Real-time Reverse Transcription-polymerase Research Articles

Quantitative CT for detecting COVID\u201119 pneumonia in suspected cases

BackgroundCorona Virus Disease 2019 (COVID-19) is currently a worldwide pandemic and has a huge impact on public health and socio-economic development. The purpose of this study is to explore the diagnostic value of the quantitative computed tomography (CT) method by using different threshold segmentation techniques to distinguish between patients with or without COVID-19 pneumonia.MethodsA total of 47 patients with suspected COVID-19 were retrospectively analyzed, including nine patients with positive real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR) test (confirmed case group) and 38 patients with negative RT-PCR test (excluded case group). An improved 3D convolutional neural network (VB-Net) was used to automatically extract lung lesions. Eight different threshold segmentation methods were used to define the ground glass opacity (GGO) and consolidation. The receiver operating characteristic (ROC) curves were used to compare the performance of various parameters with different thresholds for diagnosing COVID-19 pneumonia.ResultsThe volume of GGO (VOGGO) and GGO percentage in the whole lung (GGOPITWL) were the most effective values for diagnosing COVID-19 at a threshold of − 300 HU, with areas under the curve (AUCs) of 0.769 and 0.769, sensitivity of 66.67 and 66.67%, specificity of 94.74 and 86.84%. Compared with VOGGO or GGOPITWL at a threshold of − 300 Hounsfield units (HU), the consolidation percentage in the whole lung (CPITWL) with thresholds at − 400 HU, − 350 HU, and − 250 HU were statistically different. There were statistical differences in the infection volume and percentage of the whole lung, right lung, and lobes between the two groups. VOGGO, GGOPITWL, and volume of consolidation (VOC) were also statistically different at the threshold of − 300 HU.ConclusionsQuantitative CT provides an image quantification method for the auxiliary diagnosis of COVID-19 and is expected to assist in confirming patients with COVID-19 pneumonia in suspected cases.

FDG PET/CT imaging features and clinical utility in COVID-19

PurposeTo determine the imaging findings and potential clinical utility of FDG PET/CT in patients with laboratory-confirmed COVID-19. MethodsWe performed a single institution retrospective review of patients diagnosed with COVID-19 using real time reverse transcription–polymerase chain reaction (RT-PCR) who underwent FDG PET/CT for routine cancer care between March 1, 2020 to April 30, 2020, during the height of the pandemic in New York City, New York, United States. PET/CT scans were retrospectively reviewed for imaging findings suspicious for COVID-19. For positive scans, PET and CT findings were recorded, including location, FDG avidity (SUVmax) and CT morphology. Patient demographics and COVID-19 specific clinical data were collected and analyzed with respect to PET/CT scan positivity, lung SUVmax, and time interval between PET/CT and RT-PCR. ResultsThirty-one patients (21 males and 10 females, mean age 57 years ± 16) were evaluated. Thirteen of 31 patients had positive PET/CT scans, yielding a detection rate of 41.9%. Patients with positive scans had significantly higher rates of symptomatic COVID-19 infection (77% vs 28%, p = 0.01) and hospitalizations (46% vs. 0%, p = 0.002) compared to patients with negative scans. Eleven of 13 patients (84.6%) with positive scans had FDG-avid lung findings, with mean lung SUVmax of 5.36. Six of 13 patients (46.2%) had extrapulmonary findings of FDG-avid thoracic lymph nodes. The detection rate was significantly lower when the scan was performed before RT-PCR versus after RT-PCR (18.8% (n = 3/16) vs. 66.7% (n = 10/15), p = 0.009). Lung SUVmax was not associated with COVID-19 symptoms, severity, or disease course. ConclusionFDG PET/CT has limited sensitivity for detecting COVID-19 infection. However, a positive PET scan is associated with higher risk of symptomatic infection and hospitalizations, which may be helpful in predicting disease severity.

