with different concentration of GTN with or without pretreatment with AM404 (the AEA transporter blocker). In this study, the activity of AEA transporter and expression of CGRP mRNA elevated with the increase in the concentration of GTN. Pretreatment of the cells with AM404 (1 μM) 2 hours before GTN reduced the GTN-induced increase in both AEA transporter activity and CGRP mRNA expression significantly, and cells with the ALDH2*1/*1 homozygote genotype showed significantly higher activity of AEA transporter and CGRP mRNA expression than carriers of the ALDH2*2 allele. Therefore, we strongly suggested that GTN can stimulate CGRP expression by elevating the AEA transporter activity, which is affected by ALDH2 Glu504Lys polymorphism.

The effect of heat stress on GHR, IGF-I, ANT, UCP and COXIII mRNA expression in the liver and muscle of high and low feed efficiency female quail

1. A study was conducted to test the hypothesis that feed efficiency (FE) correlated with the expression of genes from the somatotropic axis and mitochondrial genes involved in energy production, and that the environment to which the birds are exposed influenced the expression of such genes.2. Quails were divided into High-FE and low-FE groups and maintained in a comfortable or heat stress (HS) (38°C for 24 h) environment to evaluate changes in insulin-like growth factor I (IGF-I), growth hormone receptor (GHR), adenine nucleotide translocator (ANT), uncoupling protein (UCP) and cytochrome oxidase subunit III (COX III) messenger ribonucleic acid (mRNA) expression in liver and muscle tissues.3. High-FE quails (0.28 g/g) presented a higher final body weight, greater weight gain and a better feed conversion ratio than low-FE birds (0.18 g/g). High-FE birds showed greater IGF-I mRNA expression in the liver and muscle and greater GHR mRNA expression in the muscle.4. Environmental effects only affected GHR expression in the liver, with quails under comfortable conditions exhibiting greater GHR expression than quails subjected to HS.5. There was a significant interaction between FE and environmental temperature on ANT mRNA expression in the liver. The greatest ANT mRNA expression was observed for high FE-birds that remained under comfortable conditions.6. In the liver, UCP mRNA expression did not differ among the quails and was not affected by environment or efficiency. However, comparisons of the low- and high-FE birds revealed higher levels of UCP mRNA in the muscle of low-FE birds.7. COX III mRNA expression in the liver was dependent on environmental temperature and FE. Higher COX III mRNA expression was observed in animals that remained under comfortable conditions, and high-FE birds exhibited higher expression levels compared to low-FE birds.8. These results suggest a correlation between IGF-I, GHR, ANT, UCP and COX III gene expression and FE and that environmental temperature could affect the expression of some of these genes.

Hypoxia-Inducible Factor and Target Gene Expression Are Decreased in Patients With Sepsis

Background Hypoxia-inducible factor-1 (HIF-1) is a molecular key player in response to hypoxemic/inflammatory conditions prevailing in sepsis. In a prospective observational study, we tested the hypotheses that sepsis affects HIF-1α messenger ribonucleic acid (mRNA) expression (primary hypothesis) and also (secondary hypotheses) the expression of the HIF-1α target genes adrenomedullin and β2-integrins. Furthermore, we tested that lipopolysaccharide administration increases HIF-1α mRNA and protein in naive and endotoxin-tolerant monocytes. Methods In 99 patients with sepsis and 48 healthy volunteers, leukocyte HIF-1α mRNA expression (real-time polymerase chain reaction), cytokine concentrations (enzymelinked immunosorbent assay), and intracellular distribution of HIF-1α protein (immunofluorescence staining) were assessed. In vitro, HIF-1α mRNA expression and protein were measured in naive or endotoxin-tolerant (48 h; 0.05 ng/ml lipopolysaccharide) monocytes, with/without additional lipopolysaccharide (6h; 1 μg/ml). Results In comparison to healthy volunteers, HIF-1α mRNA expression (-67%; P = 0.0001) and HIF-1α protein positive cells (-66.7%; P = 0.01) were decreased in sepsis. mRNA expression of adrenomedullin (-75%), CD11a (-85%), and CD11b (-86%; all P = 0.0001) was also decreased. In contrast, interleukin 6 (P = 0.0001), interleukin 10 (P = 0.0001), and tumor necrosis factor-α (P = 0.0002) concentrations were increased. Of note, HIF-1α mRNA expression was inversely associated with illness severity (Simplified Acute Physiology Score II; r = -0.29; P = 0.0001). In vitro, acute lipopolysaccharide administration of naive monocytic cells increased HIF-1α mRNA expression, whereas HIF-1α mRNA and protein (-60%; P = 0.001) were decreased in endotoxin-tolerant cells, which still up regulated cytokines. Conclusions In sepsis, HIF-1α mRNA expression was suppressed and inversely associated with illness severity. While acute lipopolysaccharide administration increased HIF-1α mRNA expression, prolonged stimulation suppressed HIF-1α expression. The clinical implications of decreased HIF-1α may include maladaption to tissue hypoxia or depressed immune function.

