Effect of Echinacea alone or in combination with silymarin in the carbon-tetrachloride model of hepatotoxicity

Echinacea preparations are widely used in the prevention or treatment of upper respiratory tract infections. The present study aimed to investigate the effect of a standardized Echinacea extract in experimentally induced liver toxicity and whether this herb would have a modulating effect on the silymarin-induced hepatoprotection in rats. Liver damage was induced by the administration of carbon tetrachloride (CCl₄). Echinacea extract (18 or 36?mg/kg) alone or combined with silymarin (25?mg/kg), silymarin only (25?mg/kg), or saline (control) was given once daily orally simultaneously with CCl₄ and for 1?week thereafter. Serum alanine aminotransferase (ALT) or aspartate aminotransferase were not significantly changed by treatment with Echinacea, but alkaline phosphatase (ALP) in serum decreased by 23.6% by the extract at 36?mg/kg. Silymarin given in combination with either dose of Echinacea resulted in 34.9% and 57.8% reduction in AST, 42.7% and 58% reduction in ALT and 41% and 60% reduction in ALP, compared with CCl₄ control group. Silymarin alone reduced ALT, AST, and ALP levels by 58.8%, 61.2%, and 62.2%, respectively. Histopathological examination revealed a mild decrease in degenerated hepatocytes after treatment with 36?mg/kg of Echinacea. Noticeable improvement in the liver damage was observed upon the addition of silymarin to Echinacea. A marked decrease of intracellular protein and glycogen staining was evident after the administration of CCl₄. Slight improvement in protein and glycogen staining was noted after 36?mg/kg of Echinacea. Increased hepatic protein and glycogen staining intensity was observed after silymarin and Echinacea co-treatment compared with Echinacea-only treated groups. Silymarin only treatment resulted in more or less normal histopathological and histochemical findings. The present results suggest that administration of Echinacea extract reduces the hepatoprotective effect of silymarin. Such finding is likely to have important clinical significance in patients with hepatic disease on silymarin treatment.     

Amino Acid and Acylcarnitine Profiles in Premature Neonates: A Pilot Study

Objective

To investigate the longitudinal changes in amino acid (AA) and acylcarnitine (AC) profiles of preterm neonates over the first 2 wk of life, and to detect any significant deviation from full term values that requires change of cut-off values used for detection of metabolic disorders in preterm neonates.

Methods

This observational analytical longitudinal study was conducted on 131 premature neonates (gestational age ranged from 27 to 36 wk) and 143 healthy full-term neonates. Dried blood spots were taken on the 5th and 14th postnatal day from the premature neonates and on day 5 from full term neonates for neonatal screening. Samples were analyzed for AA and AC using tandem mass spectrometer.

Results

Most AA significantly decreased on day 14 compared to day 5 among preterm neonates (p?<?0.05). The combined values of total carnitine (TC), total acylcarnitine (tAC) and short-chain acylcarnitines on day 5 among preterm neonates were statistically significantly higher compared to the day 14 sample (p 0.0001), whereas no statistically significant difference was found regarding the values of medium-, long-chain acylcarnitines, tAC/FC, and FC/TC (p?>?0.05). The levels of AA of preterm neonates were statistically significantly higher than that of the controls (p?<?0.05). The values of TC, tAC, short-, medium- and long-chain acylcarnitines, were significantly higher than those of the controls (p?<?0.05). The reference ranges for preterm neonates were determined using the 1st and 99.9th percentiles.

Conclusions

AA and AC showed an age-related distribution of their concentrations. This underlines the importance of using appropriate reference values when working with a prematurely born population.     

Correlation Between Vocal Functions and Glottal Measurements in Patients With Unilateral Vocal Fold Paralysis

Observations and analysis of glottal characteristics are critical in choosing the best modality for surgery in patients with unilateral vocal fold paralysis (UVP). This study suggests that multiple glottal characteristics influence the vocal product in patients with UVP. In addition to the horizontal position of the paralyzed vocal fold (deviation from the midline), the glottal area, degree of bowing of the paralyzed and contralateral vocal folds, maximum separation between vocal folds, compensatory glottal maneuvers, and the vertical glottic closure plane significantly influenced the quality of the voice. Clinicians should be aware of these observations to facilitate treatment planning and assessment of the results of surgical procedures used to improve voice quality in cases of UVP.     

