An efficient process for the extraction of lutein and chemical characterization of other organic volatiles from marigold (Tagetes erecta L.) flower.

Marigold (Tagetes erecta L.) petals are the primary industrial source of lutein, which is used as a colouring agent and nutrient supplement to foods. This research extracted marigold petals using different solvents, covering conventional and non-toxic green solvents. The oleoresin, free lutein, and recrystallized lutein yields varied from 8.47-16.67%, 2.56-9.62%, and 1.11-1.61%, respectively. The purity of lutein was achieved up to 92.57% and 97.64% in conventional and newly established green methods, respectively. The present study described an efficient green process to isolate lutein with significantly improved yield (2.56%) and purity (97.33%) over the conventional methods. Based on the results, 2-methyltetrahydrofurancould be a practical green alternative to the traditional toxic solvents for the processing of lutein. Further, the chemical analysis of the essential oil of the residual receptacles obtained after removing petals revealed the presence of important organic volatiles, including piperitone (54.7%) and piperitenone oxide (6.5%), indicating its usefulness for value-addition.

Vitamin D supplementation in childhood asthma: a systematic review and meta-analysis of randomised controlled trials.

BACKGROUND: There is conflicting evidence for vitamin D supplementation in childhood asthma. We aimed to systematically synthesise the evidence on the efficacy and safety of vitamin D supplementation in childhood asthma. METHODS: We searched electronic databases (Medline, Embase and Web of Science) and a register (CENTRAL) for randomised controlled trials (RCTs) published until 30 September 2021. RCTs enrolling asthmatic children (1-18 years old) and comparing vitamin D against placebo/routine care were included if they met at least one of the endpoints of interest (asthma attacks, emergency visits or hospitalisation). We used the Risk of Bias 2 tool for risk of bias assessment. Random-effects meta-analysis with RevMan 5.3 software was performed. The Grading of Recommendations Assessment, Development and Evaluation approach was used to assess the level of certainty of the evidence. RESULTS: 18 RCTs (1579 participants) were included. The pooled meta-analysis did not find a significant effect of vitamin D supplementation on asthma attacks requiring rescue systemic corticosteroids (six studies with 445 participants; risk ratio (RR) 1.13, 95% CI 0.86-1.48; I2=0%) (moderate-certainty evidence). In addition, there was no significant difference in the proportion of children with asthma attacks of any severity (11 trials with 1132 participants; RR 0.84, 95% CI 0.65-1.09; I2=58%) (very low-certainty evidence). Vitamin D does not reduce the need for emergency visits (three studies with 361 participants; RR 0.97, 95% CI 0.89-1.07; I2=0%) and hospitalisation (RR: 1.38, 95% CI 0.52-3.66; I2=0%) (low-certainty evidence). CONCLUSION: Very low- to moderate-certainty evidence suggests that vitamin D supplementation might not have any protective effect in childhood asthma.

The Induction of Alpha-1 Antitrypsin by Vitamin D in Human T Cells Is TGF-ß Dependent: A Proposed Anti-inflammatory Role in Airway Disease.