Prolonged viral shedding of SARS-CoV-2 in two immunocompromised patients, a case report

BackgroundThe duration of viable Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) shedding in immunocompromised patients is still unknown. This case report describes the duration of viable SARS-CoV-2 in two immunocompromised patients with completely different clinical courses and further addresses the immunological aspects.Case presentationsOropharyngeal swaps were collected continuously during hospitalization for two immunocompromised patients infected with SARS-CoV-2 and sent for analysis to real timereverse transcription polymerase chain reaction (RT-PCR), viral culture assessed by plaque assay and full genome sequencing. Blood samples for flow cytometry and further immunological analysis were taken once during admission. One patient was without symptoms of Coronavirus disease 2019 (COVID-19) whereas the other had severe respiratory symptoms requiring a stay at an intensive care unit (ICU) and treatment with remdesivir and dexamethasone. Despite their difference in clinical courses, they both continuously shed SARS-CoV-2 with high viral loads in culture. Both patients had undetectable anti SARS-CoV-2 IgG levels about 2 weeks after the first positive real time RT-PCR test of SARS-CoV-2, marked expansions of virus reactive CD8+ T cells but cellular markers indicative of attenuated humoral immunity.ConclusionsOur case illustrates the importance of distinguishing isolation guidelines for patients infected with SARS-CoV-2 according to their immunological status. Furthermore, it demonstrates the need for immune markers relating to viral shedding in immunocompromised patients.

Identification of lncRNA NR_028138.1 as a biomarker and construction of a ceRNA network for bipolar disorder

The pathogenesis of bipolar disorder (BD), a chronic mood disorder, is largely unknown. Noncoding RNAs play important roles in the pathogenesis of BD. However, little is known about the correlations of long noncoding RNAs (lncRNAs) with BD. Illumina high-throughput sequencing in BD patients and normal controls was used to identify differentially expressed (DE) genes. Two-step real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to validate DE-RNAs in the first cohort (50 BD and 50 control subjects). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and lncRNA-mRNA coexpression and lncRNA-microRNA (miRNA)-messenger RNA (mRNA) competing endogenous RNA (ceRNA) network analyses were used to predict the functions of DE-RNAs. Receiver operating characteristic (ROC) curve analysis and logistic regression were applied to evaluate diagnostic performance in an additional testing group (80 BD and 66 control subjects). A total of 576 significantly DE-lncRNAs and 262 DE-mRNAs were identified in BD patients, and 95 lncRNA-miRNA-mRNA interactions were used to construct a ceRNA regulatory network. Analysis of the first cohort showed that six RNAs (NR_028138.1, TCONS_00018621, TCONS_00002186, TNF, PID1, and SDK1) were differentially expressed in the BD group (P < 0.01). NR_028138.1 was used to establish a BD diagnostic model (area under the ROC curve 0.923, P < 0.004, 95% CI: 0.830–0.999). Verification in the second cohort revealed uniformly significant differences in NR_028138.1 (P < 0.0001). This study constructed a ceRNA regulatory network and provided a hypothesis for the pathogenesis of BD. NR_028138.1 was identified as a central element involved in the transcriptional regulation in BD and a potential biomarker.

Genomic insights into the antibiotic resistance pattern of the tetracycline-degrading bacterium, Arthrobacter nicotianae OTC-16

Although many bacteria have the potential to remove antibiotic residues from environmental niches, the benefits of using antibiotic-degrading bacteria to manage antibiotic pollution should be assessed against the risk of the potential expansion of antimicrobial resistance. This study investigated the antibiotic resistance pattern of the bacterium Arthrobacter nicotianae OTC-16, which shows substantial biodegradation of oxytetracycline (OTC)/tetracycline. The results showed that this strain could be resistant to at least seven categories of 15 antibiotics, based on antimicrobial susceptibility testing. The genome of A. nicotianae OTC-16 contains one chromosome (3,643,989 bp) and two plasmids (plasmid1, 123,894 bp and plasmid2, 29,841 bp). Of the 3,561 genes isolated, eight were related to antibiotic resistance. During OTC degradation by the strain OTC-16, the expression of ant2ia, sul1, tet33, and cml_e8 in the plasmid, and one gene (tetV) in the chromosome were tracked using real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Only the plasmid-derived resistance genes were up-regulated in the presence of OTC. The presence of OTC increased the tolerance of strain OTC-16 to streptomycin sulphate. The findings of this study can help deepen our understanding of the behavioural characteristics of resistance genes and adaptive evolution of drug-resistant bacteria.

