Τρίτη 10 Νοεμβρίου 2020

Plasma Proteome Profiling of Coronary Artery Disease Patients: Downregulation of Transthyretin—An Important Event

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Coronary artery disease (CAD) is a prevalent chronic inflammatory cardiac disorder. An early diagnosis is likely to help in the prevention and proper management of this disease. As the study of proteomics provides the potential markers for detection of a disease, in the present investigation, attempt has been made to identify disease-associated differential proteins involved in CAD pathogenesis. For this study, a total of 200 selected CAD patients were considered, who were recruited for percutaneous coronary intervention (PCI) treatment. The proteomic analysis was performed using two-dimensional gel electrophoresis (2-DE) and MALDI-TOF MS/MS. Samples were also subjected to Western blot analysis, enzyme-linked immunosorbent assay (ELISA), peripheral blood mononuclear cells isolation immunofluorescence (IF) analys is, analytical screening by fluorescence-activated cell sorting (FACS), and in silico analysis. The representative data were shown as of at least three experiments. A total of 19 proteins were identified. Among them, the most abundant five proteins (serotransferrin, talin-1, alpha-2HS glycoprotein, transthyretin (TTR), fibrinogen-α chain) were found to have altered level in CAD. Serotransferrin, talin-1, alpha-2HS glycoprotein, and transthyretin (TTR) were found to have lower level, whereas fibrinogen-α chain was found to have higher level in CAD plasma compared to healthy, confirmed by Western blot analysis. TTR, an important acute phase transport protein, was validated low level in 200 CAD patients who confirmed to undergo PCI treatment. Further, in silico and in vitro studies of TTR indicated a downexpression of CAD in plasma as compared to the plasma of healthy individuals. Lower level of plasma TTR was determined to be an important risk marker in the atherosclerotic-approved CAD patients. We suggest that the TTR lower level predicts disease severity and hence may serve as an important marker tool for CAD screening. However, further large-scale studies are required to determine the clinical significance of TTR.
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Evaluation of the Antibacterial and Antioxidant Activity of Mimosa rubicaulis and Reinwardtia indica

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Many plants have the property of wound healing, but most of the people are using costly allopathic medicine for the wound. This might be due to lack of awareness about the traditional uses and lack of scientific study of ethnomedicinal plants. So, this study aimed to carry out the antibacterial and antioxidant activity of two medicinal plants which are used traditionally for wound healing activity, i.e.,Mimosa rubicaulis and Reinwardtia indica. Different parts of Mimosa rubicaulis (root, stem, and leaves) and Reinwardtia indica (flower and leaves) were used for the study. Extractions were done by maceration using ethanol as solvent. Antibacterial activity was carried out by the well diffusion method, and antioxidant activities were screened by DPPH radical scavenging and NO scavenging assays. Extract of M. rubicaulis has shown a weak zone of inhibition towards S. aureus and P. vulgaris whereas R. indica has shown no zone of inhibition towards selected bacterial strains. Leaf, root, and stem extracts of M. rubicaulis have shown potent antioxidant activity, i.e., IC50 value of 9.8 µg/ml, 10.19 µg/ml, and, 13.64 µg/ml, respectively. Similarly, leaf extract of R. indica exhibited antioxidant activity with an IC50 value of 19.73 μg/ml. The percentage inhibition of NO radicals of root and stem of M. rubicaulis and leaves of R. indica was 31%, 21%, and 22%, respectively. Out of these two plant species, M. rubicaulis had shown antibacterial activity towards selected microorganism, but antioxidant activity was shown by both plant species. These properties on above mentioned two plant species might help for the development of a marketed formulation for antibacterial and wound-healing agent since wound healing is promoted by antibacterial and antioxidant activities.
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Hypoglycemic Effect of a Combined Andrographis paniculata and Caesalpinia sappan Extract in Streptozocin-Induced Diabetic Rats

