Current Pharmaceutical Biotechnology - Volume 22, Issue 15, 2021

Background: Autopsy is a valuable tool for understanding the physiopathology of any disease, and it is the gold standard to assess the cause of death. The clinical autopsy is the ultimate medical service for a patient and plays a crucial role in the context of quality control, education of physicians and other medical personnel, as well as mitigation of risk of malpractice claims. Objective: This study aims to demonstrate the importance of improving an autopsy service and the relevance of this investigation procedure in daily clinical practice by evaluating the rate of major discrepancies between the assumed cause of death and the ascertained cause of death after a complete post mortem investigation. A further aim is to classify these discrepancies as class I or class II discrepancies according to the Goldman’s criteria in order to assess performance quality. Methods: A retrospective study of the hospital autopsies performed from June 2018 to March 2020 was conducted by considering a diversified dataset, including age and sex of the deceased as well as the clinical and pathological causes of death. Results: 362 cases were taken into consideration. Major discrepancies were found in 71.3% of cases, with a class I error of 22.7% and a class II error of 48.6%. The most frequent misdiagnosis were cardiovascular disorders, embolism, and aneurism rupture. Discussion: The rate of major discrepancies and the rate of class I and class II errors are way above the rate found in the literature. Despite the high rate of a major discrepancy, evidence collected from hospital autopsies (i.e., certainty of the cause of death, unknown comorbidities) has strengthened the legal defense in cases of medical malpractice litigation. In our experience, by accurately determining the cause of death, revealing new or unexpected findings, and any possible diagnostic or technical errors, post-mortem examinations can significantly contribute to the improvement of team performance and quality of care. Conclusion: The presence of clinicians during an autopsy and the early sharing of results can be considered a new auditing strategy for hard clinical cases. Finally, by providing a clearer understanding of the nature and cause of the illness, the autopsy results assist in the grieving process by reassuring family members that action or inaction on their part had not contributed to the death.

Background: In recent decades, in the field of healthcare, awareness of the problems inherent to the quality has steadily increased. Currently, the evaluation of healthcare activities is one of the ways in which health systems regulate internal relationships and define strategic decisions. Objective: The study aims to describe in detail the entire process of developing a group of Key Performance Indicators for monitoring and implementing the management of litigation due to medical liability. Particularly, the objective is to centralize and standardize the indicators to provide scientifically reliable data on claims management to hospital professionals responsible for strategic choices. Methods: The study was conducted to analyze data relating to the claims management at Umberto I General Hospital in Rome from 2012 to 2018. All claims reported were classified according to a selection of the categories coded in the International Classification for Patient Safety system, the economic features, and the chronological references of the main management extrajudicial and judicial phases. The Process Analysis Method was followed to develop significant indicators for measuring the performance and the quality of claims management. Results and Conclusion: The results obtained demonstrate how the assessment of performance in claims management can potentially lead to greater risk control with significant repercussions in terms of reduction of disputes, speed in settling claims, reduction of management times, planning of loss prevention measures, and implementation of quality of care.

Introduction: In recent decades, the number of medical professional liability disputes has grown exponentially, becoming a well-studied aspect of public healthcare. Legal medicine is an essential tool in managing this phenomenon. Methods: This article reports the results of the analysis of disputes for medical professional liability in a Level III University Hospital in Italy. The study covers the time period from 01.01.2003 to 31.12.2019. Results: About 33% of claims have been compensated. Those claims fall within the following categories: 37% in the surgical field, 17% in the field of internal medicine, and 35% in the emergency care field. As for the types of a medical mistake, compensation was awarded in 30% of diagnostic error cases, 26% of therapeutic error cases, 47% of execution error cases, and 55% of organizational deficiency cases. Discussion: The difference in the rate of compensation between the various medical fields or types of error depends on specific medico-legal characteristics. The aim of advanced healthcare systems is to prevent medical liability disputes by analyzing this phenomenon and improving clinical risk management programs. In particular, according to our study, events related to organizational deficiencies are the most preventable. In addition, through the use of a dedicated reporting procedure, medico-legal analysis of malpractice cases may be the key to risk reduction. Conclusion: Every major hospital should set up a medico-legal watchdog responsible for collecting and analyzing information on professional medical liability disputes in order to prevent and manage such events.