GPR110 ligands reduce chronic optic tract gliosis and visual deficit following repetitive mild traumatic brain injury in mice

BackgroundRepetitive mild traumatic brain injury (mTBI) can result in chronic visual dysfunction. G-protein receptor 110 (GPR110, ADGRF1) is the target receptor of N-docosahexaenoylethanolamine (synaptamide) mediating the anti-neuroinflammatory function of synaptamide. In this study, we evaluated the effect of an endogenous and a synthetic ligand of GPR110, synaptamide and (4Z,7Z,10Z,13Z,16Z,19Z)-N-(2-hydroxy-2-methylpropyl) docosa-4,7,10,13,16,19-hexaenamide (dimethylsynaptamide, A8), on the mTBI-induced long-term optic tract histopathology and visual dysfunction using Closed-Head Impact Model of Engineered Rotational Acceleration (CHIMERA), a clinically relevant model of mTBI.MethodsThe brain injury in wild-type (WT) and GPR110 knockout (KO) mice was induced by CHIMERA applied daily for 3 days, and GPR110 ligands were intraperitoneally injected immediately following each impact. The expression of GPR110 and proinflammatory mediator tumor necrosis factor (TNF) in the brain was measured by using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) in an acute phase. Chronic inflammatory responses in the optic tract and visual dysfunction were assessed by immunostaining for Iba-1 and GFAP and visual evoked potential (VEP), respectively. The effect of GPR110 ligands in vitro was evaluated by the cyclic adenosine monophosphate (cAMP) production in primary microglia isolated from adult WT or KO mouse brains.ResultsCHIMERA injury acutely upregulated the GPR110 and TNF gene level in mouse brain. Repetitive CHIMERA (rCHIMERA) increased the GFAP and Iba-1 immunostaining of glia cells and silver staining of degenerating axons in the optic tract with significant reduction of N1 amplitude of visual evoked potential at up to 3.5 months after injury. Both GPR110 ligands dose- and GPR110-dependently increased cAMP in cultured primary microglia with A8, a ligand with improved stability, being more effective than synaptamide. Intraperitoneal injection of A8 at 1 mg/kg or synaptamide at 5 mg/kg significantly reduced the acute expression of TNF mRNA in the brain and ameliorated chronic optic tract microgliosis, astrogliosis, and axonal degeneration as well as visual deficit caused by injury in WT but not in GPR110 KO mice.ConclusionOur data demonstrate that ligand-induced activation of the GPR110/cAMP system upregulated after injury ameliorates the long-term optic tract histopathology and visual impairment caused by rCHIMERA. Based on the anti-inflammatory nature of GPR110 activation, we suggest that GPR110 ligands may have therapeutic potential for chronic visual dysfunction associated with mTBI.

Co(II) coordination polymer: crystal structure and treatment activity against chronic obstructive pulmonary disease

By employment of a semi-flexible quadritopic N-donor ligand, 5,5′-bipyrimidine (bpym), a new azide-based coordination polymer (CP) with the chemical formula of {[Co3(bpym)3(N3)6]·2H2O} n has been successfully prepared. Its application values on the chronic obstructive pulmonary disease (COPD) were assessed and the specific mechanism was explored as well. The levels of the inflammatory cytokines released by the COPD animal was measured with enzyme linked immunosorbent assay (ELISA) detection assay. The activation level of the Nuclear factor kappa beta (NF-κB) signaling pathway in the lung epithelial cells was determined with real time Reverse Transcription Polymerase Chain Reaction (RT-PCR) assay. Above all, the compound exhibited excellent treatment activity on COPD. The Co complex was found to form multiple binding interactions within the docking pocket, all those binding interactions were formed by the pyrimidine rings. Above all, the compound has excellent application values on the COPD treatment.

