Applications of Surface Plasmon Resonance (SPR) to the Study of Diverse Protein-Ligand Interactions.

Functional characterization of enzymes/proteins requires determination of the binding affinity of small molecules or other biomolecules with the target proteins. Several available techniques, such as proteomics and drug discovery strategies, require a precise and high-throughput assay for rapid and reliable screening of potential candidates for further testing. Surface plasmon resonance (SPR), a well-established label-free technique, directly measures biomolecular affinities. SPR assays require immobilization of one interacting component (ligand) on a conductive metal (mostly gold or silver) and a continuous flow of solution containing potential binding partner (analyte) across the surface. The SPR phenomenon occurs when polarized light excites the electrons at the interface of the metal and the dielectric medium to generate electromagnetic waves that propagate parallel to the surface. Changes in the refractive index due to interaction between the ligand and analyte are measured by detecting the reflected light, providing real-time data on kinetics and specificity. A prominent use of SPR is identifying compounds in crude plant extracts that bind to specific molecules. Procedures that utilize SPR are becoming increasingly applicable outside the laboratory setting, and SPR imaging and localized SPR (LSPR) are cheaper and more portable alternative for in situ detection of plant or mammalian pathogens and drug discovery studies. LSPR, in particular, has the advantage of direct attachment to test tissues in live-plant studies. Here, we describe three protocols utilizing SPR-based assays for precise analysis of protein-ligand interactions. © 2024 Wiley Periodicals LLC. Basic Protocol 1: SPR comparison of binding affinities of viral reverse transcriptase polymorphisms Basic Protocol 2: SPR screening of crude plant extract for protein-binding agents Basic Protocol 3: Localized SPR-based antigen detection using antibody-conjugated gold nanoparticles.

SRX3177, a CDK4/6-PI3K-BET inhibitor, in combination with an RdRp inhibitor, Molnupiravir, or an entry inhibitor MU-UNMC-2, has potent antiviral activity against the Omicron variant of SARS-CoV-2.

Despite considerable progress in developing vaccines and antivirals to combat COVID-19, the rapid mutations of the SARS-CoV-2 genome have limited the durability and efficacy of the current vaccines and therapeutic interventions. Hence, it necessitates the development of novel therapeutic approaches or repurposing existing drugs that target either viral life cycle, host factors, or both. Here, we report that SRX3177, a potent triple-activity CDK4/6-PI3K-BET inhibitor, blocks replication of the SARS-CoV-2 Omicron variant with IC50 values at sub-micromolar concentrations without any impact on the cell proliferation of Calu-3 cells at and below its IC50 concentration. When SRX3177 is combined with EIDD-1931 (active moiety of a small-molecule prodrug Molnupiravir) or MU-UNMC-2 (a SARS-CoV-2 entry inhibitor) at a fixed doses matrix, a synergistic effect was observed, leading to the significant reduction in the dose of the individual compounds to achieve similar inhibition of SARS-CoV-2 replication. Herein, we report that the combination of SRX3177/MPV or SRX3177/UM-UNMC-2 has the potential for further development as a combinational therapy against SARS-CoV-2 and in any future outbreak of beta coronavirus.

"Mpox in MSM: Tackling Stigma, Minimizing Risk Factors, Exploring Pathogenesis, and Treatment Approaches".

Mpox is a zoonotic disease caused by the monkeypox virus (MPV), primarily found in Central and West African countries. The typical presentation of the disease before the 2022 mpox outbreak includes a febrile prodrome 5-13 days post-exposure, accompanied by lymphadenopathy, malaise, headache, and muscle aches. Unexpectedly, during the 2022 outbreak, several cases of atypical presentations of the disease were reported, such as the absence of prodromal symptoms and the presence of genital skin lesions suggestive of sexual transmission. As per the World Health Organization (WHO), as of March 20, 2024, 94,707 cases of mpox were reported worldwide, resulting in 181 deaths (22 in African endemic regions and 159 in non-endemic countries). The United States Centers for Disease Control and Prevention (CDC) reports a total of 32,063 cases (33.85% of total cases globally), with 58 deaths (32.04% of global deaths) due to mpox. Person-to-person transmission of mpox can occur through respiratory droplets and sustained close contact. However, during the 2022 outbreak of mpox, a high incidence of anal and perianal lesions among MSMs indicated sexual transmission of MPV as a major route of transmission. Since MSMs are disproportionately at risk for HIV transmission, this review discusses the risk factors, transmission patterns, pathogenesis, vaccine, and treatment options for mpox among MSM and people living with HIV (PLWH). Furthermore, we provide a brief perspective on the evolution of the MPV in immunocompromised people like PLWH.

