Potential impact of rotavirus vaccination on reduction of childhood diarrheal disease in India: An analysis of National Family Health Survey-5.

Rotavirus is one of the leading causes of diarrhea in infants and young children worldwide. In this study, we investigated the impact of rotavirus vaccination on the prevalence of diarrheal disease among children under five years of age in India. Research on the impact of the rotavirus vaccine on reducing diarrheal disease is therefore important in contributing to the growing body of evidence on the effectiveness of this intervention in improving child health outcomes. We adopted multivariate logistic regression and propensity score matching analysis to examine the association between diarrhea and the rotavirus vaccine. The bivariate analysis finding shows that the prevalence of diarrhea was remarkably higher (9.1%) among children who had not received rotavirus and the prevalence was 7.5%, 7.5%, and 7.2% among children who received one dose, two doses, and three rotavirus doses (all) respectively. The result of multivariate logistic regression shows that children who received all three doses of the rotavirus vaccine were 16% less likely to experience diarrhea compared to those who did not receive any rotavirus vaccine. Our analysis also found that the prevalence of diarrhea decreased significantly in the years following the introduction of the vaccine. The results of this study suggest that the rotavirus vaccine has a significant impact on reducing childhood diarrheal disease in India. These results have the potential to inform policy decisions and enable healthcare professionals to concert their efforts in reducing the diarrheal disease burden and its timely prevention in children. The study will also contribute to the existing literature on the impact of rotavirus vaccination in reducing the prevalence of diarrhea among children in India.

Exploring the Pattern of Immunization Dropout among Children in India: A District-Level Comparative Analysis.

The dropout rate is one of the determinants of immunization coverage and program performance, program continuity, and follow-up. The dropout rate refers to the proportion of vaccine recipients who did not finish their vaccination schedules, and it is determined by comparing the number of infants who started the schedule to the number who completed it. It is the rate difference between the first and final dosage or the rate difference between the first vaccination and the last vaccine dropout; thus, it denotes that the first recommended dose of vaccine was received, but that the subsequently recommended dose was missed. In India, immunization coverage has shown significant improvements over the last two decades, but full immunization coverage has remained stagnant at 76.5%, of which 19.9% are partially immunized, and 3.6% are children who have been left out. In India, the Universal Immunization Programme (UIP) is challenged with cases related to dropout in immunization. Although immunization coverage in India is improving, the program is challenged by vaccination dropouts. This study provides an analysis of the determinants of vaccination dropout in India using data from two rounds of the National Family Health Survey. The finding shows that the mother's age, education, family wealth, antenatal care visit, and place of delivery were some of the variables that significantly contributed to reducing the dropout rate of immunization among children. The findings of this paper show that the dropout rate has reduced over a certain period of time. The overall improvement in the rates of dropout and increase in full immunization coverage could be attributed to various policy measures taken in the last decade in India, which brought structural changes with a positive impact on full immunization coverage and its components.

Postprandial insulin and triglycerides after different breakfast meal challenges: use of finger stick capillary dried blood spots to study postprandial dysmetabolism.

BACKGROUND: High levels of insulin and lipids following a meal are recognized risk factors for atherosclerosis. Monitoring such risk factors in the general population is hampered by the inconvenience of venipuncture blood collection, particularly for both premeal and postmeal analyses. This study examined insulin and triglyceride levels in dried blood spots (DBSs) collected after different breakfast meal challenges to assess the potential of this method for risk assessment. METHODS: Glucose levels were measured using a glucose meter, and insulin and triglycerides were determined in DBS samples collected from 19 healthy volunteers before and at four time points up to 2.5 h after consuming each of five typical breakfast meals varying in nutritional composition. RESULTS: At 2 h, glucose was within normal postprandial values (<140 mg/dl) for all meals; significantly lower glucose was seen after meal 2 (the lowest carbohydrate content) compared to the other meals. Insulin returned to normal fasting levels (<15 microIU/ml) in significantly more subjects (90%) after meal 2 and significantly fewer subjects (31%) after meal 4 (highest carbohydrate content) than the other meals. Triglycerides were elevated to a similar extent in all subjects, with no significant differences between meals; levels were still rising at 2.5 h. CONCLUSIONS: Subjects were able to collect blood spots with minimum disruption to their normal daily activities. Relative ease of collection, analyte stability in dried blood, and the close correlation with serum levels that we have previously demonstrated makes DBS a convenient and simple tool for assessing the individual impact of different diets on postprandial dysmetabolism.

