Optimization studies on batch extraction of phenolic compounds from Azadirachta indica using genetic algorithm and machine learning techniques.

Phenolic compounds play a crucial role as secondary metabolites due to their substantial biological activity and medicinal value. These compounds are present in various parts of plant species. This study focused on solid-liquid batch extraction to recover total phenolic compounds from Azadirachta indica leaves. The experimental design was based on the Taguchi L16 array, considering four independent factors: extraction time, temperature, particle size, and solid-to-solvent ratio. Among these factors, the particle size exerted the maximum influence. Particle size inversely affects the yield of total phenolic content (TPC), while temperature, time, and solid-to-liquid ratio have a direct impact. The process factors concerned were investigated both experimentally and through machine learning techniques. Support vector regression (SVR) and random forest method (RFM) algorithms were utilized for predicting TPC, while a genetic algorithm (GA) was employed to derive optimal process parameters. The GA predicts the optimal extraction factors, yielding the maximum TPC. During this study, these factors were the following: particle size of 0.15 mm, extraction time of 40 min, solid-to-liquid ratio of 1:25 g/mL, and a temperature of 55 °C, with a predicted value of 23.039 mg GAE/g of plant material. Notably, in this study, the SVR values of TPC yield closely matched the experimental values for the training and test data set when compared with the random forest method values.

Polianthes tuberosa-Mediated Silver Nanoparticles from Flower Extract and Assessment of Their Antibacterial and Anticancer Potential: An In Vitro Approach.

Plant-mediated metallic nanoparticles have beenreported for a diversified range of applications in biological sciences. In the present study, we propose the Polianthes tuberosa flower as a reducing and stabilizing agent for the synthesis of silver nanoparticles (PTAgNPs). The PTAgNPs were exclusively characterized using UV-Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), atomic force microscopy, zeta potential, and transmission electron microscopy (TEM) studies. In a biological assay, we investigated the antibacterial and anticancer activity of silver nanoparticles in the A431 cell line. The PTAgNPs demonstrated a dose-dependent activity in E. coli and S. aureus, suggesting the bactericidal nature of AgNPs. The PTAgNPs exhibited dose-dependent toxicity in the A431 cell line, with an IC50 of 54.56 µg/mL arresting cell growth at the S phase, as revealed by flow cytometry analysis. The COMET assay revealed 39.9% and 18.15 severities of DNA damage and tail length in the treated cell line, respectively. Fluorescence staining studies indicate that PTAgNPs cause reactive oxygen species (ROS) and trigger apoptosis. This research demonstrates that synthesized silver nanoparticles have a significant effect on inhibiting the growth of melanoma cells and other forms of skin cancer. The results show that these particles can cause apoptosis or cell death in malignant tumor cells. This suggests that they could be used to treat skin cancers without harming normal tissues.

The Characteristics of Green-synthesized Magnesium Oxide Nanoparticles (MgONPs) and their Biomedical Applications.

In the current epoch, noble metals/metal oxides with precise structures are needed to develop sustainable products to improve the welfare of human beings and the environment. Nanomaterials in the regime 1 -100 nm scale are a promising material for the research fraternities owing to their stupendous properties. The metallic/metal oxide nanoparticles (silver, gold, copper oxide, iron oxide, magnesium oxide) are gaining significant momentum and need to be extensively studied. Magnesium oxide nanoparticles (MgONPs) are a periclase, white hygroscopic material consisting of Mg2+ ions and O-2 ions in lattice arranged. These nanoparticles can be fabricated through physical, chemical and biological methods. The development of green synthesized MgONPs needs to be ascertained and explored its ultimate in medicine, health, cosmetics, environmental protection, chemical industries, and energy. Therefore, the present review manifests the green synthetic approaches of MgONPs and their impact on crystalline structure and shape. Further, we have provided the antibacterial and anticancer activities of MgONPs thoroughly reported in various kinds of literature. Overall, the unique MgONPs can be plausibly used as safe biomaterials in biomedical applications.

Early cytopenias and infections after standard of care idecabtagene vicleucel in relapsed or refractory multiple myeloma.