Inhibitors against DNA Polymerase I Family of Enzymes: Novel Targets and Opportunities.

DNA polymerases replicate cellular genomes and/or participate in the maintenance of genome integrity. DNA polymerases sharing high sequence homology with E. coli DNA polymerase I (pol I) have been grouped in Family A. Pol I participates in Okazaki fragment maturation and in bacterial genome repair. Since its discovery in 1956, pol I has been extensively studied, primarily to gain deeper insights into the mechanism of DNA replication. As research on DNA polymerases advances, many novel functions of this group of polymerases are being uncovered. For example, human DNA polymerase θ (a Family A DNA pol) has been shown to synthesize DNA using RNA as a template, a function typically attributed to retroviral reverse transcriptase. Increased interest in drug discovery against pol θ has emerged due to its roles in cancer. Likewise, Pol I family enzymes also appear attractive as drug-development targets against microbial infections. Development of antimalarial compounds targeting apicoplast apPOL, an ortholog of Pol I, further extends the targeting of this family of enzymes. Here, we summarize reported drug-development efforts against Family A polymerases and future perspective regarding these enzymes as antibiotic targets. Recently developed techniques, such as artificial intelligence, can be used to facilitate the development of new drugs.

Cell-specific polymerization-driven biomolecular condensate formation fine-tunes root tissue morphogenesis.

Formation of biomolecular condensates can be driven by weak multivalent interactions and emergent polymerization. However, the mechanism of polymerization-mediated condensate formation is less studied. We found lateral root cap cell (LRC)-specific SUPPRESSOR OF RPS4-RLD1 (SRFR1) condensates fine-tune primary root development. Polymerization of the SRFR1 N-terminal domain is required for both LRC condensate formation and optimal root growth. Surprisingly, the first intrinsically disordered region (IDR1) of SRFR1 can be functionally substituted by a specific group of intrinsically disordered proteins known as dehydrins. This finding facilitated the identification of functional segments in the IDR1 of SRFR1, a generalizable strategy to decode unknown IDRs. With this functional information we further improved root growth by modifying the SRFR1 condensation module, providing a strategy to improve plant growth and resilience.

Determinants of cancer screenings participation in Queensland: a scoping review.

Introduction Cancer screening programmes for cervical, breast, and colorectal cancer have successfully reduced mortality rates among target groups. However, a large proportion of women and men are unscreened. Aim This review aims to provide an overview of the literature regarding the determinants of cancer screening participation among target groups in Queensland. Methods Electronic databases were searched for studies on determinants of cancer screening participation in Queensland. Retrieved studies were screened, and eligible articles were selected for data extraction. Both peer-reviewed and grey literature studies were included. The determinants of cancer screening participation were classified according to the I-Change model. Results Sixteen out of 75 articles were selected and analysed. Information factors, such as the lack of tailored strategies, determined cancer screening participation. Age, gender, cultural beliefs, fear and past experiences were the most reported predisposing factors to cancer screening participation. Lack of knowledge, misconceptions, low awareness, timely access to service, privacy and confidentiality were mainly reported awareness and motivation factors. Encouragement from health professionals, providing more information and interactions with communities would result in different effects on cancer screening participation among the target groups. Discussion The I-Change model is a valuable tool in mapping the current determinants of cancer screening participation programs. Further research may be needed to fully understand the barriers and facilitators of cancer screening programs.

The contribution and therapeutic implications of IGHMBP2 mutations on IGHMBP2 biochemical activity and ABT1 association.