Introduction: In recent decades, the number of professional liability disputes has increased, especially in the civil sector. In these liability assessments, medico-legal experts have become the linchpin. Law No. 24/2017, concerning professional liability in Italy, requires that guidelines and best practices be established to identify the necessary elements for writing expert reports in the legal sector. Materials and Methods: The authors have created a numerical indicator to be applied to 150 legal expert reports on professional liability, enabling them to evaluate the methodological quality of those reports. The results are then compared with the outcome of the legal proceedings in order to establish the suitability of this quality index for evaluating the work of medico-legal experts. Results: Of the 150 reports considered, 14 were scored inadequate, 75 adequate and 31 good. These inadequate (according to the indicator) reports presented a higher probability of being scored inadequate in court proceedings, compared to the risk of inadequacy of all the reports; OR 4.6 (95% CI 1.25-16.90). This probability significantly increased on comparing the inadequate reports with the adequate; OR 5.6 (95% CI 1.28-24.41), and the inadequate with the good; OR 7.73 (95% CI 1.50- 39.87). Discussion: Application of the proposed indicator is simple and produces a high-quality result, thus ensuring an accurate and tenable appraisal of methodological quality. Conclusion: The indicator proposed serves as a useful starting point for creating the necessary methodological standards for medico-legal experts in the field of professional liability as recommended by Law No. 24/2017.

Background: The liver disease problem prompts investigators to search for new methods of liver treatment. Introduction: Silymarin (Sil) protects the liver by reducing the concentration of free radicals and the extent of damage to the cell membranes. A particularly interesting method to increase the bioavailability of Sil is to use synthesized gold nanoparticles (AuNPs) as reagents. The study considered whether it was possible to use the silymarin-AuNP conjugate as a potential liver-protecting drug. Methods: AuNPs were conjugated to Sil and the liver-protecting activity of the conjugate was examined. Experimental hepatitis and hepatocyte cytolysis after carbon tetrachloride action were used as a model system, and the experiments were conducted with laboratory animals. Results: For the first time, silymarin was conjugated to colloidal gold nanoparticles (AuNPs). Electron microscopy showed that the resultant preparations were monodisperse and that the mean conjugate diameter was 18-30 nm ± 0.5 nm (mean diameter of the native nanoparticles, 15 ± 0.5 nm). In experimental hepatitis in mice, conjugate administration interfered with glutathione depletion in hepatocytes in response to carbon tetrachloride. It also was conducive to an increase in energy metabolism and stimulated the monocyte-macrophage function of the liver. The results were confirmed by the high respiratory activity of the hepatocytes in cell culture. Conclusion: We conclude that the silymarin-AuNP conjugate holds promise as a liver-protecting agent in acute liver disease caused by carbon tetrachloride poisoning.

Atherosclerosis (AS) is an important pathological basis for the occurrence of Coronary Atherosclerotic Disease (CAD), stroke and other adverse cardiovascular events. AS is an inflammatory disease, and macrophages are the main inflammatory cells in AS lesions, playing a leading role in the formation of atherosclerotic plaques and the development and regression of AS. Various proinflammatory and anti-inflammatory factors act on macrophages to regulate AS. Pro-inflammatory factors recruit monocytes to accumulate in the inflammatory site and promote the transformation of monocytes to macrophages. A large number of aggregated macrophages secrete various inflammatory mediators to promote AS. Pro-inflammatory factors can induce the polarization of M1-type macrophages to start and maintain inflammation, promote the accumulation of lipids in macrophages, and accelerate the formation of foam cells. Anti-inflammatory factors can not only induce M2-type macrophages polarization, promote tissue remodeling and repair, and reduce the occurrence of AS, but also promote the metabolism of fatty acid oxidation and oxidative phosphorylation of macrophages, regulate lipid metabolism, stabilize plaques, and induce the transformation of helper T cells of type 1/2 (Th1/Th2) to Th2 cells, thus reducing inflammation. This review summarizes the effect and underlying regulatory mechanism of macrophages in the development of AS, which can provide new ideas for the diagnosis and treatment of AS targeting macrophages.