A liquid chromatography/tandem mass spectrometry method for determination of 25-hydroxy vitamin D2 and 25-hydroxy vitamin D3 in dried blood spots: a potential adjunct to diabetes and cardiometabolic risk screening.

BACKGROUND: Now emerging as an important risk factor for type 1 diabetes, vitamin D deficiency is also associated with obesity, metabolic syndrome, and type 2 diabetes and has been identified as a potential cardiometabolic risk factor. A simple, accurate screening test for 25-hydroxy vitamin D [25(OH)D] deficiency is needed. We developed a liquid chromatography/tandem mass spectrometry assay for 25-hydroxy vitamin D(2) [25(OH)D(2)] and 25-hydroxy vitamin D(3) [25(OH)D(3)] in dried blood spots. METHOD: Blood spots were collected by finger stick simultaneously with serum samples obtained by venipuncture from healthy volunteers. Disks punched from the dried blood spots were sonicated with an internal standard solution of deuterated 25(OH)D(3) (26,26,26,27,27,27-d(6)). Methanol was added to precipitate proteins prior to extraction with hexane. The extracted samples were dried and reconstituted in 50:50 methanol:H(2)O before injection into a Varian 320-MS TQ mass spectrometer. RESULTS: BLOOD SPOT ASSAY PRECISION WAS GOOD OVER THE REPORTABLE RANGE: interassay coefficients of variation were 13, 13, and 11% at concentrations of 14, 26, and 81 ng/ml, respectively, for 25-hydroxy vitamin D(3) and 12% at 23 ng/ml for 25(OH)D(2). The 25(OH)D(3) assay was linear from 3.5 to 75 ng/ml (R > 0.99). Blood spot and serum values showed excellent correlation for 25(OH)D(2) (R=0.90, n=54) and 25(OH)D(3) (R=0.91, n=83). CONCLUSIONS: This blood spot assay for 25(OH)D(2) and 25(OH)D(3) provides a convenient and cost-effective alternative to serum assays and can be automated. This may be valuable in large-scale screening for risk of type 1 diabetes, for cardiometabolic risk screening, and for monitoring vitamin D supplementation.

Cardiometabolic risk factors assessed by a finger stick dried blood spot method.

BACKGROUND: Cardiovascular and metabolic risk factors are gaining attention as potential indicators for intervention in the prevention and treatment of cardiovascular disease (CVD) and type 2 diabetes mellitus (T2DM). Blood spot technology offers an efficient, convenient method to measure these risk factors in an at-risk population. Simple and convenient methods to assess cardiometabolic risk factors will allow clinicians to formulate treatment strategies for effective prevention and management of these conditions. METHOD: Insulin and high sensitivity C-reactive protein (hs-CRP) were measured in dried blood spot and corresponding serum samples using conventional commercial serum kits based on a direct sandwich ELISA technique. The triglyceride assay involved enzymatic hydrolysis of triglycerides by lipase to glycerol and free fatty acids. The glycerol produced was then measured by coupled enzyme reactions. Blood spot assays were modified from previously published methods to give improved accuracy and turnaround time. RESULTS: Blood spot levels correlated well with serum levels of hs-CRP (r = 0.99), triglycerides (fasting, r = 0.95; nonfasting, r = 0.94), and insulin (fasting, r = 0.93; nonfasting, r = 0.97). CONCLUSION: Blood spot testing for insulin, hs-CRP, and triglycerides may be helpful in the cardiometabolic screening or monitoring of patients at risk of developing CVD, T2DM, or metabolic syndrome.

Cellular apoptosis and cytotoxicity of phenolic compounds: a quantitative structure-activity relationship study.

In this comprehensive study on the caspase-mediated apoptosis-inducing effect of 51 substituted phenols in a murine leukemia cell line (L1210), we determined the concentrations needed to induce caspase activity by 50% (I50) and utilized these data to develop the following quantitative structure-activity relationship (QSAR) model: log 1/I50 = 1.06 B5(2) + 0.33 B5(3) - 0.18pi(2,4) - 0.92. B5(3) and B5(2) represent steric terms, while pi(2,4) represents the hydrophobic character of the substituents on the ring. The strong dependence of caspase-mediated apoptosis on mostly steric parameters suggests that the process is a receptor-mediated interaction with caspases or mitochondrial proteins being the likely targets. Conversely, cytotoxicity studies of 65 electron-releasing phenols in the L1210 cell line led to the development of the following equation: log 1/ID50 = -1.39sigma+ - 0.28 B5(2,6) + 0.16 log P - 0.58I(2) - 1.04I(1) + 3.90. The low coefficient with log P may pertain to cellular transport that may be enhanced by a modest increase in overall hydrophobicity, while the presence of sigma+ is consistent with the suggestion that radical stabilization is of prime importance in the case of electron-releasing substituents. On the other hand, the QSAR for the interactions of 27 electron-attracting phenols in L1210 cells, log 1/ID50 = 0.56 log P - 0.30 B5(2) + 2.79, suggests that hydrophobicity, as represented by log P is of critical importance. Similar cytotoxicity patterns are observed in other mammalian cell lines such as HL-60, MCF-7, CCRF-CEM, and CEM/VLB. The significant differences between the cytotoxicity and apoptosis QSAR for electron-releasing phenols suggest that cytotoxicity involves minimal apoptosis in most of these substituted monophenols.

