Organocatalytic access to 3-pyridylphosphonates from vinyl phosphonates and aldehydes.

The preparation of 3-phosphorylated pyridines has remained the most challenging compared to the corresponding 2- or 4-functionalized pyridines. Herein, an unprecedented amino-organocatalyzed direct synthesis of 3-pyridylphosphonates from vinylphosphonates and aldehydes has been achieved. This allows access to a wide range of multi-substituted phosphorylated pyridines in excellent yields under transition metal-free conditions.

Long non-coding RNAs in cancer: multifaceted roles and potential targets for immunotherapy.

Cancer remains a major global health concern with high mortality rates mainly due to late diagnosis and poor prognosis. Long non-coding RNAs (lncRNAs) are emerging as key regulators of gene expression in human cancer, functioning through various mechanisms including as competing endogenous RNAs (ceRNAs) and indirectly regulating miRNA expression. LncRNAs have been found to have both oncogenic and tumor-suppressive roles in cancer, with the former promoting cancer cell proliferation, migration, invasion, and poor prognosis. Recent research has shown that lncRNAs are expressed in various immune cells and are involved in cancer cell immune escape and the modulation of the tumor microenvironment, thus highlighting their potential as targets for cancer immunotherapy. Targeting lncRNAs in cancer or immune cells could enhance the anti-tumor immune response and improve cancer immunotherapy outcomes. However, further research is required to fully understand the functional roles of lncRNAs in cancer and the immune system and their potential as targets for cancer immunotherapy. This review offers a comprehensive examination of the multifaceted roles of lncRNAs in human cancers, with a focus on their potential as targets for cancer immunotherapy. By exploring the intricate mechanisms underlying lncRNA-mediated regulation of cancer cell proliferation, invasion, and immune evasion, we provide insights into the diverse therapeutic applications of these molecules.

Unified Identification Protocol for Cross-Border Healthcare.

The Unified Identification Protocol (UIP) is an innovation which empowers patients and legal guardians to generate their unique digital identity for cross-border healthcare. This digital identity seamlessly links to local identifiers across different territories and organizations, bridging the gap between disparate systems. Combined with the International Patient Summary (IPS) - endorsed by the G7 and the EU - UIP is pioneering a new paradigm in telehealth services. Championing a user-centric approach in line with Web 3.0 principles, UIP places data control directly in the hands of patients and their legal guardians. This ensures accurate identification, streamlined access to health data, and robust privacy protection. When harmonized with tools like the SMART-On-FHIR API, FHIR Contract, DID Documents, and blockchain certification, UIP lays down transparent, user-approved guidelines for sharing healthcare data across borders. This framework guarantees that data is securely exchanged, encrypted specifically for the intended recipients upon user consent, adhering to international standards, and in full compliance with prevailing regulations. Furthermore, UIP facilitates certification of health courses and competences for patients, caregivers, and practitioners, enhancing healthcare understanding and management.

Exploring salivary exosomes as early predictors of oral cancer in susceptible tobacco consumers: noninvasive diagnostic and prognostic applications.

BACKGROUND: Salivary exosome analysis provides a noninvasive and comprehensive approach with potential applications in oral cancer diagnosis and prognosis. The early detection of oral cancer has remained a critical concern in enhancing the quality of life, especially for individuals who consume tobacco and are at greater risk of developing the disease. The current study investigates the potential of salivary exosomes in screening smokers for early signs and transformations of oral cancer. METHODS: Morphological characterization of salivary exosomes among three study groups (non-smokers as control, smokers as high-risk tobacco consumers, and Oral cancer) (n = 120) was carried out through dynamic light scattering, and nanoparticle tracking analysis. For molecular characterization, EXOCET and Fourier transform infrared spectroscopy methods were utilized. The expression of the exosomal surface protein CD63 was evaluated using Western blotting. RESULTS: Salivary exosomes exhibit noticeable differences in size between control group and tobacco consumers. The differentiation extended beyond exosome size and included variations in concentration and bio-molecular composition, as determined by FTIR screening. Tobacco consumers and oral cancer groups showed significantly larger and more concentrated exosomes compared to the healthy group. CONCLUSION: Our study provides strong evidence that the properties of salivary exosomes can serve as reliable noninvasive biomarkers for distinguishing tobacco consumers from non-smokers and oral cancer patients. Our results underscore the potential of exosome-based diagnostics in early oral cancer detection for high-risk individuals. The larger size and higher concentration of exosomes in tobacco consumers indicate early changes in cell secretions associated with the transformation from healthy to abnormal cells.