Aims: Globally, scientists are working to find more efficient antimicrobial drugs to treat microbial infections and kill drug-resistant bacteria. Background: Despite the availability of numerous antimicrobial drugs, bacterial infections still pose a serious threat to global health. A constant decline in the effectiveness of antibiotics owing to their repeated exposure as well as a short-lasting antimicrobial activity led to the demand for developing novel therapeutic agents capable of controlling microbial infections. Objective: In this study, we report the antimicrobial activity of chemically synthesized silver nanoparticles (cAgNPs) augmented with ampicillin (amp) in order to increase antimicrobial response against Escherichia coli (gram –ve), Staphylococcus aureus (gram +ve) and Streptococcus mutans (gram +ve). Methods: Nanostructure, colloidal stability, morphology and size of cAgNPs before and after functionalization were explored by UV-vis spectroscopy, FT-IR, zeta potential and TEM. The formation and functionalization of cAgNPs were confirmed from UV-vis spectroscopy and FT-IR patterns. From TEM, the average sizes of cAgNPs and cAgNP-amp were found to be 13 and 7.8 nm, respectively, and change in colloidal stability after augmentation was confirmed from zeta potential values. The antimicrobial efficacies of cAgNP-amp and cAgNPs against E. coli S. aureus and S. mutans were studied by determining Minimum Inhibitory Concentrations (MICs), zone of inhibition, assessment of viable and non-viable bacterial cells and quantitative assessment of biofilm. Results & Discussion: Our results revealed cAgNP-amp to be highly bactericidal compared to cAgNPs or amp alone. The nano-toxicity studies indicated cAgNP-amp to be less toxic compared to cAgNPs alone. Conclusion: This study manifested that cAgNPs show synergistic antimicrobial effects when they get functionalized with amp suggesting their application in curing long-term bacterial infections.

Background: L-Asparaginase is an antineoplastic agent used in the treatment of acute myeloid and acute lymphoblastic leukemia. The present study deals with the production of this chemotherapeutic enzyme drug from Aspergillus flavus NCIM 526. The production of enzymes was carried out using oil-extracted cakes in a shake flask culture. Process parameters like carbon and nitrogen sources were also taken into account. Methods: A total of six isolates were used to screen out efficient microorganisms for enzyme production. Aspergillus flavus NCIM 526 exhibited 138 IU/ml of enzyme activity in oil extracted mix cake after 96 hours of the incubation period. Molasses and l-asparagine were proved to be the best carbon and nitrogen sources for enzyme production. The enzyme was purified by column chromatography and the finest enzyme exhibited specific activity of 28 IU/mg. Results and Discussion: The fungal enzyme exhibited low Km values as compared with standard E. coli L-asparaginase, proving more substrate affinity of fungal enzyme than bacterial enzymes. Conclusion: The study explored the Aspergillus flavus NCIM 526 as a potential fungal source for high yield production of antileukemic enzyme drugs.

Aims: This research aimed at exploring potential new compounds to be used in the treatment of osteoporosis by Connectivity Map (CMap) and determining the role of fisetin in osteoporosis according to its effects on the PI3K-AKT signaling pathway in MC3T3-E1 pre-osteoblastic cells. Methods: Microarray analysis was used to obtain the differentially expressed genes in published gene expression data. Potent compounds for osteoporosis therapy were discovered by CMap analysis. DAVID and Gene Set Enrichment Analysis (GSEA) were used to discover signaling pathways that connected to osteoporosis disease. Cell viability was evaluated by a CCK-8 assay. Quantitative realtime Polymerase Chain Reaction (qRT-PCR) and western blot analysis were used to test the mRNA and protein expressions related to the PI3K-AKT signaling pathway in MC3T3-E1 cells, respectively. Results: CMap analysis identified fisetin as a promising compound for anti-osteoporosis treatment. DAVID and GSEA analysis showed that the PI3K-AKT signaling pathway was inactivated in osteoporosis. Cell experiments revealed that fisetin caused an elevation of cell viability, up-regulated the mRNA levels of the Runt-related transcription factor-2 (Runx2), Osterix (Osx), collagen type I 1 (Col1a1) and Osteoprotegerin (OPG) while down-regulated the nuclear factor-ΚB ligand (RANKL) mRNA level. Discussion: The protein levels of Runx2, Col1a1 and Osteocalcin (OCN) were also increased by fisetin. Furthermore, fisetin activated the phosphoinositide-3-kinase/protein kinase B (PI3K-AKT) signaling pathway, and blocking this pathway by the inhibitor LY-294002 could impair fisetin’s functions on proliferation, differentiation and OPG/RANKL expression ratio in the MC3T3-E1 cells. Conclusion: Our results demonstrated that fisetin could promote MC3T3-E1 cell proliferation, differentiation and increase OPG/RANKL expression ratio through activating the PI3K-AKT pathway, which has potential for the treatment of osteoporosis.

