Emerging trends and future challenges of advanced 2D nanomaterials for combating bacterial resistance.

The number of multi-drug-resistant bacteria has increased over the last few decades, which has caused a detrimental impact on public health worldwide. In resolving antibiotic resistance development among different bacterial communities, new antimicrobial agents and nanoparticle-based strategies need to be designed foreseeing the slow discovery of new functioning antibiotics. Advanced research studies have revealed the significant disinfection potential of two-dimensional nanomaterials (2D NMs) to be severed as effective antibacterial agents due to their unique physicochemical properties. This review covers the current research progress of 2D NMs-based antibacterial strategies based on an inclusive explanation of 2D NMs' impact as antibacterial agents, including a detailed introduction to each possible well-known antibacterial mechanism. The impact of the physicochemical properties of 2D NMs on their antibacterial activities has been deliberated while explaining the toxic effects of 2D NMs and discussing their biomedical significance, dysbiosis, and cellular nanotoxicity. Adding to the challenges, we also discussed the major issues regarding the current quality and availability of nanotoxicity data. However, smart advancements are required to fabricate biocompatible 2D antibacterial NMs and exploit their potential to combat bacterial resistance clinically.

Synthesis, characterization and anti-breast cancer potential of an incensole acetate nanoemulsion from Catharanthus roseus essential oil; in silico, in vitro, and in vivo study.

The characteristics of phytocompounds and essential oils have undergone extensive research in the medical and pharmaceutical sectors due to their extensive usage. In spite of the fact that these molecules are widely used, terpenes, terpenoids, and their derivatives have not yet been well characterized. This study intends to evaluate the prospective activity of incensole acetate (IA), a compound identified and isolated from Catharanthus roseus essential oil by GC/MS analysis and column chromatography, and to analyze the anticancer effect of an IA biosynthesized nanoemulsion against breast cancer. The in silico activity of IA against breast cancer targets was observed by molecular docking, ADMET assessment and molecular dynamics simulations. The IA-mediated nanoformulation exhibited cytotoxicity against breast cancer cell lines at an effective concentration when analyzed by MTT and crystal violet assay. The increased interleukin serum indicators were significantly improved as a result of nanoemulsion treatment in a DMBA-induced rat model. In addition, the anticancer properties of IA biosynthesized nanoemulsion are supported due to their potential effects on biochemical parameters, oxidative stress markers, proinflammatory cytokines, and upon tumor growth profiling in cancer-induced rats.

Determination of Nutritional and Biochemical Composition of Selected Pleurotus spps.

The global demand for good quality food is going to be increased gradually. Mushrooms are broadly used as healthy nutritious meals. The nutritional values of extracts from four distinct Pleurotus species-Pleurotus ostreatus, Pleurotus sajor-caju, Pleurotus sapidus, and Pleurotus columbinus-were determined in the current study. Firstly, proximate analysis of selected Pleurotus species was performed followed by the Bradford assay to analyze the protein spectrophotometrically; high-performance liquid chromatography (HPLC) was performed for sugar determination while GC-MS was done to determine fatty acids on organic extracts of selected mushrooms. Descriptive statistics were used to calculate the percentages while significance was determined by SPSS statistics. The results depicted that fat, protein, ash, fiber, energy contents, and total carbohydrate were in the range of 0.64-2.02%, 16.07-25.15%, 2.1-9.14%, 6.21-54.12%, 342.20-394.30 kcal/100 g, and 65.66-82.47%, respectively. The protein's maximum concentration was observed in P. ostreatus followed by P. columbinus>P. sajor-caju>P. sapidus, sequentially. Various sugars may or may not be present in selected Pleurotus spps. Among the fatty acids, the prevalence of UFA was more than that of saturated fatty acids among all selected mushrooms. From this study, it is concluded that all four Pleurotus spps. have excellent nutritional composition and can be used as valuable food and a great source of biochemical compounds.

Assessment of Hepatic Profile in Acquired Aplastic Anemia: An Experience From Pakistan.

