Differential Iron Status and Trafficking in Blood and Placenta of Anemic and Non-anemic Primigravida Supplemented with Daily and Weekly Iron Folic Acid Tablets.

ABSTRACT: The molecular mechanism of iron transfer across placenta in response to maternal anemic status/ iron supplementation is not clear. We hypothesized that maternal iron/ anemia status during early trimesters can be utilized as a biomarker tool to get estimates of placental iron status. Early interventions can be envisaged to maintain optimum placental/ foetal iron levels for healthy pregnancy outcomes. One hundred twenty primigravida were recruited and divided into non-anemic and anemic group on the basis of hemoglobin levels. The groups were randomly allocated to receive daily and weekly iron folic acid (IFA) tablets till six weeks postpartum. Hematological and iron status markers in blood and placenta were studied along with the delivery notes. Weekly IFA supplementation in anemic primigravidas resulted in significantly reduced levels of hematological markers (p < 0.01); whereas non-anemic primigravidas showed lower ferritin and iron levels, and higher soluble transferrin receptor levels (p < 0.05). At baseline, C-reactive protein and cortisol hormone levels were also significantly lower in non-anemic primigravidas (p < 0.05). A significantly decreased placental ferritin expression (p < 0.05); and an increased placental transferrin expression was seen in anemic primigravidas supplemented with weekly IFA tablets. A significant positive correlation was observed between serum and placental ferritin expression in anemic pregnant women (r = 0.80; p < 0.007). Infant weight, gestational length and placental weight were comparable in both the supplementation groups. To conclude, mother's serum iron / anemia status switches the modulation in placental iron transporter expression for delivering the optimum iron to the foetus for healthy pregnancy outcomes. TRIAL REGISTRATION: Clinical Trial Registry-India: CTRI/2014/10/005135.

Role of Trace Elements, Oxidative Stress and Immune System: a Triad in Premature Ovarian Failure.

The risk of premature ovarian failure (POF) increases in association with alteration in immunological parameters and oxidative stress (OS). Adequate intake of trace elements is required for antioxidant property and immune defense mechanism. The aim of this study was to explore the involvement of trace elements, OS, and immunological parameters in POF. This was a cross-sectional, case-control study, involving 65 participants divided into the POF (n = 35) and control (n = 30) groups. Serum levels of Se, Zn, and Cu were determined along with hormonal, OS, and immunological markers. POF group had significantly lower levels of Zn, Cu, Se, and Zn:Cu ratio. However, Se:Cu ratio was not significant between the groups. FSH and LH levels were negatively correlated with Zn and Cu levels and positively correlated with Se levels. Estrogen levels were negatively correlated with all the studied trace elements. Inter-element association between Zn and Se was significant in POF (r = - 0.39, p = 0.02) compared to control group (r = - 0.078, p = 0.65). In all the POF patients, SOD and GPx activities were significantly (p < 0.05) lower and MDA level was higher (p > 0.05) than control group. B cell marker CD19 was significantly (p < 0.0001) high in POF group. There are involvement of trace elements in hormonal regulation and antioxidant defense mechanism, which once gets altered leads to high ROS generation and affect functions of the immune system. Exaggereative immune system causing higher expression of B cell associated markers (CD19) leading to autoimmune condition in POF.

Micronutrients Drift During Daily and Weekly Iron Supplementation in Non-anaemic and Anaemic Pregnancy.

