Effect of lead exposure and nutritional iron-deficiency on immune response: A vaccine challenge study in rats.

The prevalence of iron (Fe) deficiency and subclinical lead (Pb) toxicity is high in developing countries like India, and information on their potential additive effects on immune responses is scant. The current study assessed immune parameters in dual Pb-exposed\Fe-deficient weanling SD rats. Rats were fed a control (CD) or Fe-deficient (ID) diet for 4 weeks and then evaluated for hemoglobin (Hb) and serum Fe status. Then, half the rats in each group began to receive daily oral Pb exposure (25 mg/4 ml/kg BW; gavage) or vehicle for a further 4 weeks (while maintained on original respective diets). After the 4-weeks of dosing, rats were assessed for Hb and serum Fe, and for blood lead level (BLL) and δ-aminolevulinic acid dehydratase (ALAD) activity. At this point, half the rats in each group (now n = 8) were then vaccinated with tetanus toxoid (TT), and then two boosters at 2-week intervals. All the time, rats stayed on their original respective diets along with exposure to Pb on alternate days. At 2 weeks after the final booster, rats were euthanized and blood collected to assess total/specific IgG and IgM levels; mucosal (intestinal) IgA levels were also determined. Spleens were taken to assess CD4+ and CD8+ cell levels and for ex vivo measures of splenocyte proliferation/TH1 and TH2 cytokine formation. The results indicated significant lowering of Hb and serum Fe levels in ID rats and increased blood Pb and decreased ALAD activity in all Pb-exposed rats. Fe-deficiency alone induced significant increases in ALAD activity, but only in an absence of Pb. While there was no impact of any regimen on total or TT-specific IgG, significant decreases in mucosal IgA and TT-specific IgM were seen in ID-fed Pb-exposed rats. CD4+ cell levels were not impacted by treatment; CD8+ levels were increased in all ID/Pb-exposed rats. Ex-vivo splenocyte proliferation was significantly higher among vaccinated rats, as well as ID-fed Pb-exposed unvaccinated rats. Cytokine formation in all cases was highly variable. The results suggest that Fe deficiency compromised cell-mediated, mucosal, and/or humoral immune response-related endpoints and that Pb exposure during the deficiency further impacted these outcomes.

Sex-specific role of CYP24A1 rs2762939 in the risk of essential hypertension based on the serum vitamin D and total renin concentrations.

CYP24A1, Vitamin D 24-hydroxylase catabolizes 25-hydroxyvitamin D (25(OH)D) and 1,25-dihydroxyvitamin D to 24-hydroxylated vitamin D products. It is widely known that low Vitamin D can lead to increased renal renin and angiotensin II production, consequently elevating blood pressure or development of essential hypertension (EH). We have conducted an investigation on hypertensives and controls to evaluate the association of the gene variant, CYP24A1 rs2762939 and 25(OH)D in an Indian population with EH. On gender-based stratification, with multivariate logistic analysis after adjustment for covariates, the CYP24A1 rs2762939 CC variant showed a higher risk of EH in males (aOR = 3.141, CI 1.164-8.478, P = .024) while females illustrated an inverse association with EH (aOR = 0.398, CI 0.172-0.092, P = .031). The 25(OH)D levels among the three genotypes of hypertensives substantiate these results. Our results clearly suggest that gender, CYP24A1 rs2762939, and Vitamin D status may play a significant role in disease susceptibility towards EH in Indian population.

The effect of lead (Pb) exposure and iron (Fe) deficiency on intestinal lactobacilli, E. coli and yeast: A study in experimental rats.