Background: Vitamin D upregulates anti-inflammatory and antimicrobial pathways that promote respiratory health. Vitamin D synthesis is initiated following skin exposure to sunlight, however nutritional supplementation can be required to address deficiency, for example during the winter months or due to cultural constraints. We recently reported that 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) treatment induced alpha-1 antitrypsin (AAT) expression in CD4+, but not CD8+ T cells, with evidence supporting an immunoregulatory role. Research Question: To understand the relationship between vitamin D, lung AAT levels and T lymphocytes further we investigated whether TGF-ß is required as a co-factor for 1,25(OH)2D3-induced upregulation of AAT by vitamin D in CD8+ T cells in vitro and correlated circulating vitamin D levels with lung AAT levels in vivo. Results: 1,25(OH)2D3 in combination with TGF-ß1 increased AAT expression by CD8+ T cells, as well as VDR and RXRα gene expression, which may partly explain the requirement for TGF-ß. CD4+ T cells may also require autocrine stimulation with TGF-ß as a co-factor since 1,25(OH)2D3 was associated with increased TGF-ß bioactivity and neutralisation of TGF-ß partially abrogated 1,25(OH)2D3-induced SERPINA1 gene expression. Neither CD4+ nor CD8+ T cells responded to the circulating vitamin D precursor, 25-hydroxyvitamin D3 for induction of SERPINA1, suggesting that local generation of 1,25(OH)2D3 is required. Transcriptional gene profiling studies previously demonstrated that human bronchial epithelial cells rapidly increased TGF-ß2 gene expression in response to 1,25(OH)2D3. Here, human epithelial cells responded to precursor 25(OH)D3 to increase bioactive TGF-ß synthesis. CD8+ T cells responded comparably to TGF-ß1 and TGF-ß2 to increase 1,25(OH)2D3-induced AAT. However, CD8+ T cells from adults with AAT-deficiency, homozygous for the Z allele of SERPINA1, were unable to mount this response. AAT levels in the airways of children with asthma and controls correlated with circulating 25(OH)D3. Conclusions: Vitamin D increases AAT expression in human T cells and this response is impaired in T cells from individuals homozygous for the Z allele of SERPINA1 in a clinic population. Furthermore, a correlation between circulating vitamin D and airway AAT is reported. We propose that vitamin D-induced AAT contributes to local immunomodulation and airway health effects previously attributed to vitamin D.

Factors leading to neonatal mortality due to neonatal purpura fulminans through the lens of a child death review.

A case of neonatal death due to neonatal purpura fulminans (NPF) was brought to community physicians' notice by the auxiliary nurse midwife in her catchment area as part of the routine demographic health surveillance. The community physician then conducted the child death review in the community. The neonate was born out of consanguineous marriage (mother married to her first-degree maternal cousin) with spontaneous conception. This neonate was fourth in the birth order. The second-order and third-order births had also suffered from NPF and died. The baby was delivered in a tertiary care setting, and the paediatric surgeon planned debridement of the affected part on the third day of the birth, as per the mother. However, due to inadequate counselling regarding the procedure, mother left the hospital without seeking care against medical advice, and the child died at home.

Vitamin-D supplementation as an adjunct to standard treatment of asthma in children: A randomized controlled trial (ViDASTA Trial).

OBJECTIVE: To determine the role of vitamin D supplementation as an adjunct to standard treatment in childhood asthma. STUDY DESIGN: In this placebo-controlled, blinded, randomized controlled trial, we enrolled 60 children aged 6 to 11 years with moderate persistent asthma and randomly assigned them into intervention (2000 IU per day of vitamin D) and placebo groups (n = 30 each). The primary outcome was asthma control as assessed by the childhood asthma control test (C-ACT) scores at 12 weeks post-randomization. The secondary outcomes were improvement in the forced expiration in 1 s (FEV1 ), fractional exhaled nitric oxide (FeNO), asthma exacerbations, use of systemic steroids, number of emergency visits, post-intervention vitamin D levels, and adverse outcomes. We analyzed by intention to treat. RESULTS: There was no significant difference between the C-ACT score in the two groups (median [first-third quartile] scores were 25 [24-26] in both groups, p = 0.7). Also, there was no significant difference between the two groups in terms of the FEV1 , FeNO, number of exacerbations, emergency visits, hospital admissions, and adverse outcomes. However, the post-intervention vitamin D levels (ng/ml) were significantly higher in the intervention group (35.5 vs. 18.8; p < 0.001). As compared to the baseline, both the groups showed better asthma control at 12 weeks post-intervention, irrespective of the type of intervention. CONCLUSION: Vitamin-D supplementation as an adjunct to standard treatment does not improve asthma control in children.

Ten-year use of recombinant parathyroid hormone for the treatment of hypoparathyroidism in a boy with partial Jacobsen syndrome.

