RESUMEN
INTRODUCTION: India has a significant TB burden, and ongoing attempts are being made to eradicate the disease. Globally, the number of TB deaths is declining, but not quickly enough to meet the End TB Goals. The National Strategic Plan (NSP) 2017-2025 in India set in motion an ambitious effort to expand the scope and efficacy of the National Tuberculosis Elimination Program (NTEP). METHODS: A descriptive retrospective study based on secondary data was conducted on information obtained from the electronic TB notification register for 2019, abstracted from Ni-kshay. Further, descriptive analysis was undertaken to identify the factors associated with deaths and successful treatment outcomes. The binomial logistic regression model estimates the crude relative risk and a 95% confidence interval to describe the association between predictor variables and TB treatment outcomes. RESULTS: After applying the eligibility criteria for the study population, a total of 1,44,643 (88%) TB patients were included in the study. 1,35,934 (94%) TB patients had completed the treatment and survived, while 8709 (6%) TB patients died. A significant association of treatment outcomes was observed in age, gender, key population, site of diseases, type of case, type of health facilities, HIV and Diabetes. When a logistic regression was applied, the model showed the association of the independent variables with the risk of death in TB patients. CONCLUSION: The epidemiological factors associated with treatment outcomes among TB patients should be audited systematically. A structure of TB death surveillance and response system should be established with a mortality audit, including a community-based death review (CBDR) and a facility-based medical audit (FBMA) in case the patient is hospitalized or discharged from a hospital.
Asunto(s)
Tuberculosis , Humanos , Estudios Retrospectivos , Tuberculosis/tratamiento farmacológico , Resultado del Tratamiento , Modelos Logísticos , India/epidemiología , Antituberculosos/uso terapéuticoRESUMEN
The presence of exorbitant arsenic contamination in the aquatic environment causes astronomically immense health quandaries affecting millions of people, which may lead to death in the case of prolonged indigestion of arsenic-containing drinking water. Herein, we are reporting porous chelating resin with an iron precursor for the removal of arsenic ions from water. Weak acid cation resin was functionalized under varying experimental conditions to get a suitable resin with high arsenic uptake. The theoretical results revealed that the maximum Langmuir adsorption capacities of 3.27 mg g-1 and 1.13 mg g-1 were achieved for As(V) and As(III), respectively. The kinetics of adsorption followed the pseudo-second-order (PSO) model with a high determination coefficient (R2) of 0.9963 and 0.9895 for As(V) and As(III), respectively. The Adams-Bohart, Thomas, Yoon-Nelson, and Pore diffusion models were used to identify the breakthrough curve in the fixed bed adsorption column. The column performance improved with a larger bed height (55 cm), low concentration of influent (0.25 mg L-1), and low flow rate of influent (80 mL min-1). Under this condition, the breakthrough time and exhaustion time were 314 min and 408 min for As(V) and 124 min and 185 min for As(III), respectively.
RESUMEN
Two-dimensional MXene with layered structure has recently emerged as a nanomaterial with fascinating characteristics and applicability. Herein, we prepared the newly modified magnetic MXene (MX/Fe3O4) nanocomposite using solvothermal approach and investigated its adsorption behavior to study the removal efficiency of Hg(II) ions from aqueous solution. The effect of adsorption parameters such as adsorbent dose, time, concentration, and pH were optimized using response surface methodology (RSM). The experimental data fitted well with quadratic model to predict the optimum conditions for maximum Hg(II) ion removal efficiency which were found to be at adsorbent dose 0.871 g/L, time 103.6 min, concentration 40.17 mg/L, and 6.5 pH respectively. To determine the adequacy of the developed model, a statistical analysis of variance (ANOVA) was used, which demonstrated high agreement between the experimental data and the suggested model. According to isotherm result, the experimental data were following the best agreement with the Redlich-Peterson isotherm model. The results of the experiments revealed that the maximum Langmuir adsorption capacity of 699.3 mg/g was obtained at optimum conditions, which was closed to the experimental adsorption capacity of 703.57 mg/g. The adsorption phenomena was well represented by the pseudo-second-order model (R2 = 0.9983). On the whole, it was clear that MX/Fe3O4 has lot of potential as a Hg(II) ion impurity removal agent in aqueous solutions.
