Current Pharmaceutical Biotechnology - Volume 21, Issue 7, 2020

Metallothioneins (MTs) are low-molecular-weight, cysteine-rich proteins that bind to heavy metals. MTs play a key role in the homeostasis of metal ions, maintaining intracellular redox equilibria and free radical scavenging. In several studies, under different conditions such as cancer development, drug therapy and heavy metal stress, the unique structural changes and functional effects of MT were studied. Although several assays are available to monitor the content and type of Metallothionein (MT) from environmental samples or in biomedical assays, Enzyme-Linked Immunosorbent Assays (ELISA) became the preferred method of MT detection. ELISA is low in cost, specific, simple, and efficient. This review evaluates the advantages and disadvantages of using different types of ELISA in the detection of metallothioneins from environmental or clinical samples as well as ways of its validation and cross-validation.

Background: Probiotics and their nutrient sources (prebiotics) have been shown to have positive effects on different organs of the host. The idea of their potential benefits on Central Nervous Systems (CNS) and the incidence of Anxiety, Schizophrenia, Alzheimer, Depression, Autism, and other mental disorders has proposed a new category of medicines called “psychobiotic” which is hoped to be of low-side effect anti-inflammatory, antidepressant, and anti-anxiety constitutes. Objective: In the current review, we present valuable insights into the complicated interactions between the GI microbiota (especially in the colon), brain, immune and central nervous systems and provide a summary of the main findings of the effects of pro- and prebiotics on important mental disorders from the potential mechanisms of action to their application in clinical practice. Methods: Google Scholar, Pub Med, Scopus, and Science Direct databases were searched using following key words: “probiotics”, “prebiotics”, “mental disorders”, “psychological disorders”, “depression”, “anxiety”, “stress”, “Alzheimer” and “autism spectrum”. The full text of potentially eligible studies was retrieved and assessed in detail by the reviewers. Data were extracted and then summarized from the selected papers. Results: The results of the provided evidence suggest that probiotic and prebiotics might improve mental function via several mechanisms. The beneficial effects of their application in Depression, Anxiety, Alzheimer and autism spectrum diseases have also been supported in clinical studies. Conclusion: Pro and prebiotics can improve mental health and psychological function and can be offered as new medicines for common mental disorders, however, more clinical studies are necessary to conduct regarding the clinical significance of the effects and their bioequivalence or superiority against current treatments.

Lactic acid bacteria are beneficial to human health. Lactic acid bacteria have wide applications in food, cosmetic and medicine industries due to being Generally Recognized As Safe (GRAS) and a multitude of therapeutic and functional properties. Previous studies have reported the beneficial effects of lactic acid bacteria, their extracts or ferments on skin health, including improvements in skin conditions and the prevention of skin diseases. Lipoteichoic acid isolated from Lactobacillus plantarum was reported to inhibit melanogenesis in B16F10 melanoma cells. In particular, lipoteichoic acid also exerted anti-photoaging effects on human skin cells by regulating the expression of matrix metalloproteinase- 1. The oral administration of Lactobacillus delbrueckii and other lactic acid bacteria has been reported to inhibit the development of atopic diseases. Additionally, the clinical and histologic evidence indicates that the topical application of lactic acid is effective for depigmentation and improving the surface roughness and mild wrinkling of the skin caused by environmental photo-damage. This review discusses recent findings on the effects of lactic acid bacteria on skin health and their specific applications in skin-whitening cosmetics.

The onset of Cardiovascular Disease (CVD) is known to be associated with multiple risk factors related to exogenous exposures on predisposed genetic makeup. Diet and lifestyle have a cascade effect on microbiota biodiversity, thus impacting inflammation and heart health. Atherosclerosis is a type of CVD where chronic inflammation contributes to plaque buildup in the arteries resulting in narrowed blood vessels, which obstruct blood flow. Polyphenolic compounds, including flavonoids, most commonly consumed in the form of plants, have been identified to have various mechanisms of action to reduce the inflammatory response in the body. Flavonoids provide a variety of nutraceutical functions including antioxidant, antimicrobial, anti-inflammatory, antiangiogenic, antitumor, and improved pharmacokinetic properties. Therefore, the medicinal use of polyphenolic compounds as an intervention for the inflammatory response, especially relating to the gut microbiome, may significantly reduce the risk of atherosclerotic plaque development and disease onset. This review addresses the role of polyphenolic compounds and gut microbiome in cardiovascular disease. Research studies conducted in cells and animals were reviewed. These studies clearly illustrate that dietary polyphenolic compounds influence resident gut microbiota thus they are associated with the prevention of atherosclerosis progression. Further research in this field is warranted to identify potential gut microbiome mediated therapeutic approaches for CVD.

