A second-generation, point-of-care immunoassay provided improved detection of Anaplasma and Ehrlichia antibodies in PCR-positive dogs naturally infected with Anaplasma or Ehrlichia species.

A validated second-generation SNAP 4Dx Plus (Idexx) incorporates new peptides for improved detection of antibodies against Anaplasma and Ehrlichia tick-borne pathogens in dogs. We compared the first- and second-generation SNAP 4Dx Plus using dogs naturally infected with Anaplasma or Ehrlichia species, or dogs seroreactive by an E. canis indirect fluorescent antibody test (IFAT). The second-generation immunoassay was more sensitive than the first-generation for dogs infected with A. phagocytophilum (51.1% and 29.2%, respectively), A. platys (63.6% and 35.3%, respectively), E. canis (96.2% and 88.3%, respectively), or E. ewingii (73.7% and 70.8%, respectively), and for dogs seroreactive by E. canis IFAT (87.3% and 83.9%, respectively). The second-generation immunoassay detected significantly more Anaplasma- or Ehrlichia-infected dogs that were Anaplasma (p < 0.001) or Ehrlichia (p = 0.031) seroreactive, respectively, than did the first-generation test. When Ehrlichia seroreactivity by E. canis IFAT and both immunoassays was compared, significantly more E. canis-infected dogs were seroreactive by E. canis IFAT than the first-generation (p = 0.006) but not the second-generation (p = 0.125) immunoassay. Significantly more E. ewingii-infected dogs were seroreactive by the first- (p = 0.011) and second-generation (p = 0.049) immunoassays than the E. canis IFAT. Medical records available for 7 dogs that were Anaplasma seroreactive by the second-generation but not the first-generation immunoassay revealed case management decisions that might have been different with an immediate anaplasmosis diagnosis, including earlier doxycycline therapy and less hospitalization. The second-generation SNAP 4Dx Plus test offered improved serologic detection of Anaplasma and Ehrlichia in naturally infected dogs.

A critical review of biogas production and usage with legislations framework across the globe.

This review showcases a comprehensive analysis of studies that highlight the different conversion procedures attempted across the globe. The resources of biogas production along with treatment methods are presented. The effect of different governing parameters like feedstock types, pretreatment approaches, process development, and yield to enhance the biogas productivity is highlighted. Biogas applications, for example, in heating, electricity production, and transportation with their global share based on national and international statistics are emphasized. Reviewing the world research progress in the past 10 years shows an increase of ~ 90% in biogas industry (120 GW in 2019 compared to 65 GW in 2010). Europe (e.g., in 2017) contributed to over 70% of the world biogas generation representing 64 TWh. Finally, different regulations that manage the biogas market are presented. Management of biogas market includes the processes of exploration, production, treatment, and environmental impact assessment, till the marketing and safe disposal of wastes associated with biogas handling. A brief overview of some safety rules and proposed policy based on the world regulations is provided. The effect of these regulations and policies on marketing and promoting biogas is highlighted for different countries. The results from such studies show that Europe has the highest promotion rate, while nowadays in China and India the consumption rate is maximum as a result of applying up-to-date policies and procedures.

Stereological and histopathological evaluation of doxorubicin-induced toxicity in female rats' ovary and uterus and palliative effects of quercetin and vitamin E.

Doxorubicin (DOX) is a widely used chemotherapeutic agent with demonstrated reproductive toxicity. This study sought to determine the DOX-induced toxicity in the ovary and uterus and the preventive effects of quercetin (QCT) and vitamin E (Vit.E). Female rats were divided into six groups as follows: control, QCT (20 mg/kg), Vit.E (200 mg/kg), DOX (accumulative 15 mg/kg), DOX/QCT, and DOX/Vit.E. After 3 weeks, the toxicity of DOX in ovarian and uterine tissues and the potential palliative effects of QCT and Vit.E were evaluated by histopathological-stereological methods. The findings indicate a dramatic decline in the number of ovarian follicles (p < 0.001), ovarian and its associated structures volume, the volume of the uterus, its layers, and related structures (p < 0.05). Coadministration of QCT and Vit.E with DOX-treated rats demonstrated an alleviative effect on most of the studied parameters. Nevertheless, few adverse effects were recognized concerning these antioxidants administration (p < 0.05). In conclusion, the findings of this study support the protective role of these dietary supplements in the prevention of DOX-induced toxicity in uterine and ovarian tissues.

Pediatric Emergency Care Applied Research Network (PECARN) prediction rules in identifying high risk children with mild traumatic brain injury.