Resveratrol Confers Protection Against Ischemia/Reperfusion Injury by Increase of Angiotensin (1-7) Expression in a Rat Model of Myocardial Hypertrophy.

Left ventricular hypertrophy (LVH) makes the heart vulnerable to ischemia/reperfusion (IR) injury. Angiotensin (Ang) (1-7) is recognized as a cardioprotective peptide. We investigated the effect of polyphenol resveratrol on myocardial IR injury after hypertrophy and examined cardiac content of Ang (1-7) and transcription of its receptor (MasR). Rats were divided into sham-operated, LVH, IR, LVH + IR, and resveratrol + LVH + IR groups. Myocardial hypertrophy and IR models were created by abdominal aortic banding and left coronary artery occlusion, respectively. To evaluate the electrocardiogram parameters and incidence of arrhythmias, electrocardiogram was recorded by subcutaneous leads (lead II). Blood pressure was measured through the left carotid artery. Infarct size was determined by the triphenyl tetrazolium chloride staining. The Ang (1-7) level was evaluated by immunohistochemistry. The Mas receptor mRNA level was assessed by the real-time real time reverse transcription polymerase chain reaction technique. QT-interval duration, infarct size, and incidence of ischemia-induced arrhythmia were significantly higher in the LVH + IR group. However, in the resveratrol-treated group, these parameters were decreased significantly. The cardiac level of Ang (1-7) was decreased in untreated hypertrophied hearts (LVH and LVH + IR groups). Pretreatment with resveratrol normalized the cardiac level of Ang (1-7). The mRNA level of Mas receptor was increased in all of hypertrophied hearts in the presence or absence of resveratrol. Resveratrol can decrease IR injury in rats with LVH. The anti-ischemic effect of resveratrol may be related to the enhancement of Ang (1-7)/MasR axis.

SARS-COV-2 (COVID-19) and role of real time Reverse Transcription Polymerase Chain Reaction (RT-PCR) in its diagnosis

The present article reviews the diagnosis of Novel Corona Virus Disease (COVID-19) by using Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR). COVID-19 is a contagious disease caused by severe acute respiratory syndrome coronavirus2 (SARS-CoV-2), which affects upper and lower respiratory tract. In present scenario COVID-19 is considered as global public health emergency by World Health Organization (WHO) so, the detection and prevention of this disease is vital to control this emergency. RT-PCR is a highly sensitive laboratory method for the detection of SARS-CoV-2. It is based on rapid diagnosis of SARS-CoV-2 by qualitative detection of its nucleic acid sequence. This test locates the primer and probe sets in different regions in the genome of SARS-CoV-2.

One-step real-time multiplex reverse transcription-polymerase chain reaction assay with melt curve analysis for detection of potato leafroll virus, potato virus S, potato virus X, and potato virus Y