INTRODUCTION:  Aplastic anemia (AA) is characterized by pancytopenia and hypocellular marrow in the absence of an abnormal infiltrate or increase in reticulin fibrosis. The diagnosis of AA is challenging at times due to decreased cellularity and overlapping morphological features with other bone marrow failure syndromes. Hepatitis-associated aplastic anemia (HAAA) is a rare variant in which patients typically present with jaundice and hepatitis followed by pancytopenia almost within 6 months. Post-hepatitis AA accounts for approximately 1-5% of cases, and invariably such cases are negative for the known hepatitis virus as well. There is limited literature available to understand the correlation of AA with hepatitis with none reported at the national level in our region. As AA is relatively more prevalent in Southeast Asia as compared to the western world and hepatitis is a prevalent disease in our population, the main purpose of this study was to assess the hepatic profile and determine the association of hepatitis in AA at the time of diagnosis. MATERIALS AND METHODS:  A cross-sectional study was carried out at the National Institute of Blood Disease and Bone Marrow Transplantation, Karachi, from November 2019 to December 2020 after the informed consent from patients. The study included all treatment-naïve patients of acquired AA with no prior history of taking steroids, immunosuppressive treatment, or chemoradiation therapy. Liver function tests, complete blood count, prothrombin time (PT), and activated partial thromboplastin time were performed, along with viral profiles (HAV, Hep B, Hep C, and HIV). SPSS version 23 (IBM Corp., Armonk, NY) was used for statistical analysis. Mean and standard deviations were computed for quantitative variables while percentages and frequencies were reported for qualitative variables. T-test was used to observe the main difference between groups and a p-value <0.05 was considered to be significant. RESULTS:  Out of a total of 351 patients, 29 (8.2%) patients with AA tested positive for viral hepatitis. Hepatitis A was the most prevalent hepatitis (4.0%), followed by hepatitis C (3.7%). The comparison of platelet counts in patients with and without hepatitis was reported to be of statistical significance (p-value < 0.05). A significant statistical difference (p-value < 0.0001) was found in platelet count and PT in patients of AA with and without hepatitis. CONCLUSION:  Overall, this study revealed that <10% of patients of AA had a positive screening for hepatitis A, B, and C and low platelet count, and PT was statistically significant when compared between the patients with and without hepatitis. Hepatitis being prevalent in our part of the world might have an important causal association with AA. Patients with AA should be screened for liver functions and viral hepatitis at the time of diagnosis. In addition to hepatitis A, B, and C and HIV, other causes of hepatitis should also be screened such as parvovirus B19, human herpes virus 16, and adenovirus which are not included in routine diagnostic viral testing panel.

Coronavirus Genomes and Unique Mutations in Structural and Non-Structural Proteins in Pakistani SARS-CoV-2 Delta Variants during the Fourth Wave of the Pandemic.

Genomic epidemiology of SARS-CoV-2 is imperative to explore the transmission, evolution, and also pathogenicity of viruses. The emergence of SARS-CoV-2 variants of concern posed a severe threat to the global public health efforts. To assess the potential consequence of these emerging variants on public health, continuous molecular epidemiology is of vital importance. The current study has been designed to investigate the major SARS-CoV-2 variants and emerging mutations in virus structural and non-structural proteins (NSP) during the fourth wave in September 2021 from the Punjab province of Pakistan. Twenty SARS-CoV-2 positive samples have been collected from major cities were subjected to next-generation sequencing. Among the 20 whole genomes (GenBank Accession SRR16294858-SRR16294877), 2 samples failed to be completely sequenced. These genome sequences harbored 207 non-synonymous mutations, among which 19 were unique to GISAID. The genome sequences were detected: Delta 21I, 21J variants (B.1.617.2). Mutation's spike_F157del, spike_P681R, spike_T478K, spike_T19R, spike_L452R, spike_D614G, spike_G142D, spike_E156G, and spike_R158del have been detected in all samples where K1086Q, E554K, and C1250W were unique in spike protein. These genomic sequences also harbored 129 non-synonymous mutations in NSP. The most common were NSP3_P1469S (N = 17), NSP3_A488S (N = 17), NSP3_P1228L (N = 17), NSP4_V167L (N = 17), NSP4_T492I (N = 17), NSP6_T77A (N = 17), NSP14_A394V (N = 17), NSP12_G671S (N = 18), and NSP13_P77L (N = 18). The mutation, F313Y in NSP12, detected in the current study, was found in a single isolate from Belgium. Numerous other unique mutations have been detected in the virus papain-like protease (NSP3), main protease (NSP5), and RNA-dependent RNA polymerase (NSP12). The most common non-synonymous mutations in the spike protein were subjected to stability analysis, exhibiting a stabilizing effect on structures. The presence of Delta variants may affect therapeutic efforts and vaccine efficacy. Continuous genomic epidemiology of SARS-CoV-2 in Pakistan may be useful for better management of SARS-CoV-2 infections.

