Investigation of Trifolium repens L. from the Indian Himalayan region as a phyto-therapeutic agent.

Trifolium repens is a well-known herbaceous, perennial herb and has been extensively used in the traditional medicine system over the years. Various parts of the plant are traditionally used as a curative agent against several health ailments such as skin problems, wound healing, stomach disorders, sedative, fever, antiseptic, analgesic, expectorant, psoriasis and eczema. To maximise the plant's potential for usage in the future, the review also aims to update information about its significant pharmacological properties. The ethnomedicinal benefits of T. repens have been well studied; however, the facets of the plant have not been explored yet. The current review outlines several bioactive compounds quantified from T. repens and a few of them namely quercetin, kaempferol, myricetin, acacetin and linamarin, have been reported to have biological activities such as antibacterial, antifungal, antileishmanial, anti-inflammatory, antiaging and anti-hepatotoxic activities. A significant number of in vitro studies have been done on the plant extract, but little is known about the isolation and efficacy of the potent natural bioactive compounds of T. repens. The bioactive compounds in T. repens can be used for advanced drug development against various health disorders.

Medicinal Plants as Therapeutic Alternatives to Combat Mycobacterium tuberculosis: A Comprehensive Review.

Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium tuberculosis (MTB) and a significant health concern worldwide. The main threat to the elimination of TB is the development of resistance by MTB to the currently used antibiotics and more extended treatment methods, which is a massive burden on the health care system. As a result, there is an urgent need to identify new, effective therapeutic strategies with fewer adverse effects. The traditional medicines found in South Asia and Africa have a reservoir of medicinal plants and plant-based compounds that are considered another reliable option for human beings to treat various diseases. Abundant research is available for the biotherapeutic potential of naturally occurring compounds in various diseases but has been lagging in the area of TB. Plant-based compounds, or phytoproducts, are being investigated as potential anti-mycobacterial agents by reducing bacterial burden or modulating the immune system, thereby minimizing adverse effects. The efficacy of these phytochemicals has been evaluated through drug delivery using nanoformulations. This review aims to emphasize the value of anti-TB compounds derived from plants and provide a summary of current research on phytochemicals with potential anti-mycobacterial activity against MTB. This article aims to inform readers about the numerous potential herbal treatment options available for combatting TB.

Recent Advances in Green Synthesis, Characterization, and Applications of Bioactive Metallic Nanoparticles.

Nanoparticles (NPs) are elements derived from a cluster of atoms with one or more dimensions in the nanometer scale in the range of 1-100 nm. The bio nanofabrication of metallic NPs is now an important dynamic area of research, with major significance in applied research. Biogenic synthesis of NPs is more desirable than physical and chemical synthesis due to its eco-friendliness, non-toxicity, lower energy consumption, and multifunctional nature. Plants outperform microorganisms as reducing agents as they contain large secondary biomolecules that accelerate the reduction and stability of the NPs. The produced NPs can then be studied spectroscopically (UV-Visible, XRD, Raman, IR, etc.) and microscopically (SEM, TEM, AFM, etc.). The biological reduction of a metallic ion or its oxide to a nanoparticle is quick, simple, and may be scaled up at room temperature and pressure. The rise in multi-drug resistant (MDR) microbes due to the immoderate use of antibiotics in non-infected patients is a major cause of morbidity and mortality in humans. The contemporary development of a new class of antibiotics with different mechanisms of action to kill microbes is crucial. Metals and their oxides are extremely toxic to microbes at unprecedentedly low concentrations. In addition, prevailing infections in plants and animals are raising significant concerns across the globe. NPs' wide range of bioactivity makes them ideal antimicrobial agents in agricultural and medical fields. The present review outlines the synthesis of metallic NPs from botanicals, which enables the metals to be in a stabilized form even after ionization. It also presents a valuable database on the biofunctionalization of synthesized NPs for further drug development.

Surgical management of endometriosis in a severe Hemophilia A female patient and the role of transfusion medicine specialist: A case report with review of literature.

Hemophilia A is an X-linked recessive bleeding disorder occurs due to deficiency of factor VIII (FVIII). The disease manifests exclusively in males though it rarely occurs in females due to complex pathophysiological mechanisms. We present a rare case of female hemophilia due to skewed X-inactivation which adversely affected the quality of patient life. She presented with recurrent abdominal pain and was diagnosed with severe endometriosis and underwent total abdominal hysterectomy with left salpingo-oophorectomy and appendicectomy. She was infused recombinant factor VIII both prophylactically and postoperatively as per the World Federation of Hemophilia guidelines. Recombinant Factor VIII was supplemented every 12th hourly and Factor VIII activity levels were monitored daily. She was discharged uneventfully on the postoperative day 21 after screened negative for acquired inhibitors.