BackgroundCertification of seed potato as free of viruses is essential for stable potato production. Among more than 30 virus species infecting potato, potato leafroll virus (PLRV), potato virus S (PVS), potato virus X (PVX), and potato virus Y (PVY) predominate worldwide and should be the targets of a high-throughput detection protocol for seed potato quarantine.ResultsWe developed an assay based on one-step real-time multiplex reverse transcription-polymerase chain reaction (mRT-PCR) with melt curve analysis for the four viruses and one internal control, potato elongation factor 1 alpha gene (EF1α). Virus-specific primers were derived from conserved regions among randomly selected representatives considering viral genomic diversity. Our assay simultaneously detected representative Japanese isolates of PLRV, O lineage of PVS, PVX, and NTN strain of PVY. The variability of melting temperature (Tm) values for each virus was confirmed using Japanese isolates, and virus species could be identified by the values of 87.6 for PLRV, 85.9 for PVX, 82.2 (Ordinary lineage) to 83.1 (Andean lineage) for PVS, and 79.4 (NA-N strain) to 80.5 (O strain and NTN strain) for PVY on average. The reliability of calculation was validated by comparing the calculated Tm values and measured Tm values and the values had a strong linear correlation (correlation of determination: R2 = 0.9875). Based on the calculated Tm values, representative non-Japanese isolates could also be identified by our assay. For removing false positives, two criteria were set for the evaluation of result; successful amplification was considered as 30.0 ≥ threshold cycle value, and the virus-specific peak higher than the EF1α-specific peak was considered as positive. According to these criteria, our assay could detect PLRV and PVS from 100-fold dilution of potato leaf homogenate and PVX and PVY from 1000-fold in a model assay.ConclusionThis new high-throughput detection protocol using one-step real-time mRT-PCR was sensitive enough to detect viruses in a 100-fold dilution of singly-virus contaminated homogenate in a model assay. This protocol can detect the four viruses in one assay and yield faster results for a vast number of samples, and greatly save the labor for seed potato quarantine and field surveys.

Two New Coordination Polymers: Selective Detection of Cu(II) Ion and Treatment Activity on Inner Ear Damage.

In the current study, on the basis of 1,3,5-tris(2-methylimidazol-1-yl)benzene (timb), a designed tripodal connector, two new transition metal coordination polymers (CPs), {[Cu4(timb)2(Br-IPA)4]·5H2O}n (1) and {[Zn(timb)0.5(NH2-IPA)]·4H2O}n (2) have been generated with the mixed ligand method by the reaction between the timb and corresponding metal salts in the existence of dissimilar functional isophthalic acid (H2IPA) ligands. Furthermore, the Zn(II)-based complex 2 displays high sensitivity in the detection of Cu(II) ion in water. The neural stem cells proliferation after treated via compounds was detected with Cell Counting Kit-8 detection assay. And the real time reverse transcription polymerase chain reaction was carried out for the investigation of the differentiation function of the neural stem cells after the compound 1 treatment and compound 2 treatment. Further, molecular docking simulations confirmed that the biological activity that has been observed from experiments were from the carboxyl group on the Cu complex, in contrast, the imidazole groups were only used for binding with the Cu metal ion to retain the complex structure.

Hypertension, renin-angiotensin-aldosterone-system-blocking agents, and COVID-19

BackgroundThere have been concerns regarding the safety of renin-angiotensin-aldosterone-system (RAAS)-blocking agents including angiotensin-converting enzyme inhibitors (ACEI) and angiotensin receptor blockers (ARB) during the coronavirus disease 2019 (COVID-19) pandemic. This study sought to evaluate the impact of hypertension and the use of ACEI/ARB on clinical severity in patients with COVID-19.MethodsA total of 3,788 patients aged 30 years or older who were confirmed with COVID-19 with real time reverse transcription polymerase chain reaction were identified from a claims-based cohort in Korea. The primary study outcome was severe clinical events, a composite of intensive care unit admission, need for ventilator care, and death.ResultsPatients with hypertension (n = 1,190, 31.4 %) were older and had higher prevalence of comorbidities than those without hypertension. The risk of the primary study outcome was significantly higher in the hypertension group, even after multivariable adjustment (adjusted odds ratio [aOR], 1.67; 95 % confidence interval [CI], 1.04 to 2.69). Among 1,044 patients with hypertensive medical treatment, 782 (74.9 %) were on ACEI or ARB. The ACEI/ARB subgroup had a lower risk of severe clinical outcomes compared to the no ACEI/ARB group, but this did not remain significant after multivariable adjustment (aOR, 0.68; 95 % CI, 0.41 to 1.15).ConclusionsPatients with hypertension had worse COVID-19 outcomes than those without hypertension, while the use of RAAS-blocking agents was not associated with increased risk of any adverse study outcomes. The use of ACE inhibitors or ARBs did not increase the risk of adverse COVID-19 outcomes, supporting current guidance to continue these medications when indicated.