PURPOSE: Pediatric Emergency Care Applied Research Network (PECARN) traumatic brain injury (TBI) prognostic rules is a scoring system for prediction of the need for computed tomography (CT) scanning in children with mild TBI. However, its validation has not been assessed in developing countries. Therefore, the present study was designed to assess the value of PECARN rule in identification of children with clinically important TBI (ciTBI). METHOD: In this prospective cross-sectional study, 594 children (mean age: 7.9 ± 5.3 years; 79.3% boys) with mild TBI brought to emergency ward of two healthcare centers in Tehran, Iran were assessed. PECARN checklist was filled for all patients and children were divided to three groups of low, intermediate and high risks. Patients were followed for 2 weeks by phone to assess their ciTBI status. At the end, discrimination power, calibration and overall performance of PECARN rule were assessed. RESULTS: PECARN had a sensitivity and specificity of 92.3 and 40.6%, respectively, in predicting ciTBI in children under 2 years and 100.0 and 57.8%, respectively, in individuals between the ages of 2 and 18. PECARN rule had a proper calibration in prediction of ciTBI and CT scan findings. Brier score (overall performance) of PECARN rule in predicting ciTBI in children under 2 and 2-18 years were 1.5 and 1.2, respectively. CONCLUSION: PECARN prediction rule has a proper validity in the prediction of ciTBI. Therefor it can be used for screening and identification of high risk children with mild TBI.

Mesenchymal stem cell-derived exosomes: A novel potential therapeutic avenue for cardiac regeneration.

Coronary artery diseases (CADs) represent a significant cause of death worldwide. During recent decades the rate of cardiovascular mortality has been declined as a result of modern medicine and surgery. However, despite the fact that cardiac cells, including cardiomyocytes (CMCs), vascular smooth muscle cells (VSMC) and vascular endothelial cells (VEC), can be regenerated by cardiac adult stem cell, the regenerative capacity of these cells are limited and inadequate to functionally regenerate heart damaged tissue. Thus, growth reserve of the heart fails to restore the structural integrity of the myocardium after infarction and healing is associated with scar formation. An explanation for this is that cardiac reside stem cells are present throughout the infarction site but die rapidly by apoptosis. Furthermore, microenvironment surrounding the damage site is not promising for the cells survival and renewal. Hence, recent advances in the stem cell therapy have emerged as an attractive approach to replace the lost cells. In this context, mesenchymal stem cells (MSCs) has considered as one of the most promising candidates for regeneration of cardiac cells, lost upon injury. The regenerative capacity of MSCs has primarily been centered on the hypothesis that these cells would engraft, differentiate and replace damaged cardiac cells. However, experimental and clinical observations so far have failed to establish if this differentiated is considerably relevant to MSCs cardiac regenerative properties. Recent reports have suggested that these therapeutic properties, at least in part, are mediated by paracrine factors released from MSCs. This review provides a concise summary of current evidences supporting the paracrine hypothesis of MSCs. In particular, the scope of this review focuses on the role of MSC-derived exosome (MSC-EXs) as a therapeutic modality for the treatment of CADs, particularly ischemic myocardial dysfunctions.

Neural stem/progenitor cell transplantation for spinal cord injury treatment; A systematic review and meta-analysis.

Despite the vast improvements of cell therapy in spinal cord injury treatment, no optimum protocol has been developed for application of neural stem/progenitor cells. In this regard, the present meta-analysis showed that the efficacy of the neural stem/progenitor cell (NSPC) transplantation depends mainly on injury model, intervention phase, transplanted cell count, immunosuppressive use, and probably stem cell source. Improved functional recovery post NSPC transplantation was found to be higher in transection and contusion models. Moreover, NSPC transplantation in acute phase of spinal injury was found to have better functional recovery. Higher doses (>3×10(6)cell/kg) were also shown to be optimum for transplantation, but immunosuppressive agent administration negatively affected the motor function recovery. Scaffold use in NSPC transplantation could also effectively raise functional recovery.

Expression analysis of BECN1 in acute myeloid leukemia: association with distinct cytogenetic and molecular abnormalities.