Identification of novel genetic loci for bone size and mechanosensitivity in an ENU mutant exhibiting decreased bone size.

UNLABELLED: Using a dominant ENU mutagenesis screen in C57BL/6J (B6) mice to reveal gene function, we identified a mutant, 917M, with a reduced bone size phenotype, which is expressed only in males. We show that mutation results in osteoblasts with reduced proliferation, increased apoptosis, and an impaired response to in vitro mechanical load. The mutation is mapped to a novel locus (LOD score of 7.9 at 10.5 cM) on chromosome 4. INTRODUCTION: Using a dominant ENU mutagenesis screen in C57BL/6J (B6) mice to reveal gene function, we identified a mutant, 917M, with a reduced bone size phenotype, which is expressed only in males. In this report, we show the chromosomal location of this mutation using linkage analysis and cellular characterization of the mutant phenotype. MATERIALS AND METHODS: The mutant mouse was bred to wildtype B6 to produce progeny for characterization of the bone size phenotype. Periosteal osteoblasts isolated from the tibia and femur of mutant and wildtype mice were studied for proliferation, differentiation, and apoptosis potential. To determine the chromosomal location of the mutation, a low-resolution linkage map was established by completing a genome-wide scan in B6C3H F2 male mice generated from intercross breeding of mutant mice. RESULTS AND CONCLUSIONS: Mutant progeny (16 weeks old) displayed a total body bone area that was 10-13% lower and a periosteal circumference that was 5-8% lower at the femur and tibia midshaft compared with wildtype B6 mice. Periosteal osteoblasts from mutant mice showed 17-27% reduced cell proliferation and 23% increased apoptosis compared with wildtype controls. In addition, osteoblasts from mutant mice showed an impaired response to shear stress-induced proliferation rate, an in vitro model for mechanical loading. Interval mapping in B6C3H F2 males (n = 69) indicated two major loci affecting bone size on chromosome 1 at 45 cM (LOD 4.9) and chromosome 4 at 10.5 cM (LOD 7.9, genome-wide p < 0.01). Interval mapping using body weight as covariate revealed only one significant interval at chromosome 4 (LOD 6.8). Alleles of the chromosome 4 interval inherited from the B6 mutant strain contributed to a significantly lower bone size than those inherited from C3H. A pairwise interaction analysis showed evidence for a significant interaction between loci on chromosome 1 with the chromosome 4 quantitative trait loci. The 917M locus on chromosome 4 seems to be novel because it does not correspond with those loci previously associated with bone size on chromosome 4 in B6 and C3H/HeJ mice or other crosses.

Covalent interaction between proform of eosinophil major basic protein (proMBP) and pregnancy-associated plasma protein-A (PAPP-A) is a cell-mediated event and required for proMBP inhibition of the catalytic activity of PAPP-A.

This study was undertaken to determine the mechanism by which proform of eosinophil major basic protein (proMBP) inhibits the IGFBP-4 proteolytic activity of pregnancy-associated plasma protein (PAPP)-A. Co-overexpression of PAPP-A with proMBP in 293T cells, or co-incubation of 293T cells, respectively, overexpressing proMBP and PAPP-A resulted in the formation of a covalent proMBP-PAPP-A complex and inhibition of IGFBP-4 proteolysis. Similar results were obtained when recombinant proMBP and PAPP-A were incubated in the presence of U2 osteosarcoma cells or when recombinant proMBP was added to the U2 cells overexpressing PAPP-A. In contrast, no formation of covalent proMBP-PAPP-A complex or inhibition of IGFBP-4 proteolysis was observed when recombinant proMBP and PAPP-A were incubated under cell-free conditions, although proMBP was able to interact with PAPP-A in a non-covalent manner. These new findings suggest that formation of covalent proMBP-PAPP-A complex is a cell-mediated event and is required for proMBP to inhibit the catalytic activity of PAPP-A.