Role of Chest Computed Tomography versus Real Time Reverse Transcription Polymerase Chain Reaction for Diagnosis of COVID-19: A Systematic Review and Meta-Analysis.

Background The current global pandemic of COVID-19 is considered a public health emergency. The diagnosis of COVID-19 depends on detection of the viral nucleic acid by real time reverse transcription polymerase chain reaction (RT-PCR). However, false-negative RT-PCR tests are reported and could hinder the control of the pandemic. Chest computed tomography could achieve a more reliable diagnosis and represent a complementary diagnostic tool. Aim To perform a meta-analysis and systematic review to find out the role of chest computed tomography versus RT-PCR for precise diagnosis of COVID-19 infection. Methods We searched three electronic databases (PubMed, ScienceDirect, and Scopus) from April 1 to April 20, 2020, to find out articles including the accuracy of chest computed tomography scan versus RT-PCR for diagnosis of SARS-CoV-2 infection. Observational studies, case series, and case reports were included. Results A total of 238 articles were retrieved from the search strategy. Following screening, 39 articles were chosen for full text assessment and finally 35 articles were included for qualitative and quantitative analysis. Chest computed tomography showed a wide range of sensitivity varied from 12%–100%. Conclusion Chest computed tomography is playing a key role for diagnosis and detection of COVID-19 infection. Computed tomography image findings may precede the initially positive RT-PCR assay.

MicroRNAs as predictors of future endometrial malignancies in endometrial hyperplasia without atypia.

e17563 Background: The histological criteria of predicting risk of progression of endometrial hyperplasia (EH) to endometrial cancer (EC) is based on the degree of architectural crowding and nuclear atypia. We aim to investigate the cumulative rate of progression to EC in simple or complex hyperplasia without atypia (SH/CH-nonA) and the molecular biomarkers for risk of developing EC in women with SH/CH-nonA. Methods: EH tissues of EC patients with preceding SH/CH-nonA (case) were compared to SH/CH-nonA patients without progression to EC (control) using miRNA array, followed by reverse transcription and quantitative real-time polymerase chain reaction (RT-qPCR), and verified in an independent cohort. Results: The 10-year and 15-year cumulative rates were 8% and 10%, respectively by the hospital medical records. The median time from EH to EC was 8.71 years (range, 0.55-19.84). Twenty miRNAs (p &lt; 0.05, fold change &gt; 4) were significantly different in SH/CH-nonA cases (n = 6) compared to SH/CH-nonA controls (n = 12) by miRNA array. Multiplex RT-qPCR validation (51 SH/CH-nonA controls and 19 SH/CH-nonA cases) selected miR30a-3p, miR141, miR200a, and miR200b. Area under the curve values for tissue miR30a-3p (p = 0.043), miR141(p = 0.003), miR-200a (p &lt; 0.001), and miR200b (p = 0.004) in discriminating cases from controls were 0.658, 0.734, 0.811, and 0.710, and the sensitivity and specificity rates were 56.9% and 76.5%, 73.7% and 74.5%, and 68.4% and 100%, 47.4% and 100%, respectively. Conclusions: MiR30a-3p, miR141, miR-200a, and miR200b are useful tissue biomarkers for predicting future risk of developing EC for patients with SH/CH-nonA.

Sophoridine Attenuates Vascular Endothelial Injury by Regulating the Src/PI3K Pathway: Molecular Modelling

Vascular endothelial injury is a major cause of cardiovascular diseases. Conventional treatments have several limitations; we thus explored whether sophoridine could protect against tumor necrosis factor-alpha induced vascular endothelial injury. We evaluated the effects of sophoridine on cell survival and apoptosis in TNF-α-treated human umbilical vein endothelial cells and in mice. Western blotting and real-time quantitative reverse transcription polymerase chain reaction were used to estimate the expression levels of relevant proteins and their mRNAs. The in vivo effects of sophoridine on vascular endothelial function were explored by immunofluorescence staining. Sophoridine enhanced vascular endothelial cell survival and reduced apoptosis after tumor necrosis factor-α-induced injury, and modulated the expression levels of Src, Akt, PI3K, and ZO-1. Sophoridine protected against tumor necrosis factor-α induced vascular endothelial injury by regulating the Src/Akt/ PI3K pathway.