Idecabtagene vicleucel (ide-cel) was FDA-approved in March 2021 for the treatment of relapsed/refractory multiple myeloma after 4 lines of therapy. On the KarMMa trial, grade ≥ 3 cytopenias and infections were common. We sought to characterize cytopenias and infections within 100 days after ide-cel in the standard-of-care (SOC) setting. This multi-center retrospective study included 52 patients who received SOC ide-cel; 47 reached day-90 follow-up. Data were censored at day 100. Grade ≥ 3 cytopenia was present among 65% of patients at day 30 and 40% of patients at day 90. Granulocyte colony stimulating factor (G-CSF) was administered to 88%, packed red blood cell transfusions to 63%, platelet transfusions to 42%, thrombopoietin (TPO) agonists to 21%, intravenous immunoglobulin to 13%, and CD34+ stem cell boosts to 8%. At day 100, 19% and 13% of patients had ongoing use of TPO agonists and G-CSF, respectively. Infections occurred in 54% of patients and were grade ≥ 3 in 23%. Earlier infections in the first 30 days were typically bacterial (68%) and severe (50%). Later infections between days 31 and 100 were 50% bacterial and 42% viral; only 13% were grade ≥ 3. On univariate analysis, high pre-CAR-T marrow myeloma burden (≥ 50%), circulating plasma cells at pre-lymphodepletion (LD), and grade ≥ 3 anemia at pre-LD were associated with grade ≥ 3 cytopenia at both days 30 and 90. Longer time from last bridging treatment to LD was the only significant risk factor for infection.

Inhibitory effect of selected Indian honey on colon cancer cell growth by inducing apoptosis and targeting the ß-catenin/Wnt pathway.

Colon cancer is the most prevalent cause of death from cancer across the globe. Although chemotherapy drugs are predominantly used, their toxicity always remains a cause of concern. As an alternative to synthetic drugs, natural compounds or nutraceuticals are comparatively less toxic. Honey is widely used across different cultures as an alternative form of medicine. It represents a prominent source of plant-phenolic compounds and there is demonstrable evidence of its anti-oxidant and anti-microbial activities. The aim of the present work was to investigate the anti-proliferative effect of some Indian honeys and analyze their mechanism of action in colon cancer. In order to establish the composition-activity relationship, we evaluated the bioactive components present in selected honey samples by GC-MS and HPLC analysis. Indian honey samples showed a significant inhibitory impact on cell growth by restricting cell proliferation, causing apoptosis, and restricting the cell cycle in the G2/M phase specifically for colon cancer cells. The apoptotic activities, as imparted by the honey samples, were established by Annexin V/PI staining, real-time PCR, and immunoblot analyses. The treated cells showed increased expressions of p53 and caspases 3, 8, and 9, thus indicating the involvement of both extrinsic and intrinsic apoptotic pathways. The honey samples were also found to inhibit the ß-catenin/Wnt pathway. In the next phase of the study, the efficacy of these honey samples was evaluated in colon carcinoma induced SD-rats. Overall, these findings demonstrated that selected Indian honeys could be established as effective nutraceuticals for the prevention as well as cure of colon cancer.

A Rapid Pharmacogenomic Assay to Detect NAT2 Polymorphisms and Guide Isoniazid Dosing for Tuberculosis Treatment.

Rationale: Standardized dosing of antitubercular drugs contributes to a substantial incidence of toxicities, inadequate treatment response, and relapse, in part due to variable drug concentrations achieved. SNPs in the NAT2 (N-acetyltransferase-2) gene explain the majority of interindividual pharmacokinetic variability of isoniazid (INH). However, an obstacle to implementing pharmacogenomic-guided dosing is the lack of a point-of-care assay. Objectives: To develop and test a NAT2 classification algorithm, validate its performance in predicting isoniazid clearance, and develop a prototype pharmacogenomic assay. Methods: We trained random forest models to predict NAT2 acetylation genotype from unphased SNP data using a global collection of 8,561 phased genomes. We enrolled 48 patients with pulmonary tuberculosis, performed sparse pharmacokinetic sampling, and tested the acetylator prediction algorithm accuracy against estimated INH clearance. We then developed a cartridge-based multiplex quantitative PCR assay on the GeneXpert platform and assessed its analytical sensitivity on whole blood samples from healthy individuals. Measurements and Main Results: With a 5-SNP model trained on two-thirds of the data (n = 5,738), out-of-sample acetylation genotype prediction accuracy on the remaining third (n = 2,823) was 100%. Among the 48 patients with tuberculosis, predicted acetylator types were 27 (56.2%) slow, 16 (33.3%) intermediate, and 5 (10.4%) rapid. INH clearance rates were lowest in predicted slow acetylators (median 14.5 L/h), moderate in intermediate acetylators (median 40.3 L/h), and highest in fast acetylators (median 53.0 L/h). The cartridge-based assay accurately detected all allele patterns directly from 25 µl of whole blood. Conclusions: An automated pharmacogenomic assay on a platform widely used globally for tuberculosis diagnosis could enable personalized dosing of INH.