OBJECTIVE: The current study investigated the additive effect of oral lead (Pb) exposure and dietary iron (Fe) deficiency on intestinal lactobacilli, E. coli, and yeast in SD rats. METHODS: Weanling rats were fed on control diet (CD) or iron deficient diet (ID) for 4 weeks, followed by oral Pb exposure for another 4 weeks. Lead exposure was withdrawn for 2 weeks, and then resumed after 2 weeks. Blood samples were collected to determine haemoglobin (Hb), serum iron, blood Pb and δ-Aminolevulenic acid dehydratase (ALAD) activity. Fecal samples were collected to enumerate the lactobacilli, E. coli and yeast population on selective agar media and determine Pb levels. RESULTS: Hb and serum Fe levels decreased significantly in iron deficient rats. Pb exposed rats had a significant increase in blood Pb levels and decreased ALAD activity. The lactobacilli population was significantly decreased (p<0.05) in ID rats compared to the CD group. Further, a significant decrease in the lactobacilli population was observed in Pb exposed rats irrespective of the dietary regimen. Upon withdrawal of Pb exposure, lactobacilli increased significantly in both the CD+Pb and ID+Pb groups, whereas re-exposure to Pb decreased lactobacilli population. The E. coli and yeast populations were inconsistent among both the ID and Pb exposed rats compared to controls. Fecal Pb levels increased significantly in Pb exposed rats irrespective of diet. CONCLUSION: An additive effect of dietary Fe deficiency and oral Pb exposure resulted in greater reductions in the intestinal lactobacilli population compared to either treatment alone. In addition, transient withdrawal of Pb exposure led to improved lactobacilli population irrespective of Fe status.

mRNA expression profile in peripheral blood mononuclear cells based on ADRB1 Ser49Gly and Arg389Gly polymorphisms in essential hypertension - a case-control pilot investigation in South Indian population.

BACKGROUND: ß1-Adrenoreceptor (ADRB1) genetic polymorphisms are widely studied for susceptibility to many cardiovascular diseases such as essential hypertension. However, the mRNA expression of ADRB1 is rarely studied. METHODS: A case-control pilot study with 292 hypertensives and 324 controls was designed to evaluate the role of the Ser49Gly and Arg389Gly, which are commonly studied single nucleotide polymorphisms (SNP), in the mRNA levels of ADRB1, in conjunction with its genetic predisposition to essential hypertension. RESULTS: Differential expression of ADRB1 mRNA was seen between hypertensives and controls (p<0.01) based on genetic variants of Ser49Gly. Among hypertensive subjects, Ser49Ser and Gly49Gly were highly expressed in comparison to Ser49Gly (p<0.05 and p<0.01, respectively), whereas genetic variants of Arg389Gly did not demonstrate any such variations. We found no association between the ADRB1 SNPs viz., Ser49Gly and Arg389Gly and essential hypertension. CONCLUSIONS: The increased mRNA levels of Gly49Gly may indicate a plausible role in the interindividual variations in drug response. Further, ADRB1 polymorphisms did not contribute to the genetic risk of essential hypertension. Studies with larger sample size are warranted to confirm these observations in the South Indian population.

Antihypertensive Prescription Pattern and Compliance to JNC 7 and JNC 8 at Tertiary Care Government Hospital, Hyderabad, India: A Cross-sectional Retrospective Study.

Background: The monitoring of hypertension treatment can provide insight into the rational drug use pattern. The objective of this study was to examine the antihypertensive medication use among South Indian adults with hypertension in compliance with the hypertension treatment guidelines (Seventh Joint National Committee [JNC 7] and JNC 8). Methods and Results: A total of 550 hypertensive people aged >25 years were included in this retrospective cross-sectional study. The order of drugs prescribed in the year 2012 was beta blockers (BB) > calcium channel blockers (CCB) > CCB + BB > angiotensin receptor blockers (ARB) > angiotensin-converting enzyme inhibitors (ACEI) > thiazide diuretics, whereas in the year 2014, the order has changed drastically, namely, ACEI > CCB > ARB > BB > thiazide diuretics (P < .001). Most notably, there was a large increase in the use of monotherapy (from 56.9% to 82.5%, P < .001). The usage of BB has simply moved from the first position to the last position in concordance with JNC 8 guidelines, whereas use of thiazide diuretics was found to be the least preferred drug in the 2012 prescriptions, thus deviating from JNC 7 guidelines. The use of generic names (28.3% vs 11.3%) and National List of Essential Medicines (NLEM) compliance (79.3% vs 60.9%) were significantly more in the calendar year 2012 than in the calendar year 2014 (P < .001). Conclusions: Antihypertensive medication use has gone through wide variations among south Indian adults with hypertension. Combination therapy regimens must be adopted as per the guidelines for achievement of blood pressure goals.