Circulating miRNA 27a and miRNA150-5p; a noninvasive approach to endometrial carcinoma.

The search for novel non-invasive biomarkers such as epigenetic molecular markers is new hope for common and burdensome cancers. We aim to assess serum expression of miRNA 27a and miRNA150-5p in endometrial cancer patients. Serum was drawn for 36 un-intervened endometrial cancer patients scheduled for hysterectomy and 35 controls. miRNA 27a and miRNA150-5p were measured by real time reverse transcription polymerase chain reaction. Significant overexpression of both miRNA in patients (p < 0.001). At cutoffs 0.2872 & > 1.02, miRNA 27a showed 100% sensitivity, specificity, positive and negative predictive values. miRNA150-5p showed 88.89% sensitivity, 100% specificity, 100% positive and 78.9% negative predictive values. Areas under curve were 1.0 for miRNA 27a, 0.982 for miRNA 150 performing much better than Ca125. miRNA 27a was significantly associated with type I endometroid endometrial cancer. Conclusion: miRNA 27a and miRNA-150-5P can be suggested as promising biomarkers of endometrial cancer possibly part of a miRNA panel for management.

A (4,12)-connected coordination polymer: photocatalytic degradation of dyes and effects on cerebral edema care by regulating superoxide dismutase activity

With the improvement of people's living standards and the development of modern industry, water pollution is becoming more and more serious in the global environmental pollution. Coordination polymers have the potential to address the above mentioned problem, but their poor water stability and weak light adsorption ability in the visible light region largely restrict their applications. In this study, a new coordination polymer (CP) formulated as {(NH4)2·[Cu11(μ4-OH)6(CN)6(trz)12]}n (1) was produced with the reaction between μ3-N1,N2,N4-triazolate (trz) and Cu(NO3)2·3H2O under the solvothermal reaction conditions. Complex 1 could be utilized as a photocatalytic catalyst for the degradation of the organic dyes of MB (Methylene Blue) and RhB (Rhodamine B) which shows better performance than that of TiO2 (20 nm). For the treatment of the cerebral edema care after ischemia reperfusion injury, the brain water content was evaluated firstly after treated with compound, and the superoxide dismutase expression in the brain cells was also measured through real time reverse transcription polymerase chain reaction (RT-PCR).

Effect of intraoperative blood transfusion on Treg and FOXP3 in patients with digestive tract malignancies and different ABO blood types

BackgroundBlood transfusion can cause immunosuppression and lead to worse outcomes in patients with digestive tract malignancies; however, the specific mechanism behind this is not completely understood. One theory is that increased numbers of regulatory CD3+CD4+CD25+FOXP3+ T cells (Tregs) and forkhead box protein-3 mRNA (FOXP3) expression in the blood after transfusion contribute to these outcomes. The effect of blood transfusion on immune function in patients with different ABO blood types is variable. This study investigates the effect of intraoperative blood transfusion on the number of Tregs and the expression of FOXP3 in the blood of patients with different ABO blood types and digestive tract malignancies.MethodsPatients with digestive tract malignancies who underwent radical resection and received intraoperative blood transfusion were divided into four groups according to their blood types:blood group A, blood group B, blood group O and blood group AB (n = 20 for each group). Blood was collected from all patients before surgery, immediately after transfusion, 1 day after transfusion, and 5 days after transfusion. The number of Tregs was measured by flow cytometry. The expression of FOXP3 was detected by real time reverse transcription polymerase chain reaction (RT-PCR).ResultsThere was no significant difference in the number of Tregs or expression of FOXP3 mRNA among patients with different blood types before surgery. However, the number of Tregs and the expression of FOXP3 increased after blood transfusion in all blood type groups. This increase was especially evident and statistically significant on the first day after blood transfusion when compared with measures obtained before the surgery. Measures returned to the preoperative level five days after surgery. There were significant differences in the increase of Tregs and expression of FOXP3 among patients with different blood types. The greatest increase was seen in patients with blood group B and the least in blood group A.ConclusionsIntraoperative blood transfusion can lead to an increase in blood Tregs and FOXP3 expression in patients with digestive tract malignancies. Increases were greatest on the first day after surgery and differed among patients with different blood types. Increases were greatest in blood type B and least in blood type A.

