Methadone-induced encephalopathy: a case series and literature review.

BACKGROUND: Accidental ingestion or consumption of supra-therapeutic doses of methadone can result in neurological sequelae in humans. We aimed to determine the neurological deficits of methadone-poisoned patients admitted to a referral poisoning hospital using brain magnetic resonance (MR) and diffusion weighted (DW) imaging. METHODS: In this retrospective study, brain MRIs of the patients admitted to our referral center due to methadone intoxication were reviewed. Methadone intoxication was confirmed based on history, congruent clinical presentation, and confirmatory urine analysis. Each patient had an MRI with Echo planar T1, T2, FLAIR, and DWI and apparent deficient coefficient (ADC) sequences without contrast media. Abnormalities were recorded and categorized based on their anatomic location and sequence. RESULTS: Ten patients with abnormal MRI findings were identified. Eight had acute- and two had delayed-onset encephalopathy. Imaging findings included bilateral confluent or patchy T2 and FLAIR high signal intensity in cerebral white matter, cerebellar involvement, and bilateral occipito-parietal cortex diffusion restriction in DWI. Internal capsule involvement was identified in two patients while abnormality in globus pallidus and head of caudate nuclei were reported in another. Bilateral cerebral symmetrical confluent white matter signal abnormality with sparing of subcortical U-fibers on T2 and FLAIR sequences were observed in both patients with delayed-onset encephalopathy. CONCLUSIONS: Acute- and delayed-onset encephalopathies are two rare adverse events detected in methadone-intoxicated patients. Brain MRI findings can be helpful in detection of methadone-induced encephalopathy.

Prevalence of the Most Common Virulence-Associated Genes among Brucella Melitensis Isolates from Human Blood Cultures in Hamadan Province, West of Iran.

Brucellosis is a widespread zoonotic disease causing considerable economic and public health problems. Despite animal vaccination, brucellosis remains endemic in some areas such as Iran, especially in the western Iranian province of Hamadan. We sought to detect some of the most common virulence-associated genes in Brucella isolated from human blood cultures to determine the prevalence of some virulence genes among Brucella isolates. Fifty-seven isolates were studied from patients with a clinical diagnosis of brucellosis who referred to the Infectious Diseases Ward of Sina Hospital in Hamadan Province, Iran, between April 2013 and July 2014. Blood samples were collected for the diagnosis of brucellosis using the BACTEC blood culture system. All of these isolates were confirmed by the bcsp31 Brucella-specific gene. We detected 11 virulence-associated genes of Brucella, namely cßg, virB, znuA, ure, bvfA, omp25, omp31, wbkA, mviN, manA, and manB, which are important for the pathogenesis of this bacterium in the intracellular environment by multiplex PCR. Totally, 149 patients with a clinical diagnosis of brucellosis were enrolled in this study. Fifty-seven (38.3%) patients had positive blood cultures. On biochemical and molecular testing, all of the isolates were Brucella melitensis. Ten of the virulence genes were detected among all of the 57 isolates, but the bvf gene was detected in 53 (93%) isolates. The high prevalence of virulence-associated genes among the Brucella isolates detected in Hamadan Province, Iran, underscores the pathogenicity of this bacterium in this region.

Long-Term Cost-Effectiveness of Continuous Glucose Monitoring Versus Self-Monitoring of Blood Glucose in Adults With Type 1 Diabetes in Iran.

OBJECTIVES: This study aimed to determine long-term cost-effectiveness of continuous glucose monitoring (CGM) technology versus self-monitoring of blood glucose (SMBG) in adults with type 1 diabetes (T1D) using multiple daily injections in Iran. METHODS: According to available data, the long-term costs and clinical outcomes of CGM and SMBG were estimated using the Sheffield Type 1 Diabetes Model, with a lifetime horizon from a payer's perspective. The primary outcome was the cost per quality-adjusted life year (QALY) gained. RESULTS: The lifetime cost-effectiveness analysis demonstrated that on average, the use of CGM increased life expectancy by 1.32 years and QALYs by 1.63, compared with SMBG. The CGM group had an average discounted total cost of $40 093 US dollars, whereas the SMBG group had an average discounted total cost of $13 366. This resulted in an incremental cost-effectiveness ratio (ICER) of $16 386 per QALY gained, which is less than the threshold of 3 times the gross domestic product (GDP) per capita of Iran ($24 561). CONCLUSIONS: Considering 3 times the GDP per capita as the threshold, CGM is likely to be cost-effective in Iran. However, for CGM to be very cost-effective (ie, have an ICER less than 1 times the GDP per capita) and presumably more accessible, the price of CGM should decrease to $40 per sensor, each with a lifespan of 14 days.