INTRODUCTION: In acute myeloid leukemia (AML), it has been shown that AML-derived cells often remain sensitive to autophagy-inducing stimuli, leading to the idea that harnessing the autophagy can be pertinent to AML cytotoxic therapy. Despite this promising notion, to date, there is no comprehensive study addressing autophagy-related genes expression status in AML. As a critical mediator, BECN1 influences the onset and advance of autophagy and several studies have pointed to the BECN1 recurrent allelic deletion and expression variation in a broad range of tumors. To explore this caveat, we chose this alteration-prone gene to investigate in our study. METHODS: We have analyzed the expression status of BECN1 in a series of 128 de novo AML patients using real-time quantitative polymerase chain reaction (qRT-PCR). RESULTS: In our favorable subgroup, BECN1 expression did not alter (P = 0.301), but in intermediate and unfavorable patients, we have had BECN1 low expression compared to the normal controls (P = 0.008 and P < 0.001, respectively). We found evidence for the association of reduced expression of BECN1 with FLT3-ITD mutation (19 of 27 patients), monosomal karyotype (all of 11 patients), higher age, and WBC count. CONCLUSION: Overall, remarkable association of reduced expression of BECN1 with FLT3-ITD mutation and monosomal karyotype and their functional relationship is interesting which should be addressed and verified in future studies.

Mutational screening of the NR5A1 in azoospermia.

Nuclear receptor subfamily 5 group A member 1 (NR5A1) encodes a nuclear receptor that regulates transcription of multiple genes involved in adrenal and gonadal development, steroidogenesis and the reproductive axis. Human mutations in NR5A1were initially found in two 46, XY female patients suffering from severe gonadal dysgenesis and primary adrenal failure. However, more recent case reports have suggested that heterozygous mutations in NR5A1 may also contribute to the male infertility aetiology. We have analysed the coding sequence of NR5A1 in a cohort of 90 well-characterised idiopathic Iranian azoospermic infertile men versus 112 fertile men. Heterozygous NR5A1 mutations were found in 2 of 90 (2.2%) of cases. These two patients harboured missense mutations within the hinge region (p.P97T) and ligand-binding domain (p.E237K) of the NR5A1 protein. In silico analysis of the mutations showed that founded mutations could be detrimental. In conclusion, findings of the current and previous studies suggest that mutations in the NR5A1 gene are not common in azoospermia, and male infertility and inclusion of NR5A1 mutation screening in the diagnostic workup of male infertility may seem unnecessary.

Targeting microRNA-122: walking on cutting edge of hepatitis C virus infection therapy.

Hepatitis C virus (HCV) infection, with an estimated 170 million carriers worldwide, remains a major cause of chronic liver disease. Current anti-HCV treatments have significant side effects and have met with only partial success. Therefore, a more effective therapeutic modality for HCV infection is needed. The stability and propagation of HCV is dependent on the interaction between its genome and a highly abundant liver microRNA (miRNA), known as microRNA-122 (miR-122). As a conserved host factor that would not be expected to evolve resistance mutations, miR-122 makes an attractive antiviral target. In this review we will discuss how targeting miR-122, using antisense oligonucleotides (ASOs), can be a new anti-HCV treatment.

An Iranian family with azoospermia and premature ovarian insufficiency segregating NR5A1 mutation.

In brief, we report an Iranian family with a history of both azoospermia and premature ovarian insufficiency with the same heterozygote mutation in the NR5A1 gene that can be transmitted. As far as we know, this is the first observation that a common mutation in NR5A1 can cause these above-mentioned phenotypes in a family.

Hepatitis B virus-induced hepatocellular carcinoma: the role of the virus x protein.

Hepatocellular carcinoma (HCC) is one of the most common malignant diseases and has the fourth highest mortality rate worldwide. Chronic hepatitis B virus (HBV) infection has been identified as a major risk factor in HCC. Currently available evidence support a critical role of hepatitis B virus x (HBx) gene and protein in the pathogenesis of HBV-induced HCC. HBx protein is a multifunctional regulator that modulates cellular signal transduction pathways, transcriptional regulations, cell cycle progress, DNA repair, apoptosis, and genetic stability by interacting with different host factors. This review describes the current state of knowledge about the biological roles of this protein in the development of HCC.

Therapeutic potential of RNA interference: a new molecular approach to antiviral treatment for hepatitis C.

Hepatitis C virus (HCV) infection remains a major cause of chronic liver disease with an estimated 170 million carriers worldwide. Current treatments have significant side effects and have met with only partial success. Therefore, alternative antiviral drugs that efficiently block virus production are needed. During recent decades, RNA interference (RNAi) technology has not only become a powerful tool for functional genomics but also represents a new therapeutic approach for treating human diseases including viral infections. RNAi is a sequence-specific and post-transcriptional gene silencing process mediated by double-stranded RNA (dsRNA). As the HCV genome is a single-stranded RNA that functions as both a messenger RNA (mRNA) and replication template, it is an attractive target for the study of RNAi-based viral therapies. In this review, we will give a brief overview about the history and current status of RNAi and focus on its potential application as a therapeutic option for treatment for HCV infection.