AIMS: Pregnancy is a phenomenon associated with dynamic changes in physical, mental and biochemical status of body and demands increased nutritional intake for developing foetus. The level of various micronutrients which act as co-factors for antioxidant enzymes or it-self as antioxidants gets altered with the progression of pregnancy. The present longitudinal study summarized the trend of selected micronutrients level in anaemic (AP) and non-anaemic primigravida (NAP) supplemented with daily and weekly oral iron folic acid (IFA) tablet during pregnancy and postpartum. METHODS: A total of 200 primigravida {N = 100; NAP (Hb > 11 g/dl) and N = 100 AP (Hb = 8-11 g/dl) assigned daily (N = 50) and weekly (N = 50) supplementation} were recruited and overnight fasting blood samples were withdrawn at 13-16 weeks, after 3 months and 6 weeks postpartum. The serum iron, copper, zinc, magnesium and manganese were estimated by inductively coupled plasma-atomic emission spectrophotometer. RESULTS: Serum manganese (p < 0.05) at baseline and magnesium (p < 0.01) at postpartum was significantly different between NAP and AP supplemented with daily IFA tablets. The trend of copper found to be increased during pregnancy and later declined at postpartum in both the groups. Daily supplementation resulted in significantly high iron (p < 0.05) in NAP during third trimester. CONCLUSIONS: Hypozincemia and hypomagnesemia was observed in anaemic pregnancy supplemented with weekly and daily IFA respectively. Clear evidence of altered micronutrients levels during healthy and anaemic pregnancy was seen. The reference values may be drawn from this study for the nutritional assessment during pregnancy for healthy pregnancy outcomes. TRIAL REGISTRATION: Clinical Trial Registry-India, http://ctri.nic.in, CTRI/2014/10/005135.

Weekly iron folic acid supplementation plays differential role in maintaining iron markers level in non-anaemic and anaemic primigravida: A randomized controlled study.

Anaemia during pregnancy is most commonly observed and highly prevalent in South-East Asia. Various effective programmes have been laid down for its management, mainly daily supplementation of iron folic acid (IFA) tablets. Following the same, standard obstetrical practice has included the IFA supplementation without requiring the determination of iron deficiency. In this study, a total of 120 primigravida (N = 60; non-anaemic (Hb > 11 g/dl) and N = 60 anaemic (Hb = 8-11 g/dl)) were selected among those attending the Antenatal Clinic in Department of Obstetrics and Gynaecology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India. They were supplemented with daily and weekly IFA tablets till 6 weeks postpartum. Corresponding changes in haemoglobin level on advance of pregnancy, side effects and compliance associated with daily and weekly IFA supplementation and its associations with iron status markers were studied. The inflammatory markers were also estimated. The statistical significance level (p < 0.05) between the groups were assessed by applying unpaired t-test using SPSS (version 16.0). The obtained results publicized the salutary role of daily IFA supplementation in improving the haemoglobin level and iron status markers in anaemic pregnant women though the levels could not reach up to the non-anaemic haemoglobin levels. However, weekly IFA supplementation seems to be a better approach in non-anaemic pregnant women where almost comparable results were obtained in terms of haematological parameters, gestation length and birth weight. CONCLUSION: Weekly IFA supplementation found to be as effective as daily supplementation in iron sufficient non-anaemic pregnant women whereas anaemic pregnant women should be prescribed daily IFA supplementation irrespective of iron replete/deplete state.

Suppressive effect of strychnos nux-vomica on induction of ovalbumin-specific IgE antibody response in mice.

UNLABELLED: Strychnos nux-vomica Linn. (SNV; Loganiaceae), a medicinal plant has been used as folk medicine for alleviating inflammation, joint pains and allergic symptoms. In the present study, we examined its possible immunomodulatory effect on induction of ovalbumin (OVA)-specific IgE antibody response in a murine model, as evaluated by passive cutaneous anaphylaxis (PCA). The OVA-specific IgE antibody response was significantly suppressed in BALB/c mice (H-2d), following intraperitoneal administration of aqueous stem extract of the plant along with OVA. Furthermore, the different doses of SNV extract were found to significantly suppress the induction of OVA-specific IgE antibody response. The anti-OVA IgE antibody response was suppressed in different haplotypes of mice viz., C57BL/6 (H-2b) and SWR/J (H-29). However, preliminary findings revealed no significant change in the total IgG antibody response against OVA, as evaluated by ELISA. These results confirm the suppressive activity of S. nux-vomica on allergen-specific IgE antibody response and suggest its possible application in allergic conditions. KEYWORDS: Strychnlos nux-vomica, Immunomodulation, Immunosuppression, IgE antibody response, Passive cutaneous anaphylaxis, ELISA

Correlation of T-cell response and lymphokine profile with RESA peptides of Plasmodium falciparum containing a universal T-cell epitope and an immunopotentiator, polytuftsin.

