Etiopathology, Epidemiology, Diagnosis, and Treatment of Fungal Keratitis.

Fungal keratitis (FK) is a severe ocular condition resulting from corneal infection that is prevalent in tropical countries, particularly in developing regions of Asia and Africa. Factors like corneal lens misuse, inappropriate steroid use, and diagnostic challenges have provoked the epidemic. FK causes significant vision impairment, scarring, and ocular deformities. Accurate pathological diagnosis is crucial for effective therapeutic intervention. Topical antifungal therapy with surface healing medications proves effective in preventing fungal-borne ulcers. Managing FK requires a comprehensive understanding of fungal pathogenesis, guiding formulation strategies and preventive measures to curb global ocular blindness. This review provides in-depth insights into FK, covering etiology, epidemiology, pathogenesis, therapeutic interventions, antifungal resistance, limitations, prevention, and future perspectives on ocular surface disease management.

Trends in antimicrobial susceptibility pattern of Salmonella species isolated from bacteremia patients at a tertiary care center in Northern India.

The study was done to assess the antimicrobial susceptibility pattern among Salmonella enterica serovars causing bacteremia in Northern India. In this observational study, blood samples positive for Salmonella enterica serovars from January 2021 to April 2023 were studied. Species identification was done using MALDI-ToF MS. Serotyping was done using slide agglutination method. Antimicrobial susceptibility was interpreted as per the CLSI guidelines. During the study period, 32 Salmonella enterica serovars were isolated. Salmonella enterica serovar Typhi was the predominant serovar, followed by Salmonella enterica serovar Paratyphi A. All isolates were susceptible to ceftriaxone, chloramphenicol, co-trimoxazole and cefotaxime. Pefloxacin showed 100% resistance. Resistance to nalidixic acid was found in 81.2% isolates. Of the isolates resistant to nalidixic acid, 19(73.08%) isolates were resistant to ciprofloxacin also. This changing susceptibility pattern necessitates continuous surveillance of antibiogram of Salmonella isolates to rationalize the treatment protocols for invasive salmonellosis and prevent emergence of resistant strains.

Anticounterfeiting Feature of a Writable and Self-Erasable Ni(II)-Metallogel Pad via Fluorescent "Turn-On" Detection of Cyanide.

A Schiff base 5-(2-hydroxy-3-methoxybenzylidieneamino)-1-H-imidazole-4-carboxamide (HL) comprising multibinding sites has been synthesized with the aim of fabricating a supramolecular gel. The gelator HL was characterized by FT-IR, 1H & 13C NMR, and ESI-MS techniques and also formed a [Ni(L)2] complex. The gelation property of HL was investigated with various metal ions, wherein Ni(II) selectively forms a mechanically and thermally stable supramolecular metallogel (MG) in the presence of a triethylamine base in DMF-MeOH media. Characterization of MG was accomplished with different spectro-analytical techniques such as FT-IR, ESI-MS, powder-XRD, SEM, rheological investigations, UV/vis, and fluorescence. The gelator HL displays moderate emission upon addition of Ni2+ and gives "turn-off" fluorescence output by forming the complex [Ni(L)2] (MG) due to the chelation-enhanced quenching of fluorescence (CHEQ). Job plot and ESI-MS data suggested a 2:1 stoichiometry between HL and Ni(II) in MG. Further, MG exhibited highly selective and ultrasensitive "turn-on" fluorescence signaling with CN- in the background presence of several cations and anions. The limit of detection (LoD) of MG was determined to be 6.9 × 10-9 M for CN- using the fluorescence technique. Notably, MG behaves as a fluorescent writable pad material explicitly with CN- under 365 nm UV light but not under ordinary light and the fluorescent text is self-erased after 15 min. Hence, MG can be used as a metallogel pad in the presence of CN- to communicate secret messages. Overall, the present work explores the fabrication of a thermo- and mechanostable Ni(II)-metallogel (MG), which selectively and ultrasensitively detects CN- both in the solution phase and in the gel form, wherein MG behaves as a writable and self-erasable pad with anticounterfeiting features for practical applications.