Involvement of DPP3 in modulating oncological features and oxidative stress response in esophageal squamous cell carcinoma.

Resistance to therapy in esophageal squamous cell carcinoma (ESCC) is a critical clinical problem and identification of novel therapeutic targets is highly warranted. Dipeptidyl peptidase III (DPP3) is a zinc-dependent aminopeptidase and functions in the terminal stages of the protein turnover. Several studies have reported overexpression and oncogenic functions of DPP3 in numerous malignancies. The present study aimed to determine the expression pattern and functional role of DPP3 in ESCC. DPP3 expression was assessed in normal and tumor tissues using quantitative real-time (qRT)-PCR and corroborated with ESCC gene expression datasets from Gene Expression Omnibus (GEO) and The cancer genome atlas (TCGA). DPP3 stable knockdown was performed in ESCC cells by shRNA and its effect on cell proliferation, migration, cell cycle, apoptosis, and activation of nuclear factor erythroid 2-related factor 2 (NRF2) pathway was assessed. The results suggested that DPP3 is overexpressed in ESCC and its knockdown leads to reduced proliferation, increased apoptosis, and inhibited migration of ESCC cells. Additionally, DPP3 knockdown leads to down-regulation of the NRF2 pathway proteins, such as NRF2, G6PD, and NQO1 along with increased sensitivity toward oxidative stress-induced cell death and chemotherapy. Conclusively, these results demonstrate critical role of DPP3 in ESCC and DPP3/NRF2 axis may serve as an attractive therapeutic target against chemoresistance in this malignancy.

Patient-Centric Interoperability and Cybersecurity for Cross-Border Healthcare.

In Web 3.0 the user owns the information. Decentralized Identity Documents (DID documents) allow users to create their own digital identity and decentralized cryptographic material resistant to quantum computing. A patient's DID document also contains a unique identifier for cross-border healthcare, endpoints for receiving DIDComm messages and for SOS services, and additional identifiers (passport, etc.). We propose a blockchain for cross-border healthcare to store the evidence of different electronic, physical identities, and identifiers, but also the rules approved by the patient or legal guardians to access patient data. The International Patient Summary (IPS) is the de facto standard for cross-border healthcare and includes an index of information classified into sections (HL7 FHIR Composition) that healthcare professionals and services can update and read on the patient's SOS service, then retrieving all the necessary patient information from the different FHIR API endpoints of different healthcare providers according to the approved rules.

Interoperable Universal Resource Identifier for Selective Disclosure of Data.

All the information stored in the different information systems is issued in a format that allows the holder (the information owner) to disclose only certain information to a third party, which will act as a requester, receiver and verifier of the information disclosed by the holder. We define the Interoperable Universal Resource Identifier (iURI) as a harmonized method of representing a claim (minimum piece of verifiable information) using disparate encoding systems, agnostic to the original encoding system and data format. Encoding systems are represented in Reverse Domain Name Resolution (Reverse-DNS) format for HL7 FHIR, OpenEHR, and other data formats. The iURI can then be used in JSON Web Token for Selective Disclosure (SD-JWT) and Verifiable Credential (VC), among others. The method enables a person to demonstrate data that already exists in different information systems in disparate data formats, and even an information system, to verify certain claims, in a harmonized way.

Quality and Traceability of Starting Materials from Supplier to Recipient.