Post-infarction myocardial rupture: a case of pericardial tamponade salvaged by auto-blood transfusion.

Left ventricular free-wall rupture (LVFWR) is a serious and lethal complication of acute myocardial infarction. Although this complication is not common, the fatality rate is high due to haemodynamic collapse in the setting of cardiac tamponade. We report a case of LVFWR in a patient with a rare blood group, who survived because of an innovative technique for pericardiocentesis and simultaneous transfusion of the aspirated blood into the femoral sheath. A video of the patient's ventriculography is provided.

Halothane: how should it be used in a developing country?

The anaesthetic agent halothane is still widely used in developing countries including the Islamic Republic of Iran because of its low price. Because of halothane-induced hepatitis, a rare complication, it has been replaced by other inhalation anaesthetics in Western countries; it has been suggested by some Iranian professionals that the Islamic Republic of Iran should do the same. We evaluated various dimensions of this replacement through a literature review to assess the incidence of halothane-induced hepatitis and costs of anaesthetics in the country. We also conducted a questionnaire survey of 30 anaesthesiology/gastroenterology experts about their views on the subject. The results indicate that the incidence of halothane hepatitis in the Islamic Republic of Iran is very low and could mostly be avoided by strict adherence to guidelines. Complete withdrawal of halothane in the Islamic Republic of Iran might not be appropriate at present. Comprehensive cost-effectiveness studies are needed before a decision is made on complete replacement of halothane with other anaesthetics.

Evaluation and optimization of chitosan derivatives-based gene delivery system via kidney epithelial cells.

PURPOSE: Non-viral vectors have been widely proposed as safer alternatives to viral vectors, and cationic polymers have gained increasing attention because they can form self-assembly with DNA. Chitosan is also considered to be a good candidate for gene delivery systems, since it is already known as a biocompatible, biodegradable, and low toxic material with high cationic potential. However, low solubility and transfection efficiency need to be overcome prior to clinical trial. In this work, we focus on alkyl modified chitosan which might be useful in DNA condensing and efficient gene delivery. METHODS: N, N- Diethyl N- Methyl (DEMC) and N- Triethyl Chitosan (TEC) were synthesized from chitosan polymer. In order to optimize the polymers for gene delivery, we used FITC-dextran (FD). Then the optimized polymer concentrations were used for gene delivery. Fluorescent microscope was used, in order to evaluate the polymers' efficiency for gene delivery to human embryonic kidney epithelial cells (HEK 293T). RESULTS: This modification increased chitosan's positive charge, thus these chitosan derivatives spontaneously formed complexes with FD, green fluorescence protein plasmid DNA (pEGFP), red fluorescence protein plasmid DNA (pJred) and fluorescent labeled miRNA .RESULTS gained from fluorescent microscope showed that TEC and DEMC were able to transfer FD, DNA and miRNA (micro RNA) to HEK cell line. CONCLUSION: We conclude that these chitosan derivatives present suitable characteristics to be used as non-viral gene delivery vectors to epithelial cells.

N-Diethylmethyl chitosan for gene delivery to pancreatic cancer cells and the relation between charge ratio and biologic properties of polyplexes via interpolations polynomial.

In gene therapy of pancreatic cancer, non-viral vectors show an important role. These vectors are modified with the aim of improvement for pancreatic cancer gene therapy. For this aim, we used N,N-diethyl N-methyl chitosan (DEMC) for gene delivery to human pancreatic cancer cells (AsPC-1). pEGFP (Enhanced green fluorescent protein plasmid) was used as a model plasmid. In order to evaluate the efficiency of this polymer for gene delivery, the DEMC/pEGFP complexes are characterized via photon correlation spectroscopy, gel electrophoresis, fluorescence microscopy, flow cytometry and MTT assay. Also cancer cells' mean fluorescence intensity (MFI) and size changes after transfection are evaluated. The enhancement in polyplexes' charge ratios from 5 to 40, results in 16.70-fold increase in transfection efficiency. Higher MFI, cell size and cytotoxicity were observed as the N/P ratio increased. Considering that mathematical models can be used to understand and predict consequences associated with nanomedicine, the relation between DEMC/pDNA complexes charge ratio, cell transfection and toxicity was evaluated for the first time with Lagrange's interpolation polynomial method.