ß-Carotene bioaccessibility from biofortified maize (Zea mays) is related to its density and is negatively influenced by lutein and zeaxanthin.

Biofortification of maize with provitamin A (pro-VA) carotenoids has been successful, but the bioavailability of carotenoids needs to be explored. In the present study, we investigated the carotenoid content, micellarization and intestinal cell uptake of carotenoids from 10 maize hybrids [normal maize, quality protein maize (QPM), pro-VA carotenoid and double biofortified QPM + pro-VA maize hybrids] using a simulated in vitro digestion/Caco-2 cell model. The pro-VA carotenoid content of biofortified maize hybrids is 2-10 fold higher compared to that of normal maize. Furthermore, the ratio of non-pro-VA carotenoids lutein (LUT) plus zeaxanthin (ZEA) to the sum of pro-VA carotenoids ß-cryptoxanthin (BCX), α-carotene (AC) and ß-carotene (BC) in biofortified maize was much lower compared to that of normal maize. The consumption of 200 g day-1 of biofortified Pusa-PV-16-3 (BC = 808.4 µg per 100 g; AC = 839.3 µg per 100 g; BCX = 59 µg per 100 g) and Pusa-APQH8 (BC = 345.9 µg per 100 g; AC = 1739 µg per 100 g; BCX = 644.2 µg per 100 g) maize would contribute to 52 and 64% of RDAs for adult Indian men, respectively, after adjusting for cooking losses and conversion factors. The mean efficiency of micellarization of LUT (62.2% ± 5.3), ZEA (65% ± 4.7), and BCX (54% ± 9.5) exceeded that of AC (43% ± 8.9) and BC (49.8% ± 7.8) from all the maize hybrids. Furthermore, the micellarization and uptake in Caco-2 cells during a 4 h incubation period showed high correlation (P < 0.05) with the concentration of carotenoids in the maize digesta and micellar fraction, respectively. However, the LUT + ZEA content in the maize digesta and micellar fraction was inversely (p < 0.05) related to the BC micellarization and intestinal cell uptake, respectively. These results together suggest that the enrichment of pro-VA carotenoids together with decreasing the oxygenated carotenoid metabolites such as LUT and ZEA will further improve the bioavailability of BC from maize hybrids.

Nutrition-pollution interaction: An emerging research area.

The impact of environmental pollution, especially chronic low exposures of heavy metals (Pb, Cd, Hg, As, Cr, etc.) on nutritional status and health of human and livestock, has become a cause of concern. It is established that malnutrition inhibits enzyme system, alters neurotransmitter levels, degenerate myelin, glial and neural elements, lowering of IQ scores as well as impairment of fine and gross motor coordination. Chronic low-level exposure to heavy metals also results in similar type of deformities at sub-clinical level. However, additive impact of undernutrition and adverse effects of heavy metal exposure is emerging as a serious threat to health in developing countries. High blood Pb/Cd levels and low nutrient levels cause subclinical damage of organ system such as haemopoietic, renal, nervous systems in neonates, children, post-partum women, and occupationally exposed population. This could be due to chronic low-level heavy metal exposures and vis-à-vis interaction between pollutants and nutrients. Our studies are focused on the utility of biomarkers for early subclinical detection of haemopoietic and rental toxicity. Lead exposure from non-conventional sources such as toys, pet/glass bottles, etc. suggest long-term investigation. The present review compiles result of studies conducted in this area highlighting the importance of pollution-nutrition interaction. This may facilitate policymakers on developing the strategies to counter the heavy metal exposure of humans/livestock and their consequences.

Evaluation of Rasna panchaka (indigenous drug) as oxidative stress down-regulator using serum-free explant culture system.