Emerging insights into mitochondria-specific targeting and drug delivering strategies: Recent milestones and therapeutic implications.

Mitochondria are a major intracellular organelle for drug targeting due to its functional roles in cellular metabolism and cell signaling for proliferation and cell death. Mitochondria-targeted treatment strategy could be promising to improve the therapeutic efficacy of cancer while minimizing the adverse side effects. Over the last decades, several studies have explored and focused on mitochondrial functions, which has led to the emergence of mitochondria-specific therapies. Molecules in the mitochondria are considered to be prime targets, and a wide range of molecular strategies have been designed for targeting mitochondria compared with that of the cytosol. In this review, we focused on the molecular mechanisms of mitochondria-specific ligand targeting and selective drug action strategies for targeting mitochondria, including those premised on mitochondrial targeting of signal peptides (MTS), cell-penetrating peptides (CPPs), and use of lipophilic cations. Furthermore, most research has concentrated on specific conjugation of ligands to therapeutic molecules to enhance their effectiveness. There are several variations for the ideal design and development for mitochondrial-targeted drugs, such as selecting a suitable ligand and linker targets. However, some challenges related to drug solubility and selectivity could be resolved using the nanocarrier system. Nanoparticles yield excellent advantages for targeting and transmitting therapeutic drugs, and they offer elegant platforms for mitochondria-specific drug delivery. We explain many of the advanced and proven strategies for multifunctional mitochondria-specific targets, which should contribute to achieving better anticancer therapies in a promising future.

Identification and Characterization of a Newly Isolated Chitinase-Producing Strain Bacillus licheniformis SSCL-10 for Chitin Degradation.

Chitinases or chitinolytic enzymes have different applications in the field of medicine, agriculture, and industry. The present study is aimed at developing an effective hyperchitinase-producing mutant strain of novel Bacillus licheniformis. A simple and rapid methodology was used for screening potential chitinolytic microbiota by chemical mutagenesis with ethylmethane sulfonate and irradiation with UV. There were 16 mutant strains exhibiting chitinase activity. Out of the chitinase-producing strains, the strain with maximum chitinase activity was selected, the protein was partially purified by SDS-PAGE, and the strain was identified as Bacillus licheniformis (SSCL-10) with the highest specific activity of 3.4 U/mL. The induced mutation model has been successfully implemented in the mutant EMS-13 (20.2 U/mL) that produces 5-6-fold higher yield of chitinase, whereas the mutant UV-11 (13.3 U/mL) has 3-4-fold greater chitinase activity compared to the wild strain. The partially purified chitinase has a molecular weight of 66 kDa. The wild strain (SSCL-10) was identified as Bacillus licheniformis using 16S rRNA sequence analysis. This study explores the potential applications of hyperchitinase-producing bacteria in recycling and processing chitin wastes from crustaceans and shrimp, thereby adding value to the crustacean industry.

Footprint of the COVID-19 Pandemic in India: A Study of Immune Landscape and Other Factors Shielding Mortality.

The impact of the SARS-CoV-2 pandemic has significantly affected global health and created a world crisis. The exponentially increasing numbers of infection and mortality have made preventive measures challenging. India being a highly populated nation has so far effectively counteracted the pandemic outbreak with a significantly lower rate of mortality despite the high infection rates. The genetic architecture of the immune response genes in the Indian population, BCG vaccination, the predominantly young age group of people, and their traditional food habits might contribute to the lower rate of mortality. Human leukocyte antigens (HLA) play a vital role in triggering T cells, and natural killer (NK) cells can immediately react to eliminate infected cells. Activation of virus-specific CD4+ T cells and CD8+ cytotoxic T cells selectively targets the infected cells and strengthens the immunoregulatory system. The checkpoint for NK cell function is the engagement of killer Ig-like receptors (KIR) molecules with their respective HLA ligands overexpressed or expressed on the compromised virus-infected cells which have shown polymorphism among different ethnic groups. Here, we explore if certain KIR-HLA motifs grant Indians a survival advantage in terms of the low rate of mortality. Additionally, enhanced immunity through BCG vaccination may favor fruitful eradication of SARS-CoV-2 and provide the way out as in therapeutic intervention and vaccination strategies.