Detection of hepatitis C virus (HCV) RNA in the peripheral blood mononuclear cells of HCV-infected patients following sustained virologic response

It is unclear whether direct-acting antiviral drugs (DAAs) result in the complete eradication of HCV infection or whether some quantities of the virus may persist after achieving a sustained virologic response (SVR). Aim The aim of this work was to study the possibility of the persistence of HCV RNA in peripheral blood mononuclear cells (PBMCs) after achieving SVR following DAA treatment. This study included 100 patients infected with HCV genotype 4, who were candidates for receiving DAAs and who achieved SVR during follow-up, as determined at 12 and/or 24 weeks following the end of treatment. All patients were subjected to demographic, biochemical and hematological assessments. Detection of HCV RNA in the serum and PBMCs and determination of the HCV genotype were performed with real-time PCR. We detected HCV RNA in the PBMCs of 20 out of 100 (20%) patients infected with HCV genotype 4, who achieved SVR. However, the persistent viral load in the PBMCs was very low (range: 400–900 U/mL; mean ± SD: 645.45 ± 153 U/mL). Multiple logistic regression analysis showed that only the higher posttreatment levels of aspartate transaminase (AST) were significantly predictive of HCV RNA persistence in the PBMCs (OR: 1.29; 95% CI: 1.08–1.55). Additionally, according to the Cox proportional hazard model, liver cirrhosis was the only significant risk factor for the persistence of HCV infection in PBMCs (HR: 5.8; 95% CI: 1.3–26.1; P < 0.02). Our results indicated the persistence of HCV RNA in some HCV patients who achieved SVR after treatment with DAAs.

     

A pharmaceutical study on chlorzoxazone orodispersible tablets: formulation,in-vitro and in-vivo evaluation

Context: Muscle spasm needs prompt relief of symptoms. Chlorzoxazone is a centrally muscle relaxant.

Objectives: The aim of this study was to prepare chlorzoxazone orodispersible tablets (ODTs) allowing the drug to directly enter the systemic circulation and bypassing the first-pass metabolism for both enhancing its bioavailability and exerting a rapid relief of muscular spasm.

Materials and methods: ODTs were prepared by direct compression method using Pharmaburst®500, Starlac®, Pearlitol flash®, Prosolv® odt and F-melt® as co-processed excipients. Three ratios of the drug to the other excipients were used (0.5:1, 1:1 and 2:1).

Results and Discussion: All ODTs were within the pharmacopeial limits for weight and content. ODTs containing Pharmaburst®500 showed the shortest wetting time (～45.33?s), disintegration time (DT) (～43.33?s) and dissolution (Q_15min 100.63%). By increasing the ratio of CLZ: Pharmaburst®500 from 0.5:1 to 1:1 and 2:1, the DT increased from 26.43 to 28.0 and 43.33?s, respectively. By using Prosolv® odt, ODTs failed to disintegrate in an acceptable time?>180?s. DT of ODTs using different co-processed excipients can be arranged as follows: Pharmaburst® 500?<?F-melt®?<Pearlitol flash®?<Starlac®?<Prosolv® odt. Pharmacokinetic study of the optimum formula F1 (50?mg CLZ) in rabbits using HPLC-UV detector revealed a shorter T_max (0.333?h) compared with Myofen® capsules (250?mg CLZ) (1.083?h) which is considered a promising treatment, especially for the rapid relief of muscle spasm.

Conclusion: It could be concluded that orodispersible tablets are a promising carrier for CLZ designed for management of muscle spasm due to the enhanced dissolution and rapid absorption of the drug through the oral mucosa.     