Mutations within immunoglobulin mu DNA binding protein (IGHMBP2), an RNA-DNA helicase, result in SMA with respiratory distress type I (SMARD1) and Charcot Marie Tooth type 2S (CMT2S). The underlying biochemical mechanism of IGHMBP2 is unknown as well as the functional significance of IGHMBP2 mutations in disease severity. Here we report the biochemical mechanisms of IGHMBP2 disease-causing mutations D565N and H924Y, and their potential impact on therapeutic strategies. The IGHMBP2-D565N mutation has been identified in SMARD1 patients, while the IGHMBP2-H924Y mutation has been identified in CMT2S patients. For the first time, we demonstrate a correlation between the altered IGHMBP2 biochemical activity associated with the D565N and H924Y mutations and disease severity and pathology in patients and our Ighmbp2 mouse models. We show that IGHMBP2 mutations that alter the association with activator of basal transcription (ABT1) impact the ATPase and helicase activities of IGHMBP2 and the association with the 47S pre-rRNA 5' external transcribed spacer. We demonstrate that the D565N mutation impairs IGHMBP2 ATPase and helicase activities consistent with disease pathology. The H924Y mutation alters IGHMBP2 activity to a lesser extent while maintaining association with ABT1. In the context of the compound heterozygous patient, we demonstrate that the total biochemical activity associated with IGHMBP2-D565N and IGHMBP2-H924Y proteins is improved over IGHMBP2-D565N alone. Importantly, we demonstrate that the efficacy of therapeutic applications may vary based on the underlying IGHMBP2 mutations and the relative biochemical activity of the mutant IGHMBP2 protein.

Islatravir: evaluation of clinical development for HIV and HBV.

INTRODUCTION: Islatravir (ISL) is a nucleoside reverse transcriptase translocation inhibitor (NRTTI) that inhibits HIV RT through multiple mechanisms. Contrary to all approved NtRTIs, islatravir retains a 3'OH group. In vitro and clinical data show that ISL is an ultrapotent investigational drug with high tolerability. AREAS COVERED: The historical development of islatravir and its mechanisms of HIV and HBV inhibition and resistance are covered. Additionally, the outcomes of Phase I and Phase II clinical trials are discussed. EXPERT OPINION: Current first-line antiretroviral therapy, preexposure, and postexposure prophylactic interventions are highly effective in maintaining low or undetectable viral load. Despite these measures, an unusually high rate of new infections every year warrants developing novel antivirals that can suppress drug-resistant HIV and improve compliance. ISL, an NRTTI once deemed a long-acting drug, was placed on a clinical hold. The outcome of ongoing clinical trials with a reduced ISL dose will decide its future clinical application. Additionally, MK-8527, which inhibits HIV via same mechanism as that of ISL may supersede ISL. Data on ISL inhibition of HBV are scarce, and preclinical data show dramatically lower ISL efficacy against HBV than currently preferred nucleos(t)ide drugs, indicating that ISL may not be a potent anti-HBV drug.

Evaluation of ganglion cell-inner plexiform layer thickness in the diagnosis of pre-perimetric glaucoma and comparison to retinal nerve fiber layers.

PURPOSE: Evaluation of ganglion cell-inner plexiform layer thickness in the diagnosis of pre-perimetric glaucoma (PPG) and comparison to retinal nerve fiber layers. METHODS: This study was a prospective hospital-based study. A total of 30 PPG and control patients were studied for retinal nerve fiber layer thickness (RNFL) and ganglion cell-inner plexiform layer complex (GC-IPL) by spectral-domain optical coherence tomography. PPG was defined as eyes with a normal visual field and one or more localized RNFL defects that were associated with a typical glaucomatous disc appearance. Diagnostic abilities of GC-IPL, optic nerve head (ONH), and RNFL parameters were computed using area under receiver-operating curve (AUROC), sensitivity, and specificity. RESULTS: GC-IPL parameters showed significant changes in PPG cases as compared to normal subjects in each region ( P value < 0.001). RNFL parameters also differed significantly from normal subjects in all quadrants ( P value 0.003 to < 0.001). Within GC-IPL parameters, the superotemporal region had the maximum area under the curve (AUC), followed by inferior, superior, and inferotemporal regions. Within RNFL parameters, the inferior quadrant had the maximum AUC, followed by superior and nasal quadrants. the GC-IPL parameters in PPG showed that the AUC of the GC-IPL parameters was much higher than those of the ONH and RNFL values. CONCLUSION: Although both the parameters RNFL and GC-IPL showed significant changes in PPG patients compared to healthy subjects, a higher AUC of GC-IPL points toward the higher sensitivity of GC-IPL than RNFL for detecting glaucoma in early stages.

Development of an all-solid-state air-cooled high-power blue diode laser for metal processing.