Asunto(s)
Mercurio , Contaminantes Químicos del Agua , Contaminantes Químicos del Agua/análisis , Mercurio/análisis , Agua/química , Magnetismo , Adsorción , Cinética , Concentración de Iones de HidrógenoRESUMEN
In the present scenario discharge of heavy-metal ions into water bodies is a global threat that is causing serious health hazards even in low concentrations. Thus, in order to remediate the heavy-metal [Hg(II) and Pb(II)] toxicity, an organic-inorganic hybrid functional porous metallo-polymeric network i. e, poly(Zirconyl methacrylate-co-1-vinyl imidazole) (pZrVIm) was fabricated via one-pot facile synthesis approach. The pZrVIm architecture has shown high removal efficiency for Hg(II) and Pb(II) aqueous medium even in extremely low quantities. Advanced instrumental techniques were used to characterize the structural and morphological characteristics of pZrVIm. Different experimental variables i.e., reaction time, pH, initial feed concentration, co-ion effects etc. were explored to examine adsorption behaviour. The maximum adsorption capacities (qmax) of pZrVIm5 were calculated as 168.06 and 162.34 mg g-1 for Hg(II) and Pb(II) respectively by the Langmuir isotherm model. Data from isotherms showed that monolayer adsorption on a homogeneous surface is the rate-limiting stage and followed pseudo-second-order kinetic process. The Artificial Neural Network (ANN) modelling was used to validate kinetics and isotherm data which revealed high accuracy of the model with correlation coefficient values (R = 0.99). Various types of isotherm models such as Langmuir, Freundlich, Dubinin-Radushkevich, Temkin, Redlich-Peterson, Toth and Koble-Corigen have been studied to determine the adsorption phenomena. The pore diffusion model revealed breakthrough time of 91 h and 84 h, Hg(II) and Pb(II) with the feed concentration of 15 mg L-1 respectively. The study revealed that pZrVIm5 has great potential for heavy metal ions remediation for water treatment.
Asunto(s)
Mercurio , Metales Pesados , Contaminantes Químicos del Agua , Plomo , Porosidad , Mercurio/química , Polímeros , Iones , Adsorción , Cinética , Contaminantes Químicos del Agua/análisis , Concentración de Iones de Hidrógeno , TermodinámicaRESUMEN
The rearrangement of dithiolanes and dithianes to access disulfide-linked-dithioesters under visible-light photoredox catalysis via controlled C-S bond cleavage has been disclosed. Unlike, the usual deprotection of dithioacetals to corresponding aldehydes under the oxidative conditions, we observed unique regioselective oxidative reactivity of five and six membered cyclic dithioacetals to form disulfide-linked-dithioesters by exchanging DMAP and imidazole bases. The generality of the protocol has been demonstrated by exploring a wide range of substrates. As an application, inâ situ generated thiyl radical has been trapped with disulfides to prepare hetero-disulfides of potential utility. The protocol proved to be practical on gram scale quantity and relied on clean energy source for the transformation. Based on the series of control experiments, cyclic voltammetry and Stern-Volmer studies the plausible mechanism has been proposed.
RESUMEN
Background: Osteodystrophia fibrosa (ODF) is a metabolic disorder affecting the skeletal system, causing progressive loss of calcified bone mass and its replacement with fibrous tissue, which may be a sequel to primary or secondary hyperparathyroidism. This report intends to document the clinicopathological findings of ODF in a flock of young goats fed primarily on a wheat bran-rich diet. Case description: In a flock of 50 stall-fed goats aged 1 to 2 years, seven were clinically presented with bilateral facial enlargement, leading to dyspnea and difficulty in prehension and mastication. Among the seven clinically affected goats, four died in 2 months. Findings/treatment and outcome: The clinical examination revealed bilateral mandibular enlargement and limb deformities. On radiography, the maxilla and mandible had decreased radiopacity. Fine needle aspiration cytology (FNAC) from the affected bones showed occasional fibroblasts and individual osteoclasts clusters. On necropsy, the enlarged mandible revealed a meaty consistency. Undecalcified histological sections of the mandible showed severe osteopenia, multiple osteoclasts, Howship's lacunae, and extensive fibroplasia. Dietary corrective measures led to the prevention of ODF in the rest of the flock. Conclusion: Excessive wheat bran feeding in stallfed goats might have led to calcium and phosphorus imbalance, resulting in nutritional secondary hyperparathyroidism and subsequent skeletal deformities. FNAC of the affected bones, gross and histological findings provide a clinicopathological diagnosis of ODF.