Objective: The current study reports a green, rapid and one-pot synthesis of FeSO4 nanoparticles using Hibiscus rosasinensis floral extract as a reducing and capping agent. 0.5M of FeSO4 was stirred with the floral extract of H. rosasinensis for around 20 minutes at 37ºC and pH 7. Methods: The development of pink color was considered as the endpoint of reduction and the nanoparticles were characterized by UV-Vis spectrum, EDAX, DLS, FTIR, FESEM, and XRD. UV-Vis spectral analysis indicated a peak at 530 nm and EDAX measurement revealed the presence of Fe, S, O and C elements in the nanoparticle sample. The FTIR analysis showed amines, alcohol and alkene groups that act as capping agents for the produced nanoparticles. FESEM and XRD determination presented FeSO4 nanoparticles of 40-60 nm in size. The synthesized nanoparticles were found to have antibacterial activity against 6 pathogenic bacteria with MIC and MBC of 40 mg/mL. Results: To determine the toxicity at the eukaryotic level, brine shrimp toxicity assay was conducted and 100% mortality was found at concentrations >0.06 mg/mL. Gel shift assay suggested the mechanism of toxicity of FeSO4 NPs by binding and degradation of DNA molecules. Conclusion: From the results, the authors demonstrate the ease of green synthesis of FeSO4 nanoparticles and its bioactivity that may have potential applications as drugs and drug delivery systems against various diseases.

Background: Rheumatoid Arthritis (RA) is an autoimmune, systemic disease mainly affecting joints. Presently, there is no specific treatment/ drug available for curing RA except few supportive medicines. Therefore, the focus has been shifted to medicinal plants for the treatment of such diseases. Choerospondias axillaris commonly known as Lupsi/Lapsi and has been reported to have several properties for the treatment of various diseases. Objective: The present study has been conducted to explore the anti-inflammatory effects of Choerospondias axillaris fruit extract on Synoviocytes (FLS) and Collagen-Induced Arthritis (CIA) rat model. Methods: Methanolic extract of the Choerospondias axillaris fruit was used for determining phytochemical, antioxidant and anti-inflammatory properties. Antioxidant activity of Choerospondias axillaris fruit was determined by free radicals scavenging assays and bioactive compounds were identified via LC-MS/MS analysis. Anti-inflammatory effect was investigated in RA and Osteo Arthritis (OA) primary cells and also in Collagen Induced Arthritis (CIA) rat models. Further, the medicinal properties of anti-inflammatory bioactive compounds were supported by docking studies. Results: In-vitro and in-vivo studies showed significant decrease in the levels of inflammatory cytokines. Docking analysis revealed that quercetin inhibits TNF-α having -9.1 kcal/mol binding energy and 10.13 μM inhibitory constant. Quercetin also inhibits IL-6 having -6.6 kcal/mol binding energy and 21.9 μM inhibitory constant. Conclusion: Observed results suggest that the underutilized fruit Choerospondias axillaris can be used to reduce the inflammation of inflammatory diseases like RA.

Objective: Frequent administrations for DPPIV-resistant GLP-1 analogs are necessary to maintain the blood concentrations due to the short half-life of less than 5 minutes. However, most delivery systems that possess the ability of sustainable release of GLP-1 have drawbacks such as low yield, high cost and undesirable side effects. Therefore, we aimed to prepare a simple and efficient delivery system that could be feasibly applied to reduce blood glucose. Methods: A novel GLP-1 delivery system (GLP-1-ELPs-SA) was prepared and characterized by circular dichroism. Furthermore, the activity and property of GLP-1-ELPs-SA were evaluated in vitro and in vivo. Results: GLP-1-ELPs-SA are easily expressed in E. coli in a soluble formulation and purified through the inverse transition cycle. GLP-1-ELPs-SA spontaneously generated depot under physiological conditions. GLP-1-ELPs-SA was also found to be dispersed in the blood vessels from the depot and showed a high affinity to bind with mice (C57BL/6J) albumin, which shows that GLP-1-ELPs-SA has a long circulation time in vivo. Conclusion: Our delivery system could markedly decrease the clearance of recombinant proteins based on serum albumin, without substantially increasing the protein molecular weight and remarkably reducing the blood glucose within 120 h.

Aims: The present study aims to determine the antimicrobial efficacy of Alphonsea madraspatana leaves extract against selected uropathogens. Background: The plant Alphonsea madraspatana is an endangered species, reported to exhibit high antimicrobial activity due to the presence of phenolic compounds. Prevalence of high UTI infection and increased cases of bacterial resistance directed for alternative approach to meet the challenge of drug resistance. Objective: Our objective is to determine antimicrobial efficacy of Alphonsea madraspatana leaves extract against selected uropathogens and subsequent in-silico analysis to predict the underlying mechanism. Methods: Phytochemicals extraction from the dried leaves of Alphonsea madraspatana was performed using solvent gradient technique. All the extracts were subjected to preliminary phytochemical screening using liquid chromatography-mass spectrometry. Antimicrobial activity of the prepared extract was determined against the selected uropathogens using agar diffusion method. Finally, molecular docking study of the selected bio-actives was performed against a representative bacterial resistance enzyme “DNA Gyrase”. Results: Methanolic extract exhibits relatively higher antimicrobial activity against the selected strains with Minimum Inhibitory Concentration (MIC) and minimum bactericidal concentration (MBC) of 1.56 ± 1 ug/mL and 6.25 ± 2 ug/mL, respectively. Phytochemical screening showed the presence of 3 flavonoids compounds such as Luteolin-7-O-glucoside, Kaempferol-3-O- rotinoside-7-O-rhamnoside and Genestein-7-O-glucoside. The results of molecular docking shows Luteolin-7-O-glucoside has best docking scores of −8.5 kcal/mol than other ligand molecules. Experimental simulation in presence of DNA Gyrase inhibitors showed lowest MIC and MBC value for E. Coli, which was found to be 1.56 ±1 ug/mL and 6.25±2 ug/mL respectively, support the docking outcomes. Conclusion: Outcomes of this study suggested that the methanolic extract of this plant shows good anti-microbial potential against resistant uropathogens.

