Automated detection of premature delivery using empirical mode and wavelet packet decomposition techniques with uterine electromyogram signals.

An accurate detection of preterm labor and the risk of preterm delivery before 37 weeks of gestational age is crucial to increase the chance of survival rate for both mother and the infant. Thus, the uterine contractions measured using uterine electromyogram (EMG) or electro hysterogram (EHG) need to have high sensitivity in the detection of true preterm labor signs. However, visual observation and manual interpretation of EHG signals at the time of emergency situation may lead to errors. Therefore, the employment of computer-based approaches can assist in fast and accurate detection during the emergency situation. This work proposes a novel algorithm using empirical mode decomposition (EMD) combined with wavelet packet decomposition (WPD), for automated prediction of pregnant women going to have premature delivery by using uterine EMG signals. The EMD is performed up to 11 levels on the normal and preterm EHG signals to obtain the different intrinsic mode functions (IMFs). These IMFs are further subjected to 6 levels of WPD and from the obtained coefficients, eight different features are extracted. From these extracted features, only the significant features are selected using particle swarm optimization (PSO) method and selected features are ranked by Bhattacharyya technique. All the ranked features are fed to support vector machine (SVM) classifier for automated differentiation and achieved an accuracy of 96.25%, sensitivity of 95.08%, and specificity of 97.33% using only ten EHG signal features. Our proposed algorithm can be used in gynecology departments of hospitals to predict the preterm or normal delivery of pregnant women.

Metabolic intervention of aflatoxin B1 toxicity by curcumin.

ETHNO PHARMACOLOGICAL RELEVANCE: Curcumin, bioactive principle of turmeric (Curcuma longa Linn) is an important constituent of Indian traditional medicine. Turmeric has been known to possess several therapeutic properties. AIM OF THE STUDY: The modulatory effect of dietary curcumin (0.05%, w/w) on drug metabolizing and general marker enzymes of liver and formation of AFB(1)-adducts (DNA and protein) due to dietary AFB(1) exposure for a period of 6 weeks in a rodent model, have been evaluated. MATERIALS AND METHODS: Drug metabolizing enzymes CYP1A1, GSHT, UGT1A and general marker enzymes (LDH, ALT, AST, ALP and gamma-GT) of liver were estimated by standardized methods. Aflatoxin adducts (DNA and protein) were quantitated by indirect competitive ELISA. RESULTS: Dietary curcumin enhanced GSHT (p<0.001) and UGT1A1 (p<0.05) activity and significantly reduced the activity of CYP1A1 (p<0.001), in rats exposed to aflatoxin B(1). Supplementation of curcumin in the diet normalized the altered activities of LDH and ALT. At molecular level, curcumin significantly reduced AFB(1)-N(7)-guanine adduct (p<0.001) excretion in the urine, DNA adduct (p<0.05) in the liver and albumin adduct (p<0.001) in the serum. CONCLUSION: The experimental results substantiates that curcumin intervention ameliorates the AFB(1) induced toxicity.

Synthesis and characterization of mercapturic acid (N-acetyl-L-cysteine)-aflatoxin B1 adduct and its quantitation in rat urine by an enzyme immunoassay.

A simple and sensitive indirect noncompetitive enzyme immunoassay to quantitate mercapturic acid-aflatoxin B1 (AFB1) adduct in rat urine is reported. A novel procedure was developed for in vitro synthesis of an immunogen, bovine serum albumen-glutathione-aflatoxin B1 (BSA-GSH-AFB1) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride. Sulphydryl group's analysis confirmed the conjugation of-SH groups to AFB1. Thin-layer chromatography and spectral analysis (absorption, fluorescence, and Fourier transform infrared) of the conjugates further confirmed the formation of the adducts. Polyclonal antibodies specific to mercapturic acid-AFB1 adduct were produced against BSA-GSH-AFB1. The assay was found to be linear in the range of 100 pg-100 ng of the analyte (y = a + bx). A 50% displacement of BSA-GSH-AFB1 antibodies was achieved at an inhibitory concentration (IC50) of 11.9 ng GSH-AFB1 (r2 = 0.98) and 1.22 ng N-acetyl-L-cysteine (NAC)-AFB1 (r2 = 0.98). Spiking 5 microg/mL of reference standard to the control rat urine showed a recovery of 98 +/- 2%. The immunoassay was validated in a rodent model exposed to a single oral dose of 1 mg/kg body mass of pure AFB1. The excretion of NAC-AFB1 adduct was quantitated at the end of 24 h. The concentration of the NAC-AFB1 adduct excreted in urine as determined by the immunoassay was found to be in the range of 3.22-5.97 microg/mg creatinine. The present method may find wide application as a biochemical tool in molecular epidemiological and intervention studies with respect to human exposure to dietary aflatoxins.