Quality of processes and products is based on traceability and review of both components, material processing and product flow throughout the manufacturing and supply chain. Blockchain technology enables cross-border audit trail and traceability while reducing costs. Donors are the providers of biological raw material (starting material). They can share their health records when donating by using an IPS document or a FHIR Questionnaire-response resource. It allows health personnel to retrieve and verify relevant clinical information when donating. Additionally, health personnel can generate an anonymized and de-identified digital twin of the donor for research purposes, and it can be updated over time. The starting material can include a reference to a digital twin of an unknown supplier, which improves the data quality and enhances research possibilities. Adverse reactions and events can be also recorded on blockchain to improve safety, transparency, traceability, medical research and product quality.

The mediating role of sleep quality on the relationship between internet gaming disorder and perceived stress and suicidal behaviour among Indian medical students.

Background: In the recent digital era, individuals with internet gaming disorder (IGD) have reported a much higher prevalence of poor sleep quality, perceived stress and suicidal behaviour. However, the underlying mechanisms for these psychological problems remain unknown. Aims: The primary aims of this study were to explore the mediating role of sleep quality on the relationship between IGD and the health outcomes of perceived stress and suicidal behaviour and to assess the prevalence and risk factors for IGD among medical students. Methods: A cross-sectional study enrolling 795 medical students from two medical colleges in a rural area of North India was conducted from April to May 2022. The study participants were chosen using a stratified random sampling approach. A self-administered questionnaire was used to collect data, including sociodemographic and personal information and gaming characteristics. The study also included the Gaming Disorder and Hazardous Gaming Scale, the Pittsburgh Sleep Quality Index, the Perceived Stress Scale-10 and the Suicide Behaviors Questionnaire-Revised to measure IGD, sleep quality, perceived stress and suicidal behaviour, respectively. Multiple logistic regression for the risk factors and Pearson's correlation test for the relationship between variables were used. Hayes' PROCESS macro for SPSS was employed to carry out mediation analysis. Results: Among the 348 gamers with a mean age of 21.03 (SD 3.27) years, the prevalence of IGD was 15.23% (95% confidence interval: 11.6% to 19.4%). In the correlational analysis, small to large (r: 0.32-0.72) significant relationships between scores of IGD and other health outcomes were established. The indirect effect (B=0.300) via sleep quality accounted for 30.62% of the total effect (B=0.982) of IGD on perceived stress (partially mediated), while sleep quality (B=0.174) accounted for 27.93% of the total effect (B=0.623) of IGD on suicidal behaviour (partially mediated). The factors of being male, living in a single-parent family, using the internet for other than academic purposes (1-3 hours and more than 3 hours/day), playing games for more than 3 hours/day and playing games with violent content were associated with IGD symptoms. Conclusions: Using a dimensional measure, the results specified the relationship between IGD and perceived stress and suicidal behaviour by demonstrating that sleep quality meditated them. This modifiable mediating factor can be addressed by psychotherapy to mitigate the risk of perceived stress and suicidal behaviour among the future medical workforce.

Heterologous expression of Chlorophytum borivilianum Squalene epoxidase in tobacco modulates stigmasterol production and alters vegetative and reproductive growth.

KEYMESSAGE: CbSE overexpression increased stigmasterol levels and altered plant morphology. The genes upstream and downstream of CbSE were found to be upregulated, which confirms its regulatory role in the saponin biosynthetic pathway. Chlorophytum borivilianum is a high-value medicinal plant with many promising preclinical applications that include saponins as a major active ingredient. Squalene epoxidase (SE) is one of the major rate-limiting enzymes of the saponin biosynthetic pathway. Here, we functionally characterized C. borivilianum SE (CbSE) by over-expressing heterologously in Nicotiana tabacum. The heterologous expression of CbSE resulted in stunted pant growth with altered leaf and flower morphology. Next, RT-qPCR analysis of transgenic plants overexpressing CbSE revealed increased expression levels of Cycloartenol synthase (CAS), Beta amyrin synthase (ßAS), and cytochrome P450 monooxygenase 51 (CYP51) (Cytochrome P450), which encode key enzymes for triterpenoid and phytosterol biosynthesis in C. borivilianum. Further, Methyl Jasmonate (MeJa) treatment upregulated Squalene synthase (SQS), SE, and Oxidosqualene cyclases (OSCs) to a significant level. GC-MS analysis of the leaf and hairy roots of the transformants showed an increased stigmasterol content (0.5-1.0 fold) compared to wild type (WT) plants. These results indicate that CbSE is a rate-limiting gene, which encodes an efficient enzyme responsible for phytosterol and triterpenoid production in C. borivilianum.