RESUMEN
The cyanation of organic compounds is an important synthetic transformation and mainly relies on a toxic CN source. Undeniably, thiocyanate salt has emerged as a very mild and environmentally benign CN source, yet its synthetic utility for cyanation is highly limited to very few types of organic compounds. Herein, we report the direct cyanation of cyclic thioacetals for accessing compounds with two different functional groups (thiocyano-thioesters) in one pot using sodium thiocyanate via photoredox catalysis. The protocol has been further extended for the direct cyanation of disulfides and diselenide to access aryl thiocyanates and aryl selenocyanate. A plausible mechanism has been proposed based on a series of control experiments, cyclic voltammetry and Stern-Volmer studies.
Asunto(s)
Nitrilos , Tiocianatos , Catálisis , Cianatos/química , Disulfuros , Compuestos de Selenio/químicaRESUMEN
Bacteria readily form resilient phenotypes to counter environmental and antibiotic stresses. Here, we demonstrate a class of small molecules that inhibit a wide range of Pseudomonas aeruginosa phenotypes and enable antibiotics to kill previously tolerant bacteria, preventing the transition of tolerant bacteria into a persistent population. We identified two proteins, type IV pili and lectin LecA, as receptors for our molecules by methods including a new label-free assay based on bacterial motility sensing the chemicals in the environment, the chemical inhibition of bacteriophage adsorption on pili appendages of bacteria, and fluorescence polarization. Structure-activity relationship studies reveal a molecule that inhibits only pili appendage and a class of chimeric ligands that inhibit both LecA and pili. Important structural elements of the ligand are identified for each protein. This selective ligand binding identifies the phenotypes each protein receptor controls. Inhibiting LecA results in reducing biofilm formation, eliminating small colony variants, and is correlated with killing previously tolerant bacteria. Inhibiting pili appendages impedes swarming and twitching motilities and pyocyanin and elastase production. Because these phenotypes are controlled by a broad range of signaling pathways, this approach simultaneously controls the multiple signaling mechanisms preventing bacteria to elude antibiotic treatments.
Asunto(s)
Pseudomonas aeruginosa , Factores de Virulencia , Antibacterianos/metabolismo , Antibacterianos/farmacología , Bacterias/metabolismo , Lectinas/metabolismo , Ligandos , Fenotipo , Pseudomonas aeruginosa/metabolismo , Factores de Virulencia/genética , Factores de Virulencia/metabolismoRESUMEN
Glycolipid, ganglio-N-tetraosylceramide (asialo-GM1), on the mammalian cells are known to be recognized by type IV pili of Pseudomonas aeruginosa. In this work, we show that asialo-GM1 can also be recognized by Lectin A (LecA), another adhesin protein of the P. aeruginosa, by a fluorescent polarization assay, a label-free bacterial motility enabled binding assay, and bacterial mutant studies. On hydrated semi-solid gel surfaces, asialo-GM1 enables swarming and twitching motilities, while on solid surfaces facilitates the bacterial adherence of P. aeruginosa. These results indicate that asialo-GM1 can modulate bioactivities, adherence, and motilities, that are controlled by opposite signaling pathways. We demonstrate that when a solution of pilin monomers or LecA proteins are spread on hydrated gel surfaces, the asialo-GM1 mediated swarming motility is inhibited. Treatment of artificial liposomes containing asialo-GM1 as a component of lipid bilayer with pilin monomers or LecA proteins caused transient leakage of encapsulated dye from liposomes. These results suggest that pili and LecA proteins not only bind to asialo-GM1 but can also cause asialo-GM1 mediated leakage. We also show that both pili and LecA mutants of P. aeruginosa adhere to asialo-GM1 coated solid surfaces, and that a class of synthetic ligands for pili and LecA inhibits both pili and LecA-mediated adherence of P. aeruginosa on asialo-GM1-coated surfaces.