Electrochemical growth of high-aspect ratio nanostructured silver chloride on silver and its application to miniaturized reference electrodes.

The sensitivity of many biological and chemical sensors is critically dependent on the stability of the potential of the reference electrode being used. The stability of a reference electrode's potential is highly influenced by the properties of its surface. In this paper, for the first time, the formation of nanosheets of silver chloride on silver wire is observed and controlled using high anodic constant potential (>0.5 V) and pulsed electrodeposition. The resulting nanostructured morphology substantially improves the electrode's potential stability in comparison with the conventional globular surface structure. The increased stability is attributed to the increase in the surface area of the silver chloride produced by the nanosheet formation.

Hepatitis B virus infection in dentistry: a forgotten topic.

More than two billion people have been infected with hepatitis B virus (HBV). Globally, 350-400 million suffer from chronic HBV infection. It is postulated that dentists and dental staff are infected and transmit the virus to their patients more than any other occupation. The aim of this article is to review the HBV incidence in dental society, the points of view of dentists and their patients regarding transmission of the virus during dental procedures, the occurrence of HBV outbreaks in dental clinics and the importance of methods of preventing HBV infection in dentistry.

Distance to water source and altitude in relation to active trachoma in Rombo district, Tanzania.

OBJECTIVES: To investigate the relationship between distance to water source, altitude and active trachoma in children in Rombo district, Tanzania. METHODS: In each of Rombo's 64 villages, 10 balozis (groups of 8-40 households) were selected at random and all resident children aged 1-9 years were examined for clinical signs of active trachoma. The households of these children and village water sources were mapped using differentially corrected global positioning system data to determine each household's altitude and distance to the nearest water supply. RESULTS: We examined 12 415 children and diagnosed 1171 cases of active trachoma (weighted prevalence=9.1%, 95% CI: 8.0, 10.2%). Active trachoma prevalence ranged from 0% to 33.7% across villages. Increasing distance to the nearest water source was significantly associated with rising trachoma prevalence (age-adjusted odds ratio for infection (OR) for highest quartile compared to lowest=3.56, 95% CI 2.47, 5.14, P for trend <0.0001). Altitude was significantly inversely associated with trachoma prevalence (age-adjusted OR for highest quartile compared to lowest=0.55, 95% CI 0.41, 0.75, P for trend <0.0001). These associations remained significant after adjustment in multivariate analysis. CONCLUSIONS: Trachoma is endemic in Rombo district, although the prevalence varies considerably between villages. Spatial mapping is a useful method for analysing risk factors for active trachoma.

Development of immunoaffinity solid phase microextraction probes for analysis of sub ng/mL concentrations of 7-aminoflunitrazepam in urine.

We report on the development of solid phase microextraction probes for drug analysis, prepared with antibodies specific for benzodiazepines covalently immobilized to the surface. In the technique, immobilized antibody probes are exposed to a sample containing the drug for 30 min. Extracted drugs are subsequently desorbed from the probes in 500 microL of methanolic desorption solution, which is dried, reconstituted in a small volume of injection solution and analysed by LC-MS/MS. The antibodies were characterized both before and after immobilization, to facilitate the rational selection of antibodies for such analyses. Polyclonal and monoclonal antibodies were compared as was the impact of affinity purification of the polyclonal antibody to isolate the drug-specific fraction. The probes were evaluated for utility in analyzing 7-aminoflunitrazepam at sub ng/mL concentrations in urine, which is expected to be found several days after a single oral dose of 2 mg of flunitrazepam. Such analyses are required in monitoring for abuse of this drug, both in terms of 'club drug' use and in cases of drug-facilitated sexual assault. In these cases drug concentrations in blood and urine are much lower than in chronic abuse cases and are difficult to analyse by conventional methods. The method developed has a limit of detection of 0.02 ng/mL, with accuracy ranging from 1% to 27% and precision (% R.S.D.) ranging from 2% to 10% between the lower and upper limits of quantitation for the analysis of 7-aminoflunitrazepam in urine. The dynamic range of the method is from 0.02 ng/mL, which is limited by the instrument sensitivity, to 0.5 ng/mL, which is approaching the capacity of the probes. This would allow for quantitative analysis of samples at concentrations below that measurable by many other methods for general benzodiazepines analysis from urine, and a highly selective screen for samples at higher concentrations. The method has similar limits of detection to the most sensitive literature methods specifically designed for such analysis but with the advantage of significantly simplified sample preparation. This simplification makes the technique more amenable for use by both professionals and non-professionals.