Genome-wide analysis of Catalase gene family reveal insights into abiotic stress response mechanism in Brassica juncea and B. rapa.

Environmental stresses affect the yield and productivity of Brassica crops. Catalases are important antioxidant enzymes involved in reducing excess hydrogen peroxide produced by environmental stresses. In the present study, nine and seven CAT family members in two oilseed Brassica species (B. juncea and B. rapa) were identified with complete characterization based on gene and protein structure. Phylogenetic classification categorized CAT proteins into three classes and differentiated the monocot and dicot-specific CAT proteins. Further, the gene and protein characterizations revealed a high degree of conservation across the CAT family members. Differences were observed in the CAT-HEME binding affinity in CAT1, CAT2, and CAT3 isozymes, which could suggest their differential enzyme activities in different conditions. Furthermore, protein-protein interaction with other antioxidant proteins suggested their coordinated role in ROS scavenging mechanisms. Notably, the differential gene expression of BjuCATs and BraCATs and CAT enzyme activities suggested their crucial roles in major abiotic stresses faced by Brassica species. Promoter analysis in BjuCATs and BraCATs suggested the presence of abiotic-stress responsive cis-regulatory elements. Gene regulatory network analysis suggested miRNA and TF mediated stress response in BjuCATs and BraCATs. CAT family screening and characterization in Brassica sp. has established a basic ground for further functional validation in abiotic and heavy-metal stresses which can help in developing stress tolerant crops.

Impact of Chronic Sodium Fluoride Toxicity on Antioxidant Capacity, Biochemical Parameters, and Histomorphology in Cardiac, Hepatic, and Renal Tissues of Wistar Rats.

The study was designed to determine the fluoride distribution after its oral exposure in drinking water and its associated impact on biochemical, antioxidant markers and histology in the liver, kidney, and heart of male Wistar rats. On 100 ppm exposure, the highest accretion of fluoride occurred in the liver followed by the kidney and heart. Fluoride exposure significantly (p˂0.05) increased the plasma levels of dehydrogenase, aminotransferases, kidney injury molecule-1 (KIM-1), and other plasma renal biomarkers but decreased the levels of total plasma proteins and albumin in a dose-dependent manner. Reduction (p˂0.05) in the activities of antioxidant enzymes viz. acetylcholinesterase, arylesterase, superoxide dismutase, catalase, glutathione peroxidase, and reductase with increased levels of protein and lipid peroxidation was recorded in the liver, kidney, and heart of fluoride-administered rats. Fluoride exposure (100 ppm) induced lipid peroxidation was highest in kidney (4.4 times) followed by liver (2.6 times) and heart (2.5 times) and as compared to their respective control. The percent rise in protein oxidation at 30% was almost equal in the kidney and liver but was 21.5% in the heart as compared to control. The histopathological alterations observed included congestion and hemorrhage along with degeneration and necrosis of parenchymal cells in hepato-renal tissues and myocardium, severity of which varied in a dose-dependent manner. Taken together, fluoride distribution in the liver, heart, and kidney after chronic fluoride intake correlated well with fluoride-induced hepatic and cardio-renal toxicity in a concentration-dependent manner. These results draw attention that chronic fluoride intake pose a significant health risk for human and animal residents of fluoride endemic areas.