Arabidopsis ACINUS is O-glycosylated and regulates transcription and alternative splicing of regulators of reproductive transitions.

O-GlcNAc modification plays important roles in metabolic regulation of cellular status. Two homologs of O-GlcNAc transferase, SECRET AGENT (SEC) and SPINDLY (SPY), which have O-GlcNAc and O-fucosyl transferase activities, respectively, are essential in Arabidopsis but have largely unknown cellular targets. Here we show that AtACINUS is O-GlcNAcylated and O-fucosylated and mediates regulation of transcription, alternative splicing (AS), and developmental transitions. Knocking-out both AtACINUS and its distant paralog AtPININ causes severe growth defects including dwarfism, delayed seed germination and flowering, and abscisic acid (ABA) hypersensitivity. Transcriptomic and protein-DNA/RNA interaction analyses demonstrate that AtACINUS represses transcription of the flowering repressor FLC and mediates AS of ABH1 and HAB1, two negative regulators of ABA signaling. Proteomic analyses show AtACINUS's O-GlcNAcylation, O-fucosylation, and association with splicing factors, chromatin remodelers, and transcriptional regulators. Some AtACINUS/AtPININ-dependent AS events are altered in the sec and spy mutants, demonstrating a function of O-glycosylation in regulating alternative RNA splicing.

Biogenic nanoparticles: a comprehensive perspective in synthesis, characterization, application and its challenges.

BACKGROUND: Translating the conventional scientific concepts into a new robust invention is a much needed one at a present scenario to develop some novel materials with intriguing properties. Particles in nanoscale exhibit superior activity than their bulk counterpart. This unique feature is intensively utilized in physical, chemical, and biological sectors. Each metal is holding unique optical properties that can be utilized to synthesize metallic nanoparticles. At present, versatile nanoparticles were synthesized through chemical and biological methods. Metallic nanoparticles pose numerous scientific merits and have promising industrial applications. But concerning the pros and cons of metallic nanoparticle synthesis methods, researchers elevate to drive the synthesis process of nanoparticles through the utilization of plant resources as a substitute for use of chemicals and reagents under the theme of green chemistry. These synthesized nanoparticles exhibit superior antimicrobial, anticancer, larvicidal, leishmaniasis, wound healing, antioxidant, and as a sensor. Therefore, the utilization of such conceptualized nanoparticles in treating infectious and environmental applications is a warranted one. CONCLUSION: Green chemistry is a keen prudence method, in which bioresources is used as a template for the synthesis of nanoparticles. Therefore, in this review, we exclusively update the context of plant-based metallic nanoparticle synthesis, characterization, and applications in detailed coverage. Hopefully, our review will be modernizing the recent trends going on in metallic nanoparticles synthesis for the blooming research fraternities.

Supporting researchers through full-service hotline and consultation services: Success in simplicity, customization, and staffing.

Navigating the research domain at an academic medical center can be challenging, even for seasoned investigators. To address this, Duke University launched two initiatives: (1) a research navigation "hotline" to provide brief assistance with a variety of research questions; and (2) researcher onboarding and consultation, a one-to-one tailored offering to ensure that researchers are equipped to navigate research resources and processes effectively. The services are provided by the myRESEARCHnavigators (MRN) team, funded by Duke's CTSA. The diverse scientific backgrounds of the six team members align well with those of the research community, allowing for a good match between the researcher and MRN team member. The MRN team answers approximately 30 questions per month, and has provided consultations to almost 400 researchers. Both services receive high satisfaction ratings (4 or 5 stars [out of 5 stars] given to 90% of hotline answers, and 99% of researcher onboarding/consultation sessions). As of July 2019, the School of Medicine has determined that the consultations are critical to their mission and have made them a requirement for new research faculty. The team will continue marketing both services to encourage adoption.