Simultaneous determination of amphotericin B, tobramycin and vancomycin in rabbit ocular biofluids and tissues by LC-MS/MS: An antimicrobial therapy for keratitis and its PK-PD application.

A rationale poly-microbial keratitis (PMK) therapy requires quick identification of pathogen (bacteria and fungi) and their efficient treatment. However, majority of healthcare providers are still having trouble finding an effective medicine to treat PMK due to constraints such as antimicrobial resistance, dose and dosing schedule. Thus, a broad spectrum anti-fungal and antibacterial having less resistance in community involving combination therapy such as amphotericin B (AmB), tobramycin (TBR) and vancomycin (VCM) is required. Hence, to characterize the pharmacokinetic (PK) and PK-pharmacodynamic (PD) indices, a rapid and sensitive simultaneous LC-MS/MS bioanalytical method was developed and validated for the quantification of AmB, TBR and VCM in rabbit ocular biofluids and tissues. Chromatographic resolution was achieved on a Zorbax C18 column with a mobile phase composed of acetonitrile and 0.4 % formic acid in deionized water using a gradient mode of elution. The calibration curves showed good linearity over the concentration range of 1.95-500 ng/mL for AmB and TBR, 3.9-800 ng/mL for VCM, respectively. The lower limit of quantification (LLOQ) was found to be 1.95 ng/mL for AmB and TBR, and 4.5 ng/mL for VCM. Analyte extraction was performed by simple protein precipitation method with minimal sample volume of 10 µL. Finally, the developed method was validated for selectivity, linearity (r2 > 0.99), precision, accuracy, matrix effects, and stability. The ocular pharmacokinetic profile of commercial AmB, TBR, and VCM formulations was further assessed using the validated method and the PK-PD indices along with dosing frequency was predicted by PK-PD modelling using Phoenix WinNonlin Software.

Idiopathic Pulmonary Fibrosis: Where do We Stand and How Far to Go?

Idiopathic pulmonary fibrosis is a progressive and incurable lung disease characterized by collagen deposition, alveolar inflammation, fibroblast proliferation, and the destruction of lung tissue structures. It is a rare yet severe condition with a high mortality rate, typically leading to death within 3-5 years of diagnosis. The clinical presentation of idiopathic pulmonary fibrosis (IPF) involves a gradual and substantial loss of lung function, ultimately resulting in respiratory failure. Despite more than half a century of intensive research, the origin of IPF remains a mystery. Despite its unknown etiology, several genetic and non-genetic factors have been linked to IPF. Recent significant advancements have been made in the field of IPF diagnosis and treatment. Two oral small-molecule drugs, pirfenidone and nintedanib, have recently gained approval for the treatment of IPF. Pirfenidone exhibits antifibrotic, antioxidant, and anti-inflammatory properties, while nintedanib is a tyrosine kinase inhibitor with selectivity for vascular endothelial growth factor (VEGF) receptors, prostaglandin F (PGF) receptors, and fibroblast growth factor (FGF) receptors. Both of these compounds are capable of slowing down the progression of the disease with an acceptable safety profile. This review provides a brief introduction, historical background, epidemiological insights, and an exploration of various environmental risk factors that may influence the lung microenvironment and contribute to the advancement of IPF. The review also delves into the diagnosis, signaling pathways, and ongoing clinical trials worldwide. A thorough review of the literature was conducted using PubMed and Google Scholar to gather information on various aspects of IPF. Numerous potential drugs are currently under investigation in clinical trials, and the completion of this process is crucial to the ultimate goal of finding a cure for IPF patients. The investigation of the role of genes, surfactant proteins, infectious agents, biomarkers, and epigenetic changes holds the promise of offering earlier and more accurate understanding and diagnosis of IPF. This information could be instrumental in the development of new therapeutic approaches for treating IPF and is expected to be of great interest to researchers.