Immunochemical detection of glycated lens crystallins and their circulating autoantibodies in human serum during aging.

PURPOSE: The aim of this investigation was to exploit lens-specific glycated crystallins as an immunogen to detect human glycated crystallins and their circulating autoantibodies in human serum during aging in relation to the development of cataract. METHODS: Polyclonal antibodies were produced against human total lens proteins (40-80 years) in rabbits. The specificity of the antibodies produced were determined by antibody capture assay using purified human lens crystallins (high molecular weight fraction [HMW]+alpha, HMW+alpha-glycated, beta, beta-glycated, gamma, and gamma-glycated) as antigens. The cross-reactivity of these lens specific antibodies against rat beta-, beta-glycated, gamma-, and gamma-glycated lens crystallins was also analyzed. A non-competitive enzyme linked immunosorbent assay (ELISA) methodology was developed for the detection of circulating lens crystallins in human sera using HMW+alpha, HMW+alpha-glycated, beta-, and beta-glycated crystallins from humans and gamma- and gamma-glycated crystallins from rats as immobilized antigens. Circulating autoantibodies were also detected in human sera by antibody capture assay. The methodology was validated by evaluating 60 human serum samples collected from cataract patients and 30 human serum samples from apparently normal subjects belonging to the same age group. RESULTS: The polyclonal antibodies raised against human total lens proteins showed 90% and 65% cross-reactivity with rat gamma- and beta-crystallins, respectively, by ELISA. Further, these polyclonal antibodies were capable of detecting both native and in vitro synthesized glycated crystallins. Their IC50 values were observed to be (i) human total lens proteins (55 ng), (ii) human HMW+alpha (16.45 ng), (iii) human HMW+alpha-glycated (273 ng), (iv) human beta- (37.82 ng), (v) human beta-glycated (260 ng), (vi) rat gamma- (105.34 ng), and (vii) rat gamma-glycated (313 ng). The immunochemical analysis of human serum indicated a significant change (p<0.001) in the levels of circulating beta-glycated and gamma-glycated crystallins in the age group of 40-80 years with respect to their control groups. However, there was no statistically significant change in the levels of HMW+alpha-glycated crystallins in the age group of 40-80 years as compared to their age-matched controls. Notably, the levels of serum gamma-glycated crystallins were found to be threefold higher than that of HMW+alpha-glycated and beta-glycated crystallins in the age group of 70-80 years. Circulating autoantibodies to HMW+alpha-glycated, beta-glycated, and gamma-glycated crystallins were detected in the serum of both apparently normal and cataract patients in the age group of 40-80 years by antibody capture assay. The levels of these autoantibodies were significantly higher at every time point compared to their respective controls. Autoantibodies to gamma-glycated crystallins were found to be twofold and 3.2 fold higher as compared to the levels of autoantibodies to beta-glycated and HMW+alpha-glycated crystallins, respectively. Western blot and immunohistochemical analysis substantiated the observations made in non-competitive ELISA. CONCLUSIONS: During the course of aging, leakage of lens crystallins (HMW+alpha, HMW+alpha-glycated, beta, beta-glycated, gamma, and gamma-glycated) elicit an immune response resulting in the formation of autoantibodies in cataract patients (40-80 years) as compared to age matched controls. This is the first experimental report where polyclonal antibodies raised against lens-specific glycated crystallins were capable of detecting the early leakage of glycated crystallins in human subjects. This immunochemical approach has implications in the early detection of senile cataract.

Immunochemical detection of glycated beta- and gamma-crystallins in lens and their circulating autoantibodies (IgG) in streptozocin induced diabetic rat.