Pediatric hypoparathyroidism (HPT) is caused by inherited or acquired defects involving the synthesis or secretion of PTH, resistance to PTH action, or inappropriate regulation of PTH. Several syndromes such as DiGeorge syndrome, HDR (hypoparathyroidism, sensorineural deafness and renal dysplasia) syndrome, HRD (hypoparathyroidism, retardation, and dysmorphism) syndrome, Kenny-Caffey syndrome etc. may have associated HPT. In the present communication, we describe, the hitherto unreported, occurrence of HPT in a child with partial Jacobsen syndrome. Chromosomal Microarray analysis showed a heterozygous deletion of 4.7 Mb at cytoband 11q24.3q25 encompassing approximately 20 genes including JAM3 and NTM genes. The child was treated with recombinant human parathyroid hormone (rhPTH1-34) for 10 years. Throughout follow up, he required several adjustments in dosages of rhPTH1-34 and oral calcium to maintain serum calcium concentrations in low normal ranges. The bone turnover markers remained normal and oral calcium supplements were completely taken off after 8 years. In conclusion, our single-case experience indicates that long-term therapy of chronic HPT with rhPTH1-34 is safe and reduces the need for additional therapies.

Complementary Proteomics, Genomics approaches identifies potential pathogenicity/virulence factors in Tilletia indica induced under the influence of host factor.

Karnal bunt disease of wheat is incited by quarantine fungal pathogen T. indica. Till date, there is little information on the pathogenic mechanisms involved in Karnal bunt. In order to understand the molecular mechanisms of disease pathogenesis, highly aggressive T. indica TiK isolate was cultured in the presence of host factor extracted from developing spikes of wheat variety WH-542. Modulation in protein profile of mycelial proteins and secretome from TiK cultured in the absence and presence of host factor was analyzed by 2-DE. Fifteen and twenty nine protein spots were up-regulated/differentially regulated in the proteome of mycelial and secreted proteins, respectively and identified using MALDI-TOF/TOF. Identified proteins are involved in suppression of host defense responses, lignin degradation of plant cell wall, penetration, adhesion of pathogen to host tissues, pathogen mediated reactive oxygen species generation, hydrolytic enzymes, detoxification of host generated reactive oxygen species. Further, integration of proteomic and genomic analysis has led to candidate pathogenicity/virulence factors identification. They were functionally annotated by sequence as well as structure based analysis. In this study, complementation of proteomics and genomics approaches resulted in novel pathogenicity/virulence factor(s) identification in T. indica.

Combined potentiating action of phytochemical(s) from Cinnamomum tamala and Aloe vera for their anti-diabetic and insulinomimetic effect using in vivo rat and in vitro NIH/3T3 cell culture system.

The present investigation was undertaken to analyze the ethanolic extracts of leaves of Cinnamomum tamala and Aloe vera for their anti-diabetic and insulinomimitic effect by determining the levels of blood sugar, glycosylated hemoglobin, and serum lipid profile (total cholesterol, triglycerides, high density lipoprotein (HDL), and low density lipoprotein (LDL)) after daily administration of each alone and in combined at 250 mg/kg in alloxan (ALX)-induced diabetic rats. Treatment of diabetic rats with the extracts restored the elevated biochemical parameters significantly. The anti-diabetic effect further potentiated the insulin signaling pathway by co-administration of both extracts. The molecular mechanisms of modulating gene expression and cellular signaling through the insulin receptor were also evaluated on specific targets of the insulin signaling pathway, including insulin receptor substrate (IRS), phosphatidylinositol 3-kinase (PI3-K), AKT, and the glucose transporter (GLUT4) on NIH/3T3 cell line by western blotting, ELISA, semiquantitative RT-PCR, and real-time PCR. The active principle of both extracts revealed insulin mimicking effect as indicated by increased expression of pIRS1 and pAKT in time-dependent manner. There was no significant difference in PI3-K content between unchallenged and challenged groups. Enhanced expression of GLUT-4 transcript further suggested that the Cinnamomum and Aloe phytochemicals could serve as a good adjuvant in the present armamentarium of anti-diabetic drugs by either mimicking or improving insulin action. This study reveals that ethanolic extracts of C. tamala and A. vera have potent therapeutic efficacy and prospect for the development of phytomedicine for diabetes mellitus.

Combating malaria with plant molecules: a brief update.