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Introduction. Researchers usually use herbal combinations to explore and develop traditional medicine to obtain additional benefits in the treatment of diseases, including diabetes. This study aims to evaluate the hypoglycemic effect of the combination of Andrographis paniculata (Burm. f.) Wall ex Nees and Caesalpinia sappan Linn extract (APCSE) on diabetes-induced rats. There has not been sufficient research on this combination; however, single extract studies of these plants have been widely conducted. Materials and Methods. Male Sprague Dawley rats (160–200 g) were induced by injecting a low dose of streptozotocin (35 mg/kg BW) twice and fed with a high-fat diet containing 25% fat, whereas control animals received only standard feed. Rats were treated with APCSE at doses of 100 mg and 200 mg/kg B W for seven days and compared to the APE and CSE groups treated with the extract at 100 mg, respectively. For the control group, rats were treated with metformin with a dose of 250 mg/kg. The antihyperglycemic and antihyperlipidemic effects were determined by measuring blood glucose levels and lipid profiles (cholesterol, triglycerides, HDL, and LDL). To assess the impact of the extract on pancreatic and adipose tissue, the number of pancreatic beta cells and adipocytes was evaluated through histopathological and immunohistochemical study. Results and Discussion. In a nonfasting state, the blood glucose change in APCSE 200 mg was 18.65% and was significantly lower from the DM group. However, a single extract of APE and CSE showed lower fasting blood glucose levels compared to the combined extract. Lipid profiles show no significant differences in cholesterol levels between groups; however, all treatment groups, including metformin, showed higher triglyceride levels. The APE-tr eated group showed significantly lower HDL and LDL, whereas CSE only showed lower LDL. The β-cell number was significantly higher after treatment with single extract CSE. The CSE and the combined extract groups showed hyperplasia adipocytes. Conclusion. The combined extract of APCSE has a moderate antihyperglycemic effect; however, a single extract may have better potential than the combined extract.
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Effects of the Glutamine Administration on T Helper Cell Regulation and Inflammatory Response in Obese Mice Complicated with Polymicrobial Sepsis

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This study investigated the impacts of GLN on inflammation and T cell dysregulation in obese mice complicated with sepsis. Mice were divided into normal control (NC) and high-fat diet groups. The high-fat diet provided 60% of energy from fat and was administered for 10 weeks to induce obesity. Mice fed with a high-fat diet were then assigned to sham (SH) and sepsis with saline (SS) or GLN (SG) groups. The SH group was subjected to laparotomy, while the sepsis group underwent cecal ligation and puncture (CLP). The SS group was intravenously injected with saline. The SG group was intravenously administered GLN after CLP. Mice were sacrificed at 12, 24, or 48 h post-CLP, respectively. Results demonstrated that in the presence of obesity, sepsis drove CD4+ T cells toward the helper T (Th)2 and Th17 lineages. Also, expressions of inflammatory cytokines and macrophage infiltration markers in adipose tissues and lungs were elevated. Treatment of obese mice with GLN after sepsis reversed Th polarization and downregulated macrophage infiltration and inflammatory cytokine, whereas the tight junction-associated protein expression increased in the lungs. These findings suggest that the intravenous administration of GLN to obese mice after sepsis modulated a more balanced Th cell lineage, alleviated inflammation, and attenuated lung injury.
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Expression and Functional Characterization of a Novel Antimicrobial Peptide: Human Beta-Defensin 118

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Purpose. β-Defensin 118 (DEFB118) is a novel host defense peptide (HDP) identified in humans. To evaluate its potentials for future utilization, the DEFB118 gene was expressed in Escherichia coli (E. coli) and the recombinant protein was fully characterized. Methods. The DEFB118 protein was obtained by heterologous expression using E. coli Rosetta (DE3). Antibacterial activity of DEFB118 was determined by using various bacterial strains. IPEC-J cells challenged by E. coli K88 were used to determine its influences on inflammatory responses. Results. The E. coli transformants yielded more than 250 μg/mL DEFB118 protein after 4 h induction by 1.0 mM IPTG. The DEFB118 was estimated by SDS-PAGE to be 30 kDa, and MALDI-TOF analysis verified that it is a human β-defensin 118. Importantly, the DEFB118 s howed antimicrobial activities against both Gram-negative bacteria (E. coli K88 and E. coli DH5α) and Gram-positive bacteria (S. aureus and B. subtilis), with a minimum inhibitory concentration (MIC) of 4 μg/mL. Hemolytic assays showed that DEFB118 had no detrimental impact on cell viability. Additionally, DEFB118 was found to elevate the viability of IPEC-J2 cells upon E. coli K88 challenge. Moreover, DEFB118 significantly decreased cell apoptosis in the late apoptosis phase and downregulated the expression of inflammatory cytokines such as IL-1β and TNF-α in IPEC-J2 cell exposure to E. coli K88. Conclusions. These results suggested a novel function of the mammalian defensins, and the antibacterial and anti-inflammatory properties of DEFB118 may allow it as a potential substitute for conventionally used antibiotics or drugs.
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A Rare Malignant Disease, Dermatofibrosarcoma Protuberans of the Breast: A Retrospective Analysis and Review of Literature