In Vitro Evaluation of Dalbergia sissoo and Acacia modesta gum as Pharmaceutical Binders for Drug Delivery System

Abstract The present study aimed to compare the crude, modified and hydrolyzed gums of Dalbergia sissoo and Acacia modesta as a biodegradable binder for drug delivery system using acetaminophen as a model drug. The physiochemical properties such as pH, fluorescence analysis and swelling index were determined. The gums were hydrolyzed and modified. Acetaminophen tablets were prepared using wet granulation technique and the gum solutions were used as a binder. Hydroxypropyl methylcellulose was used as a synthetic binder. Different properties of granules and tablets were evaluated. Results showed that both gums were acidic in nature, while D. sissoo and A. modesta showed light brown and creamy color in fluorescence analysis. The swelling ratio was the highest in water followed by 0.1N HCl and least in phosphate buffer. The prepared tablets showed faster and slower dissolution profiles in the same dissolution system. The crude gums have the highest dissolution rate, and this rate was decreased in the case of modified and hydrolyzed gums samples. The crude gums showing slower release can be useful in sustained-release tablets, while the modified gums having faster release rate are helpful in conventional tablet formulation. Taken together, the selected gums could be a good model for evaluation as a binder or hydrophilic polymer in tablet formulation.

Identification of Peptides as Novel Inhibitors to Target IFN-γ, IL-3, and TNF-α in Systemic Lupus Erythematosus.

Autoimmune disorder is a chronic immune imbalance which is developed through a series of pathways. The defect in B cells, T cells, and lack of self-tolerance has been greatly associated with the onset of many types of autoimmune complications including rheumatoid arthritis, systemic lupus erythematosus (SLE), multiple sclerosis, and chronic inflammatory demyelinating polyneuropathy. The SLE is an autoimmune disease with a common type of lupus that causes tissue and organ damage due to the wide spread of inflammation. In the current study, twenty anti-inflammatory peptides derived from plant and animal sources were docked as ligands or peptides counter to proinflammatory cytokines. Interferon gamma (IFN-γ), interleukin 3 (IL-3), and tumor necrosis factor alpha (TNF-α) were targeted in this study as these are involved in the pathogenesis of SLE in many clinical studies. Two docking approaches (i.e., protein-ligand docking and peptide-protein docking) were employed in this study using Molecular Operating Environment (MOE) software and HADDOCK web server, respectively. Amongst docked twenty peptides, the peptide DEDTQAMMPFR with S-score of -11.3018 and HADDOCK score of -10.3 ± 2.5 kcal/mol showed the best binding interactions and energy validation with active amino acids of IFN-γ protein in both docking approaches. Depending upon these results, this peptide could be used as a potential drug candidate to target IFN-γ, IL-3, and TNF-α proteins to control inflammatory events. Other peptides (i.e., QEPQESQQ and FRDEHKK) also revealed good binding affinity with IFN-γ with S-scores of -10.98 and -10.55, respectively. Similarly, the peptides KHDRGDEF, FRDEHKK, and QEPQESQQ showed best binding interactions with IL-3 with S-scores of -8.81, -8.64, and -8.17, respectively.

Biochemical Stress Markers, Antioxidants, and Infectious Wound-Healing Potential of UV Irradiation and Salt Stress Effects on the Pre-Treated Seed of Bitter Melon (Momordica charantia L.).