We present the design and development of an all-solid-state (fluid/refrigerant-free) 100 W scale blue-laser system and show its applications in precision copper works. We combine powerful laser-diode arrays with Peltier chips on a compact laser head to achieve stable thermal and optical performance. Good agreement between the thermal simulation of the 3D laser head and experiments validates stable thermal performance. The laser system emits 40-100 W continuous wave at λ = 452.2 ± 2.5 nm with 98% power stability and ∼24% wall-plug efficiency inside a portable enclosure. This is the first, to the best of our knowledge, all-solid-state air-cooled laser with a 100 W class output. We achieved kW/cm2 intensity level on an mm-size focus with this source and demonstrated cutting, bending, and soldering copper on a battery pack. Furthermore, the copper-solder joints have nanoscale adhesion without cracks. Additionally, we unveil that 0.5-4 kW/cm2 intensity laser annealing scan makes copper strips mechanically resilient to withstand extreme loading cycles without nanoscale cracks.

Smoking cessation and its significant role in the Indian scenario.

Given the increased health dangers of tobacco use, particularly in developing countries, smoking cessation intervention is crucially important. The aim of this study is to determine and assess the effectiveness of a comprehensive smoking cessation intervention program, incorporating behavior modification, counseling, and pharmacologic treatments, in the context of the Indian scenario. The process of initiating smoking or tobacco cessation begins with the evaluation of the distinct stages that smokers undergo as part of their journey toward behavioral change. There are five different levels of preparation for quitting smoking, i.e., i) not prepared (pre-contemplation); ii) unsure (contemplation); iii) prepared (preparation); iv) action; and v) maintenance. Behavior modification and counseling are essential. The "5 A's"-based intervention uses ask, advise, assess, assist, and arrange as part of its strategy. First-line treatments such as nicotine replacement therapy, bupropion, and varenicline, as well as second-line treatments such as clonidine, cytisine, and nortriptyline, are the foundation of pharmacologic care. Every healthcare professional has a duty to help smokers stop using tobacco, and the intervention should be both therapeutic and diagnostic. Combining behavioral and social support yields the best results, along with pharmacotherapy whenever needed.

A Review of miRNA Regulation in Japanese Encephalitis (JEV) Virus Infection.

Japanese encephalitis (JE) is a mosquito-borne disease that causes neuronal damage and inflammation of microglia, and in severe cases, it can be fatal. JE infection can resist cellular immune responses and survive in host cells. Japanese encephalitis virus (JEV) infects macrophages and peripheral blood lymphocytes. In addition to regulating biological signaling pathways, microRNAs in cells also influence virus-host interactions. Under certain circumstances, viruses can change microRNA production. These changes affect the replication and spread of the virus. Host miRNAs can contain viral pathogenicity by downregulating the antiviral immune response pathways. Simultaneous profiling of miRNA and messenger RNA (mRNA) could help us detect pathogenic factors, and dual RNA detection is possible. This work highlights important miRNAs involved in human JE infection. In this study, we have shown the important miRNAs that play significant roles in JEV infection. We found that during JEV infection, miRNA-155, miR-29b, miRNA-15b, miR-146a, miRNA-125b-5p, miRNA-30la, miR-19b-3p, and miR-124, cause upregulation of human genes whereas miRNA-432, miRNA-370, microRNA-33a-5p, and miRNA-466d-3p are responsible for downregulation of human genes respectively. Further, these miRNAs are also responsible for the inflammatory effects. Although several other miRNAs critical to the JEV life cycle are yet unknown, there is currently no evidence for the role of miRNAs in persistence.

Potential therapeutic targets for Mpox: the evidence to date.

INTRODUCTION: The global Mpox (MPX) disease outbreak caused by the Mpox virus (MPXV) in 2022 alarmed the World Health Organization (WHO) and health regulation agencies of individual countries leading to the declaration of MPX as a Public Health Emergency. Owing to the genetic similarities between smallpox-causing poxvirus and MPXV, vaccine JYNNEOS, and anti-smallpox drugs Brincidofovir and Tecovirimat were granted emergency use authorization by the United States Food and Drug Administration. The WHO also included cidofovir, NIOCH-14, and other vaccines as treatment options. AREAS COVERED: This article covers the historical development of EUA-granted antivirals, resistance to these antivirals, and the projected impact of signature mutations on the potency of antivirals against currently circulating MPXV. Since a high prevalence of MPXV infections in individuals coinfected with HIV and MPXV, the treatment results among these individuals have been included. EXPERT OPINION: All EUA-granted drugs have been approved for smallpox treatment. These antivirals show good potency against Mpox. However, conserved resistance mutation positions in MPXV and related poxviruses, and the signature mutations in the 2022 MPXV can potentially compromise the efficacy of the EUA-granted treatments. Therefore, MPXV-specific medications are required not only for the current but also for possible future outbreaks.