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Dermatofibrosarcoma protuberans (DFSP) is a rare low-grade fibroblastic mesenchymal tumor derived from the dermis. The aim of this retrospective analysis was to summarize the clinicopathological data from our cases and published cases to offer more evidence for the recognition of dermatofibrosarcoma protuberans (DFSP). A total of 6 breast DFSP patients who had received treatment in our hospital were retrospectively enrolled, and detailed clinicopathological data were gathered for analysis. The median age was 29.5 years (ranging from 17 to 42 years). Most cases presented a red or brown-red, mobile, well-circumscribed, protruding, breast mass (ranging from 1 to 3 cm). For histopathology, all cases (6/6) showed a storiform pattern of spindle cells that were positive for CD34 (6/6) and Vimentin (5/6) and neg ative for smooth muscle actin (0/6) and S-100 protein (0/6). The majority of patients (5/6) underwent wide local excision, with 2 cases treated with radiotherapy. With a median follow-up of 36 months, all 6 patients survived without recurrence or metastasis. The PubMed database was used to search for similar cases. Eventually, 36 cases were included in this review, while cases without detailed clinical information or not reported in English were excluded from the analysis. To summarize, DFSP of the breast is an extremely rare malignancy characterized by spindle tumor cells arranged in a storiform pattern and positivity for CD34. The core needle biopsy is one of the crucial methods for its preoperative diagnosis. Management of DFSP is mainly based on surgical excision. It is prone to local recurrence, so long-term follow-up is required.
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Evaluation of Heavy Metal and Microbial Contamination in Green Tea and Herbal Tea Used for Weight Loss in the Palestinian Market

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The use of green tea and herbal tea for weight loss is increasing worldwide owing to the rising rates of obesity. There are concerns about the safety and quality of these herbal products owing to their increased consumption worldwide. In this study, we evaluated randomly collected samples of green tea and herbal tea and tested them for heavy metal and microbial contamination. Eighteen samples of green tea or herbal tea of widely used brands in Palestine were tested for heavy metal and microbial contamination. The results showed that 7 of the samples had toxic heavy metals such as chromium (Cr) and lead (Pb), and their concentrations were above the allowable limits set by the World Health Organization (WHO). Moreover, 6 of the samples that were tested had microbial contamination with high total aerobic micr obial count (TAMC) and total yeast and mold count (TYMC). This could be due to improper handling and storage conditions of these herbal products. This study is the first of its kind in Palestine, and its results are forewarning to all the responsible authorities, including the Ministry of Health (MoH), to take immediate corrective actions such as quality control testing, auditing, and registration of marketed tea products.
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Effect of Coptis chinensis on Biofilm Formation and Antibiotic Susceptibility in Mycobacterium abscessus

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Mycobacterium abscessus infections are notoriously difficult to be treated and newer treatment options are required. Coptis chinensis (C. chinensis) and its main compound berberine are frequently used to treat bacterial and viral infections. In this study, the susceptibility of M. abscessus to C. chinensis extract and berberine was assessed by minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) evaluation. The effects of C. chinensis and berberine on biofilm formation and antibiotic susceptibility in M. abscessus were observed. C. chinensis at concentrations of MIC (1.5 mg/mL) and 2 × MIC (3.0 mg/mL) and berberine at ½ × MIC (0.125 mg/mL) demonstrated a strong inhibition of biofilm formation. Concentration of C. chinensis at ½ × MIC resulted in a signi ficant reduction in MICs of trimethoprim/sulfamethoxazole (TMP/SXT), clarithromycin (CLA), and linezolid (LZD). Similarly, ½ × MIC berberine had a significant effect on the MIC reductions of nine antibiotics including TMP/SXT, CLA, and LZD. Notably, the resistance level MIC of LZD against M. abscessus was reversed to a susceptible level by treatment with either C. chinensis or berberine. Therefore, C. chinensis and berberine have the potential to produce a synergistic antimycobacterial effect, reduce biofilm formation, and decrease antibacterial resistance to LZD in M. abscessus.
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The Therapeutic Properties of Plants Used Traditionally to Treat Gastrointestinal Disorders on Groote Eylandt, Australia

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The First Australians had well-developed healing systems. Groote Eylandt inhabitants used a variety of plant species to treat diarrhoea and other gastrointestinal illnesses. This study was undertaken to test, identify, and evaluate traditional medicines to treat these conditions against gastrointestinal bacterial, protozoal, and viral pathogens, as well as against cancer cell proliferation. Six plant species (Buchanania obovata Engl., Casuarina equisetifolia L., Eucalyptus tetrodonta F. Muell., Planchonia careya (F. Muell.) R. Knuth, Terminalia carpentariae C. T. White, and Vigna vexillata (L.) A. Rich.) were selected from a survey of a panel of elders from the Warnindhilyagwa tribe and compared with the published literature. Decoctions prepared according to traditional methods were screened for growth inh ibitory activity of a panel of diarrhoea-causing bacterial pathogens by disc diffusion and liquid dilution MIC assays. Inhibitory activity against the gastrointestinal protozoal parasite Giardia duodenalis and antiproliferative activity against human colorectal (Caco2) and cervical (HeLa) cancer cell lines were evaluated using MTS-based colorimetric cell proliferation assays. Preliminary antiviral screening was accomplished using an MS2 bacteriophage plaque reduction assay. Toxicity was evaluated using Artemia franciscana nauplii mortality and HDF cell viability bioassays. All traditional medicines tested inhibited bacterial growth, often with MIC values substantially
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In Vitro Infrared Thermographic Assessment of Temperature Change in the Pulp Chamber during Provisionalization: Effect of Remaining Dentin Thickness