Culture Duration-, Donor-, and Medium-Dependent Changes in OATP1B3- Mediated Telmisartan Uptake in Human Hepatocytes

Human hepatic organic anion-transporting polypeptides (OATPs), including OATP1B1, 1B3, and 2B1, are expressed at the basolateral membrane of hepatocytes and mediate the uptake of a variety of compounds from blood into hepatocytes. The liver-specific OATPs are increasingly recognized as playing important roles in the pharmacokinetic (PK) of many drugs and thus, involved in the clinically significant drug-drug interactions (DDIs). However, the evaluation of the specific roles of individual OATPs in hepatocytes is challenging because of the lack of selective inhibitors and probe substrates for each OATP member. In the present study, the uptake activity of OATP1B3 was examined in human hepatocytes cultured up to 14 days using an in vitro uptake assay. The results showed that OATP-mediated uptake of rosuvastatin, a substrate for OATPs declined substantially in cultured human hepatocytes. In contrast, the uptake of OATP1B3-selective substrate telmisartan was not measureable at earlier culture periods, but became detectable on Day 7 and showed culture duration-dependent changes from Day 7 to 14. Quantitative polymerase chain reaction (qPCR) analyses illustrated that the OATP functional change was not correlated with messenger ribonucleic acid (mRNA) expression alteration in hepatocytes cultured for 3 hours or 7 days. The OATP1B3-mediated telmisartan uptake was also culture medium- and donor-dependent, and only observed in 3 of 5 lots of hepatocytes cultured in 2 of 3 media tested. These results show that using human hepatocytes cultured in certain conditions may provide an excellent addition to transfected cell lines as a way to distinguish OATP1B3 from other hepatic OATP family members, such as OATP1B1, to provide more understanding of OATP-mediated clinical DDI.

Detection of Bladder Cancer by Measuring CD44v6 Expression in Urine With Real-time Quantitative Reverse Transcription Polymerase Chain Reaction

To examine urinary CD44v6 total ribonucleic acid (RNA) expression in patients with bladder cancer using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and evaluate its potential as a novel marker of bladder cancer. We used the bladder cancer cell line T24 and determined CD44v6 expression in cancer cells using in situ hybridization and immunohistochemistry. Subsequently, we obtained urine samples from 21 patients with bladder cancer and 25 patients without bladder cancer (controls). We extracted total RNA from the urine samples, measured CD44v6 total RNA expression in both groups using qRT-PCR, and compared the expression between groups. We also compared the sensitivity, specificity, and concordance rate between CD44v6 total RNA expression analysis by qRT-PCR and cytologic analysis, UroVysion fluorescent in situ hybridization, bladder tumor antigen identification, and nuclear matrix protein 22 measurements. We observed increased CD44v6 expression in bladder cancer cells using in situ hybridization and immunohistochemistry. CD44v6 total RNA expression was significantly higher in the urine samples of patients with bladder cancer than in those of controls. We calculated the cutoff value from the receiver operating characteristic curve and obtained sensitivity and specificity values of 85.7% and 72.0%, respectively, for qRT-PCR analysis. Our results suggest that CD44v6 total RNA levels in urine can serve as a potential noninvasive biomarker of bladder cancer.