Cloning and functional expression of a novel Gi protein-coupled receptor for adenine from mouse brain

An orphan G protein-coupled receptor from the rat has recently been demonstrated to act as a transmembrane receptor for the nucleobase adenine. The receptor is possibly involved in nociception. Here we report the cloning and functional expression of an additional G(i)-coupled receptor for adenine (Genbank accession code DQ386867). mRNA for this receptor was obtained from mouse brain and the mouse neuroblastoma x rat glioma hybrid cell line NG108-15. The new mouse protein sequence shares only 76% identity with that of the rat adenine receptor, suggesting that the receptors are not species homologs but distinct receptor subtypes. In human 1321N1 astrocytoma cells stably expressing the new mouse receptor, adenine and 2-fluoroadenine inhibited the isoproterenol-induced cAMP formation with IC(50) concentrations of 8 and 15 nM, respectively. The adenosine receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 1 muM) as well as the P2 receptor antagonist suramin (300 muM) failed to change the responses to adenine. In contrast, pretreatment of cells with pertussis toxin abolished the effect of adenine. When the novel adenine receptor was expressed in Sf21 insect cells, a specific binding site for [(3)H]adenine was detected. In competition assays, the rank order of potency of selected ligands was identical to that obtained in membranes from NG108-15 cells and rat brain cortex (adenine > 2-fluoroadenine > 7-methyladenine > 1-methyladenine > N(6)-dimethyladenine). In summary, our data show that a second mammalian DNA sequence encodes for a G(i)-coupled GPCR activated by low, nanomolar concentrations of adenine.     

Aspartate transaminase to platelet ratio index in hepatitis C virus and Schistosomiasis coinfection

AIM: To assess the diagnostic accuracy,of aminotransferase-to-platelet ratio index(APRI) alone and with antischistosomal antibody(Ab) in patients with hepatitis C virus(HCV) and schistosomiasis coinfection. METHODS: This retrospective study included medical records of three hundred and eighty three Egyptianmen patients who had undergone percutaneous liver biopsy between January 2006 to April 2014 in tertiary care hospital in Qatar for diagnosis or monitoring purpose were selected. Data of patients 18 years of age were included in the study. The values of HCV RNA titer and antischistosomal antibody titer were also taken into consideration. Patients were excluded from the study if they had any other concomitant chronic liver disease,including; history of previous antiviral or interferon therapy,immunosuppressive,therapy,chronic hepatitis B infection,human immunodeficiency virus co-infection,autoimmune hepatitis,decompensated liver disease,hepatocellular carcinoma,prior liver transplantation,and if no data about the liver biopsy present. RESULTS: Median age of patients was 46 years. About 7.1% had no fibrosis,whereas 30.4%,37.5%,20.4%,and 4.6% had fibrosis of stage Ⅰ,Ⅱ,Ⅲ,and Ⅳ respectively. In bivariate analysis,APRI score,levels of AST,platelet count and age of patient showed statistically significant association with liver fibrosis(P 0.0001); whereas antischistosomal antibody titer(P = 0.52) and HCV RNA titer(P = 0.79) failed to show a significant association. The respective AUC values for no fibrosis,significant fibrosis,severe fibrosis and cirrhosis of APRI score were 63%,73.2%,81.1% and 88.9% respectively. This showed good sensitivity and specificity of APRI alone for grading of liver fibrosis. But the inclusion of anti-Schistosoma antibody did not improve the prediction of fibrosis stage. CONCLUSION: The study results suggest that noninvasive biochemical markers like APRI are sensitive and specific in diagnosing the degree of fibrosis and cirrhosis in patients with coinfection of HCV and schistosomiasis as compared to biopsy. The addition of antischistosomal Ab to APRI did not improve sensitivity for predicting the degree of cirrhosis.     

Prevalence and treatment patterns of ranibizumab and photodynamic therapy in a tertiary care setting in Malaysia

AIM: To describe the prevalence and changes in treatment patterns of ranibizumab and photodynamic therapy (PDT) among retinal disease patients who attended the Ophthalmology Clinic in the tertiary care Hospital Selayang from 2010 to 2014. METHODS: Study subjects were recruited retrospectively using the Electronic Medical Record (EMR) database software in Hospital Selayang. Demographic data, medical history, diagnostic procedure, treatments and diagnosis of patients were recorded. RESULTS: The five-year analysis included 821 patients with a mean age of 65.9±11.73y. Overall, there were a higher number of males (63.1%) and a higher number of Chinese (47.4%) patients. Among the 821 patients, 62.9% received ranibizumab injection followed by 19.2% PDT therapy and 17.9% had ranibizumab combined with PDT therapy. Age-related macular degeneration (AMD) and polypoidal choroidal vasculopathy (PCV) were the most common retinal eye diseases reported, recording prevalence of 25.0% and 45.6%, respectively. The trend in ranibizumab treatment was reported to increase while PDT showed a decrease in trend from year 2010 to 2014. In terms of treatment, following multiple logistic regression, AMD was associated with the subjects being more likely to have received ranibizumab monotherapy (P<0.001) while PCV was associated with more likely to have received PDT (P<0.001) and PDT combined with ranibizumab therapy (P<0.001). CONCLUSION: The tertiary care setting in Malaysia is consistent with management of patients from other countries whereby ranibizumab is the most common treatment given to patients with AMD, while PCV patients most commonly receive PDT and ranibizumab combined with PDT therapy.     