CONTEXT: The importance of phytochemicals/natural products as potential therapeutic agents in the present context is gaining a lot of importance. India with a rich heritage of such preparations needs evaluation as potent drugs. Explant culture system is a method, which is sensitive, reliable, reproducible and is capable of mimicking the in situ conditions maintaining the tissue in sufficiently high level of integration. AIM: The current study aimed to test the antioxidant activity of test compounds, namely, traditional aqueous (4212) and aqueous-methanolic (4308) extracts of Rasna panchaka using liver explant cultures. MATERIALS AND METHODS: Dose-response optima of extracts (0.2-10 µg/mL) were determined using mouse liver explant culture system up to 48 h. The antioxidant property of extracts was assessed by primary oxidative defense parameters, namely, superoxide-dismutase (SOD), catalase, reduced glutathione (GSH), and malondialdehyde (MDA). RESULTS: The results indicated that the cellular architecture of the cultured tissue was well conserved in the first 6 h with a gradual display of specific changes in the next 24 h. There was a significant increase in MDA levels in experimental groups indicating the oxidative stress induction in explants. A dose of 2.0 µg/mL extracts have shown statistically significant (P < 0.05) protection against oxidative stress. MDA levels, a measure of lipid peroxidation, were significantly (P < 0.01) reduced by 50% in extract treated explants compared to control. This effect was accompanied by the increase in the first defense enzymes SOD (50%) and catalase (18%) with no change in reduced GSH levels. CONCLUSION: The study enforces the importance of "explant culture system," as it not only reduces the use of nonclinical/animal model but also is rapid and sensitive. Further, results of the current study also suggest that aqueous-methanolic extract of Rasna panchaka is having superior antioxidant activity compared to traditional water extract.

Validation of therapeutic anti-inflammatory potential of Arjuna Ksheera Paka - A traditional Ayurvedic formulation of Terminalia arjuna.

Arjuna Ksheera Paka (AKP), a traditional Ayurvedic formulation of Terminalia arjuna (T. arjuna) bark powder is used for its cardioprotective effects. However, its anti-inflammatory efficacy remained unexplored. In the present study, AKP was prepared in cow milk (as per standard Ayurvedic procedure) and compared with standard hydroalcoholic extract (HA) of T. arjuna. The extracts were analyzed for gross phytoconstituents levels, and their antioxidant activity was assayed by DPPH free radical scavenging activity and inhibition of lipid peroxidation. The in vivo anti-inflammatory activity of AKP and HA was studied in carrageenan-induced hind paw biphasic edema in C57BL/6 mice (at 200, 400 and 800 mg/kg BW). The percentage extraction yield of AKP was two folds higher than HA implying that the phytoconstituents in AKP were diluted by a factor of 0.5. The total polyphenol content of HA was (3.8 times) higher than AKP and the antioxidant activity of HA was also higher compared to AKP. Both the extracts, however, showed significant (p < 0.05) anti-inflammatory activity in reducing paw edema in mice. The efficacy of HA was more than AKP at early phase of inflammation, whereas, in the late phase of inflammation AKP was more efficacious and equipotent to HA. Thus, regardless of low in vitro antioxidant activity, AKP exhibited potential in vivo anti-inflammatory activity. The higher efficacy of AKP could be due to the presence of milk solids. These milk solids may act as adjuvants to T. arjuna's phytoconstituents, contributing to their sustained bioavailability, leading to higher in vivo anti-inflammatory efficacy at lower drug concentrations.

Switching of antihypertensive drugs at Tertiary Care Government Hospital, Hyderabad, India: A cross-sectional retrospective investigation.