External validation of prediction models for patient-reported outcome measurements collected using the selfBACK mobile app.

BACKGROUND: External validation is essential in examining the disparities in the training and validation cohorts during the development of prediction models, especially when the application domain is healthcare-oriented. Currently, the use of prediction models in healthcare research aimed at utilising the under-explored potential of patient-reported outcome measurements (PROMs) is limited, and few are validated using external datasets. OBJECTIVE: To validate the machine learning prediction models developed in our previous work [29] for predicting four pain-related patient-reported outcomes from the selfBACK clinical trial datasets. METHODS: We evaluate the validity of three pre-trained prediction models based on three methods- Case-Based Reasoning, Support Vector Regression, and XGBoost Regression-using an external dataset that contains PROMs collected from patients with non-specific neck and or low back pain using the selfBACK mobile application. RESULTS: Overall, the predictive power was low, except for prediction of one of the outcomes. The results indicate that while the predictions are far from immaculate in either case, the models show ability to generalise and predict outcomes for a new dataset. CONCLUSION: External validation of the prediction models presents modest results and highlights the individual differences and need for external validation of prediction models in clinical settings. There is need for further development in this area of machine learning application and patient-centred care.

Exploratory application of machine learning methods on patient reported data in the development of supervised models for predicting outcomes.

BACKGROUND: Patient-reported outcome measurements (PROMs) are commonly used in clinical practice to support clinical decision making. However, few studies have investigated machine learning methods for predicting PROMs outcomes and thereby support clinical decision making. OBJECTIVE: This study investigates to what extent different machine learning methods, applied to two different PROMs datasets, can predict outcomes among patients with non-specific neck and/or low back pain. METHODS: Using two datasets consisting of PROMs from (1) care-seeking low back pain patients in primary care who participated in a randomized controlled trial, and (2) patients with neck and/or low back pain referred to multidisciplinary biopsychosocial rehabilitation, we present data science methods for data prepossessing and evaluate selected regression and classification methods for predicting patient outcomes. RESULTS: The results show that there is a potential for machine learning to predict and classify PROMs. The prediction models based on baseline measurements perform well, and the number of predictors can be reduced, which is an advantage for implementation in decision support scenarios. The classification task shows that the dataset does not contain all necessary predictors for the care type classification. Overall, the work presents generalizable machine learning pipelines that can be adapted to other PROMs datasets. CONCLUSION: This study demonstrates the potential of PROMs in predicting short-term patient outcomes. Our results indicate that machine learning methods can be used to exploit the predictive value of PROMs and thereby support clinical decision making, given that the PROMs hold enough predictive power.

Characterization of APX and APX-R gene family in Brassica juncea and B. rapa for tolerance against abiotic stresses.

KEY MESSAGE: APX and APX-R gene families were identified and characterized in two important oilseed species of Brassica. Gene expression under abiotic stress conditions, recombinant protein expression, and analysis further divulged their drought, heat, and salt-responsive behavior. Ascorbate peroxidases (APX) are heme-dependent enzymes that rid the cells of H2O2 and regulate diverse biological processes. In the present study, we performed APX gene family characterization in two Brassica sp. (B. juncea and B. rapa) as these are commercially important oilseed crops and affected severely by abiotic stresses. We identified 16 BjuAPX and 9 BraAPX genes and 2 APX-R genes each in B. juncea and B. rapa genomes, respectively. Phylogenetic analysis divided the APX genes into five distinct clades, which exhibited conservation in the gene structure, motif organization, and sub-cellular location within the clade. Structural analysis of APX and APX-R proteins revealed the amino acid substitutions in conserved domains of APX-R proteins. The expression profiling of BjuAPX and BraAPX genes showed that 3 BjuAPX, 7BraAPX, and 2 BraAPX-R genes were drought and heat responsive. Notably, BjuAAPX1a, BjuAPX1d, BjuAAPX6, BraAAPX1a, BraAAPX2, and BraAAPX3b showed high expression levels in RT-qPCR. Cis-regulatory elements in APX and APX-R gene promoters supported the differential behavior of these genes. Further, two stress-responsive genes BjuAPX1d and BraAAPX2 were cloned, characterized, and their roles were validated under heat, drought, salt, and cold stress in bacterial expression system. This study for the first time reports the presence of APX activity in dimeric and LMW form of purified BraAAPX2 protein. The study may help pave way for developing abiotic stress-tolerant Brassica crops.