Prevalence and risk factors of gestational diabetes mellitus in Asia: a systematic review and meta-analysis

Background

Gestational diabetes mellitus (GDM) is a of the major public health issues in Asia. The present study aimed to determine the prevalence of, and risk factors for GDM in Asia via a systematic review and meta-analysis.

Methods

We systematically searched PubMed, Ovid, Scopus and ScienceDirect for observational studies in Asia from inception to August 2017. We selected cross sectional studies reporting the prevalence and risk factors for GDM. A random effects model was used to estimate the pooled prevalence of GDM and odds ratio (OR) with 95% confidence interval (CI).

Results

Eighty-four studies with STROBE score?≥?14 were included in our analysis. The pooled prevalence of GDM in Asia was 11.5% (95% CI 10.9–12.1). There was considerable heterogeneity (I² >?95%) in the prevalence of GDM in Asia, which is likely due to differences in diagnostic criteria, screening methods and study setting. Meta-analysis demonstrated that the risk factors of GDM include history of previous GDM (OR 8.42, 95% CI 5.35–13.23); macrosomia (OR 4.41, 95% CI 3.09–6.31); and congenital anomalies (OR 4.25, 95% CI 1.52–11.88). Other risk factors include a BMI ≥25?kg/m² (OR 3.27, 95% CI 2.81–3.80); pregnancy-induced hypertension (OR 3.20, 95% CI 2.19–4.68); family history of diabetes (OR 2.77, 2.22–3.47); history of stillbirth (OR 2.39, 95% CI 1.68–3.40); polycystic ovary syndrome (OR 2.33, 95% CI1.72–3.17); history of abortion (OR 2.25, 95% CI 1.54–3.29); age?≥?25 (OR 2.17, 95% CI 1.96–2.41); multiparity ≥2 (OR 1.37, 95% CI 1.24–1.52); and history of preterm delivery (OR 1.93, 95% CI 1.21–3.07).

Conclusion

We found a high prevalence of GDM among the Asian population. Asian women with common risk factors especially among those with history of previous GDM, congenital anomalies or macrosomia should receive additional attention from physician as high-risk cases for GDM in pregnancy.

Trial registration

PROSPERO (2017: CRD42017070104).

     

Diagnostic Value of Claudin-4 and EZH2 Immunohistochemistry in Effusion Cytology

Background: Metastatic adenocarcinoma (MAC) accounts for most cases of malignant effusions. Sometimes, it can be difficult to distinguish MAC from reactive mesothelial cells (RMC) in cytologic specimens. Our aim was to assess the diagnostic performance of a novel immunohistochemical panel composed of claudin-4 and EZH2 in differentiating MAC from RMC in effusion cytology. Methods: A total of 80 cases of serous effusions (48 MAC and 32 RMC) were included. Immunohistochemistry using claudin-4 and EZH2 was performed on cell block sections of these cases. Assessment of staining patterns, intensity and percentage of target cells stained was done. Results: Claudin-4 showed membranous staining in 46/48 of MAC and 1/32 of RMC. High EZH2 (≥ 50% of target cells) was detected in 42/48 MAC and 2/32 RMC. For the discrimination between MAC and RMC, claudin-4 exhibited 95.8% sensitivity and 96.9% specificity, high-EZH2 exhibited 87.5% sensitivity and 93.8% specificity, while the combination of both claudin-4 and high EZH2 showed 100% sensitivity and 90.6% specificity. Conclusion: Claudin-4 shows high sensitivity and specificity in differentiation between MAC and RMC in effusion cytology, and might be useful as a solitary marker for MAC. Adding EZH2 to claudin-4 increases the sensitivity to 100%. However, the interpretation of EZH2 results can be challenging due to its focal expression in RMC and inflammatory cells.