Rani Kumar

Role of oxidative stress markers and antioxidants in the placenta of preeclamptic patients

Aim:  The present study aimed to evaluate and compare the placental variables of oxidative stress markers in preeclamptic women.

Surgical importance of arterial segments of human kidneys: an angiography and corrosion cast study.

OBJECTIVE Variations in the arterial supply of human kidney have been observed frequently, either in routine dissections or surgical practice. The main objective of the present study was to describe the arterial segmental pattern of human kidneys and its variation by angiography and corrosion cast techniques. MATERIALS AND METHODS Forty kidneys were washed and a plastic cannula was inserted into renal artery and the omnipaque dye was injected into it and X-ray was taken. The corrosion casts were prepared by injecting coloured acetate butyrate (CAB) granules solution. RESULTS Five vascular segments of kidney were seen based on the branching pattern of the renal artery by angiography and corrosion cast techniques. The renal artery was divided into anterior and posterior branches. The anterior branch further divided into four branches viz. apical segmental artery (ASA), Upper segmental artery (USA), middle segmental artery (MSA), lower segmental artery (LSA) while the posterior branch continue as posterior segmental artery. The origins of segmental arteries were variable. In 60% cases apical segmental artery (ASA) had common origin with upper segmental artery (USA) while in 40% cases it took origin directly from the main renal artery. Similarly the variations in the origin of the other branches of anterior division of renal artery were observed. The posterior segmental artery (PSA) however was single and comparatively small and supplied the posterior surface of the kidney. CONCLUSION The knowledge of the vascular pattern of the kidney is thus important for the purpose of angiography and surgical procedures especially for nephrectomy and kidney transplantation.

Evaluation of Organic and Conventional Rice Production Systems for their Productivity, Profitability, Grain Quality and Soil Health

Considering the importance of organic farming and growing demand for organically produced foods, field studies were conducted for 5 years (2004-05 to 2009-10) on a black clayey vertisol soil at the Directorate of Rice Research, Hyderabad, to study the influence of organic and conventional farming systems on productivity, grain quality, soil health and economic returns of super fine rice varieties. Two main plot treatments, with and without plant protection, and four sub plot treatments viz., Control; 100% inorganics; 100% organics; and 50% inorganics+50% organics (integrated nutrient management, INM) were imposed. During wet season, grain yields under 100% inorganics and INM were near stable (4.7-5.5 t/ha) and superior to organics by 15-20% during the first two years, which improved with organics (4.8-5.2 t/ha) in the later years to comparable levels with inorganics, while it had taken five years during dry season. Moderate improvement in nutritional quality was recorded with organics, especially in brown rice. There was a significant improvement in soil physical, fertility and biological properties with organics, which resulted in further improvement in soil quality indices. The sustainability index of the soil was maximum with organics (1.63) compared to inorganics (1.33), after five years of study. The soil organic carbon (SOC) stocks were higher with organics by 44 and 35%, compared to conventional system during wet and dry seasons, respectively, after five years of study. The carbon sequestration rate was also positive with organics (0.97 and 0.57 t/ha/yr during wet and dry seasons, respectively), compared to conventional system that recorded negative SOC sequestration rate (-0.21 and -0.33 t/ha/yr during wet and dry seasons, respectively). Benefit cost ratio was less with organics in the initial years and improved later over inorganics by fifth year.

Presence of embryotoxic factor in the sera of neonates affected by myelomeningocele: A study on chick embryo

An experimental study on chick embryos was conducted where 0.02 mL of serum, taken from five neonates (1 to 7 days of age) with myelomeningocele was injected separately into the fertile white leghorn eggs, at zero hour of incubation. A total of 150 experimental and 135 control eggs were injected. It was observed that the embryos of chick exposed to experimental serum had an increased frequency of embryonic death and/or gross abnormalities as compared with the control group (chi 2 = 32.07; P less than .001). The anomalies observed were those of neural tube, gastrointestinal system, and musculoskeletal system. In addition, there was a generalized growth retardation among experimental embryos. These malformations are probably due to an embryotoxic factor, which is present in the serum of the baby with myelomeningocele and has been transmitted transplacentally.