Covalent organic frameworks (COFs) and metal-organic frameworks (MOFs) as electrochemical sensors for the efficient detection of pharmaceutical residues.

Pharmaceutical residues are the undecomposed remains from drugs used in the medical and food industries. Due to their potential adverse effects on human health and natural ecosystems, they are of increasing worldwide concern. The acute detection of pharmaceutical residues can give a rapid examination of their quantity and then prevent them from further contamination. Herein, this study summarizes and discusses the most recent porous covalent-organic frameworks (COFs) and metal-organic frameworks (MOFs) for the electrochemical detection of various pharmaceutical residues. The review first introduces a brief overview of drug toxicity and its effects on living organisms. Subsequently, different porous materials and drug detection techniques are discussed with materials' properties and applications. Then the development of COFs and MOFs has been addressed with their structural properties and sensing applications. Further, the stability, reusability, and sustainability of MOFs/COFs are reviewed and discussed. Besides, COFs and MOFs' detection limits, linear ranges, the role of functionalities, and immobilized nanoparticles are analyzed and discussed. Lastly, this review summarized and discussed the MOF@COF composite as sensors, the fabrication strategies to enhance detection potential, and the current challenges in this area.

Nanodiamonds enable femtosecond-processed ultrathin glass as a hybrid quantum sensor.

The quantum properties of fluorescent nanodiamonds offer great promise for fabricating quantum-enabled devices for physical applications. However, the nanodiamonds need to be suitably combined with a substrate to exploit their properties. Here, we show that ultrathin and flexible glass (thickness 30 microns) can be functionalized by nanodiamonds and nano-shaped using intense femtosecond pulses to design cantilever-based nanomechanical hybrid quantum sensors. Thus fabricated ultrathin glass cantilevers show stable optical, electronic, and magnetic properties of nitrogen-vacancy centers, including well-defined fluorescence with zero-phonon lines and optically detected magnetic resonance (ODMR) near 2.87 GHz. We demonstrate several sensing applications of the fluorescent ultrathin glass cantilever by measuring acoustic pulses, external magnetic field using Zeeman splitting of the NV centers, or CW laser-induced heating by measuring thermal shifting of ODMR lines. This work demonstrates the suitability of the femtosecond-processed fluorescent ultrathin glass as a new versatile substrate for multifunctional quantum devices.

Understanding Sociocultural Influences on Physical Activity in Relation to Overweight and Obesity in a Rural Indigenous Community of Fiji Islands.

INTRODUCTION: Given health disparities and increased rates of obesity and non-communicable diseases seen in Indigenous populations worldwide and the evidence connecting sociocultural knowledge with physical activity, health, and wellbeing, this research was undertaken to understand the social and cultural components contributing to obesity in the Indigenous Fijian rural areas. METHODS: This research is a community-based participatory research (CBPR) project, which engaged community members from a rural iTaukei village in the Fiji Islands. Data collection was carried out through community consultation and semi-structured interviews. The data was analysed using descriptive thematic analysis. RESULTS: Four major themes emerged associated with sociocultural, economic, political, and physical environmental factors. Males emphasised sports and working on farmlands as preferred types of physical activity, while females focused on family activities and daily activities and support for females' separate playgrounds. There was a focus on previous health promotion programs that did not incorporate the cultural values, cultural competence beliefs, and traditional ways of the rural Indigenous Fijian community. CONCLUSION: The healthcare providers and policymakers need to recognise the iTaukei community culture and appreciate traditional methods to promote equitable community participation in decision-making for health promotion. These findings should inform future research and community-based health programs to address the physical activity levels of the rural Indigenous community and may be relevant to other Indigenous peoples.

ABT1 modifies SMARD1 pathology via interactions with IGHMBP2 and stimulation of ATPase and helicase activity.