Objective: The aim of this work is to identify and purify bioactive compounds from 70132;… aqueous methanolic extract (AME) of B. spectabilis leaves collected in Cairo, Egypt and assessed their rheumatoid arthritis activity. Methods: The methanolic extract of B. spectabilis leaves was fractionated and subjected to different chromatographic techniques to isolate pure new compounds which were identified by one dimensional and two dimensional nuclear magnetic resonance (NMR) spectroscopic analyses and mass spectrometric methods. The isolated compounds were evaluated for their anti-inflammatory activity on adjuvant induced chronic rheumatoid arthritis. Results & Discussion: Seven bioactive compounds were purified and identified for the first time from the methanolic extract of Bougainvillea spectabilis leaves; secologanin dimethyl acetal, α- and β-amyrin, α- and β-amyrin acetate, Kaempferol and kaempferol-3-O-rhamnoside. The previously mentioned compounds had a significant effect against rheumatoid arthritis. Conclusion: The seven compounds isolated from the methanolic extract of B. spectabilis leaves showed strong anti-rheumatoid arthritis activity.

Background: In December 2019, an outbreak of a pneumonia-like illness, Corona virus disease 2019 (COVID-19), originating from Wuhan, China, was linked to novel coronavirus, now termed SARS-CoV-2. Unfortunately, no effective drugs or vaccines have been reported yet. The main protease (M^PRO) remains the most validated pharmacological target for the design and discovery of inhibitors. Objective: The purpose of the study was to find a prospective natural scaffold as an inhibitor for M^PRO main protease in SARS-CoV-2 and compare it with repurposed antiviral drugs lopinavir and nelfinavir. Methods: Natural compound libraries were screened for potential scaffold against M^PRO main protease. Molecular dynamics simulation, MM-GBSA and principal component analyses of enzyme- ligand complexes were carried out with the top-ranking hits and compared with the repurposed antiviral drugs lopinavir and nelfinavir. Results: The structure-based virtual screening indicated phenylbenzopyrone of flavonoids as one of the top-ranking scaffolds that have the potential to inhibit the main protease with the Oglycosidic form, performing better than the corresponding aglyconic form. Simulation studies indicated that glycosidic form of flavonoid is a more suitable inhibitor with compounds rutin, procyanidin B6, baicalin and galloylquercetin, demonstrating high affinity and stability, and rutin, emerging as one of the best candidate compounds. Interestingly, rutin was reported to have inhibitory activity against similar protease (3Cprotease of enterovirus A71) and implicated in lung fibrosis. Conclusion: The present study on flavonoids, possessing a potential scaffold for inhibiting main protease activity for all betacoronavirus is an attempt to provide new and safe drug leads within a reasonably short period.

Background: Human glutathione S-transferases (hGSTs) are phase-II detoxification enzymes that catalyze the conjugation of electrophilic compounds and glutathione. Anomalous excess production of NO in the cellular environment under diseased or stressed condition results in lethal effects to the cell. Studies have reported that the evolution of tyrosine-based GSTs as a defense mechanism by the cell to mitigate Nitric Oxide (NO) toxicity. The dual role of hGSTP1 as NO carrier and scavenger is a prelude for the research forthwith. Objective: A plausible role of hGSTM1 as NO carrier is considered. Being a prominent cellular messenger and secondary metabolite, excess production of NO is lethal to the cell. Moreover, hGSTM1 polymorphisms lead to diminished catalytic activity that promotes a diseased state. Hence, it is compelling to generate hGSTM1 mutants that have more catalytic efficacy compared to Wild Type (WT). Methods: hGSTM1 mutants with enhanced efficiency were generated using in silico and in vitro Site-Directed Mutagenesis (SDM). WT and mutant proteins were overexpressed and purified using affinity chromatography. The catalytic activity and binding efficiency of WT and mutant proteins towards CDNB (1-chloro-2, 4-dinitrobenzene) & NO were determined. Results: NO assay reveals the probable interaction of WT hGSTM1 with NO. In silico, SDM studies provided E129K and Q109K mutants with superior NO binding efficiency as compared to WT. The catalytic activity (GST and NO assays) of the mutants corroborate the in silico results. Conclusion: WT hGSTM1 is recognized as a positive NO carrier. The novel mutant enzymes E129K and Q109K are inferred to possess superior NO carrying capacity.