Assessing the Prevalence of Pulmonary Embolism and the Clot Burden in Hospitalized Patients with Chronic Obstructive Pulmonary Disease Exacerbation.

OBJECTIVE: This prospective cohort study aimed to assess the pulmonary embolism (PE) rate and clot burden in patients with chronic obstructive pulmonary disease (COPD) exacerbation. MATERIAL AND METHODS: A total of 248 patients entered the study, and their clinical probability of PE was assessed using the Geneva score. Patients with high clinical probability underwent computed tomographic pulmonary angiography, while those with low or intermediate probability underwent a d-dimer test. RESULTS: Among the patients analyzed, 14 individuals (5.6%) were confirmed to have PE using computed tomographic pulmonary angiography. A 3-month follow-up revealed 3 cases of PE out of 232 patients initially deemed PE-free. Mortality rates were higher among patients with venous thromboembolism at admission than those diagnosed with PE during follow-up. Pulmonary embolism (PE) prevalence among patients with COPD exacerbation was 5.6%. CONCLUSION: The results of this study show the importance of screening for PE in patients with COPD presenting with dyspnea. Not all of them are due to COPD exacerbation; a small minority of them can be due to PE, which needs prompt screening, confirmation, and therapy. However, further research with larger cohorts is required to understand better the potential benefits and implications of systematic screening for pulmonary embolism in this specific patient population.

Does Adding the Pulmonary Infarction and Right Ventricle to Left Ventricle Diameter Ratio to the Qanadli Index (A Combined Qanadli Index) More Accurately, Predict Short-Term Mortality in Patients with Pulmonary Embolism?

Background The Qanadli index can be used to assess the severity of pulmonary arterial involvement in patients with acute pulmonary embolism. However, it seems that considering pulmonary infarction and right ventricle/left ventricle (RV/LV) ratio along with this index (called the combined Qanadli index) can provide a more accurate view of changes in cardiovascular parameters in these patients and help predict mortality in a better manner. In this regard, we evaluated the ability of the combined Qanadli index versus the Qanadli index in predicting short-term mortality in patients with pulmonary embolism. Methods This retrospective study enrolled 234 patients with acute pulmonary embolism. Patients were divided into two groups: those who expired in 30 days and who survived. Then they were evaluated by computed tomography angiography of pulmonary arteries. The RV/LV diameter ratio and also pulmonary artery obstruction index (PAOI) were calculated. The patient's computed tomography scans were reviewed for pulmonary infarction. By adding the RV/LV ratio and pulmonary infarction to PAOI, a new index called the modified Qanadli score was made. Univariable and multivariable logistic regression was done for finding predictors of mortality. Results Nine cases (40%) of patients in the mortality group and 42 (20%) of survivors had ischemic heart disease and the difference was significantly meaningful. The mean Qanadli index in the mortality group was 16.8 ± 8.45 and in survivors was 8.3 ± 4.2. By adding the pulmonary infarction score and PAOI score to RV/LV ratio score, the odds ratio (OR) for predicting mortality increased significantly to 13 and 16, respectively, which were significantly meaningful. Based on our findings, the highest OR for predicting short-term mortality was obtained through a combined Qanadli index (PAOI score + pulmonary infarction score + RV/LV score) that was 17 in univariable and 18 in multivariable logistic regression analysis ( p -value = 0.015). Conclusion The new combined Qanadli index has more ability than the Qanadli index and RV/LV ratio for predicting changes in cardiovascular parameters and short-term mortality in patients with pulmonary embolism.

Delivery of LNA-antimiR-142-3p by Mesenchymal Stem Cells-Derived Exosomes to Breast Cancer Stem Cells Reduces Tumorigenicity.