Asunto(s)
Gangliósido G(M1) , Pseudomonas aeruginosa , Animales , Adhesión Bacteriana/fisiología , Proteínas Fimbrias/genética , Proteínas Fimbrias/metabolismo , Fimbrias Bacterianas/metabolismo , Gangliósido G(M1)/metabolismo , Lectinas/metabolismo , Lectinas/farmacología , Ligandos , Liposomas/metabolismo , Mamíferos/metabolismo , Pseudomonas aeruginosa/metabolismoRESUMEN
A visible light-mediated, metal-free, regioselective dihydrothionation of terminal aromatic as well as heteroaromatic alkynes has been achieved using Eosin Y as a photoredox catalyst at room temperature. The protocol gives direct access to different 1,3-dithiolanes under neutral and mild reaction conditions without the use of any base or additives. The electron-donating, electron-withdrawing and electron-deactivating groups tolerated the photocatalytic reaction conditions. The control experiments, cyclic voltammetry, and Stern-Volmer experiment were carried out to gain an insight into the mechanistic pathway. The protocol proved to be scalable at the gram level and also for practicality the deprotection of 1,3-dithiolanes has been demonstrated. The method uses clean energy under sustainable conditions.
RESUMEN
NaMgF3 -based phosphors have been described frequently in the literature. Their synthesis faces difficulties typical of fluoride materials. A simple precipitation synthesis for NaMgF3 -based phosphors is described in this paper. This consisted of mixing aq. NaF and MgCl2 /MgSO4 solutions. Various activators could be incorporated by adding the required salts during this process. Characteristic emission of the activators was observed in the prepared phosphors. As-prepared samples exhibited predominantly trivalent lanthanide emission. After thermal treatment in a reductive atmosphere, europium-doped samples showed the intense emission of Eu2+ . By virtue of the intense nature of the emission, lifetime measurements could be made for this sample. Notably, intense thermoluminescence and optically stimulated luminescence were observed in NaMgF3 :Eu. A simple, fast method for the synthesis of NaMgF3 was therefore developed.
Asunto(s)
Luminiscencia , Sustancias Luminiscentes , Europio , FluorurosRESUMEN
Bacteria in general can develop a wide range of phenotypes under different conditions and external stresses. The phenotypes that reside in biofilms, overproduce exopolymers, and show increased motility often exhibit drug tolerance and drug persistence. In this work, we describe a class of small molecules that delay and inhibit the overproduction of alginate by a non-swarming mucoid Pseudomonas aeruginosa. Among these molecules, selected benzophenone-derived alkyl disaccharides cause the mucoid bacteria to swarm on hydrated soft agar gel and revert the mucoid to a nonmucoid phenotype. The sessile (biofilm) and motile (swarming) phenotypes are controlled by opposing signaling pathways with high and low intracellular levels of bis-(3',5')-cyclic diguanosine monophosphate (cdG), respectively. As our molecules control several of these phenotypes, we explored a protein receptor, pilin of the pili appendages, that is consistent with controlling these bioactivities and signaling pathways. To test this binding hypothesis, we developed a bacterial motility-enabled binding assay that uses the interfacial properties of hydrated gels and bacterial motility to conduct label-free ligand-receptor binding studies. The structure-activity correlation and receptor identification reveal a plausible mechanism for reverting mucoid to nonmucoid phenotypes by binding pili appendages with ligands capable of sequestering and neutralizing reactive oxygen species.
Asunto(s)
Alginatos/metabolismo , GMP Cíclico/análogos & derivados , Proteínas Fimbrias/antagonistas & inhibidores , Pseudomonas aeruginosa/efectos de los fármacos , Sitios de Unión/efectos de los fármacos , GMP Cíclico/química , GMP Cíclico/farmacología , Proteínas Fimbrias/metabolismo , Ligandos , Oxidación-Reducción , Fenotipo , Pseudomonas aeruginosa/metabolismo , Relación Estructura-ActividadRESUMEN
The application of lipid-based drug delivery technologies for bioavailability enhancement of drugs has led to many successful products in the market for clinical use. Recent studies on amine-containing heterolipid-based synthetic vectors for delivery of siRNA have witnessed the United States Food and Drug Administration (USFDA) approval of the first siRNA drug in the year 2018. The studies on various synthetic lipids investigated for delivery of such nucleic acid therapeutics have revealed that the surface pK a of the constructed nanoparticles plays an important role. The nanoparticles showing pK a values within the range of 6-7 have performed very well. The development of high-performing lipid vectors with structural diversity and falling within the desired surface pK a is by no means trivial and requires tedious trial and error efforts; therefore, a practical solution is called for. Herein, an attempt to is made provide a solution by predicting the statistically significant pK a through a predictive quantitative structure-activity relationship (QSAR) model. The QSAR model has been constructed using a series of 56 amine-containing heterolipids having measured pK a values as a data set and employing a partial least-squares regression coupled with stepwise (SW-PLSR) forward algorithm technique. The model was tested using statistical parameters such as r 2, q 2, and pred_r 2, and the model equation explains 97.2% (r 2 = 0.972) of the total variance in the training set and it has an internal (q 2) and an external (pred_r 2) predictive ability of â¼83 and â¼63%, respectively. The model was validated by synthesizing a series of designed heterolipids and comparing measured surface pK a values of their nanoparticle assembly using a 2-(p-toluidino)-6-napthalenesulfonic acid (TNS) assay. Predicted and measured surface pK a values of the synthesized heterolipids were in good agreement with a correlation coefficient of 93.3%, demonstrating the effectiveness of this QSAR model. Therefore, we foresee that our developed model would be useful as a tool to cut short tedious trial and error processes in designing new amine-containing heterolipid vectors for delivery of nucleic acid therapeutics, especially siRNA.
