The Suppressive Effect of Mamiran Cream on Atopic Dermatitis-Like Skin Lesions In Vivo

Background. The Chinese herbal formula Mamiran cream (MMC) has been known for its ameliorative effects on diverse skin diseases, such as eczema. Atopic dermatitis (AD; eczema) is a chronic recurrent skin disease dominated by T-helper type 2-driven inflammation (Th2). Objective. In this study, the inhibitory effect of MMC on AD was investigated in vivo. Methods. An animal model was established by sensitization with 2,4-dinitrochlorobenzene (DNCB) on the skin of SD rats. Cutaneous administration of MMC was applied, and its mechanism of action was investigated via RT-PCR and IHC assay. Result. Our data showed that topical application of MMC reduced the skin severity scores and alleviated the histological changes. Furthermore, immunohistochemical analysis demonstrated that MMC significantly decreased the levels of Th2 cytokine IL-5 and IL-4Ra in the skin lesion. In addition, it was demonstrated that MMC downregulated the mRNA expression of TNF-α, IL-1β, IL-6, IL-10, and TLR4. Moreover, MMC inhibited the activation of NF-κB, JNK1, and STAT6 pathways in skin lesions. Conclusions. Our findings suggest that MMC exhibits the inhibitory effect on AD, suggesting that MMC may be a potential therapeutic agent for this atopic disorder.

Scutellarin Reduces Cerebral Ischemia Reperfusion Injury Involving in Vascular Endothelium Protection and PKG Signal

Flavonoid glycoside scutellarin (SCU) has been widely applied in the treatment of cerebral ischemic diseases in China. In this article, we conducted research on the working mechanisms of SCU in hypoxia reoxygenation (HR) injury of isolated cerebral basilar artery (BA) and erebral ischemia reperfusion (CIR) injury in rat models. In isolated rat BA rings, HR causes endothelial dysfunction (ED) and acetylcholine (ACh) induces endothelium-dependent vasodilation. The myography result showed that SCU (100 µM) was able to significantly improve the endothelium-dependent vasodilation induced by Ach. However, SCU did not affect the ACh-induced relaxation in normal BA. Further studies suggested that SCU (10–1000 µM) dose-dependently induced relaxation in isolated BA rings which were significantly blocked by the cGMP dependent protein kinase (PKG) inhibitor Rp-8-Br-cGMPs (PKGI-rp, 4 µM). Pre-incubation with SCU (500 µM) reversed the impairment of endothelium-dependent vasodilation induced by HR, but the reversing effect was blocked if PKGI-rp (4 µM) was added. The brain slice staining test in rats’ model of middle cerebral artery occlusion (MCAO) induced CIR proved that the administration of SCU (45, 90 mg/kg, iv) significantly reduced the area of cerebral infarction. The Western blot assay result showed that SCU (45 mg/kg, iv) increased brain PKG activity and PKG protein level after CIR surgery. In conclusion, our findings suggested that SCU possesses the ability of protecting brain cells against CIR injury through vascular endothelium protection and PKG signal. Graphic Abstract

Synthesis, in Silico Study, and Anti-Cancer Activity of Thiosemicarbazone Derivatives

Thiosemicarbazones are known for their biological and pharmacological activities. In this study, we have synthesized and characterized 3-Methoxybenzaldehyde thiosemicarbazone (3-MBTSc) and 4-Nitrobenzaldehyde thiosemicarbazone (4-NBTSc) using IR, 1HNMR and 13C NMR. The compound’s in vitro anticancer activities against different cell lines were evaluated. Molecular docking, Insilco ADMET, and drug-likeness prediction were also done. The test compounds showed a comparative IC50 and growth inhibition with the standard drug Doxorubicin. The IC50 ranges from 2.82 µg/mL to 14.25 µg/mL in 3-MBTSc and 2.80 µg/mL to 7.59 µg/mL in 4-NBTSc treated cells. The MTT assay result revealed, 3-MBTSc inhibits 50.42 and 50.31 percent of cell growth in B16-F0 and EAC cell lines, respectively. The gene expression showed that tumor suppressor genes such as PTEN and BRCA1 are significantly upregulated in 7.42 and 5.33 folds, and oncogenes, PKC, and RAS are downregulated −7.96 and −7.64 folds, respectively in treated cells. The molecular docking performed on the four targeted proteins (PARP, VEGFR-1, TGF-β1, and BRAFV600E) indicated that both 4-NBTSc and 3-MBTSc potentially bind to TGF-β1 with the best binding energy of −42.34 Kcal/mol and −32.13 Kcal/mol, respectively. In addition, the test compound possesses desirable ADMET and drug-likeness properties. Overall, both 3-MBTSc and 4-NBTSc have the potential to be multitargeting drug candidates for further study. Moreover, 3-MBTSc showed better activity than 4-NBTSc.