Concurrent determination of anti-microbial and anti-inflammatory drugs in lachrymal fluid and tissue by LC-MS/MS: A potential treatment for microbial keratitis and its PK-PD evaluation.

Unforeseen surfacing of microbial keratitis (MKT) over the years has led to a requisite for promising treatment strategy involving combination of antifungal and antibacterial agents. Subsequently, symptoms associated with MKT including inflammation and watery eyes require treatment with anti-inflammatory agents. Thus, a requirement of functional clinical treatment strategy involving combination of anti-inflammatory corticosteroids (Betamethasone) with antifungal polyene (Amphotericin B, AmB) and antibacterials macrolide (Azithromycin, AZT) and aminoglycoside (Neomycin, NEO). In the ensuing pursuit, a sensitive and fast simultaneous LC-MS/MS method of four drastically different analytes in rabbit tear fluid and cornea was developed and validated as per US-FDA guidelines. The gradient LC set-up was used with C18 column and flow rate of 0.55 mL/min along with short run time of 7 min. The calibration curves showed good linearity over the concentration range of 0.07-300 ng/mL, 1.00-400 ng/mL, 3.00-600 ng/mL and 8.00-900 ng/mL for AZT, AmB, NEO and BEM respectively. The bioanalytical method requires only 10 µL of ocular sample and analytes were extracted with fast protein precipitation with acidic methanol. Finally, the developed method was validated for selectivity, linearity (r2 > 0.99), precision, accuracy, matrix effects, and stability. PK-PD indices and dosing frequency was predicted using Phoenix WinNonlin Software, based on single dose ocular pharmacokinetics and MIC values of AmB, AZT and NEO. According to the PK-PD simulation, S. aureus and E. coli required 6 and 12 instillations of AZT per 24 h, respectively whereas 12 instillation of NEO requires per 24 h for S. aureus. The result suggests that to minimize antimicrobial resistance; drug, dose and dosing schedule depend upon the pathogen as well as the strain.

Short- and Long-Term Retentivity of Knowledge by Various Teaching Methods in Medical Education and Perception of Students Towards Them: A Comparative Study in a Medical University Hospital of Northern India.

BACKGROUND: To develop doctors with appropriate knowledge of health and diseases, reasonable medical abilities, and a positive attitude toward patients and their families, it is important to reexamine the methods used to educate and train medical school students. To establish which is best for both medical students and professors, the various teaching and learning methodologies must be compared and analyzed. This study attempts to determine the preferred medical education techniques among medical students as well as the caliber of the classes they attend. METHODS: This is a before-and-after study conducted among 480 first- (240) and second-year (240) undergraduate students. Students were divided into three groups. Each group was assigned a teacher who was responsible for teaching four short topics according to the common understanding and knowledge level of both year students in four different ways: traditional blackboard method, offline PowerPoint presentation, online PowerPoint presentation, and online annotative. Application-based learning and self-learning were the other two teaching methods conducted in a monitored environment. An MCQ-based pre- and post-test were taken to assess the improvement, and a feedback form was filled out by each student to assess their perception. To assess long-term retention, a surprise follow-up test was conducted after 15 days. RESULTS: For all the teaching methods except for traditional blackboard and online presentation, there was a significant improvement in the post-test scores as compared to the pre-test scores (p<0.05). Retentivity was more remarkable in online application-based and self-learning methods. 77.2% of the study participants preferred offline presentation as the mode of teaching. CONCLUSION: Retention was found to be highest in self-directed and application-based learning. So, students should be encouraged and motivated for self-study after every lecture, whatever the teaching method used by teachers.

Inflammatory Mediators and GBM Malignancy: Current Scenario and Future Prospective.