RESUMEN
BACKGROUND: Amyloid-ß42 (Aß42) is associated with plaque formation in the brain of patients with Alzheimer's disease (AD). Studies have suggested the potential utility of plasma Aß42 levels in the diagnosis, and in longitudinal study of AD pathology. Conventional ELISAs are used to measure Aß42 levels in plasma but are not sensitive enough to quantitate low levels. Although ultrasensitive assays like single molecule array or immunoprecipitation-mass spectrometry have been developed to quantitate plasma Aß42 levels, the high cost of instruments and reagents limit their use. OBJECTIVE: We hypothesized that a sensitive and cost-effective chemiluminescence (CL) immunoassay could be developed to detect low Aß42 levels in human plasma. METHODS: We developed a sandwich ELISA using high affinity rabbit monoclonal antibody specific to Aß42. The sensitivity of the assay was increased using CL substrate to quantitate low levels of Aß42 in plasma. We examined the levels in plasma from 13 AD, 25 Down syndrome (DS), and 50 elderly controls. RESULTS: The measurement range of the assay was 0.25 to 500âpg/ml. The limit of detection was 1âpg/ml. All AD, DS, and 45 of 50 control plasma showed measurable Aß42 levels. CONCLUSION: This assay detects low levels of Aß42 in plasma and does not need any expensive equipment or reagents. It offers a preferred alternative to ultrasensitive assays. Since the antibodies, peptide, and substrate are commercially available, the assay is well suited for academic or diagnostic laboratories, and has a potential for the diagnosis of AD or in clinical trials.
Asunto(s)
Enfermedad de Alzheimer/sangre , Péptidos beta-Amiloides/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Mediciones Luminiscentes/métodos , Fragmentos de Péptidos/sangre , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Análisis Costo-Beneficio , Síndrome de Down/sangre , Ensayo de Inmunoadsorción Enzimática/economía , Femenino , Humanos , Mediciones Luminiscentes/economía , Masculino , Persona de Mediana Edad , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: The nucleotide excision repair pathway is a sophisticated DNA repair mechanism that reduces DNA damage caused by exogenous factors. Excision repair cross complementation group 1 (ERCC1) is a prominent member of this pathway that maintains the genomic stability. The aim of this study is to determine the association between the immunohistochemical expression of ERCC1 and the clinical and pathological features of oral cavity squamous cell carcinomas. MATERIALS AND METHODS: The sections of formalin fixed paraffin embedded tissue blocks of oral squamous cell carcinomas (n = 60) were immunohistochemically stained with anti-ERCC1 antibody. The association between the nuclear expression of ERCC1 and the clinicopathological parameters of the tumors and the patient outcomes was evaluated using the chi-square test. RESULTS: ERCC1 expression was evident in all studied cases of the oral squamous cell carcinomas. A high ERCC1 expression was associated with smaller tumors, tumors without lymph node involvement and well-differentiated tumors (p < 0.001). Better outcomes were associated with higher expression of ERCC1 (p = 0.028). CONCLUSION: ERCC1 seems to be an efficient biomarker for prognostication of oral squamous cell carcinomas. High expression of ERCC1 indicates more favorable course of the disease.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Endonucleasas/metabolismo , Neoplasias de la Boca/metabolismo , Neoplasias de la Boca/patología , Carcinoma de Células Escamosas de Cabeza y Cuello/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/patología , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/metabolismo , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Nicotiana , Adulto JovenRESUMEN
Synthesis of concave and vaulted 2H-pyran-fused BINOLs has been achieved. A regioselective, path-breaking concerted cascade route allows the placement of six-membered heteroaromatic rings at the sterically crowded 7,8 and 7',8' positions of BINOL. DFT studies with relative energetics that support the kinetically controlled reaction pathway are preferred, matching the experimental results. The new BINOLs exhibit smaller dihedral angle than BINOL on the diol part; this structural feature can be an assisting factor for better ligation with metals in the metal-catalyzed reactions. Corresponding C2 symmetric [5] and [7]-oxa-helicenoids have an overlapping, sterically crowded geometry.