Hibernoma: A missed diagnosis!!

Hibernoma is a rare, benign soft tissue tumor described in <200 case reports/case series. It is slow-growing, painless, and commonly mistaken for lipoma or liposarcoma. Histopathological diagnosis is must for confirmation. Total excision is the treatment of choice. We present a rare case of hibernoma of thigh, which was initially misdiagnosed as atypical lipomatous tumor/well-differentiated liposarcoma on imaging, which turned out to be a hibernoma on histopathological examination.

Autopsy in dengue encephalitis: An analysis of three cases.

Dengue viral infection with unusual presentations has been described in literature, but autopsy is rarely done and is only limited to some cases, based on the literature review. Here, we present the autopsy findings in three cases of dengue encephalitis. All the three patients clinically presented with signs of meningoencephalitis and were positive for dengue non-structural (NS) 1 antigen (Ag) in the serum and cerebrospinal fluid. The postmortem findings revealed cerebral edema, inflammation, hemorrhage, and microinfarcts in all the three cases with herniation of the brain in one case. Sub-massive hepatocellular necrosis was seen in one case. The renal findings included hemorrhage into the Bowman's capsule with red cell cast in two of three cases. The pulmonary findings included a diffuse destruction of the alveoli and hemorrhage into the alveolar spaces in all the three cases.

Investigation into a community outbreak of Salmonella Typhi in Bengaluru, India.

BACKGROUND & OBJECTIVES: Outbreaks of infection due to Salmonella enterica servovar Typhi (S. Typhi) are a great threat to public health. A rapid molecular typing method to characterize strains implicated in an outbreak is critical in implementing appropriate control measures. This study was done to demonstrate the power of a PCR-based method to provide rapid insights into the genetic relatedness amongst the Salmonella isolates implicated in a suspected typhoid fever outbreak. METHODS: Forty two S. Typhi isolates originating from three geographically distinct areas, with one area suspected to have a single-source outbreak were included in the study. The genetic fingerprint of all isolates was generated using enterobacterial repetitive intergenic consensus sequence based-PCR (ERIC-PCR). The antimicrobial susceptibility profiles were also evaluated. RESULTS: ERIC-PCR was found to be rapid and reproducible with a discriminatory index of 0.766. The dendrogram constructed based on ERIC-PCR fingerprinting revealed the existence of 12 distinct genotypes. The location suspected to have an outbreak displayed two genotypes amongst the 24 isolates. The other two locations (18 isolates) displayed genetic heterogeneity. The clonality of the outbreak isolates from the time-matched control isolates was established. The observed antimicrobial susceptibility profiles did not have any discriminatory power to subtype the isolates compared to the genetic fingerprints. INTERPRETATION & CONCLUSIONS: Our study demonstrated the discriminatory power and value of ERIC-PCR in the typing of S. Typhi isolates and providing valuable epidemiological insights.

A Study of Pap Smear in HIV-Positive Females.

BACKGROUND: HIV-positive females are more likely to have abnormal Pap smears than HIV-negative women. These abnormal Pap smears are usually associated with low CD4 cell counts and human papilloma virus infection. MATERIALS AND METHODS: This was a prospective hospital-based study from April 2013 till March 2014. A total of 250 (both symptomatic and asymptomatic) HIV-positive females were examined in Gynaecology OPD at R.C.S.M. G.M.C and C.P.R. Hospital, Kolhapur, and their cervical smears were taken. They were categorized as per modified Bethesda system 2001. The findings in HIV-positive women were correlated with risk factors (age, disease duration, CD4 count and ART use). AIMS: To study the spectrum of cytological abnormalities on Pap smear in HIV-positive females and classify precancerous and cancerous lesions in HIV-positive females according to Bethesda system 2001 and to be familiar with terminology and morphological criteria of Bethesda system 2001. To study the association of Pap smears abnormalities among HIV-positive women with their immune status (CD4 count). RESULTS: NILM is the commonest finding (83.2 %) which is subdivided into non-inflammatory, non-specific and specific inflammatory and atrophic smears. Candida vaginitis was the commonest cause of specific inflammatory condition accounted for (2.52 %) of all inflammatory smears. The percentage of squamous cell abnormalities was 12 %: ASCUS + ASC-H-6.22 %, LSIL-2.10 %, HSIL-3.4 % and SCC-0.8 %. The highest incidence of intraepithelial lesions in HIV-positive females was in the age group 31-40 years. There is no association of Pap smear abnormalities among HIV-positive women with their immune status (CD4 count) and duration of ART. CONCLUSION: Periodic, regular gynaecologic and Pap smear examination would help in early detection of intraepithelial lesions and their treatment so as to prevent invasive malignancy and mortality.