Two RESA repeat sequences, (EENVEHDA)2 and (DDEHVEEPTVA)2, were chemically linked to a universal T-cell epitope, CS.T3 and polytuftsin, and a natural immunopotentiator, was physically mixed with these conjugates. The immunogens were studied for in vitro antigen-induced T-cell proliferation, and cytokine levels were measured in the culture supernatants. The RESA peptide(s)-CS.T3 conjugate containing polytuftsin showed the highest stimulation index (SI) as compared to the RESA peptide-CS.T3 conjugates or RESA peptides alone. Spleen cells from mice primed with either RESA peptide(s)-CS.T3 conjugate or RESA peptide-CS.T3 conjugate containing polytuftsin, when pulsed in vitro with the respective RESA peptide, showed a higher proliferation index as compared to spleen cells primed and pulsed in vitro with the respective RESA peptides. This observation has an important relevance during natural reinfection for boosting the immune response. The culture supernatants from the cells primed and pulsed in vitro with RESA peptide-CS.T3 conjugate and RESA peptide-CS.T3 conjugate containing polytuftsin showed higher IL-2 and IFN-gamma levels as compared to the RESA peptides alone. Very low IL-4 levels were detected with the above formulations. The cytokine profile is suggestive of a CD4+ TH1 type of immune response, which is ideal for the killing of intracellular pathogens like the malarial parasite.

Potentiation of immune response against the RESA peptides of Plasmodium falciparum by incorporating a universal T-cell epitope (CS.T3) and an immunomodulator (polytuftsin), and delivery through liposomes.

Synthetic peptides representing repeat sequences of ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum have shown poor immunogenicity and protection. In this study, the RESA peptides [(EENVEHDA)2 and (DDEHVEEPTVA)2] were chemically linked to a universal T-cell determinant, CS.T3, derived from the CS protein of P. falciparum. Polytuftsin (TKPR)40, a polymer of naturally occurring immunomodulator "tuftsin," was physically mixed with these conjugates. These preparations in alum and liposomes were immunized in four inbred strains of mice with different genetic backgrounds to study the humoral response. In the case of liposome-entrapped preparations, a 10 microg dose of antigen showed the optimum antibody response. Mice immunized with liposome containing RESA peptide(s)-CS.T3 conjugate along with polytuftsin showed the highest antibody levels in all the strains, whereas the RESA peptide(s) alone, adsorbed on alum or entrapped in liposomes, showed either poor or moderate antibody levels. The antibodies raised against liposome-entrapped preparations in both high-responder strain (SJL/J H-2s) and low-responder strain (FVB/J H-2q) showed 2 4-fold lower Kd values as compared to the alum adsorbed preparations, suggestive of high affinity antibodies. All the antigen preparations predominantly induced IgG2a and IgG2b isotype response, suggesting that the T-helper response involved is of the CD4 Thl type. The in vitro merozoite reinvasion inhibition assay showed 50-92% inhibition with sera raised against different antigen formulations. The highest percentage inhibition was observed with the RESA peptide-CS.T3 conjugate containing polytuftsin in liposomes. Thus, the incorporation of peptide antigens inside liposomes not only reduced the antigen dose by 5-fold but also elicited a high titre with high affinity antibodies and the inhibition of merozoites to RBC in vitro. Therefore, we conclude that the incorporation of these synthetic constructs in liposomes could be a useful strategy for the development of a subunit immunogen against malaria.

Increase in the immunogenicity of HIV peptide antigens by chemical linkage to polytuftsin (TKPR40).