PURPOSE: The secondary metabolites in plants are the basis of defense and stress balance, which is an important aspect in plant growth. The UV-B treatment (a biotic stress) and salt stress on bitter melon (Momordica charantia L.) were studied, and the impact of pre-sowing seed treatment was evaluated on the basis of biochemical and enzymatic biomarkers, antioxidants, and wound-healing potential during early growth stages. METHODS: The UV-B treatment for 5 and 10 min and salt stress 250 mM and 500 mM treatments were applied, and 21-day seedling tissue were collected for total phenolic contents (TPC), total flavonoid contents (TFC), antioxidant, chlorophyll contents, hydrogen peroxide, total soluble sugar, enzymes activities, and wound-healing potential studies. RESULTS: The TPC, TFC, diphenyl picrylhydrazyl (DPPH), chlorophyll contents, and total soluble sugar were recorded higher at 5 min treatment with UV-B and salt stress at 250 mM concentration. Antioxidant enzymes activities were recorded higher for 10 min UV-B treatment and 500 mM salt treatment. Wound-healing potential was found significant at 5 min treatment with UV-B radiation, which was studied in vivo in rabbits. The LC-MS analysis revealed a variety of phenolic compounds in the seedlings. CONCLUSION: The study concluded that treatments significantly affect the biological activities of bitter melon seeds at the seedling stage, and the seeds contain important phenolic compounds responsible for its antioxidant potential and enzymatic activities. Future studies could be focused on the later stages of growth, development, and yield characteristics subjected to salt stress along with UV-B radiation treatment.

Association of Viruses in the Development of Cardiovascular Diseases.

Cardiovascular diseases (CVD), primarily inflammatory cardiomyopathy, are characterized by the infiltration of inflammatory cells into the myocardium. It has a relatively high risk of deteriorating heart function and has heterogeneous etiologies. Inflammatory cardiomyopathy is mainly mediated by viral infections but can also be mediated by protozoa, fungal or bacterial infections. Besides that, there are a wide variety of drugs, toxic substances, and systemic immune-mediated diseases that result in the development of cardiovascular diseases (CVDs). Despite broad research, inflammatory cardiomyopathy has a poor prognosis. The roles of the pathogens, host genomic counterparts and environmental triggers in the progression of disease are still under consideration, including the role of some viruses as active inducers and others as bystanders. In this review article, we review the available evidence on the types, pathogenesis and treatment of myocarditis, inflammatory cardiomyopathy, and atherosclerosis with a particular focus on virus-associated cardiac diseases.

The emerging role and significance of circular RNAs in viral infections and antiviral immune responses: possible implication as theranostic agents.

Circular RNAs (circRNAs) are ubiquitously expressed, covalently closed rings, produced by pre-mRNA splicing in a reversed order during post-transcriptional processing. Circularity endows 3'-5'-linked circRNAs with stability and resistance to exonucleolytic degradation which raises the question whether circRNAs may be relevant as potential therapeutic targets or agents. High stability in biological systems is the most remarkable property and a major criterion for why circRNAs could be exploited for a range of RNA-centred medical applications. Even though various biological roles and regulatory functions of circRNAs have been reported, their in-depth study is challenging because of their circular structure and sequence-overlap with linear mRNA counterparts. Moreover, little is known about their role in viral infections and in antiviral immune responses. We believe that an in-depth and detailed understanding of circRNA mediated viral protein regulations will increase our knowledge of the biology of these novel molecules. In this review, we aimed to provide a comprehensive basis and overview on the biogenesis, significance and regulatory roles of circRNAs in the context of antiviral immune responses and viral infections including hepatitis C virus infection, hepatitis B virus infection, hepatitis delta virus infection, influenza A virus infection, Epstein-Barr virus infection, kaposi's sarcoma herpesvirus infection, human cytomegalovirus infection, herpes simplex virus infection, human immunodeficiency virus infection, porcine epidemic diarrhoea virus infection, ORF virus infection, avian leukosis virus infection, simian vacuolating virus 40 infection, transmissible gastroenteritis coronavirus infection, and bovine viral diarrhoea virus infection. We have also discussed the critical regulatory role of circRNAs in provoking antiviral immunity, providing evidence for implications as therapeutic agents and as diagnostic markers.

Simultaneous fluorometric determination of the DNAs of Salmonella enterica, Listeria monocytogenes and Vibrio parahemolyticus by using an ultrathin metal-organic framework (type Cu-TCPP).