Exosomes, nano-sized cell-derived vesicles, have been employed as non-synthetic carriers of various pharmaceutics in numerous studies. As higher expression levels of miR-142-3p and miR-150 in breast cancer stem cells (BCSCs) are associated with their clonogenic and tumorigenic capabilities, the present study aims to exploit the mesenchymal stem cells-derived exosomes (MSCs-Exo) to deliver LNA-antimiR-142-3p into MCF7-derived cancer stem-like cells to suppress expression levels of miR-142-3p and miR-150 in order to reduce clonogenicity and tumorigenicity. Our results indicated that the MSCs-Exo can efficiently deliver the LNA-antimiR-142-3p to breast cancer stem-like cells to reduce the miR-142-3p and miR-150 expression levels. Furthermore, the inhibition of the oncomiRs with the delivery of LNA-antimiR-142-3p resulted in a significant reduction of clone-formation and tumor-initiating abilities of the MCF7-derived cancer stem-like cells. In conclusion, we showed that MSCs-derived exosomes could be used as a feasible nanovehicles to deliver RNA-based therapeutics into BCSCs to improve the cancer treatment. HIGHLIGHTS: Exosomes secreted by bone marrow-derived mesenchymal stem cells efficiently transfer the LNA-antimiR-142-3p to breast cancer stem cells. Exosomes-mediated delivery of LNA-antimiR-142-3p to the breast cancer stem cells leads to downregulation of miR-142-3p and miR-150 and the overexpression of target genes. Delivery of LNA-antimiR-142-3p by the exosomes reduces the colony formation capability of breast cancer stem cells in vitro. Inhibition of miR-142-3p and miR-150 by the LNA-antimiR-142-3p loaded exosomes reduces the tumorigenicity of breast cancer stem cells in vivo.

Polymorphisms in Promoter Region of the Interferon-Gamma Receptor-1 Gene and its Relation with Susceptibility to Brucellosis.

BACKGROUND & OBJECTIVE: Brucellosis is one of the most prevalent bacterial zoonotic diseases which afflicts both humans and animals. Genetic factors play an important role in susceptibility to brucellosis. One of these factors is interferon-gamma (IFN-), which is vital in the defense mechanism against infectious diseases such as brucellosis. The purpose of this study was to evaluate the relationship between two single nucleotide polymorphisms (SNPs) at positions -611 and -56 within the promoter region of interferon-gamma receptor-1 gene (IFN- R1) and brucellosis. METHODS: In this research, the genomic DNA was collected from 60 peripheral blood samples infected with brucellosis and 68 healthy volunteers. DNA was extracted by salting out method. Then, DNA genotypes were analyzed using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP). RESULTS: The results showed that there is a significant difference in -611 SNP frequencies between control and patient groups. At position -611, CC genotype was related to patient group (P=0.024) and TT genotype was related to the control group. According to the results, males had a higher frequency of Brucella infection. CONCLUSION: The presence of C allele in position -611 in IFNγ R1 gene promoter was related to a higher risk of disease and susceptibility to brucellosis. Moreover, the presence of T allele in position γ.

Exosome-mediated delivery of functionally active miRNA-142-3p inhibitor reduces tumorigenicity of breast cancer in vitro and in vivo.

BACKGROUND: Exosomes, widely recognized natural nanovesicles, represent one of the recently discovered modes of intercellular communication due to their ability to transmit crucial cellular information that can be engineered to have robust delivery and targeting capacity. MiR-142-3p, one of the upregulated microRNAs (miRNAs) in many types of breast cancer, activates the canonical Wnt signaling pathway and transactivates the miR-150 expression, and results in the hyperproliferation of cancer cells in vitro and mammary glands in vivo. MATERIALS AND METHODS: In this study, we exploited the exosomes isolated from bone marrow-derived mesenchymal stem cells (MSCs-Exo) to deliver LNA (locked nucleic acid)-modified anti-miR-142-3p oligonucleotides to suppress the expression level of miR-142-3p and miR-150 in 4T1 and TUBO breast cancer cell lines. RESULTS: The in vitro results showed that the MSCs-Exo can efficiently deliver anti-miR-142-3p to reduce the miR-142-3p and miR-150 levels and increase the transcription of the regulatory target genes, APC and P2X7R. We also evaluated in vivo distribution of the MSCs-Exo in tumor-bearing mice. The in vivo result indicated that MSCs-Exo can penetrate the tumor site and are suitable nanovehicles to deliver the inhibitory oligonucleotides into the tumor tissues to downregulate the expression levels of miR-142-3p and miR-150. CONCLUSION: We showed that MSCs-derived exosomes could be used as a feasible nanovehicle to deliver drug molecules like LNA-anti-miR-142-3p in both in vitro and in vivo studies.