Evaluation of process excellence tools in improving donor flow management in a tertiary care hospital in South India.

BACKGROUND: Process Excellence is a value based approach and focuses on standardizing work processes by eliminating the non-value added processes, identify process improving methodologies and maximize capacity and expertise of the staff. AIM AND OBJECTIVES: To Evaluate the utility of Process Excellence Tools in improving Donor Flow Management in a Tertiary care Hospital by studying the current state of donor movement within the blood bank and providing recommendations for eliminating the wait times and to improve the process and workflow. MATERIALS AND METHODS: The work was done in two phases; The First Phase comprised of on-site observations with the help of an expert trained in Process Excellence Methodology who observed and documented various aspects of donor flow, donor turn around time, total staff details and operator process flow. The Second Phase comprised of constitution of a Team to analyse the data collected. The analyzed data along with the recommendations were presented before an expert hospital committee and the management. RESULTS: Our analysis put forward our strengths and identified potential problems. Donor wait time was reduced by 50% after lean due to better donor management with reorganization of the infrastructure of the donor area. Receptionist tracking showed that 62% of the total time the staff wastes in walking and 22% in other non-value added activities. Defining Duties for each staff reduced the time spent by them in non-value added activities. Implementation of the token system, generation of unique identification code for donors and bar code labeling of the tubes and bags are among the other recommendations. CONCLUSION: Process Excellence is not a programme; it's a culture that transforms an organization and improves its Quality and Efficiency through new attitudes, elimination of wastes and reduction in costs.

Optimising assessment intervals improves visual outcomes in ranibizumab-treated age-related neovascular degeneration: using the stability phase as a benchmark.

BACKGROUND: To observe visual acuity change in the stability phase when follow-up intervals are decreased in ranibizumab-treated neovascular age-related macular degeneration (nvAMD). METHODS: Selection of patients was based on a review of a cohort of 189 eyes of 154 patients with nvAMD treated with intravitreal ranibizumab in routine clinical practice. Patients were transferred from a base hospital with a 8-week follow-up interval to a community eye clinic, enabling a new follow-up interval of 4 weeks. Staff, assessment, and treatment protocols were equivalent in the two centres. Patients were included when they were in the stability phase of treatment defined 1 month after having completed their three initiation treatments with ranibizumab. Each patient was required to have attended at least a further 12 visits; this means a follow-up time for a year or longer, consisting of six visits at the base hospital followed by six visits at the new eye clinic. The best-corrected visual acuity (BCVA), follow-up intervals and injection numbers were collected. RESULTS: Seventy-two eyes of 62 patients were included. The mean follow-up interval for the six visits in the base hospital was 56.81 days, and in the new eye centre 31.81 days. The BCVA loss in the base hospital was -1.13 letters, compared to a gain of +4.61 letters in the community eye clinic over the six visits. The number of ranibizumab injections was 3.67 in the base hospital, compared with 3.91 in the other centre over the respective periods. CONCLUSION: Visual acuity improves and severe visual loss decreases when follow-up intervals reduce from approximately 8 weeks to 4 weeks. Furthermore, using the stability phase to evaluate the outcome and effectiveness of our treatments for age-related macular degeneration appeared to be an efficient tool.

Reduction of estrogen-induced transformation of mouse mammary epithelial cells by N-acetylcysteine.

A growing number of studies indicate that breast cancer initiation is related to abnormal estrogen oxidation to form an excess of estrogen-3,4-quinones, which react with DNA to form depurinating adducts and induce mutations. This mechanism is often called estrogen genotoxicity. 4-Catechol estrogens, precursors of the estrogen-3,4-quinones, were previously shown to account for most of the transforming and tumorigenic activity. We examined whether estrogen-induced transformation can be reduced by inhibiting the oxidation of a 4-catechol estrogen to its quinone. We demonstrate that E6 cells (a normal mouse epithelial cell line) can be transformed by a single treatment with a catechol estrogen or its quinone. The transforming activities of 4-hydroxyestradiol and estradiol-3,4-quinone were comparable. N-Acetylcysteine, a common antioxidant, inhibited the oxidation of 4-hydroxyestradiol to the quinone and consequent formation of DNA adducts. It also drastically reduced estrogen-induced transformation of E6 cells. These results strongly implicate estrogen genotoxicity in mammary cell transformation. Since N-acetylcysteine is well tolerated in clinical studies, it may be a promising candidate for breast cancer prevention.