Ultrathin (<10 nm) nanosheets of a metal-organic framework (MOF-NSs) were prepared in high-yield and scalable production by a surfactant-assisted one-step method. The MOF-NSs possess distinguished affinity for ssDNA but not for dsDNA. This causes the fluorescence of the labeled DNA to be quenched. On binding to the target DNA (shown here for Salmonella enterica, Listeria monocytogenes and Vibrio parahemolyticus), the labeled duplex is released and the fluorescence of the label is restored. The labels Texas Red, Cy3 and FAM were used and give red, red or green fluorescence depending on the kind of pathogen. The detection limits are 28 pM, 35 pM and 15 pM for the gene segments of Salmonella enterica, Listeria monocytogenes and Vibrio parahemolyticus, respectively. Graphical abstract Schematic of an ultrasensitive fluorescent biosensor for multiplex detection of pathogenic DNAs based on ultrathin MOF nanosheets (type Cu-TCPP).

Anticancer, antithrombotic, antityrosinase, and anti-α-glucosidase activities of selected wild and commercial mushrooms from Pakistan.

Mushrooms have been accepted as nutraceutical foods because of their high nutritional and functional values. They have also gained interest due to their medicinal properties, economic importance, and organoleptic merit. In this study, wild Ganoderma lucidum and four commercial mushrooms, that is, Pleurotus ostreatus, Volvariella volvacea, Hericium erinaceus, and Lentinus edodes from Pakistan were screened for their biological activities such as anticancer, antityrosinase, anti-α-glucosidase, and antithrombotic activities from their methanol, ethanol, and water extracts. Enzyme inhibition assay showed that selected mushrooms are potent inhibitors with %age inhibition ranging from 19.00% to 80.91%, and 32.85% to 83.38% for tyrosinase and α-glucosidase, respectively. The best tyrosinase inhibition was shown by P. ostreatus whereas L. edodes was found best as α-glucosidase inhibitor. These mushrooms were tested against cancer cell lines including HT-29 colon and H-1299 lungs carcinoma cell lines. G. lucidum showed 29% and 24% viability of cells against HT-29 and H-1299 cell lines, respectively. This antiproliferative effect was in dose-dependent manner, and the maximum inhibition was observed at 200 µg/ml. Mushrooms extracts were also found effective against clot lysis. The percentage of clot lysis was in the range of 27%-29%. The research would provide knowledge to the people of Pakistan about the importance of locally available commercial mushrooms and wild mushrooms for health improvement and prevention against different kinds of diseases.

Solution-Phase Synthesis of Platinum Nanoparticle-Decorated Metal-Organic Framework Hybrid Nanomaterials as Biomimetic Nanoenzymes for Biosensing Applications.

The synthesis of nanomaterials with specific properties and functions as biomimetic nanoenzymes has attracted extensive attention in the past decades due to their great potential to substitute natural enzymes. Herein, a facile and simple method for the preparation of platinum nanoparticle (PtNP)-decorated two-dimensional metal-organic framework (MOF) nanocomposites was developed. A ligand with heme-like structure, Fe(III) tetra(4-carboxyphenyl)porphine chloride (TCPP(Fe)), was applied to synthesize MOF nanosheets (denoted as Cu-TCPP(Fe) nanosheets) in high yield. Ultrathin Cu-TCPP(Fe) nanosheets with thickness less than 10 nm were used as a novel template for the growth of ultrasmall and uniform PtNPs. Significantly, the obtained hybrid nanomaterials (PtNPs/Cu-TCPP(Fe) hybrid nanosheets) exhibit enhanced peroxidase-like activity compared to PtNPs, Cu-TCPP(Fe) nanosheets, and the physical mixture of both due to the synergistic effect. On account of the excellent peroxidase-like activity of PtNPs/Cu-TCPP(Fe) hybrid nanosheets, we established a colorimetric method for sensitive and rapid detection of hydrogen peroxide. Furthermore, by combining with glucose oxidase, a cascade colorimetric method was established to further detect glucose with excellent sensitivity and selectivity.

Structural and functional annotation of citrate synthase from Aspergillus niger ANJ-120.