Molecular Profiles of Pyramidal Neurons in the Superior Temporal Cortex in Schizophrenia

Disrupted synchronized oscillatory firing of pyramidal neuronal networks in the cerebral cortex in the gamma frequency band (i.e., 30–100 Hz) mediates many of the cognitive deficits and symptoms of schizophrenia. In fact, the density of dendritic spines and the average somal area of pyramidal neurons in layer 3 of the cerebral cortex, which mediate both long-range (associational) and local (intrinsic) corticocortical connections, are decreased in subjects with this illness. To explore the molecular pathophysiology of pyramidal neuronal dysfunction, we extracted ribonucleic acid (RNA) from laser-captured pyramidal neurons from layer 3 of Brodmann's area 42 of the superior temporal gyrus (STG) from postmortem brains from schizophrenia and normal control subjects. We then profiled the messenger RNA (mRNA) expression of these neurons, using microarray technology. We identified 1331 mRNAs that were differentially expressed in schizophrenia, including genes that belong to the transforming growth factor beta (TGF-β) and the bone morphogenetic proteins (BMPs) signaling pathways. Disturbances of these signaling mechanisms may in part contribute to the altered expression of other genes found to be differentially expressed in this study, such as those that regulate extracellular matrix (ECM), apoptosis, and cytoskeletal and synaptic plasticity. In addition, we identified 10 microRNAs (miRNAs) that were differentially expressed in schizophrenia; enrichment analysis of their predicted gene targets revealed signaling pathways and gene networks that were found by microarray to be dysregulated, raising an interesting possibility that dysfunction of pyramidal neurons in schizophrenia may in part be mediated by a concerted dysregulation of gene network functions as a result of the altered expression of a relatively small number of miRNAs. Taken together, findings of this study provide a neurobiological framework within which specific hypotheses about the molecular mechanisms of pyramidal cell dysfunction in schizophrenia can be formulated.

Molecular Profiles of Parvalbumin-Immunoreactive Neurons in the Superior Temporal Cortex in Schizophrenia

Dysregulation of pyramidal cell network function by the soma- and axon-targeting inhibitory neurons that contain the calcium-binding protein parvalbumin (PV) represents a core pathophysiological feature of schizophrenia. In order to gain insight into the molecular basis of their functional impairment, we used laser capture microdissection (LCM) to isolate PV-immunolabeled neurons from layer 3 of Brodmann's area 42 of the superior temporal gyrus (STG) from postmortem schizophrenia and normal control brains. We then extracted ribonucleic acid (RNA) from these neurons and determined their messenger RNA (mRNA) expression profile using the Affymetrix platform of microarray technology. Seven hundred thirty-nine mRNA transcripts were found to be differentially expressed in PV neurons in subjects with schizophrenia, including genes associated with WNT (wingless-type), NOTCH, and PGE2 (prostaglandin E2) signaling, in addition to genes that regulate cell cycle and apoptosis. Of these 739 genes, only 89 (12%) were also differentially expressed in pyramidal neurons, as described in the accompanying paper, suggesting that the molecular pathophysiology of schizophrenia appears to be predominantly neuronal type specific. In addition, we identified 15 microRNAs (miRNAs) that were differentially expressed in schizophrenia; enrichment analysis of the predicted targets of these miRNAs included the signaling pathways found by microarray to be dysregulated in schizophrenia. Taken together, findings of this study provide a neurobiological framework within which hypotheses of the molecular mechanisms that underlie the dysfunction of PV neurons in schizophrenia can be generated and experimentally explored and, as such, may ultimately inform the conceptualization of rational targeted molecular intervention for this debilitating disorder.