The role of polycyclic aromatic hydrocarbon-DNA adducts in inducing mutations in mouse skin.

Polycyclic aromatic hydrocarbons (PAH) form stable and depurinating DNA adducts in mouse skin to induce preneoplastic mutations. Some mutations transform cells, which then clonally expand to establish tumors. Strong clues about the mutagenic mechanism can be obtained if the PAH-DNA adducts can be correlated with both preneoplastic and tumor mutations. To this end, we studied mutagenesis in PAH-treated early preneoplastic skin (1 day after exposure) and in the induced papillomas in SENCAR mice. Papillomas were studied by PCR amplification of the H-ras gene and sequencing. For benzo[a]pyrene (BP), BP-7,8-dihydrodiol (BPDHD), 7,12-dimethylbenz[a]anthracene (DMBA) and dibenzo[a,l]pyrene (DB[a,l]P), the codon 13 (GGC to GTC) and codon 61 (CAA to CTA) mutations in papillomas corresponded to the relative levels of Gua and Ade-depurinating adducts, despite BP and BPDHD forming significant amounts of stable DNA adducts. Such a relationship was expected for DMBA and DB[a,l]P, as they formed primarily depurinating adducts. These results suggest that depurinating adducts play a major role in forming the tumorigenic mutations. To validate this correlation, preneoplastic skin mutations were studied by cloning H-ras PCR products and sequencing individual clones. DMBA- and DB[a,l]P-treated skin showed primarily A.T to G.C mutations, which correlated with the high ratio of the Ade/Gua-depurinating adducts. Incubation of skin DNA with T.G-DNA glycosylase eliminated most of these A.T to G.C mutations, indicating that they existed as G.T heteroduplexes, as would be expected if they were formed by errors in the repair of abasic sites generated by the depurinating adducts. BP and its metabolites induced mainly G.C to T.A mutations in preneoplastic skin. However, PCR over unrepaired anti-BPDE-N(2)dG adducts can generate similar mutations as artifacts of the study protocol, making it difficult to establish an adduct-mutation correlation for determining which BP-DNA adducts induce the early preneoplastic mutations. In conclusion, this study suggests that depurinating adducts play a major role in PAH mutagenesis.

Improved measurement of dibenzo[a,l]pyrene-induced abasic sites by the aldehyde-reactive probe assay.

Dibenzo[a,l]pyrene (DB[a,l]P) induces abundant amounts of depurinating adducts that spontaneously dissociate to form abasic sites in DNA. However, several previous studies that used the aldehyde-reactive probe (ARP) assay, could not verify abasic site formation by DB[a,l]P. Therefore, we examined whether a modification of the ARP assay would allow greater quantification of abasic sites. A previous study indicated that the abasic site quantification is improved by letting ARP trap the nascent abasic sites in cells, before extracting DNA for the assay. To test whether the addition of ARP to the DB[a,l]P-DNA adduct-forming reaction would improve abasic site quantification, we treated calf thymus DNA (0.625 mg/mL) with DB[a,l]P (80 microM) and 3-methylcholanthrene-treated rat liver microsomes with or without ARP (3 mM). The inclusion of ARP in the adduct-forming reaction resulted in significantly greater detection of abasic sites (62 lesions/10(6) bp versus 3.7 lesions/10(6) bp). DB[a,l]P also induces DNA strand breaks. The strand breaks may occur at abasic sites and by other mechanisms, such as oxidative damage. ARP/O-methoxyamine-abasic site conjugates are refractory to strand breakage, however, ARP or O-methoxyamine (3-10 mM) could only partially protect DB[a,l]P-induced DNA degradation, presumably by protecting the abasic sites, but not the other strand breaks. These results suggest that if DNA strand breakages occur at the abasic sites or at bases flanking them, and the fragments are lost during DNA extraction, abasic site estimation could be compromised. To obtain an independent line of evidence for abasic site formation in DB[a,l]P-treated cells, mouse Mbeta16 fibroblasts were treated with DB[a,l]P and O-methoxyamine. O-Methoxyamine is known to potentiate cytotoxicity of abasic site-inducing chemicals by forming abasic site conjugates, which partially inhibits their repair. O-Methoxyamine was found to increase DB[a,l]P cytotoxicity in these cells, supporting the idea that DB[a,l]P formed abasic sites. In summary, the inclusion of ARP in the DB[a,l]P-DNA adduct-forming reaction traps and protects the nascent abasic sites, allowing an improved quantification of abasic sites.