Background: Metabolic Syndrome (MS) is a clinical condition consisting of risk factors associated with type two diabetes and developing cardiovascular disease. It has been suggested that resistin is a linkage between obesity, inflammation and type two diabetes. This study aims to investigate whether Resistin Gene (RETN) polymorphism (+62G>A) is linked to MS and resistin levels among the Egyptian population. Methods: This study was performed with 310 Egyptian volunteers: 160 MS subjects and 150 controls. Anthropometric parameters and biochemical variables were determined. The RETN +62G>A polymorphism was genotyped by PCR-RFLP technique. Results: The resistin levels of the MS group were significantly higher than those of the control group. Resistin levels were positively correlated with anthropometric parameters and liver biomarkers in the MS group. According to RETN +62G>A polymorphism, carriers with the A allele (GA/AA) had significantly increased resistin levels than subjects with the GG genotype, consequently, the RETN +62G >A polymorphism was found to be related to MS, biochemical parameters and anthropometric variables. Conclusion: These findings propose that the RETN +62G>A polymorphism has a great impact on the circulating resistin concentrations, and that resistin levels are strongly related to MS. Therefore, this RETN polymorphism is related to the risk of the prevalence of MS in the Egyptians.

Background: Endometrial cancer is one of the most common gynecological cancer in the developed countries and occurs mainly in postmenopausal women. Angiogenesis is important for cancer formation as it provides nutrients for growing tumor mass. Most tumors do not show detectable Homeobox A5 (HOXA5 level), suggesting its potential role as a cancer suppressor. It was demonstrated that HOXA5 is involved in the progression of various types of cancer and the loss of its expression correlates with higher pathological grade and poorer outcome. Objective: The aim of the study was to evaluate HOXA5 expression at transcriptome and protein levels. Materials and Methods: The study enrolled 45 women diagnosed with endometrial cancer and 15 without neoplastic changes. The histopathological examination allowed us to divide cancer tissue samples according to the degree of histological differentiation: G1, 17; G2, 15; G3, 13. The expression of the HOXA5 protein was determined by immunohistochemistry. Microarray and RT-qPCR techniques were used to assess HOXA5 expression at the mRNA level. Results: The reaction to the HOXA5 protein was only visible in glandular cells in G1 endometrial cancer and was lower compared to the control. In grades 2 and 3, reactions were noted at the limit of the method’s sensitivity. In addition, reduced HOXA5 expression was observed at the transcriptome level. Conclusion: HOXA5 may become a potential complementary molecular marker, allowing early detection of neoplastic changes in the endometrium. It also seems that detection of HOXA5 at the mRNA and protein levels may be helpful in improving the accuracy of diagnosis and planning effective oncological therapy.

Background: Changes in the cellular behavior depend on environmental and intracellular interactions. Cancer treatments force the changes, first on the molecular level, but the main visible changes are macroscopic. During radiotherapy, cancer cell’s adhesion, proliferation and migration should be well monitored. In over 60% of diagnosed cancers cases, patients are given treatments with different protocols of radiotherapy, which result in possible metastasis and acute whole body response to toxic radiation. Objective: Effectiveness of the therapy used depends on the sensitivity/resistance of irradiated cancer cells. Cellular mechanisms of cancer protection, such as the activation of DNA damage and repair pathways, antioxidants production and oxidative stress suppression during treatments are not desirable. Cancer cells monitoring require the development of novel techniques, and the best techniques are non-invasive and long-term live observation methods, which are shown in this study. Methods: In cancers, invasive and metastatic phenotypes could be enhanced by stimulation of proliferation rate, decreased adhesion with simultaneous increase of motility and migration potential. For such reasons, the Ionizing Radiation (IR) stimulated proliferation; migration with lowered adhesiveness of cancer Me45 and normal fibroblasts NHDF were studied. Using impedance measurements technique for live cells, the adhesion of cells after IR exposition was assessed. Additionally proliferation and migration potential, based on standard Wound Healing assay were evaluated by timelapse microscopic observations. Results: We found simulative IR dose-ranges (0.2-2 Gy) for Me45 and NHDF cells, with higher proliferation and adhesion rates. On the other hand, lethal impact of IR (10-12 Gy) on both the cell lines was indicated. Conclusion: Over-confluence cell populations, characterized with high crowd and contact inhibition could modulate invasiveness of individual cells, convert them to display migration phenotype and advance motility, especially after radiotherapy treatments.