The use of synthetic peptide antigens in human prophylaxis still suffers from the very important problem of finding suitable carriers devoid of side effects. A desirable carrier for use in humans would be poorly immunogenic by itself, yet it would enhance the immune response to the peptide antigen. In the study reported herein, we examined the role of polytuftsin (TKPR40), a synthetic polymer of the natural immunomodulator tuftsin, as a carrier for synthetic peptides of HIV derived from the gp41 and gp120 proteins. Chimeric immunogens were constructed by chemical linkage between synthetic peptides of HIV and polytuftsin. These were employed for immunization of mice of different MHC haplotypes, and the humoral and cellular immune responses developed against the peptides were assessed by measuring total IgG, IgG, subclasses, T-cell proliferation, and in vitro cytokine release. A significantly stronger immune response was observed in mice immunized with the peptide-polytuftsin conjugates than in mice receiving the peptide dimers (peptide-peptide). Peptide-polytuftsin conjugates induced IgG2a and IgG2b isotype switching after both primary and secondary immunization. In addition, there was a positive correlation between the amounts of cytokines and the shift in the IgG isotypes. These data suggest that the use of polytuftsin as a carrier may increase the immune response against poorly immunogenic synthetic peptides.

Polytuftsin: its possible effects and mechanism during macrophage activation.

Polytuftsin (PT) a 35-40 repeat unit of tuftsin (TKPR), when administered as a conjugate with the malarial peptide, ring-infected erythrocyte surface antigen (RESA), enhanced antigen-induced lymphoproliferation and antibody levels in mice as compared to RESA alone. This enhancement was unrelated to the H-2 background of the animals. The present study was undertaken with a view to understanding the mechanism(s) responsible for this immune enhancement. Peritoneal adherent cells (PAC) from H-2b and H-2d mice were incubated with RESA alone, PT-conjugated RESA, a physical mixture of RESA + PT and PT alone. They were subsequently evaluated for I-A expression using monoclonal antibodies and flow cytometry as well as cell-ELISA. Significant increase in I-A expression on PAC was observed in all 4 groups as compared to untreated cells. Whereas cells treated with PT-conjugated RESA showed highly significant increase in I-A (P < 0.001), the other groups showed moderate increase (P < 0.05). This enhancement was attributable to increase in the number of I-A-positive cells rather than I-A molecules per cell. Moreover, IL-1 release, as assayed by bioassay, was significantly higher in cells treated with conjugated RESA as compared to cells treated with RESA or PT alone (P < 0.05). Thus, it would appear that PT-conjugated RESA peptide of the malarial antigen selectively enhances major histocompatibility complex (MHC) class II molecules on antigen-presenting cells (APC) and may therefore improve immune functions by stimulating better antigen presentation and proliferation of T cells.

Comparative assessment of pedal pressing rates of self-stimulation of hypothalamus and midbrain with both square wave and sine wave stimulus parameters.

In Wistar rats, the regional differences of pedal pressing rates of self-stimulation (SS) of lateral hypothalamus (LH) and substantia nigra-ventral tegmental area (SN-VTA) were assessed with electrodes implanted in both regions in each subject. Average of SS rates of SN-VTA sites was significantly higher than that of LH sites, tested with both sine wave and square wave types of stimuli. There was no significant difference in SS rates between males and females, and also in the females between different days of oestrus cycles. The high rates of robust SS observed in this study relative to SS rates reported in past literature were probably due not only to the placements of electrodes in the main substrates of SS, but also to the parameters of stimulus used (0.25 sec trains of sine waves through bipolar electrodes).

Physiological responses of Vicia faba plants to sulfur dioxide.

Exposure of broad bean (Victa faba L.) plants to 270 +/- 32 and 670 +/- 45 micrograms m 3SO2 for 1.5 hr daily between 40 and 85 days of their ages resulted in an increase in their transpiration rate, water saturation deficit, phenol content, and peroxidase activity and a decrease in protein content. With the increase in number of exposures of plants to SO2, chlorotic and brown, necrotic visible injury signs were also developed in leaves. It was further noted that the magnitude of undesirable biochemical changes, which possibly helped in the formation of new pigment characteristic of necrotic tissue of SO2-exposed plants, was not totally dependent on the pollutant concentration.