Studies on regulation of IGF (insulin-like growth factor)-binding protein (IGFBP) 4 proteolysis by pregnancy-associated plasma protein-A (PAPP-A) in cells treated with phorbol ester.

PAPP-A (pregnancy-associated plasma protein-A) is produced by hSFs (human skin fibroblasts) and hOBs (human osteoblasts) and enhances the mitogenic activity of IGFs (insulin-like growth factors) by degradation of IGFBP-4 (insulin-like growth factor-binding protein 4). PKC (protein kinase C) activation in these cells led to reduction in IGFBP-4 proteolysis. This study was undertaken to determine the mechanism by which activation of PKC suppresses IGFBP-4 proteolysis. Treatment of hSFs/hOBs with TPA (PMA; 100 nM) reduced IGFBP-4 proteolysis without significantly decreasing the PAPP-A level in the CM (conditioned medium). Immunodepletion of the proform of eosinophil major basic protein (proMBP), a known PAPP-A inhibitor, from CM of TPA-treated cells (TPA CM) failed to increase IGFBP-4 proteolytic activity. Transduction of hSFs with proMBP retrovirus increased the concentration of proMBP up to 30 ng/ml and led to a moderate reduction in IGFBP-4 proteolysis. In contrast, TPA treatment blocked IGFBP-4 proteolysis but failed to induce a detectable amount of proMBP in the CM. While proMBP overexpression led to the formation of a covalent proMBP-PAPP-A complex and reduced the migration of PAPP-A on SDS/PAGE, TPA treatment dose- and time-dependently increased the conversion of a approximately 470 kDa PAPP-A form (PAPP-A470) to a approximately 400 kDa PAPP-A form (PAPP-A400). Since unreduced PAPP-A400 co-migrated with the 400 kDa recombinant PAPP-A homodimer and since PAPP-A monomers from reduced PAPP-A470 and PAPP-A400 co-migrated on SDS/PAGE, conversion of PAPP-A470 to PAPP-A400 is unlikely to be caused by proteolytic cleavage of PAPP-A. Consistent with the data showing that the increase in the ratio of PAPP-A400/PAPP-A470 is correlated with the extent of reduction in IGFBP-4 proteolysis, partially purified PAPP-A400 exhibited a 4-fold reduction in IGFBP-4 proteolytic activity compared with PAPP-A470. These data suggest that a novel mechanism, namely conversion of PAPP-A470 to the less-active PAPP-A400, could account for the TPA-induced suppression of PAPP-A activity.

Synthesis, cytotoxicity, and QSAR analysis of X-thiophenols in rapidly dividing cells.

In this study, the cytotoxicities of a series of X-thiophenols vs rapidly growing mouse leukemia cells in vitro are determined. The resulting ID(50) values are then used to formulate a quantitative structure-activity relationship, which is well-correlated by the Brown variation of the Hammett electronic parameter, sigma-plus (sigma(+) ), such that Log 1/ID(50) = -0.93 (+/-0.18) sigma(+) + 0.86 (+/-0.24) I(H) + 3.99 (+/-0.13). I(H) represents an indicator variable that calls attention to the unusual activity of halogens and pseudohalogens. In lieu of sigma(+), homolytic bond dissociation energies (BDE) are also used successfully to correlate the cellular cytotoxicities of thiophenols. The nature of substituent effects on cellular toxicity is examined, and they reveal that electron-releasing substituted thiophenols such as 4-amino thiophenol and the 4-alkoxy thiophenols are highly cytotoxic and effective at inhibiting cellular proliferation at physiological pH. On the other hand, electron-attracting substituted thiophenols such as the 4-cyano and 4-halogen analogues show a reduced ability to inhibit the cell growth of this cell line. Thus, there is a clear parallel between enhanced biological activity and electron releasing ability as measured by sigma(+) constants or BDE values. The susceptibility of the cellular interaction to electronic effects as delineated by the coefficient with the sigma(+) term (also called the Hammett rho value) is high (-0.96), suggesting that substantial energetic assistance is provided by the substituents and that a weak initiating radical reactant such as superoxide radical may be involved. Previous cytotoxicity studies of a large diverse data set of X-phenols in this cell line and embryo cells have also revealed a more pronounced dependence on sigma(+) and deltaBDE. A comparison of reaction constants obtained from thiophenoxy radical formation reactions and phenoxy radical formation reactions in organic media suggests radical-mediated involvement in cell cytotoxicity. Such cells could be more vulnerable to the effects of reactive thiyl species on their metabolism and subsequent proliferation.