Prevalence of anaemia among women of reproductive age group in a rural block of Northern India

Background: Nutritional anaemia is a major public health problem in India and is primarily due to iron deficiency. The National Family Health Survey-3 (NFHS-3) data suggests that anaemia is widely prevalent among all age groups, and is particularly high nearly 55.3% amongst the most vulnerable in all women (15-49 years) Aims & Objectives: 1. To determine prevalence of Anaemia among women of reproductive age group in rural block of Haryana. 2. Effects of anaemia on mean height and weight of women of reproductive age group. Material Methods: Cross-sectional, descriptive. All the women of reproductive age group (15-45 years) of CHC Sample block were included as study participants. Results: The overall prevalence of anaemia was 48.9%. 4302 out of 8590 females had varying severity of anaemia while anaemia was absent in 51.1% of the study participants. Out of the 8590 females, 1612 (18.8%) were mildly anaemic, 2374 (27.6%) were moderately anaemic and 217 (2.5%) were severely anaemic. The study revealed that mean weight and mean height in non anaemic females was more than that of varying degree (severe, moderate, mild) of anaemic females. Conclusion: the present study revealed anaemia to be a major health problem among the women of reproductive age group in rural areas in Haryana affecting their health status

Phytonanotechnology: Recent applications and the role of Biocorona

405-414Phytonanotechnology is lately gaining increased interest owing to its potential to modernize agriculture for better yield and nutritional quality. Consequently, Nano-Agri products like nano-biosensors, nano-carriers, and growth augmenters are being developed and applied. However, the limited knowledge of molecular interactions taking place at nano-bio interface remains a major concern. The nanotechnological interventions for healthier crops could rather turn out tobe risky and inefficient in the absence ofclear understanding of molecular mechanisms of nano-bio interactions. Upon entry into tissues or cells, nanoparticles (NPs) adsorb biomolecules forming a biocorona which determines NP uptake, translocation, and reactivity. The composition of biocorona is dependent on the physicochemical characteristics of the NPs, their surroundings, and the interaction time. Recent nascent studies in plants showed the potential of biocorona to influence major cellular pathways or plant responses like energy synthesis, pathogenesis, stress tolerance, and leaf senescence. This mini-review aims at summarizing the recent application of phytonanotechnology, the current status of biocorona studies with an overview of research bottlenecks and future prospects

Antisense expression of a gene encoding a calcium-binding protein in transgenic tobacco leads to altered morphology and enhanced chlorophyll

Entamoeba histolytica contains a novel calcium-binding protein like calmodulin, which was discovered earlier, and we have reported the presence of its homologue(s) and a dependent protein kinase in plants. To understand the functions of these in plants, a cDNA encoding a calcium-binding protein isolated from Entamoeba histolytica (EhCaBP) was cloned into vector pBI121 in antisense orientation and transgenic tobacco plants were raised. These plants showed variation in several phenotypic characters, of which two distinct features, more greenness and leaf thickness, were inherited in subsequent generations. The increase in the level of total chlorophyll in different plants ranged from 60% to 70%. There was no major change in chloroplast structure and in the protein level of D1, D2, LHCP and RuBP carboxylase. These morphological changes were not seen in antisense calmodulin transgenic tobacco plants, nor was the calmodulin level altered in EhCaBP antisense plants

Biochemical and immunochemical characterization of Brassica juncea glyoxalase I

A homogenous preparation of glyoxalase I (S-lactoylglutathione-lyase, EC 4.4.1.5) was obtained from Brassica juncea seedlings. The enzyme is a heterodimer with 27,000 and 29,000 M<SUB>r</SUB> subunits and native M<SUB>r</SUB> of 56,000. The circular dichroic spectra of the protein showed characteristics of a distinctly helical protein, and magnesium affected the secondary structure. It is a zinc metalloenzyme. Amino acid modification studies suggested the involvement of histidine residues in catalysis. Apo-glyoxalase I was reactivated by divalent cations Mn<SUP>2+</SUP> (0.5 Mm)&gt;Mg<SUP>2+</SUP> (5 Mm)&gt;Zn<SUP>2+</SUP> (0.05 Mm) and Ca<SUP>2+</SUP> (0.01 Mm). Monospecific, polyclonal anti-glyoxalase I antibodies were raised, which showed its presence in seeds, roots, hypocotyl, cotyledon and different flower parts. They showed varied degree of cross reactivity with the extracts from various plants, yeast, bacteria and animal system

A simple chromatographic procedure for co-purification of calreticulin and calmodulin from <em>Brassica juncea</em> seedlings

281-290A 60 kDa heat stable calcium buffer protein, calreticulin, is co-purified with 17 kDa calmodulin from seedlings of Brassica juncea using anion exchange chromatography. The proteins were identified by MALDI-TOF/MS and LCMS, respectively. Like other plant calreticulins, partially purified Brassica calreticulin had an affinity for Ca2+ as it stained blue with stains-all staining and is glycosylated as it stained pink with PAS staining. Further, the two were separated on Phenyl sepharose chromatography due to their differential affinity. BjCaM stimulated cAMP phosphodiesterase activity and showed Ca2+-dependent mobility shifts on one- and two-dimension SDS-PAGE. It stained blue with stains-all confirming it to be Ca2+ binding protein. BjCaM had cysteine residue as confirmed by Biotin Switch technique, but it did not nitrosylate. Co-purification of the two proteins indicates their possible interaction, which needs to be confirmed