Malaria is one of the deadliest diseases on the planet affecting about 50% of the population worldwide. It is a leading cause of morbidity and mortality in the developing world. Plasmodium falciparum, a tiny parasite is the major cause of malaria and is possibly the most dangerous stow-away in history. Malaria has become a major economic concern to some of the tropical and sub-tropical countries. Though a number of antimalarials have been developed from plants as such or their semi-synthetic analogues, there is again an alarming situation of drug resistance against most of the antimalarial drugs. Plants have been an excellent source of antimalarial compounds. There are several plant leads exhibiting antimalarial activity better than the existing drugs. A systematic evaluation of these plant based leads is the need of the time to develop safe, effective and affordable new antimalarials. The present review is an update of plant based antimalarial agents.

Direct analysis in real time by mass spectrometric technique for determining the variation in metabolite profiles of Cinnamomum tamala Nees and Eberm genotypes.

Cinnamomum tamala Nees & Eberm. is an important traditional medicinal plant, mentioned in various ancient literatures such as Ayurveda. Several of its medicinal properties have recently been proved. To characterize diversity in terms of metabolite profiles of Cinnamomum tamala Nees and Eberm genotypes, a newly emerging mass spectral ionization technique direct time in real time (DART) is very helpful. The DART ion source has been used to analyze an extremely wide range of phytochemicals present in leaves of Cinnamomum tamala. Ten genotypes were assessed for the presence of different phytochemicals. Phytochemical analysis showed the presence of mainly terpenes and phenols. These constituents vary in the different genotypes of Cinnamomum tamala. Principal component analysis has also been employed to analyze the DART data of these Cinnamomum genotypes. The result shows that the genotype of Cinnamomum tamala could be differentiated using DART MS data. The active components present in Cinnamomum tamala may be contributing significantly to high amount of antioxidant property of leaves and, in turn, conditional effects for diabetic patients.

Induction of MAP kinase homologues during growth and morphogenetic development of Karnal bunt (Tilletia indica) under the influence of host factor(s) from wheat spikes.

Signaling pathways that activate different mitogen-activated protein kinases (MAPKs) in response to certain environmental conditions, play important role in mating type switching (Fus3) and pathogenicity (Pmk1) in many fungi. In order to determine the roles of such regulatory genes in Tilletia indica, the causal pathogen of Karnal bunt (KB) of wheat, semi-quantitative and quantitative RT-PCR was carried out to isolate and determine the expression of MAP kinase homologues during fungal growth and development under in vitro culture. Maximum expression of TiFus3 and TiPmk1 genes were observed at 14th and 21st days of culture and decreased thereafter. To investigate whether the fungus alters the expression levels of same kinases upon interaction with plants, cultures were treated with 1% of host factors (extracted from S-2 stage of wheat spikes). Such treatment induced the expression of MAPks in time dependent manner compared to the absence of host factors. These results suggest that host factor(s) provide certain signal(s) which activate TiFus3 and TiPmk1 during morphogenetic development of T. indica. The results also provides a clue about the role of host factors in enhancing the disease potential due to induction of MAP kinases involved in fungal development and pathogenecity.

Relationship between serum vitamin D, disease severity, and airway remodeling in children with asthma.

RATIONALE: Little is known about vitamin D status and its effect on asthma pathophysiology in children with severe, therapy-resistant asthma (STRA). OBJECTIVES: Relationships between serum vitamin D, lung function, and pathology were investigated in pediatric STRA. METHODS: Serum 25-hydroxyvitamin D [25(OH)D(3)] was measured in 86 children (mean age, 11.7 yr): 36 with STRA, 26 with moderate asthma (MA), and 24 without asthma (control subjects). Relationships between 25(OH)D(3), the asthma control test (ACT), spirometry, corticosteroid use, and exacerbations were assessed. Twenty-two of 36 children with STRA underwent fiberoptic bronchoscopy, bronchoalveolar lavage, and endobronchial biopsy with assessment of airway inflammation and remodeling. MEASUREMENTS AND MAIN RESULTS: 25(OH)D(3) levels (median [IQR]) were significantly lower in STRA (28 [22-38] nmol/L) than in MA (42.5 [29-63] nmol/L) and control subjects (56.5 [45-67] nmol/L) (P < 0.001). There was a positive relationship between 25(OH)D(3) levels and percent predicted FEV(1) (r = 0.4, P < 0.001) and FVC (r = 0.3, P = 0.002) in all subjects. 25(OH)D(3) levels were positively associated with ACT (r = 0.6, P < 0.001), and inversely associated with exacerbations (r = -0.6, P < 0.001) and inhaled steroid dose (r = -0.39, P = 0.001) in MA and STRA. Airway smooth muscle (ASM) mass, but not epithelial shedding or reticular basement membrane thickness, was inversely related to 25(OH)D(3) levels (r = -0.6, P = 0.008). There was a positive correlation between ASM mass and bronchodilator reversibility (r = 0.6, P = 0.009) and an inverse correlation between ASM mass and ACT (r = -0.7, P < 0.001). CONCLUSIONS: Lower vitamin D levels in children with STRA were associated with increased ASM mass and worse asthma control and lung function. The link between vitamin D, airway structure, and function suggests vitamin D supplementation may be useful in pediatric STRA.