RP-HPLC and HPTLC Identification and Quantification of Flavonoids in the Leaves Extract of Bauhinia acuminata Linn. (Caesalpiniaceae)

Aims: The present investigation is aimed at the identification and quantification of flavonoids in the methanol extract of leaves of Bauhinia acuminata Linn. (Caesalpiniaceae) by a validated HPTLC method and confirmation of the same by RP-HPLC. Methodology: CAMAG HPTLC system with VisionCATS software and HPLC Agilent Infinity 1260 were employed for the study. Pre-coated silica gel 60 F254 plates were used as stationary phase and Toluene : Ethyl acetate : Formic acid (5:4:0.2, v/v/v) was used as mobile phase in HPTLC while in HPLC Thermo hypersil BDS C18 column and Methanol and 0.4% Phosphoric acid (65 : 35) were used as stationary and mobile phase respectively. Results: Among different standards used in HPTLC, only quercetin was found to be present and further quantitatively analyzed. The method shows good correlation coefficient (R2 ≥ 0.99) and linearity in the range of 0.5 to 5.0 µg/band. The concentration of quercetin in the test extract was found to be 0.514 µg/mg of the extract. The LOD and LOQ values were 0.84 µg/band and 2.59 µg/band respectively. The RP-HPLC study also represented good % RSD of Retention time (0.025) and Area (0.09) which signifies the high precision and repeatability. The assay result (4.99 %) also indicated good presence of the quercetin in the extract. Conclusion: The methods were rapid, cost effective and may be employed for quality-control and standardization of quercetin in different plant extracts.

Evaluation of antioxidant activity of unpurified and purified datura seed

Aim: Evaluation of effect of shodhana process of datura seed (Datura stramonium) antioxidant activity. Methods: The datura was purified with two different processes viz. cow’s urine, and cow’s milk by Dolayantra method. Antioxidant activity was evaluated by in-vitro 1,1-Diphenyl-2-picryl hydrazyl (DPPH) radicals scavenging activity and Ferric reducing power ability (FRPA) assay. Result: cow urine purified datura (CUD) showed a significant effect in inhibiting DPPH, reaching up to 85.96% at concentration 1000 mcg/ml and its IC50 was 314.3 mcg/ml while Unpurified Datura Seed (UD) reaching up to 86.06% at concentration 1000 mcg/ml and its IC50 was 171.73 mcg/ml. The Cow Milk Purified Datura Seed (CMD) extract reach 69.72% at concentration 1000 mcg/ml and its IC50 value was 640. The IC50 value of Ascorbic acid was 23.35 mcg/ml. CUD showed a significant effect in reduction of ferric ion, reaching up to 85.59% reduction at concentration 1000 mcg/ml and its EC50 was 304.46 mcg/ml while UD reaching up to 86.11% at concentration 1000 mcg/ml and its EC50 was 280.28 mcg/ml. CMD extract reach 52.28% at concentration 1000 mcg/ml and its EC50 value was 1091.3mcg/ml.

Simultaneous Determination and Validation of Flupirtine Maleate and Paracetamol in Combined Dosage Form by Chromatographic Technique

Objective: The focus of this research was to establish a validated high-performance thin layer chromatographic (HPTLC) method for analysing Flupirtine maleate and Paracetamol in a combined dosage form. Method: Paracetamol and Flupirtine maleate were measured using a mobile phase of Ethyl acetate: Chloroform (7:5 v/v) at 286 nm. This technique was validated in accordance with the International Conference on Harmonization (ICH) guidelines. Results: The Rf value for paracetamol was 0.31 and 0.52 for Flupirtine maleate in this existing technique. Paracetamol's linearity was found to be in the range of 3250-6500 ng/band, while Flupirtine maleate's linearity was found to be in the range of 1000-2000 ng/band. The method's accuracy was determined by recovery experiments, which revealed a percent recovery of 98 to 102 percent. The % RSD was determined to be less than 2 in the Precision investigation, and the assay result for both compounds was within the limit.

The Correlation between Gene Expression and DNA Methylation Levels of RAS p21 Protein Activator 3 (RASA3) Gene in Saudi Autistic Children

The potential role of DNA methylation pattern in autism has been provided by revealing the differences in methylation level of multiple genes which are significantly associated with their expression and implicated in ASD pathogenesis. RASA3 is a member of GTPase-activating proteins, RASA3 is highly expressed in brain tissues and can be deregulated by different epigenetic mechanisms. Many studies reported that differentially expressed RASA3 is correlated with its aberrant methylation. Accordingly, this has been suggested that deferentially expression of RASA3 may be correlated with its methylation levels which could play a role in ASD which brought our attention to identify differentially-expressed genes that could be associated with their methylation level of ASD in Saudi population, by performing comparative gene expression of RASA3 then investigate its relation to methylation level. This study was conducted on 18 Saudi autistic children as well as their healthy-control siblings. Relative expression of a candidate gene (RASA3) was measured using RT-qPCR. Furthermore, MethyLight assay was performed to estimate methylation level and evaluate its impact on RASA3 expression. Interestingly, RASA3 expression has found to be dysregulated in ASD cases. In contrast, MethyLight assay result showed no differences in the methylation patterns among ASD cases in the candidate region. However, it remains an open question whether these dysregulations of RASA3 expression could be a biomarker for early screening/detection of some cases which may also suggest a role for RasGAPs in autistic brain function.