Citrate synthase (CS) is involved in citric acid biosynthesis which is a well-established metabolic pathway. The condensation of acetyl-CoA with oxaloacetate is catalyzed by CS. Citric acid (CA) has a number of applications in pharmaceutical industry. CA in combination with bicarbonates is used as an effervescent in the preparations of tablets and powders. It has also been used as an anticoagulant and acidulant to form mild astringent. In current study, detailed structural and functional analyses of CS protein were carried out using various bioinformatics tools. Structural modeling was also done by building 3D model of CS from Aspergillus niger ANJ-120 using Modeller 9.16 software. The 3D Model was then evaluated using different online approaches. Furthermore, superimposition of query and template structures, Root Mean Squared Deviation and visualization of generated model were done through UCSF Chimera 1.5.3. Even though various roles of CS protein were already known and verified experimentally, here we presented a structural analysis of CS protein. The structural investigation of CS protein will be helpful for protein engineering strategies and understanding the interactions among proteins. Due to large number of applications, the production of citric acid by A. niger and its bioinformatics studies will offer substantial improvement in commercial scale intensification of this useful product.

Wild Mushrooms: A Potential Source of Nutritional and Antioxidant Attributes with Acceptable Toxicity.

This paper describes in detail proximate composition, nutritional profile, phytochemical constituents, antioxidant activities, antimicrobial potential, and antihemolytic activity (towards human erythrocytes) of various fractions of wild Ganoderma lucidum. Proximate analysis established that wild G. lucidum comprises about 87.02±5.45% of moisture, and the remaining part is a rich source of proteins (8.59±0.37%), crude fiber (54.21±1.2%), and carbohydrate (35.16%) with smaller fat content (3.33 %). Similarly, phytochemical screening revealed the presence of flavonoids (217.51±0.30 mg/g), ascorbic acid (116±7.32 mg/g), phenolics (360.72±34.07 mg/g), ß-carotenes (0.42±0.04 µg/g), and lycopene (0.05±0.00 µg/g). Extracts of wild G. lucidum in various solvents provided first line protection against Escherichia coli and Pasteurella multocida in the order of ethyl acetate> ethanol> methanol> n-hexane> water. Furthermore, aqueous and methanolic extracts of wild G. lucidum were found to be safe towards human erythrocytes. Overall, wild mushroom (G. lucidum) was found to be a good source of dietary supplements, antimicrobial and antioxidant agents in the pursuance of its commercial utilization in food and pharmaceutical industries.

Clinicohematological and cytogenetic profile of myelodysplastic syndromes in Pakistan-compare and contrast.

BACKGROUND: Myelodysplastic syndromes (MDS) are clonal stem cell disorders exhibiting cytopenias, ineffective hematopoiesis and morphological dysplasia. Bone marrow cytogenetics, inspite of being incorporated as mandatory tool in diagnosis are done less frequently due to limited availability of this technique in Pakistan. The aim of the study was to study baseline clinicohematological and cytogenetic characteristics of patients presenting with de novo MDS. RESULTS: A retrospective cross sectional study was done at National Institute of Blood Diseases and Bone Marrow Transplantation, Karachi, Pakistan from 2010 to 2016. Total of 177 patients were included in the study having median age 51 years and male to female ratio of 3:1. Pancytopenia was observed in 80 (45%) patients and bicytopenia in 74 (42%). Mean Hb% was 7.8 ± 2.18 g/dl, total leukocyte count (TLC) 8.8 ± 13.6 × 109/l, platelet count was 82 ± 95.7 × 109/l. Of total 170 (96%) were transfusion dependent. Refractory cytopenias with multilineage dysplasia (RCMD) was the most common world health organization (WHO) category. Karyotype was done in 98 (55%) patients out of which 44 (45%) had abnormal karyotype, complex karyotype (CK) was most commonly observed in 12 (12.2%) followed by monosomy 7 in 7 (7.1%). CONCLUSIONS: We found younger median age at diagnosis, higher mean TLC and no significant history of recurrent infections. CK and monosomy 7 carry bad prognostic implications and early disease transformation to acute myeloid leukemia (AML). Monosomy 7 being associated with bad overall survival, such patients must be identified early with close clinical follow up and offered stem cell transplant. This is the largest cohort of patients of MDS evaluated for baseline clinical and cytogenetic characteristics in our country.