CRISPR-Cas enzymes: The toolkit revolutionizing diagnostics.

The programmable nature of sequence-specific targeting by CRISPR-Cas nucleases has revolutionized a wide range of genomic applications and is now emerging as a method for nucleic acid detection. We explore how the diversity of CRISPR systems and their fundamental mechanisms have given rise to a wave of new methods for target recognition and readout. These cross-disciplinary advances found at the intersection of CRISPR biology and engineering have led to the ability to rapidly generate solutions for emerging global challenges like the COVID-19 pandemic. We further discuss the advances and potential for CRISPR-based detection to have an impact across a continuum of diagnostic applications.

Bioactive compounds and their libraries: An insight into prospective phytotherapeutics approach for oral mucocutaneous cancers.

Oral mucocutaneous cancers (OMCs) are cancers that affect both the oral mucosa and perioral cutaneous structures. Common OMCs are squamous cell carcinoma (SCC), basal cell carcinoma (BCC) and malignant melanoma (MM). Anatomical similarities and conventions which categorizes these lesions blur the magnitude of OMCs in diverse populations. The burden of OMC is high in the sub-Saharan Africa and Indian subcontinents, and the cost of management is prohibitive in the resource-limited, developing world. Hence, there is a pressing demand for the use of cost-effective in silico approaches to identify diagnostic tools and treatment targets for diseases with high burdens in these regions. Due to their ubiquitousness and accessibility, the use of therapeutic efficacy of plant bioactive compounds in the management of OMC is both appropriate and plausible. Furthermore, screening known mechanistic disease targets with well annotated plant bioactive compound libraries is poised to improve the routine management of OMCs provided that the requisite access to database resources are available and accessible. Using natural products minimizes the side effects and morbidities associated with conventional therapies. The development of innovative treatments approaches would tremendously benefit the African and Indian populace and reduce the mortalities associated with OMCs in the developing world. Hence, we discuss herein, the potential benefits, opportunities and challenges of using bioactive compound libraries in the management of OMCs.

Overcoming challenges in implementing measures across multiple centers of a chain of hospitals to combat COVID-19 pandemic.

PURPOSE: Coronavirus outbreak was first reported towards the end of 2019 and has now been declared a pandemic by the World Health Organization. Post declaration, India announced lockdown and various guidelines were issued by Health authorities worldwide for the organizations to function in this emergency aeon. Different organizations strategized differently to respond to COVID 19 pandemic. The aim was to implement COVID 19 guidelines across all the centres of a chain of Eye hospitals in India in short span of time overcoming the challenges. METHODS: A SMART strategy was confabulated within the team. There were numerous challenges like lack of time, rapidly changing guidelines, no travelling scenarios, training and educating the staff in huge numbers, rapidly evolving situation, change management, need of increased vigilance, and language barrier at certain centres, sustaining quality assurance and motivating the staff in stressful times. The challenges were surpassed using a power packed strategy, dedicated efforts and team work grasping the available tools and technology from a single location. This template can serve us and other hospitals to respond in a better way for similar future challenges. RESULTS: The COVID 19 protocols were implemented across all the centres successfully. The entire Quality assurance system drifted to online platform and education and awareness methods took the digital thread. Teamwork synergized with the aforementioned things and we came out with flying colours achieving the goals in short span of time overcoming all the barriers. CONCLUSION: Difficult times bring challenges and challenges teach us the innovative ways to convert them into opportunities. A well planned strategy, best utilization of available resources, team work and continuous monitoring can bring the best outcomes forward.