Quality of Different Brands of Metronidazole Benzoate Oral Suspensions Available at Jimma Town, Southwest Ethiopia: Pharmaceuticals Quality Study

Background: Poor quality drugs include substandard and counterfeit medicines. Poor quality antibiotics selectively kill the susceptible strain and leaves resistant strain to multiply. Metronidazole is a broad spectrum antibiotic whose effectiveness depends up on a dose administered to the patient. Objective: The aim of the study is to assess the quality of different brands of metronidazole Benzoate oral suspension available in Jimma town, West Ethiopia. Methods: Cross-sectional study was conducted in Jimma town, West Ethiopia from Feb 08 – Mar 28, 2018. The assay result of all the seven brands of Metronidazole Benzoate oral suspensions was entered to statistical package for social sciences software version 24.0 for windows. Then, one way analysis of variance was performed using Tukey test to determine whether there exists significant difference in assay result of the brands (p<0.05). Result: All the seven Metronidazole benzoate oral suspensions assessed in this study passed British Pharmacopoeia 2013 specification of identity test of the drug. All the brands passed the assay test and total aerobic microbial count specification United States Pharmacopoeia 2015. The highest percentage of drug content was obtained for Metrolag, 105.56%, while the least content for Mizel, 93.12%. However, statistical comparison of drug contents at 95% confidence interval indicates that there is significant difference in drug content within and among the seven brands of Metronidazole benzoate oral suspensions (p<0.05). The pH of all the brands was with in United States Pharmacopoeia 2015 specification limit.

Mass spectrometric analysis of digesta does not improve the allergenicity assessment of GM crops

An investigation of the potential allergenicity of newly expressed proteins in genetically modified (GM) crops comprises part of the assessment of GM crop safety. However, allergenicity is not completely predictable from a definitive assay result or set of protein characteristics, and scientific opinions regarding the data that should be used to assess allergenicity are continuously evolving. Early studies supported a correlation between the stability of a protein exposed to digestive enzymes such as pepsin and the protein’s status as a potential allergen, but over time the conclusions of these earlier studies were not confirmed. Nonetheless, many regulatory authorities, including the European Food Safety Authority (EFSA), continue to require digestibility analyses as a component of GM crop risk assessments. Moreover, EFSA has recently investigated the use of mass spectrometry (MS), to make digestion assays more predictive of allergy risk, because it can detect and identify small undigested peptides. However, the utility of MS is questionable in this context, since known allergenic peptides are unlikely to exist in protein candidates intended for commercial development. These protein candidates are pre-screened by the same bioinformatics processes that are normally used to identify MS targets. Therefore, MS is not a standalone allergen identification method and also cannot be used to predict previously unknown allergenic epitopes. Thus, the suggested application of MS for analysis of digesta does not improve the poor predictive power of digestion assays in identifying allergenic risk.

Comparative assessment of SARS-CoV-2 serology in healthcare workers with Abbott Architect, Roche Elecsys and The Binding site ELISA immunoassays.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) serology testing is key for assessing seroprevalence and antibody response post-vaccination in immunocompromised patients. Here we performed a comparison between two high-throughput nucleocapsid assays (Abbott SARS-CoV-2 IgG and Roche Elecsys Anti-SARS-CoV-2) and The Binding Site (TBS) anti-Spike IgG/A/M- SARS-CoV-2 ELISA kit. 236 samples were collected across 2 sites, Portsmouth Hospital University NHS Trust (PHU) and The Dudley Group NHS Trust. We derived concordance, agreement and assay performance as well as using receiver operating characteristic (ROC) curves to redefine the assay threshold of the Abbott assay. Result concordance between the Abbott and TBS was 66%. Discrepant samples were analysed using the Roche assay which showed 100% agreement with the TBS assay. In samples analysed >58 days post-PCR, the sensitivity of Abbott and Roche was 100%. In samples analysed >100 days post-PCR the sensitivity of the Abbott assay dropped to 77.2% but remained at 100% for the Roche assay. A redefined Abbott threshold of 0.64 increased the sensitivity to 90% giving results similar to the Roche and TBS assays. In conclusion, this study demonstrated Abbott assay had a lower sensitivity in comparison to TBS and Roche. This study established TBS can be implemented as a viable alternative for SARS-CoV-2 serology testing where high-throughput assays are not available on site. Furthermore, anti-spike assays, such as TBS, could be used to monitor vaccination responses to deduce SARS-CoV-2 population-immunity. Further optimization studies are required to evaluate the performance characteristics of these assays which could facilitate widescale sero-epidemiological surveillance.