In vivo evaluation of hypoglycemic activity of Aloe spp. and identification of its mode of action on GLUT-4 gene expression in vitro.

The present study evaluated the hypoglycemic activity of Aloe extract on streptozotocin-induced diabetic mice and focuses its effect on GLUT-4 gene expression under in vitro cell-culture system. Administration of extract at the dosage of 130 mg/kg body weight per day for 4 weeks resulted in significant decrease in blood glucose and total cholesterol in streptozotocin (60 mg/kg body weight) induced diabetic mice. The hypoglycemic effect was compared with metformin. The activities of carbohydrate metabolizing enzymes were brought back to near normal level after the treatment and glucose homeostasis was maintained. Lyophilized aqueous Aloe extract (1 mg/ml) upregulated the GLUT-4 mRNA synthesis in mouse embryonic NIH/3T3 cells.

Repeatability of the low-dose ACTH test in asthmatic children on inhaled corticosteroids.

AIM: To assess the repeatability of low-dose Synacthen test (LDST) in asthmatic children receiving high-dose fluticasone propionate (FP). METHODS: Low-dose Synacthen test was performed on 18 children with stable chronic asthma treated with FP at a constant daily dose of > or =500 microg and repeated 1 month later. Repeatability was assessed using the Kappa statistic for categorical variables. RESULTS: Fifteen patients had consistent results (either two normal or two abnormal responses) and three patients had inconsistent results (one normal and one abnormal response). The Kappa statistic was 0.56 indicating fair to good agreement between the tests. CONCLUSION: The results of adrenal function testing in patients on inhaled steroids can have major implications for patient management, making it important to use a test with excellent repeatability. The LDST conducted using our protocol does not fulfil this criterion.

Randomized, double-blind, placebo-controlled clinical trial of the efficacy of treatment with zinc or vitamin A in infants and young children with severe acute lower respiratory infection.

BACKGROUND: Acute lower respiratory infection (ALRI) is a leading cause of childhood death. Zinc supplementation prevents ALRI. Vitamin A supplementation reduces childhood mortality, but its benefit concerning ALRI-specific mortality is unproven. OBJECTIVE: The objective was to evaluate the effect of zinc and vitamin A on the clinical recovery of children with severe ALRI. DESIGN: In a controlled trial with a factorial design, 153 children aged 2-24 mo who were hospitalized with severe ALRI were randomly assigned to receive 10 mg zinc as acetate (twice daily for 5 d) plus vitamin A placebo, 10 000 micro g retinol equivalents vitamin A (twice daily for 4 d) plus zinc placebo, zinc plus vitamin A, or zinc and vitamin A placebos. The main outcome variable was the time for resolution of very ill status; other outcomes were resolution of fever, tachypnea, and feeding difficulty. RESULTS: Recovery rates from very ill status and from fever in zinc-treated boys were 2.6 times (P = 0.004) and 3 times (P = 0.003) those in non-zinc-treated children; feeding difficulty and tachypnea were not significantly different between groups after an adjusted analysis. Recovery rates were not significantly different between groups on the basis of vitamin A treatment. At discharge, serum zinc was 6.06 micro mol/L higher (P = 0.001) in the zinc-treated children, and serum retinol was 0.387 micro mol/L higher (P = 0.001) in the vitamin A-treated children. CONCLUSION: Zinc treatment significantly reduces duration of fever and very ill status in boys, but not in girls, with severe ALRI. Vitamin A treatment of children with severe ALRI had no significant beneficial effect.