Glioblastoma multiforme is one of the most widespread and dangerous forms of brain tumor with high inflammation. The tumor microenvironment comprises diverse tumor cells, different types of immune cells, and the extracellular matrix. Inflammatory mediators like chemokines, cytokines, and growth factors possibly serve as a capable therapeutic target to quash their tumor-promoting properties in glioblastoma multiforme (GBM). Cytokines are a heterogeneous group of soluble functional proteins which are also associated with the induction and progression of tumors. These are supposed to have both pro-inflammatory (such as tumor necrosis factor-α (TNF-α), interleukin-17A (IL-17A), interferon-γ (IFN-γ), IL-4, IL-2, IL-6, IL-12, IL-13) and anti-inflammatory (such as transforming growth factor-ß (TGF-ß), IL-10, and granulocyte-macrophage colony-stimulating factor (GM-CSF)) actions and are the crucial communications channels in the tumor microenvironment. In the present minireview we discuss the tumor microenvironment and inflammatory mediators and focus on the involvement of cytokines in establishing communication with the tumor microenvironment. The presented data highlight the possible roles of cytokines in communication between glioblastoma cells and tumor microenvironment. Cytokines formed by immune cells protect the host organs while cytokines secreted by tumor cells are used for their advantage. Though the clinical trials with a number of immunotherapeutic agents are going on around the globe, there is still a requirement for thorough investigation of the regulatory mechanism managing GBM growth, recurrence, and tumor response to the therapy.

Liquid Biopsies Poorly miRror Renal Ischemia-Reperfusion Injury.

Acute kidney injury (AKI) is the rapid reduction in renal function. It is often difficult to detect at an early stage. Biofluid microRNAs (miRs) have been proposed as novel biomarkers due to their regulatory role in renal pathophysiology. The goal of this study was to determine the overlap in AKI miRNA profiles in the renal cortex, urine, and plasma samples collected from a rat model of ischemia-reperfusion (IR)-induced AKI. Bilateral renal ischemia was induced by clamping the renal pedicles for 30 min, followed by reperfusion. Urine was then collected over 24 h, followed by terminal blood and tissue collection for small RNA profiling. Differentially expressed (IR vs. sham) miRs within the urine and renal cortex sample types demonstrated a strong correlation in normalized abundance regardless of injury (IR and sham: R2 = 0.8710 and 0.9716, respectively). Relatively few miRs were differentially expressed in multiple samples. Further, there were no differentially expressed miRs with clinically relevant sequence conservation common between renal cortex and urine samples. This project highlights the need for a comprehensive analysis of potential miR biomarkers, including analysis of pathological tissues and biofluids, with the goal of identifying the cellular origin of altered miRs. Analysis at earlier timepoints is needed to further evaluate clinical potential.

Rotablation-Assisted Staged Culottes Bifurcation Stenting of Left Main Chronic Total Occlusion.

Left main coronary artery chronic total occlusion is an unusual finding discovered on coronary angiography. Historically, coronary artery bypass graft has been the preferred treatment. However, recent studies have revealed the role of left main percutaneous coronary intervention in selected patients. This is a case of staged left main coronary artery chronic total occlusion percutaneous coronary intervention. (Level of Difficulty: Advanced.).

A multi-cation responsive Ni(II)-supramolecular metallogel mimics a molecular keypad lock via reversible fluorescence switching.