A fires novel report of exosomal electrochemical sensor for sensing micro RNAs by using multi covalent attachment p19 with high sensitivity.

Exosomes are natural spherical phospholipids vesicles derived from cells. Dropping MCF-7 exosome (with high biomarkers as exosomal case) covalently onto the SCPE-GNP causes the sensor to be stable due to the electrochemical induction of positive charges of different biomarkers with [Fe(CN)6] -3/-4 reactions. In the following, the covalent p19 connection with the biomarkers of exosome turns off the sensor. After adding the hybrid of miR21-probe, its tight coupling to p19 has reestablished the system. As a result, this sensor has been able to detect miR21 with high sensitivity and specificity. For the first time, the exosomal electrochemical properties were proven as the electrochemical amplifier bed. The limit of detection (LOD) was 1 a.M. due to the existence of various biomarkers for connecting covalent to p19. Electrochemical impedance (EIS) and differential pulse voltammetry (DPV) are used for miR sensing on the MCF-7 exosome-p19 composite. Transmission Electron Microscopy (TEM), Atomic Force Microscopy (AFM), Dynamic Light Scattering (DLS) and Ultraviolet Visible (UV) spectroscopic techniques are used to understand the interactions between each layer. These vesicles can be used as a natural source for biocompatible devices that are used in transfection using an electrochemical method at a cost-effective and high-performance basis.

Molecular detection of Neospora caninum infection in ovine aborted foetuses in the Mashhad area, Iran

N. caninum could cause abortion in small ruminants. The aim of the study was to detect N. caninum infection in ovine aborted foetuses in the Mashhad area by PCR examination. During the period 2009 to 2013, 71 ovine aborted foetuses were collected and their brain samples examined by PCR. Of the 71 brains of the aborted foetuses, N. caninum DNA was detected in seven (9.8%) samples. In conclusion, it seems that N. caninum may act as a causative agent of abortion in sheep in the Mashhad area.

Clinical and Diagnostic Aspects of Brucellosis and Antimicrobial Susceptibility of Brucella Isolates in Hamedan, Iran.

Current drug regimens for brucellosis are associated with relatively high rates of therapeutic failure or relapse. Reduced antimicrobial susceptibility of Brucella spp. has been proposed recently as a potential cause of therapeutic failure. The aim of this study was to evaluate the antibiotic resistance pattern of Brucella melitensis clinical isolates by E-test method in Hamadan, west of Iran. In a 15-month period, all patients with suspected brucellosis were enrolled. Blood specimens were collected for diagnosis of brucellosis by BACTEC system and serological tests. Antimicrobial susceptibility of clinical isolates to 7 antibiotics was assessed by the E-test method. One hundred forty-nine patients with brucellosis were evaluated. 38.3% of cultures of clinical samples were positive for BACTEC system, of which 91.2% were associated with a positive serological test result. No significant associations were found between serology and the culture method. All Brucella isolates were susceptible to doxycycline, streptomycin, gentamicin, ciprofloxacin, and moxifloxacin. However, decreased sensitivity to rifampin and trimethoprim-sulfamethoxazole was found in 35.1% and 3.5% of isolates, respectively. Because of the high rates of intermediate sensitivity to rifampin among Brucella isolates, this drug should be prescribed with caution. We recommend restricting the use of rifampin for treatment of brucellosis except as an alternative drug for special situations.

The Optimization of Molecular Detection of Clinical Isolates of Brucella in Blood Cultures by eryD Transcriptase Gene for Confirmation of Culture-Negative Samples.