Asunto(s)
Infecciones por Coronavirus/veterinaria , Virus de la Bronquitis Infecciosa/aislamiento & purificación , Enfermedades de las Aves de Corral/virología , Animales , Pollos , Infecciones por Coronavirus/epidemiología , Infecciones por Coronavirus/virología , India/epidemiología , Enfermedades de las Aves de Corral/epidemiologíaRESUMEN
Antibiotics are known to promote bacterial formation of enhanced biofilms, the mechanism of which is not well understood. Here, using biolayer interferometry, we have shown that bacterial cultures containing antibiotics that target cell walls cause biomass deposition on surfaces over time with a linear profile rather than the Langmuir-like profiles exhibited by bacterial adherence in the absence of antibiotics. We observed about three times the initial rate and 12 times the final biomass deposition on surfaces for cultures containing carbenicillin than without. Unexpectedly, in the presence of antibiotics, the rate of biomass deposition inversely correlated with bacterial densities from different stages of a culture. Detailed studies revealed that carbenicillin caused faster growth of filaments that were seeded on surfaces from young bacteria (from lag phase) than those from high-density fast-growing bacteria, with rates of filament elongation of about 0.58 and 0.13â µm min-1 , respectively. With surfaces that do not support bacterial adherence, few filaments were observed even in solution. These filaments aggregated in solution and formed increased amounts of biofilms on surfaces. These results reveal the lifestyle of antibiotic-induced filamentous bacteria, as well as one way in which the antibiotics promote biofilm formation.
Asunto(s)
Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Carbenicilina/farmacología , Pared Celular/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Pseudomonas aeruginosa/efectos de los fármacos , Adhesión Bacteriana/efectos de los fármacos , Escherichia coli/citología , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa/citología , Propiedades de SuperficieRESUMEN
25-Hydroxyvitamin D (25-(OH)D) deficiency is recently been described as one of the multiple factors responsible for pediatric seizures. 25-Hydroxyvitamin D3 and 25-Hydroxyvitamin D2 are the well-known markers to determine Vitamin D status. In this work we report the development of a sensitive and cost effective HPLC technique for the quantification of the vitamin D metabolites from dried blood spot samples (DBS). The metabolites were extracted using acetonitrile-methanol-0.1% formic acid (60 : 20 : 20 (v/v)) and analyzed on an Acclaim C18 column (150 × 4.6 mm i.d., 3 µm) at a flow rate of 1 mL/min. The method was linear in the range of 10-80 ng/mL. Limit of detection and limit of quantification (LOQ) of the method were 5 and 10 ng/mL respectively. Extensive stability studies demonstrated the analytes to be stable in stock and matrix with a percent change within the acceptable range of ±15%. Comparison of the newly developed HPLC-DBS method with the reported LC-MS-DBS and electrochemiluminescence immunoassay (ECLIA) methods followed by Bland-Altman analysis demonstrated a bias of 0.08 and -0.14, respectively proving the methods are comparable. Application of the developed method to a pediatric seizure cohort depicted 46.6% of cases as deficient and 26.6% as insufficient for 25-(OH)D. Among deficient cases 8 samples were below 10 ng/mL and exact amount was not calculated since these were below the LOQ levels. The mean ± standard deviation (S.D.) in the remaining 6 deficient cases was 13.22 ± 2.80 ng/mL. The levels in healthy infants were 33.9 ± 6.11 ng/mL. The method can be used routinely for assessing 25-(OH)D deficiency in newborn.