Spinal anesthesia using Taylor's approach helps avoid general anesthesia in short stature asthmatic patient.

The case history of a 35-year-old female patient with short stature is presented. She was posted for rectopexy in view of rectal prolapse. She was a known case of bronchial asthma. She had crowding of intervertebral spaces, which made administration of spinal anesthesia via the normal route very difficult. Taylor's approach for administration of the same was tried and proved successful, thus saving the patient from receiving general anesthesia in the presence of bronchial asthma, for a perineal surgery. The possible cause for the difficulty in administration of spinal anesthesia and the Taylor's approach are discussed, and reports of similar cases reviewed.

A 36-year-old man with vomiting, pain abdomen, significant weight loss, hyponatremia, and hypoglycemia.

Diagnosis of Strongyloides stercoralis hyperinfection can be a challenge. The key to a timely diagnosis is to have a high index of suspicion. We present a rare case of a 36-year-old human immunodeficiency virus negative male patient, who was on multidrug therapy for lepromatous leprosy and was treated for type 2 lepra reactions with steroids in the past. The patient presented with vomiting and pain abdomen, persistent hyponatremia, and terminal hypoglycemia. He had features of malnutrition and had a rapid downhill course following admission. A diagnosis of S. stercoralis hyperinfection with sepsis and multiorgan failure, adrenal hemorrhage, and syndrome of inappropriate antidiuretic hormone secretion was established on a postmortem examination.

Blueprinting in assessment: A tool to increase the validity of undergraduate written examinations in pathology.

CONTEXT/BACKGROUND: Written examinations are the most commonly employed method for assessment of cognitive skills in medical education. The few disadvantages of essay questions are less number of questions, limited sampling, unfair distribution of questions over topics, vague questions etc., Blueprinting overcomes these issues, increasing the validity of examinations. OBJECTIVES: To describe the process of developing a blueprint for undergraduate written examinations in pathology; and to evaluate its effect as a tool to increase the content validity of assessment. METHODOLOGY: A workshop was conducted in the Department of Pathology to sensitize the faculty about the importance of blueprinting. A blueprint was prepared for written examinations in pathology, question papers were set accordingly and administered in preliminary examinations. Feedback was collected from the students and faculty to know their perceptions about the question papers with reference to blueprinting. RESULTS: The students and faculty felt that there was appropriate distribution of questions across topics (77% and 89%, respectively), appropriate weightage given to topics of public health importance (65% and 100%), examinations were fair (86% and 89%). All the faculty felt that blueprints aligns assessment with objectives and helps as a guide and to paper construction. CONCLUSIONS: Students were satisfied as blueprinting helped them to attempt examination better. The faculty who validated the blueprint felt that it helps in distribution of appropriate weightage and questions across the topics and blueprinting should be an integral part of assessment.

Nevus lipomatosus cutaneous superficialis: a rare case report.

Nevus lipomatosus cutaneous superficialis is a rare idiopathic hamartomatous anomaly of the skin characterized by the presence of clusters of mature fat cells among the collagen bundles of the dermis. The classic, or solitary type, presents with asymptomatic soft, skin-colored to yellow papules or nodules. We report the case of a 12-year-old boy with congenital, classical nevus lipomatosus cutaneous superficialis that started as a single painless nodule. Over time, the nodule developed into lesions that presented as skin-colored, well-defined, soft sessile growths with a cerebriform surface centered by comedo-like plugs that increased in size and spread over a large area (approximately 12×4 cm) over the right gluteal region. We report this case as it is an uncommon condition with the intent to highlight its clinical and histopathological features, and differential diagnosis.