BACKGROUND: Brucellosis is a zoonosis disease which is widespread across the world. OBJECTIVES: The aim of the present study is the evaluation of culture-negative blood samples. MATERIALS AND METHODS: A total of 100 patients with suspected brucellosis were included in this experimental study and given positive serological tests. Diagnosis was performed on patients with clinical symptoms of the disease, followed by the detection of a titer that was equal to or more than 1:160 (in endemic areas) by the standard tube agglutination method. Blood samples were cultured by a BACTEC 9050 system, and subsequently by Brucella agar. At the same time, DNA from all blood samples was extracted by Qiagen Kit Company (Qia Amp Mini Kit). A molecular assay of blood samples was carried out by detection of eryD transcriptase and bcsp 31 genes in specific double PCR reactions. The specificity of the primers was evaluated by DNA from pure and approved Brucella colonies found in the blood samples, by DNA from other bacteria, and by ordinary PCR. DNA extraction from the pure colonies was carried out by both Qiagen Kit and Chelex 100 methods; the two were compared. RESULTS: 39 cases (39%) had positive results when tested by the BACTEC system, and 61 cases (61%) became negative. 23 culture-positive blood samples were randomly selected for PCR reactions; all showed 491 bp for the eryD gene and 223 bp for the bcsp 31 gene. Interestingly, out of 14 culture-negative blood samples, 13 cases showed positive bonds in PCR. The specificity of the PCR method was equal to 100%. DNA extraction from pure cultures was done by both Chelex 100 and Qiagen Kit; these showed the same results for all samples. CONCLUSIONS: The results prove that the presented double PCR method could be used to detect positive cases from culture-negative blood samples. The Chelex 100 method is simpler and safer than the use of Qiagen Kit for DNA extraction.

Effects of short-term over-supplementation of copper in milk on hematology, serum proteins, weight gain, and health in dairy calves.

Thirty-six calves were used in the present study. The animals were divided equally into three groups (control, test 1, and test 2). The three groups of calves were homogeneous for parity of dams, sex, and month of birth. From 14 days of age, in the test 1 group copper as copper sulfate (Merck Co, Germany) was added to each meal of milk at a rate of 10 mg/kg of milk for 14 days and in test 2 group copper as copper sulfate was added to each meal of milk at a rate of 20 mg/kg of milk for 14 days. Blood samples were taken by jugular venipuncture using disposable syringes at 14 (before Cu supplementation), 30, 60, and 80 days of age. Anticoagulated blood was used for CBC determination. Plane tubes were used for harvesting of serum and the amounts of total serum protein, albumin, iron, and copper were measured. Calves were weighted at birth and at the end of trial (day 80) and total gain and mean daily gain were calculated. Days of treatment for ill calves were also recorded during experiment. Group (treatment) had no significant effect on the amounts of measured parameters except MCH values (p < 0.05) which were significantly lower in test 1 group than other trial groups. Age (sampling time) had significant effects on the values of most measured parameters (p < 0.05) except WBC, lymphocyte, total protein, and fibrinogen. Significant interactions between sampling time and group were not seen for any of measured parameters. No significant differences were seen for total weight gain and mean daily gain between trial groups. Chi-square test revealed no significant difference for the days of treatment between trials groups.

Ileal relaxation induced by Mentha longifolia (L.) leaf extract in rat.

The effect of Mentha longifolia (L.) leaf hydroalcoholic extract (MLE) was examined on rat ileal smooth muscle contractions. Last portion of ileum from male adult Wistar rat was mounted in an organ bath containing Tyrode solution. The tissue was contracted by carbachol (CCh, 10 microM), KCl (60 mM) and BaC12 (4 mM) and then MLE (0.0625-1 mg mL(-1)) was added to the bath cumulatively. The effect of MLE on KCl-induced contraction was examined after tissue incubation with propranolol (1 microM), naloxone (1 microM) and N omega-nitro-L-arginine methyl ester (L-NAME, 100 microM). The effect of MLE on CaCl2-induced ileal contraction in Ca(2+)-free with high potassium Tyrode solution was also evaluated. The role of potassium channels was examined by ileum incubation (5 mim) with tetraethylammonium (TEA, 1 mM). The results showed that KCl-, CCh and BaCl2-induced ileal contractions were inhibited (p < 0.001) by cumulative concentrations of MLE with the same potency. In addition, MLE (0.25-1 mg mL(-1)) inhibited (p < 0.01) ileal contractions induced by CaCl2 (0.45-2.7 mM) in a concentration-related manner. The antispasmodic effect of MLE was affected neither by propranolol, L-NAME nor by naloxone. The MLE concentration-response curve was shifted to the right (p < 0.05) by tissue incubation with TEA. From results it may be suggested that Mentha longifolia hydroalcoholic leaf extract induces its spasmolytic activity mainly through disturbance in calcium mobilization and partly by potassium channels activation. Present results show that Mentha longifolia leaf extract exerts relaxant effects on intestinal smooth muscle, consistent with the traditional use of the plant to treat gastrointestinal disorders such as diarrhea and colic.