Association between human papillomavirus (HPV) and the oral squamous cell carcinoma: a systematic review

The human papillomavirus (HPV) is an epitheliotropic agent whose high-risk genotypes have a well-established link with the development of cervical cancer. Although the relation of HPV to the oral squamous cell carcinoma (OSCC) has been studied since the beginning of the 1980s, its role in the oral carcinogenesis and the probable underlying molecular mechanisms are still not fully elucidated. We performed a systematic review of the worldwide scientific literature, published until the preparation of the present paper, concerning the association of HPV with OSCC, scrutinizing the samples, prevalence levels, the techniques utilized and relevant findings of the studies. The results showed that HPV is associated with approximately one quarter of OSCCs. Another interesting feature is the distinct pattern of infection in these oral tumors, including the participation of genotypes that are uncommon in cervical malignant lesions, such as HPV-38, 44, 53 and 70. Equally interesting is the possibility of carcinogenic action without the occurrence of viral integration, verified by the high expression of messenger ribonucleic acid (mRNA) of E6 and E7 from high-risk genotypes in cases whose virus remain in the episomal form. These findings support the assumption of HPV involvement in the genesis of OSCC, whereas warn about the possibility of unexpected viral behaviors that sometimes are not perceived or understood due to the technological limitations of the time and to the shortage of studies with the adequate approaches.

Changes in electrical response function and myosin heavy chain isoforms following denervation and reinnervation of bilateral posterior cricoarytenoid muscles in dogs

Conclusions: Both electrical response function and mRNA expression of myosin heavy chain (MyHC) types 2X, 1, and Neonatal of bilateral posterior cricoarytenoid (PCA) muscle changed after denervation or reinnervation in canines. Objectives: There is a need to investigate the electrical response function MyHC alteration of denervation or reinnervation in the bilateral PCA muscle of large animals. Methods: MyHC isoforms expression profile and PCA muscle function outcome were detected by real-time reverse transcribed-polymerase chain reaction and muscle response to functional electrical stimulation, 9 weeks after denervation and reinnervation with ansa–recurrent laryngeal nerve anastomosis in dogs. Results: Denervation produced up-regulation of MyHC-1 and MyHC-Neonatal messenger ribonucleic acid (mRNA) expression. Reinnervation caused a decrease of MyHC-2X mRNA expression. The electrical voltage threshold of vocal fold movement and maximum abduction of denervation were greater than that of the reinnervated or control group. The denervated vocal abduction maximum of response to electrical stimulation was less than that in reinnervation or control groups.

The Effect of Simvastatin on the Expression of Catalase in Human Retinal Pigment Epithelial Cells

Purpose: To evaluate the effects of simvastatin on the catalase expression in human retinal pigment epithelium. Methods: Retinal pigment epithelial (RPE) cells were incubated for 6 hours and 24 hours with various concentrations of simvastatin. In addition, RPE cells were incubated with 200 μM of H2O2 and various concentrations of simvastatin. After incubation, real-time polymerase chain reaction (PCR) was performed to examine the catalase messenger ribonucleic acid (mRNA) expression and a catalase assay was performed to examine the catalase activity in RPE. Intracellular reactive oxygen species (ROS) was measured using a fluorescence activated cell sorter (FACS). Results: Simvastatin increased the amount of catalase mRNA and catalase activity at 10 μM in RPE cells. Under oxidative stress (200 μM of H2O2), 2.5 μM of simvastatin increased the catalase mRNA expression and 5 μM of simvastatin increased catalase activity in RPE cells. In addition, simvastatin reduced free radical formation but this effect was diminished in the presence of an irreversible catalase inhibitor, 3-amino-1,2,4-triazole (3-AT). Conclusions: Simvastatin exhibits anti-oxidative effects by inducing the catalase expression in human RPE cells. This anti-oxidative effect may be beneficial for preventing age-related macular degeneration induced by oxidative stress. J Korean Ophthalmol Soc 2014;55(10):1535-1542

Estrogen receptor modulatory effects of germinated brown rice bioactives in the uterus of rats through the regulation of estrogen-induced genes