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Interim crowns and partial fixed dental prosthesis materials generate exothermic heat during polymerization. The amount of heat transmitted to the pulp chamber can be a function of several factors, including the thickness and quality of the remaining dentin after crown preparation. The aim of this in vitro study was to measure with infrared thermography the temperature changes on the adjacent surface of the chamber roof of premolar teeth extracted from young and old patients (having different thicknesses of remaining dentin after crown preparation) during fabrication of provisional resinous restorations. Twenty extracted human first and second maxillary premolar teeth (10 from young patients, with a relatively large pulp chamber, and 10 from older patients, with a relatively small pulp chamber) were used. The roots were sectioned to expose the inner side of the chamber roof, and the crowns were provisionalized after preparation for a metal-ceramic crown. Two provisional materials, Turbo Temp 2 and Luxatemp Fluorescence, were used. Temperature changes on the inner side of the chamber roof were measured at 2-second intervals using an infrared thermal imaging camera. After completion of the temperature recordings, the teeth were sectioned and the remaining dentin thickness was determined. The older group (mean thickness: 2.82 mm) and younger group (mean thickness: 1.9 mm) differed significantly in dentin thickness (). The mean greatest temperature increases recorded on the chamber roof of teeth with less remaining dentin were 4.07°C for Turbo Temp 2 and 3.94°C for Luxatemp Fluorescence, while increases in the premolars with greater dentin thickness were 1.69°C for Turbo Temp 2 and 1.64°C for Luxatemp Fluorescence. Significant interactions were found between tooth groups (for Turbo Temp 2 and for Luxatemp Fluorescence). No significant differences were found between assessed materials regardless of the thickness of the remaining dentin (for the older group and for the younger group). Dentin had a significant effect in limiting the temperature increase generated during polymerization of provisional materials, indicating good thermal insulating properties of this tissue. A remaining dentin thickness of 1.9 mm or more is sufficient to protect the pulp from any temperature increase during provisionalization using tested materials.
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Improving Stem Cell Clinical Trial Design and Conduct: Development of a Quality Assessment Tool for Stem Cell Clinical Trials

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Background. Clinical trials are at the cornerstone of evidence-based stem cell therapies, but the quality assessment for designing and conduct these sometimes-complex studies are scarce of evidence. This study is aimed at developing a handy quality assessment tool for stem cell clinical trials, enhancing capacity of the self-regulate overall quality, and participating protection. Methods. The framework of quality assessment tool was based on the PQRS (progress-quality-regulation-scientific) quality assessment tool, and detailed quality indicators were developed by leader group discussion, expert consulting, and literature review. Stem cell clinical trials were retrieved from the International Clinical Trials Registry Platform, and corresponding quality indicators were assessed and extracted. The validity and feasibility of conceptual quality assessment tool were further evaluated by using structural equation modeling. Results. The quality assessment tool for stem cell clinical trials contains four critical quality attributes, including participant protection, scientific value, quality control, and stem cell products, and 9 observed quality indicators. From 11 primary clinical trial registries in the International Clinical Trials Registry Platform, 9410 stem cell trial registrations were identified, and 1036 studies were eligible for publications and protocols screening. After reviewed full text, 37 studies were included in the validity and feasibility evaluation: 32 studies were completed, and 3 studies terminated early. Most of the studies (83.79%) were in the early phase, and 63.16% of the studies were investigator-initiated trial. To further tested for validity, the critical quality attributes and quality indicators (QIs) between expertise further validated by the SEM method, which s howed a good fit for the model (;;;;). Compared with exploratory trials, evaluating using the quality assessment tool, confirmatory trials performed similarly in participant protection, scientific value, and quality control, but lower in stem cell products. Conclusions. The results of critical quality attributes and quality indicators between expertise and confirmatory validation analysis are basically consistent, indicating the feasibility and validity of applying this quality assessment tool for overall quality evaluation of stem cell trials.
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