SMA with respiratory distress type 1 (SMARD1) and Charcot-Marie-Tooth type 2S (CMT2S) are results of mutations in immunoglobulin mu DNA binding protein 2 (IGHMBP2). IGHMBP2 is a UPF1-like helicase with proposed roles in several cellular processes, including translation. This study examines activator of basal transcription 1 (ABT1), a modifier of SMARD1-nmd disease pathology. Microscale thermophoresis and dynamic light scattering demonstrate that IGHMBP2 and ABT1 proteins directly interact with high affinity. The association of ABT1 with IGHMBP2 significantly increases the ATPase and helicase activity as well as the processivity of IGHMBP2. The IGHMBP2/ABT1 complex interacts with the 47S pre-rRNA 5' external transcribed spacer and U3 small nucleolar RNA (snoRNA), suggesting that the IGHMBP2/ABT1 complex is important for pre-rRNA processing. Intracerebroventricular injection of scAAV9-Abt1 decreases FVB-Ighmbp2nmd/nmd disease pathology, significantly increases lifespan, and substantially decreases neuromuscular junction denervation. To our knowledge, ABT1 is the first disease-modifying gene identified for SMARD1. We provide a mechanism proposing that ABT1 decreases disease pathology in FVB-Ighmbp2nmd/nmd mutants by optimizing IGHMBP2 biochemical activity (ATPase and helicase activity). Our studies provide insight into SMARD1 pathogenesis, suggesting that ABT1 modifies IGHMBP2 activity as a means to regulate pre-rRNA processing.

Quantification of Indoor Respirable Suspended Particulate Matters (RSPM) and Asthma in Rural Children of Delhi-NCR.

OBJECTIVES: To assess the exposure of indoor respirable suspended particulate matters (PM10, PM2.5, and PM1) and their association with asthma in children in a rural area of Delhi-NCR. METHODS: It was a cross-sectional study. Fifty children with asthma from both biomass fuel users in group A and liquefied petroleum gas (LPG) fuel users in group B households were enrolled along with 50 healthy control subjects. The diagnosis of asthma was done as per the Global Initiative for Asthma (GINA), 2014. The 24-h levels of PM from all three groups of households were measured and compared. The level of PM with confounding factors like smoking and room occupancy was also compared between the groups. RESULTS: The 24-h concentrations of PM10, PM2.5, and PM1 were found significantly higher in the households of group A and group B as opposed to group C (p < 0.001). The number of smokers with a mean pack year and a lack of an exhaust fan was highest in group A and lowest in group C, while diesel and kerosene machines were highest in group B. The PMs were highest in group A even with different confounding factor (p < 0.001). The level of all PM was higher in group B than in group C, despite the presence of both types of fuel in group C households. The level of all PM was highest during the cooking hour. CONCLUSION: The level of 24-h PM was highest in group-A households. However, the level of PM was higher in group-B households than group C despite the presence of biomass fuel users in group C. This may be due to the higher number of smokers, poor room-occupancy and lack of exhaust fans.

Teaching clinical anatomy to medical students by flipped classroom.

Background We aimed to introduce the flipped classroom approach for teaching clinical anatomy and to assess its perception and feasibility among medical undergraduate students. Methods Our study included 151 MBBS students of the first professional. We took written informed consent after the study was approved by the Ethics Committee. Selected topics of clinical anatomy were taught using the flipped classroom (FCR) method. It involved pre-class, in-class and post-class activities. In pre-class activity, pre-reading material was given to the students 1 week before the class. An assignment was given 2 days before the class in the form of solving multiple- choice questions, drawing well-labelled diagrams, etc. In- class activity included a pre-test followed by a discussion of the topic in the form of problem-based questions in the class on the principle of Think-Pair-Share. Post-class activity included summarization of the topic by students in the form of a group activity followed by a feedback session. Feedback was collected using a pre-validated feedback questionnaire on Google forms. Data collected were analysed using SPSS 21.0 and Microsoft office 2010. The response to close-ended questions was expressed as percentage. Open-ended questions were analysed by grouping of qualitative responses. Results We found FCR to be a feasible, interesting and necessary pedagogical tool for medical education. Most students (95%) agreed that FCR is a useful technique for better understanding of clinical anatomy. They learnt better from FCR than other teaching methods and their in-depth understanding of the topic improved with FCR. Conclusion FCR is an established teaching-learning tool but it needs to be implemented in undergraduate teaching of clinical anatomy for better understanding of the topics. Didactic lectures do not touch upon clinical correlations in a case-based manner, which can be easily discussed in FCR.