PurposeThe expression of genes regulated by estrogen in the uterus was studied in ovariectomized (OVX) rats treated with germinated brown rice (GBR) bioactives, and compared to Remifemin or estrogen at different doses to identify the regulation of these genes in the uterus and their molecular mechanisms.MethodsRats were treated orally with GBR bioactives (phenolics), acylated steryl glucosides (ASG), γ-amino butyric acid (GABA), and γ-oryzanol (ORZ) at 100 and 200 mg/kg, Remifemin (REM) at 10 mg/kg and 20 mg/kg, or estrogen (EST) at 0.2 mg/kg. Ribonucleic acid (RNA) was extracted from the uterus, and messenger (m)RNA expression of selected genes encoding estrogen receptor-beta (ER-β), calcium-binding protein (CaBP9k), complement protein (C3), heat shock protein 70 kDa (HSP70), and interleukin (IL)-4 receptor were quantified. Similarly, serum steroid hormone concentration was monitored at 2, 4, and 8 weeks after treatments. ER-β antibody binding to the uterus sections was also studied using immunohistochemistry.ResultsThe group treated with EST (0.2 mg/kg) upregulated ER-β, C3, and IL-4 receptor genes compared to other groups (P<0.001). GBR phenolics (200 mg/kg) treatment upregulated the ER-β gene almost to the level of the sham non-treated group. The CaBP9k gene showed upregulation in groups treated with ASG (200 mg/kg), EST (0.2 mg/kg), and ORZ (200 mg/kg) (P<0.05). Estrogen levels increased in groups treated with EST, ASG, and ORZ (200 mg/kg) compared to the OVX untreated group (P<0.05), and there was a slight non-significant decrease (P>0.05) in the progesterone levels in the OVX untreated group compared to the sham and other treated groups. There was a significant increase at 8 weeks in the level of FSH (P<0.05) in the treated groups compared to the OVX untreated group. There was no significant difference (P>0.05) in serum luteinizing hormone (LH) between the OVX untreated group and other groups. The sham and GBR phenolics treated group showed ER-β reactivity at the glandular epithelium, while the group treated with EST showed immunoreactivity at the glandular, luminal, and stromal epithelium.ConclusionGBR phenolics moderately regulate the expression of ER-β, HSP70, and IL-4 receptor genes, and gave a positive immunoreaction to ER-β antigen in the uterus. ASG regulates the expression of CaBP9k and IL-4 receptor genes, and ORZ regulates the expression of the CaBP9k gene, while GABA at 100 mg/kg regulates the expression of the HSP70 gene. GBR and its bioactives might have an effect on estrogen-regulated genes in the uterus of rats.

Expression of prolactin receptor messenger ribonucleic acid on peripheral blood mononuclear cells in patients with Hashimoto's thyroiditis

[Summary] To investigate the role played by serum prolactin and prolactin receptor in the pathogenesis of Hashimoto's thyroiditis(HT).33 newly diagnosed patients with HT with euthyroidism and 32 healthy subjects were enrolled as HT group and normal control group respectively.The expression of prolactin receptor mRNA on peripheral blood mononuclear cells (PBMC) were detected by realtime fluorescence quantitative PCR,serum prolactin was determinated by immunochemiluminescence assay and interleukin 2 was detected by Enzyme-linked immunosorbent.Compared with the controls,the level of prolactin receptor mRNA on PBMC,serum prolactin,interleukin 2 in HT patients were significantly higher[1.190 ± 0.913 vs 0.149 ± 0.130,(13.941 ± 7.109 vs 11.022 ±3.461)ng/ml,(102.165 ±43.716 vs 81.862 ± 32.128) pg/ml,P＜0.05] ; it implies that prolactin may be involved in the body's immune defences through combing with its receptor,promoting the activation of lymphocytes,and participating in the development of Hashimoto's thyroiditis. Key words: Prolactin receptor; Hashimoto's thyroiditis; Reverse transcription-polymerase chain reaction; Realtime fluorescence quantitative PCR