Synthesis of a bidentate N,O-donor Schiff base fluorescent ligand 5-(diethylamino-2-((4-(diethylamino-2-((4-(diethylamino)phenylimino)ethyl)phenol) (HL) adopting a new preparation procedure and its complexes with Ni(II) (1) and Zn(II) (2) has been illustrated. Structures of HL and 1 have been elucidated using X-ray single crystal analysis. Moreover, HL leads to the formation of a mechanically stable Ni(II)-gel (MG) upon treatment with Ni(NO3)2·6H2O in the presence of triethylamine (TEA) using THF/MeOH (1 : 1) solvents at rt. The gelator HL, complexes 1-2 and MG have been characterized by different spectroscopic and microscopic techniques including NMR (1H & 13C), FT-IR, ESI-MS, SEM, powder-XRD, rheology, UV/vis and fluorescence analysis. Rheological studies suggested good mechanical and thermal stability, whereas SEM analysis reveals a porous earth crust-like morphology of MG. Notably, 1 : 1 complexation between HL and Ni(II) forms a stable gel (MG), whereas 2 : 1 (HL : Ni2+) complexation leads to partial gelation. Formation of the Ni(II)-MG leads to slight "Turn-OFF" fluorescence relative to HL with a limit of detection (LOD) of 7.76 × 10-9 M; however, MG is considered as the "ON" state due to moderate emission. Remarkably, Ni(II)-MG further displayed reversible "ON-OFF-ON" fluorescence switching behavior through detection of Zn2+, Cu2+ and Hg2+. The emission intensity of MG is quenched with Cu2+/Hg2+ but enhances with Zn2+ in 1 : 1 (MG : M2+) stoichiometry. Therefore, MG mimics a sequence dependent molecular keypad lock for Cu2+ (C), Hg2+ (H) and Zn2+ (Z) to give the maximum output. Association and quenching constants were calculated by the Benesi-Hildebrand method, and from the Stern-Volmer plot the LOD was determined to be 4.2 × 10-6 M, 5.8 × 10-6 M and 7.8 × 10-6 M for MG with Zn(II), Cu(II) and Hg(II), respectively. To date, Ni(II) based MGs have been explored only toward electrochemical, thermal and conduction studies; however, the present work demonstrates the fluorescent reversible cation detection behavior of Ni(II)-MG to act as a molecular keypad lock for development of password protection devices.

Development of highly selective fluorescent ferrocenyl-iminopyridine chemosensor for biologically relevant Fe3.

Design, synthesis, characterization, and ion detection studies of two ferrocene-appended Schiff bases namely N-(2-[ferrocenylamino]ethyl)-5-nitropyridin-2-amine (1) and ferrocenylamino-1H-imidazole-4-carboxamide (2) been reported. Both the chemosensors have been thoroughly characterized using Fourier transfer infrared, 1 H and 13 C nuclear magnetic resonance, high resolution mass spectrometry, and ultraviolet/visible (UV/visible) and fluorescence spectral techniques. Probes 1 and 2 were designed with the aim of appending the ferrocenyl group with pyridine ring having an amine substitution (for 1) and imidazole ring with an amide substitution (for 2). Interaction of these probes with a series of cations and anions was examined through UV/vis and fluorescence spectral techniques. Probe 2 exhibited an insignificant response towards anions and loss of selectivity for cations, whereas 1 displayed highly selective detection towards biologically important Fe3+ in 2:1 (probe:cation) stoichiometry. Notably, none of the cations and anions could interfere the selectivity of Fe3+ ensured by 1 in aqueous medium. The limit of detection for Fe3+ detection using 1 was determined to be 0.2 ppm. The results strongly suggest that 1 could find promising future application as a chemosensor for Fe3+ in biological systems for quantification and qualitative analysis.

Utility of the 2018 revised ISN/RPS thresholds for glomerular crescents in childhood-onset lupus nephritis: a Pediatric Nephrology Research Consortium study.