PURPOSE: This study used an immunochemical approach aimed to detect the glycated crystallins (beta- and gamma-crystallin) in rat lens and their circulating specific autoantibodies in serum during the course of cataractogenesis. METHODS: Streptozocin (STZ; 55 mg/kg body mass) induced diabetic male Wistar/NIN rats (2-3 months old) and control nondiabetic rats were used for this study. Plasma glucose, glycated hemoglobin and body weight were evaluated on day zero, and at the interval of every two weeks up to the eighth week of post-injection in both the groups. Other biochemical parameters, such as the levels of nonprotein sulfhydryl (-SH) groups and the activity of gamma-glutamyl transpeptidase (gamma-GT) in lens proteins were also estimated. Cataract progress was monitored by measuring the advanced glycation end product (AGE)-like fluorophores in both intact lens as well as in lens homogenate employing digital based image analysis and spectrofluorimetric methods. Similarly, the polyclonal antibodies specific to beta-glycated-, gamma-glycated-, beta-, and gamma-crystallins were used to determine the concentration of respective immunogens in lens by noncompetitive ELISA and their respective circulating antibodies by antibody capture assay. The profile of glycated lens protein (soluble and insoluble fractions) during the course of cataractogenesis was assessed by the western blot technique. RESULTS: STZ induced diabetic rats showed typical signs of diabetes (hyperglycemia, increased water and food intake with no increase in body weight). Biochemical analysis of total lens protein showed a significant (p = <0.001) decrease in the levels of nonprotein -SH groups. The activity of lenticular gamma-GT in diabetic rats was found to be unaltered as compared to the control group. Digital analysis of intact lens illustrated a positive correlation (r(2)=0.888) with the formation of AGE-like fluorophores during the course of cataractogenesis. A similar trend was also observed in the levels of AGE-like fluorophores in the total lens homogenate of diabetic animals during the course of cataractogenesis. The concentration of beta- and gamma-glycated-crystallins in the rat lens (soluble and insoluble fractions) was analyzed by non-competitive ELISA. The concentration of beta- and gamma-glycated-crystallins were found to be enhanced by the end of week eight, as compared to the control group. Concomitantly, crystallin-specific (beta- and gamma-glycated-crystallin) autoantibodies were also detected in the serum of the diabetic rats from week two onwards. Western blot analysis indicated the formation of enhanced glycated lens crystallins (beta- and gamma-crystallin) in the insoluble fraction. CONCLUSIONS: The following was observed during the course of cataractogenesis: (1) there was an enhanced formation of AGEs-like fluorophores in intact lens; (2) beta- and gamma-glycated-crystallin levels increased in the rat lens (insoluble fraction) by the end of week eight; and (3) release of these glycated lens proteins into peripheral circulation resulted in the production of autoantibodies to beta- and gamma-glycated-crystallins that could be detected as early as week two, after induction of diabetic status in experimental rats.

Seizure anticipation in pediatric epilepsy: use of Kolmogorov entropy.

The purpose of this paper is to demonstrate feasibility of using trends in Kolmogorov entropy to anticipate seizures in pediatric patients with intractable epilepsy. Surface and intracranial recordings of preseizure and seizure activity were obtained from five patients and subjected to time series analysis using Kolmogorov entropy. This metric was compared with correlation dimension and power indices, both known to predict seizures in some adult patients. We used alarm levels and introduced regression analysis as a quantitative approach to the analysis of trends. Surrogate time series evaluated data nonlinearity, as a precondition to the use of nonlinear measures. Seizures were anticipated before clinical or electrographic seizure onset for three of the five patients from the intracranial recordings, and in two of five patients from the scalp recordings. Anticipation times varied between 2 and 40 minutes. This is the first report in which simultaneous surface and intracranial recording are used for seizure prediction in children. We conclude that the Kolmogorov entropy and power indices were as effective as the more commonly used correlation dimension in anticipating seizures. Further, regression analysis of the Kolmogorov entropy time series is feasible, making the analysis of data trends more objective.

Development of an immunoanalytical method for the detection of beta- and gamma-crystallins and anti-crystallin antibodies. A molecular biomarker for cataract.

PURPOSE: To develop and evaluate an immunoanalytical method for the detection of beta- and gamma-crystallins and anti-crystallin antibodies. MATERIALS AND METHODS: Beta and gamma-crystallins isolated from rat lens were used as immunogens to raise polyclonal antibodies in rabbits. Antibody capture assay and western blot analysis showed that the antibodies to beta- and gamma-crystallins were specific. An indirect competitive enzyme linked immunosorbent assay (ELISA) developed to quantitate beta- and gamma-crystallin showed an IC50 value of 70 ng and 65 ng, respectively, based on regression analysis. Spiking studies with purified beta-crystallin antibodies showed that 33 ng of the purified antibody gave an absorbance of 1.1 at 450 nm, indicating the sensitivity of the method. RESULTS: Antibodies to beta- and gamma-crystallins were not detected in serum samples of the cataractous CFY/NIN rats (used as an animal model for induction of experimental cataract by feeding high galactose diet). However, the cataractous rat serum samples effectively displaced beta- and gamma-crystallin antibodies, indicating that these crystallins leak during cataract formation. The concentration of beta- and gamma-crystallins in the rat serum, as analysed by indirect competitive ELISA, was found to be in the range of 17.6-81.6 micrograms/ml [corrected] and 12.4-19.6 micrograms/ml, respectively. CONCLUSIONS: The methodology developed in the present study may find application as a biochemical tool in molecular epidemiology of cataract.