Comparison of the Modified Centers for Disease Control and Prevention 2019-Novel Coronavirus Real-Time RT-PCR Method for Detection of Infectious and Heat-Inactivated Virus on Stainless Steel.

BackgroundInfectious Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) was used in the validation of methods for detection of SARS-CoV-2 on stainless steel surfaces in the AOAC Research Institute Emergency Response Validation project. Handling infectious virus requires Biosafety Level (BSL)-3 facilities.ObjectiveTo compare the recovery and detection of infectious and heat-inactivated (65 °C for 30 min) SARS-CoV-2 from stainless steel by the modified US Centers for Disease Control and Prevention (CDC) 2019-Novel Coronavirus Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR) Diagnostic Panel.MethodsViral stocks were diluted in viral transport medium and deposited onto stainless steel test areas at 2 x 103 and 2 x 104 genomic copies for low and high, respectively. Test areas were sampled, and aliquots of the resulting test solutions analyzed by RT-qPCR according to the CDC method. Results were analyzed by Probability of Detection (POD) statistics.ResultsThe low level, where fractional positive results (25–75%) are expected, yielded PODI = 0.80 (0.58, 0.92) for the infectious virus and PODHI = 0.15 (0.05, 0.36) for the heat-inactivated virus. The bias, dPODHI = -0.65 (-0.80, -0.35), demonstrated a statistical difference between infectious and heat-inactivated virus detection. No difference was observed at the high inoculation level.ConclusionDespite the statistical difference observed, the use of the heat-inactivated virus is a viable alternative for matrix extension studies using a method comparison study design.HighlightsThe use of heat-inactivated SARS-CoV-2 can mitigate the need for a BSL-3 facility for matrix extension validation of alternative methods in SARS-CoV-2 studies.

Detection of Israel variant 2 (IS/1494/06) genotype of Infectious Bronchitis Virus in a layer chicken flock

The aim of this study was to determine an infectious bronchitis (IB) infection, caused by an Israel variant 2 (IS/1494/06)-like IBV, in a layer chicken flock regularly vaccinated with vaccines containing IBV H120 and 4/91 strains. Mild respiratory symptoms, drop in egg production and soft-shelled eggs and eventually death were observed in a layer chicken flock. Clinical samples from four diseased chickens were examined for the detection and genotyping of IBV by virus isolation, a commercial real time reverse transcription polymerase chain reaction (rRT-PCR) and nucleotide sequencing. Both Israel variant 2 (IS-Var2) and 793/B serotypes were detected from samples by rRT-PCR, but sequencing results of a 345 bp part of S1 gene revealed that our IBV isolate, HFT-IBV, was IS/1494/06 (IS-Var2)-like with the 97.7% genetic similarity. These results suggested that immunity against vaccination with a combination of different genotypes (H120 and 4/91) could not be protective for IS-Var2 IBV field infection. In addition, identification of genotypes from the clinical samples, such as swabs and organ samples by commercial rRT-PCR assays failed to find correct IBV genotype responsible for the IB infection, whereas analysis of nucleotide sequencing of S1 gene of IBV as a gold standard test undoubtedly detected causative genotype of the infection. Also, the findings indicated that there is an urgent need for consider genotype- or protectotype-match vaccination strategies in the field to prevent vaccine- and IB-dependent economic losses of the poultry sector and logically protect chickens from IBV infection.