Refolding of <i>β</i>-Stranded Class I Chitinases of <i>Hippophae rhamnoides</i> Enhances the Antifreeze Activity during Cold Acclimation

<div><p>Class I chitinases hydrolyse the <i>β</i>-1,4-linkage of chitin and also acquire antifreeze activity in some of the overwintering plants during cold stress. Two chitinases, HrCHT1a of 31 kDa and HrCHT1b of 34 kDa, were purified from cold acclimated and non-acclimated seabuckthorn seedlings using chitin affinity chromatography. 2-D gels of HrCHT1a and HrCHT1b showed single spots with pIs 7.0 and 4.6 respectively. N-terminal sequence of HrCHT1b matched with the class I chitinase of rice and antifreeze proteins while HrCHT1a could not be sequenced as it was N-terminally blocked. Unlike previous reports, where antifreeze activity of chitinase was cold inducible, our results showed that antifreeze activity is constitutive property of class I chitinase as both HrCHT1a and HrCHT1b isolated even from non-acclimated seedlings, exhibited antifreeze activity. Interestingly, HrCHT1a and HrCHT1b purified from cold acclimated seedlings, exhibited 4 and 2 times higher antifreeze activities than those purified from non-acclimated seedlings, suggesting that antifreeze activity increased during cold acclimation. HrCHT1b exhibited 23–33% higher hydrolytic activity and 2–4 times lower antifreeze activity than HrCHT1a did. HrCHT1b was found to be a glycoprotein; however, its antifreeze activity was independent of glycosylation as even deglycosylated HrCHT1b exhibited antifreeze activity. Circular dichroism (CD) analysis showed that both these chitinases were rich in unusual <i>β</i>-stranded conformation (36–43%) and the content of <i>β</i>-strand increased (∼11%) during cold acclimation. Surprisingly, calcium decreased both the activities of HrCHT1b while in case of HrCHT1a, a decrease in the hydrolytic activity and enhancement in its antifreeze activity was observed. CD results showed that addition of calcium also increased the <i>β</i>-stranded conformation of HrCHT1a and HrCHT1b. This is the first report, which shows that antifreeze activity is constitutive property of class I chitinase and cold acclimation and calcium regulate these activities of chitinases by changing the secondary structure.</p></div

Purification of HrCHT1a and HrCHT1b.

<p>(A) Silver stained 15% SDS-PAGE gel showing apoplastic proteins isolated from non-acclimated and 3 weeks cold acclimated seedlings. (B) SDS-PAGE and (C) 10% native gel showing purified class I chitinase from 3 weeks cold acclimated apoplastic extracts using chitin affinity chromatography. (D) Two dimensional gel electrophoresis of HrCHT1a and HrCHT1b (E). Purified HrCHT1a and HrCHT1b (70 µg) were loaded on 3–10 non-linear IPG strips and subjected to iso-electric focusing. After IEF, proteins were resolved on 15% SDS-PAGE for the second dimension.</p

Low Temperature Stress Modulated Secretome Analysis and Purification of Antifreeze Protein from <i>Hippophae rhamnoides</i>, a Himalayan Wonder Plant

Plants' distribution and productivity are adversely affected by low temperature (LT) stress. LT induced proteins were analyzed by 2-DE-nano-LC–MS/MS in shoot secretome of <i>Hippophae rhamnoides</i> (seabuckthorn), a Himalayan wonder shrub. Seedlings were subjected to direct freezing stress (−5 °C), cold acclimation (CA), and subzero acclimation (SZA), and extracellular proteins (ECPs) were isolated using vacuum infiltration. Approximately 245 spots were reproducibly detected in 2-DE gels of LT treated secretome, out of which 61 were LT responsive. Functional categorization of 34 upregulated proteins showed 47% signaling, redox regulated, and defense associated proteins. LT induced secretome contained thaumatin like protein and Chitinase as putative antifreeze proteins (AFPs). Phase contrast microscopy with a nanoliter osmometer showed hexagonal ice crystals with 0.13 °C thermal hysteresis (TH), and splat assay showed 1.5-fold ice recrystallization inhibition (IRI), confirming antifreeze activity in LT induced secretome. A 41 kDa polygalacturonase inhibitor protein (PGIP), purified by ice adsorption chromatography (IAC), showed hexagonal ice crystals, a TH of 0.19 °C, and 9-fold IRI activity. Deglycosylated PGIP retained its AFP activity, suggesting that glycosylation is not required for AFP activity. This is the first report of LT modulated secretome analysis and purification of AFPs from seabuckthorn. Overall, these findings provide an insight in probable LT induced signaling in the secretome

Glycosylation analysis of HrCHT1a and HrCHT1b.

<p>Figure showing Ponceau-S staining (A) and Con-A peroxidase staining (B) of HrCHT1a and HrCHT1b purified from non-acclimated and cold acclimated seedlings to detect their glycosylation.</p

Secondary structure analysis of HrCHT1a and HrCHT1b.

<p>Far UV range CD spectra of HrCHT1a (A, B) and HrCHT1b (C, D) purified from non-acclimated (NA) and cold acclimated (CA) seedlings with 0.4 mM calcium and 0.2 mM EGTA. Each spectra presented in the figure are averaged from three spectra.</p