A Comparative Study of Apoptosis in Placentas of Normal and Preeclamptic Indian Pregnant Women by TUNEL Assay and M30 Immunostaining

Preeclampsia is a unique life‐threatening disorder of human pregnancy associated with the abnormal placentation caused by the inadequate trophoblastic invasion due to altered apoptosis of these cells. The aim of the present study was to assess and compare the apoptosis in trophoblastic cells in various zones (villous and extravillous) of placentas of preeclamptic and normotensive nonproteinuric pregnant women.

sFlt-1 commutes unfolded protein response into endoplasmic reticulum stress in trophoblast cells in preeclamptic pregnancies

Preeclampsia (PE) and its subtypes (early and late onset) are serious concerns all across the globe affecting about 8% of total pregnancies and accounts for approximately 60,000 deaths annually with a predominance in developing under-developed and countries. The two-stage model in the progression of this disease, deficient spiral artery remodelling and an imbalance between angiogenic (VEGF) and anti-antigenic factor(s) (sFlt-1) are well established facts pertaining to this disease. The presence of increased sFlt-1, high oxidative stress and Endoplasmic reticulum stress (ER stress) have been proposed in preeclamptic pregnancies. Recently, the role of endoplasmic reticulum stress in the onset of the variant forms of PE highlighted a new window to explore further. In our previous studies, we demonstrated that sFlt-1 can induce apoptosis and oxidative stress in trophoblast cells. However the role of sFlt-1, in inducing ER stress is not known so far. In the present study, we for the first time demonstrated significant ER stress in the placental cells (BeWo Cells) (in vitro) when exposed to sera from preeclamptic pregnancies having increased concentration of sFlt-1. The expression of ER stress markers (GRP78, eIF2α, XBP1, ATF6 and CHOP) at both transcript and protein levels were compared (between preeclamptic and normotensive non-proteinuric women) at three different time points (8h, 14h and 24hrs), analyzed and found to be significant (p<0.05). Conclusion Our results suggested that sFlt-1, released from placental cells in preeclampsia may be one of the various factors having potential to induce endoplasmic reticulum stress in BeWo cells.

sFlt-1 (sVEGFR1) induces placental endoplasmic reticulum stress in trophoblast cell: implications for the complications in preeclampsia - an in vitro study

Background The concentration of sFlt-1, a major anti-angiogenic protein in maternal circulation has been seen to be raised in preeclamptic pregnancies. Endoplasmic reticulum (ER) stress represents one of the three (immunological, oxidative and ER stress) major stresses which placenta undergoes during pregnancies. The present study is designed to investigate the role of sFlt-1 in induction of ER stress in trophoblast cells. Materials and Methods Maternal serum levels of anti-angiogenic protein sFlt-1 and central regulator of unfolded protein response GRP78 was measured using sandwich ELISA. The expression of various ER stress markers (GRP78, eIF2α, XBP1, ATF6 and apoptotic protein CHOP) were analyzed depending on various treatments given to the trophoblast cells using Immunofluorescence, western blot and q-RT PCR. Results Increased expression of ER stress markers (GRP78, eIF2α, XBP1 ATF6 and apoptotic protein CHOP) was detected in the placental trophoblast cells treated with raised concentration of sFlt-1. Conclusion Significant upregulated expression of ER stress markers in trophoblast cells exposed with increased concentration of sFlt-1 suggested that it may be one of the anti-angiogenic factors present in maternal sera which not only contributes to oxidative stress but also may cause endoplasmic reticulum stress.

Value of Quantitative assessment of Myeloid Nuclear Differentiation Antigen expression and other flow cytometric parameters in the diagnosis of Myelodysplastic syndrome

The diagnosis of myelodysplastic syndrome (MDS) based on morphology is particularly difficult in low‐grade MDS. Thus, the role of myeloid nuclear differentiation antigen (MNDA) and other flow cytometric (FCM) parameters in MDS was evaluated.

Absence of CD9 expression in acute myeloid leukemia: possible correlation with t(8;21).