BACKGROUND: The revised 2018 ISN/RPS Classification System for lupus nephritis (LN) includes calculations for both activity index (A.I.) and chronicity index (C.I.). Unchanged were the thresholds of < 25%, 25-50%, and > 50% crescents to distinguish between mild, moderate, and severe activity/chronicity. We aimed to evaluate these thresholds for percent crescents in childhood-onset LN. METHODS: Eighty-six subjects < 21 years of age were enrolled from the Pediatric Glomerulonephritis with Crescents Registry, a retrospective multi-center cohort sponsored by the Pediatric Nephrology Research Consortium. Thresholds of 10%, 25%, and 50% for both cellular/fibrocellular and fibrous crescents were interrogated for primary outcomes of kidney failure, eGFR, and eGFR slope. RESULTS: Median age at time of initial biopsy was 14 years (range 1-21). Median follow-up time was 3 years (range 1-11). Cumulative incidence of kidney failure was 6% at 1 year and 10% at latest follow-up. Median eGFR slope was - 18 mL/1.73 m2/min (IQR - 51 to + 8) at 1 year and - 3 mL/min/1.73 m2/year (IQR - 19 to + 6) at latest follow-up. We found no difference in kidney failure at the proposed < 25% and 25-50% cellular crescents thresholds, and thus added a new provisional threshold of 10% that better predicted outcomes in children. Moreover, use of 10% and 25% thresholds for fibrous crescents showed a fourfold and sevenfold increase in risk of kidney failure. CONCLUSIONS: In children with crescentic LN, use of 10% and 25% thresholds for cellular crescents better reflects disease activity, while these thresholds for fibrous crescents better discriminates kidney disease outcomes. A higher resolution version of the Graphical abstract is available as Supplementary information.

Renal Survival in Children with Glomerulonephritis with Crescents: A Pediatric Nephrology Research Consortium Cohort Study.

There is no evidence-based definition for diagnosing crescentic glomerulonephritis. The prognostic implications of crescentic lesions on kidney biopsy have not been quantified. Our objective was to determine risk factors for end-stage kidney disease (ESKD) in patients with glomerulonephritis and crescents on kidney biopsy. A query of the Pediatric Nephrology Research Consortium's Pediatric Glomerulonephritis with Crescents registry identified 305 patients from 15 centers. A retrospective cohort study was performed with ESKD as the primary outcome. Median age at biopsy was 11 years (range 1-21). The percentage of crescents was 3-100% (median 20%). Etiologies included IgA nephropathy (23%), lupus (21%), IgA vasculitis (19%) and ANCA-associated GN (13%), post-infectious GN (5%), and anti-glomerular basement membrane disease (3%). The prevalence of ESKD was 12% at one year and 16% at last follow-up (median = 3 years, range 1-11). Median time to ESKD was 100 days. Risk factors for ESKD included %crescents, presence of fibrous crescents, estimated GFR, and hypertension at biopsy. For each 1% increase in %crescents, there was a 3% decrease in log odds of 1-year renal survival (p = 0.003) and a 2% decrease in log odds of renal survival at last follow-up (p < 0.001). These findings provide an evidence base for enrollment criteria for crescentic glomerulonephritis in future clinical trials.

Trade-off expression of melatonin receptor subtypes (Mel1a and Mel1b) and androgen receptor in lung of a tropical bird, Perdicula asiatica.

The role of circulatory steroid hormone along with melatonin in lung of any seasonally breeding bird has never been explored so far. This could be interesting because steroid hormones are immunosuppressive while melatonin is immunostimulatory in nature. In our present study, we report the effect of exogenous melatonin and testosterone on expression of melatonin receptor subtypes (Mel1a and Mel1b ) and androgen receptor in lung of a tropical bird Perdicula asiatica. Birds were collected from vicinity of Varanasi and acclimatized in laboratory with sufficient food and water. The birds were treated with melatonin and testosterone at dose of 25 µg/100 g B.wt./day and 1 mg/100 g B.wt./day, respectively, for 28 days. At the end of the experiment, the birds were sacrificed and lung tissue and blood sample were collected for immunohistochemistry, Western blot analysis and hormonal assay. Testosterone treatment increased circulatory testosterone and upregulated expression of androgen receptors whereas downregulated expression of melatonin receptor subtypes Mel1a and Mel1b . Melatonin administration increased peripheral melatonin and upregulated expression of melatonin receptor subtypes Mel1a and Mel1b while downregulated androgen receptor. Thus, our results suggest that a trade-off relationship between melatonin and testosterone exists in regulation of their receptors in lung of Perdicula asiatica.