OBJECTIVE: Switching of antihypertensive drugs is attributed to uncontrolled blood pressure (BP) which imposes a great burden on health economics. But again, switching leads to accomplishment of the goal BP, thereby improving the health status. Such studies are well documented in developed nations but rarely reported in developing countries, especially in India. Therefore, the aim of this study was to evaluate various factors associated with switching of antihypertensive drugs. METHODS: A cross-sectional retrospective investigation was performed using a standardized schedule adapting the World Health Organization indicators for drug utilization in a tertiary care government hospital, Hyderabad, India. A total of 429 prescriptions were monitored for a switchover to a different antihypertensive drug in 180 days. RESULTS: The results revealed that the duration of hypertension (HTN) >5-10 years (adjusted odds ratio [aOR] = 3.73, P < 0.05), two or more symptoms of HTN (aOR = 3.42, P < 0.05), 2014 prescriptions (aOR = 4.54, P < 0.001), polytherapy (aOR = 2.85, P < 0.001), noncompliance to National List of Essential Medicine (NLEM) (aOR = 1.631, P < 0.05), and systolic BP (SBP) (aOR = 1.77, P < 0.05) were the predictors, which were highly likely to switch (38.5%) the antihypertensive drugs. Diuretics (0.7%) were poorly prescribed, the first line of therapy suggested by Seventh Joint National Committee (JNC VII). Stepwise logistic regression analysis revealed, the calendar year 2014 (odds ratio [OR] = 3.23, P < 0.001), polytherapy (OR = 2.5, P < 0.001), and the level of SBP ≥140 mmHg (OR = 1.82, P < 0.01) as the three major predictors which showed a likelihood of switching medication. CONCLUSIONS: Findings of the study reveals predictors of the switchover like uncontrolled SBP, duration of HTN, compliance with the list of NLEM drugs, polytherapy, enabling the clinicians to critically analyze the patients' profile, and hence, reach target BP soon, i.e., decreased cardiovascular risk.

Evaluation of Bar, Barnase, and Barstar recombinant proteins expressed in genetically engineered Brassica juncea (Indian mustard) for potential risks of food allergy using bioinformatics and literature searches.

The potential allergenicity of Bar, Barnase, and Barstar recombinant proteins expressed in genetically engineered mustard for pollination control in plant breeding was evaluated for regulatory review. To evaluate the potential allergenicity of the Bar, Barnase and Barstar proteins amino acid sequence comparisons were made to those of known and putative allergens, and search for published evidence to the sources of the genes using the AllergenOnline.org database. Initial comparisons in 2012 were performed with version 12 by methods recommended by the Codex Alimentarius Commission and the Indian Council of Medical Research, Government of India. Searches were repeated with version 15 in 2015. A literature search was performed using PubMed to identify reports of allergy associated with the sources of the three transgenes. Potential open reading frames at the DNA insertion site were evaluated for matches to allergens. No significant sequence identity matches were identified with Bar, Barnase or Barstar proteins or potential fusion peptides at the genomic-insert junctions compared to known allergens. No references were identified that associated the sources of the genes with allergy. Based on these results we conclude that the Bar, Barnase and Barstar proteins are unlikely to present any significant risk of food allergy to consumers.

Combination of spices and herbal extract restores macrophage foam cell migration and abrogates the athero-inflammatory signalling cascade of atherogenesis.

The trapping of lipid-laden macrophages in the arterial intima is a critical but reversible step in atherogenesis. However, information about possible treatments for this condition is lacking. Here, we hypothesized that combining the polyphenol-rich fractions (PHC) of commonly consumed spices (Allium sativum L (Liliaceae), Zingiber officinale R (Zingiberaceae), Curcuma longa L (Zingiberaceae)) and herbs (Terminalia arjuna (R) W & A (Combretaceae) and Cyperus rotundus L (Cyperaceae)) prevents foam cell formation and atherogenesis. Using an in vitro foam cell formation assay, we found that PHC significantly inhibited lipid-laden macrophage foam cell formation compared to the depleted polyphenol fraction of PHC (F-PHC). We further observed that PHC attenuated the LDL and LPS induced CD36, p-FAK and PPAR-γ protein expression in macrophages and increased their migration. NK-κB-DNA interaction, TNF-α, ROS generation, and MMP9 and MMP2 protein expression were suppressed in PHC-treated macrophages. The anti-atherosclerotic activity of PHC was investigated in a high fat- and cholesterol-fed rabbit model. The inhibition of foam cell deposition within the aortic intima and atheroma formation confirmed the atheroprotective activity of PHC. Therefore, we conclude that the armoury of polyphenols in PHC attenuates the CD36 signalling cascade-mediated foam cell formation, enhances the migration of these cells and prevents atherogenesis.