Sir, Recognition of novel immunophenotypic associations with the genetically and prognostically distinct subgroups of acute leukemia is desirable as they can provide valuable addition to the diagnostic algorithms and also help in minimal residual disease monitoring. CD9 is a tetraspanin molecule that regulates integrin-mediated functions such as adhesion and motility [1]. It has been shown earlier that low expression or absence of CD9 on leukemic cells fairly accurately predicts t(12;21) (ETV6RUNX1) in B-cell acute lymphoblastic leukemia (B-ALL) [2]. Similar to ALL with t(12;21), RUNX1-RUNX1T1 (previously AML1-ETO) fusion in acute myeloid leukemia (AML) with t(8;21) results in repression of normal RUNX1 gene [3]. A possibility of RUNX1 having a regulatory role in CD9 expression has been raised earlier [4]. We analyzed our AML cases to ascertain whether differential CD9 expression can discriminate AML with t(8;21), which is associated with a favorable outcome, from other AML subtypes. Sixty consecutive cases of AML over a 6-month period were retrospectively analyzed for immunophenotypic expression of CD9 and correlated with the cytogenetic and molecular analysis data. The patient age ranged from 2 months to 74 years (median 11 years). The median age of 11 years in our study group may indicate referral bias from our pediatric oncology unit. There were 40 males and 20 females (M : F 2 : 1). Routine immunophenotyping was performed for CD9 expression on blasts along with CD13, CD33, CD117, MPO, CD15, CD65, CD11b, CD36, CD19, CD10, CD79a, CD38, cCD3, CD2, CD4, CD7, CD34, CD45, HLA-DR, CD56, CD14, and CD64 using FC-500 (Beckman Coulter) flow cytometer. Additional markers such as CD18, CD41, and CD61 were used in selected cases. Blasts were gated as CD45 dim events with low side scatter (SSC) on CD45/SSC plot [5]. CD9 median fluorescence intensity (MFI) in the blasts was documented. CD9 positivity was defined as expression in at least 20% gated cells, using internal negative controls (lymphocytes gated on CD45/SSC plot). As per the AML classification (WHO, 2008) [3], 17 of 60 cases were AML with recurrent genetic abnormalities. This included 8 cases with t(8;21)(q22;q22), seven cases with t(15;17) (q22;q12), and two cases with inv(16)(p13.1q22). One case was classified as AML with myelodysplasia-related changes. The remaining 42 cases belonged to AML, not otherwise specified category. Cytogenetic and/or molecular analysis regarding common recurrent cytogenetic abnormalities—PML-RARA, RUNX1-RUNX1T1, and CBFBMYH11—was performed as per the methods described earlier [6]. Cases with absence of common recurrent cytogenetic abnormalities mentioned above were grouped along with the patients with normal cytogenetics. So finally, the cases were divided as normal cytogenetics (n = 37), t(8;21)(q22;q22) (n = 8), t(15;17)(q22; q12) (n = 7), inv(16)(p13.1q22) (n = 2), -Y (n = 2), trisomy 8 (n = 1), t(7;11)(p15;p15) (n = 1), hyperdiploidy (n = 1), and complex karyotype (n = 1). Statistical analysis was performed using SPSS, version 20, Fischer’s exact test, and Mann–Whitney U test, and P < 0.05 was considered significant. Among the various cytogenetic subgroups, CD9 was absent in 6/8 (75%) cases (P = 0.011) with t(8;21)(q22; q22) (Figure 1), 9/37 (24.3%) patients with normal cytogenetics, 1/7 (14.3%) with t(15;17)(q22;q12), and none of the two cases with inv(16)(p13.1q22). Two of the other six cases (33.3%) were also negative for CD9 (Table 1). CD9 MFI difference between cases with t(8;21) (q22;q22) and normal cytogenetics was not significant (Table 1). Fusion protein ETV6-RUNX1 in B-ALL with t(12;21) exerts a dominant negative effect on normal RUNX1 function [7]. These cases also have absence of CD9 expression, possibly indicating a direct regulatory role of RUNX1 in CD9 expression. The differential CD9 expression in B-ALL with t(12;21) has also formed the basis of proposed diagnostic algorithm to classify childhood B-ALL into genetically distinct entities in an accurate, cost-effective, and less time-consuming manner [8]. Similar to precursor B-ALL with t(12;21), AML with the t (8;21) has RUNX1 fused with RUNX1T1, resulting in suppression of RUNX1 function. In the study by Racke et al.,