Aim: The aim of present investigation is to identify the potential targets for Thymidylate Synthase and Amp-C β-lactamase from non-alkaloidal fractions of Moringa oleifera leaves. Background: Bioactive constituents from medicinal plants, either as pure compounds or as crude forms, provide vast opportunities for new drug discoveries. Due to an increasing demand for chemical diversity in screening programs, seeking therapeutic drugs from natural products, mainly from edible plants, has grown throughout the world. Moringa oleifera has an impressive range of medicinal uses with high nutritional value. Therefore, this medicinal plant has been used widely in traditional Indian medicine for anti-inflammation, anticancer and antibacterial infections. Objectives: The primary objective is to identify the phytoconstituents present in the maximum proportion in non-alkaloidal fractions of ethanolic leaf extract of Moringa oleifera. Then, the identified phytoconstituents were used to ensure the potential target molecules for binding affinity towards the target proteins viz. Thymidylate Synthase (1HVY) and Amp-C beta-lactamase (1FSY) by docking analysis. Methods: In present investigation, ethanolic extract of Moringa leaves was prepared and then fractionated on the basis of presence/absence of alkaloids. The antimicrobial activity of different fractions of ethanolic leaf extract was evaluated against various pathogens. Later, after this, bioactive molecules present in the non-alkaloidal fractions of ethanolic leaf extract were accomplished through GC-MS analysis, and finally, the identified phytocompounds were analyzed through docking studies to evaluate their affinity for target proteins viz. Thymidylate Synthase (1HVY) and Amp-C β-lactamase (1FSY). Results: The antimicrobial activity of non-alkaloidal fractions of ethanolic leaf extract was evaluated against various pathogens which exhibited significant antimicrobial activity. Twenty phytocompounds were identified as gas chromatogram of non-alkaloidal fractions (chloroform and ethyl acetate) of leaf extract of M. oleifera; Four most prominent compounds having highest peak area percentage were identified as Ethane, 1,1,2,2-tetrachloro, (46.45%) 2-Propanone, 1,1,3-trichloro, (13.77%) Heptasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13-tetradecamethyl (17.87%) and 2,4-Dichlorodiphenylsulfone (17.64%). Other notable compounds were 9,12-Octadecadienoic acid (Z,Z) (14.06%), Oleic acid, 3- (octadecyloxy)propyl ester (12.41%), Fluoranthene (6.98%), Phenol, 2,4-bis( 1,1-dimethylethyl) (4.16%) and Phthalic acid, butyl nonyl ester (3.47%). Only, five compounds viz. 2,6-Bis(1,1- dimethylethyl)phenol(C1), Dodecamethylcyclohexasiloxane(C2), Chlorodimethylethylsilane(C3), Fluoranthene(C4) and Hexadecanoic acid, methyl ester(C5) showed the maximum interaction with 1HVY with highest docking score of -178.51Kcal/mol, - 231.65Kcal/mol, -129.18Kcal/mol, - 173.10Kcal/mol and -220.78Kcal/mol, respectively. In addition, three compounds viz. Dodecamethylcyclohexasiloxane( C2), Fluoranthene(C4) and Hexadecanoic acid, methyl ester(C5) showed the maximum interaction with 1FSY with highest docking score of -137.23Kcal/mol, -54.34Kcal/mol and -153.84Kcal/mol, respectively. Conclusion: Moringa plant may provide incredible capabilities to develop pharmacological products. The present finding demonstrated that Moringa oleifera is an excellent plant candidate to be used for improving the health of communities.