Defect in mitochondrial functions in damaged human mitral valve

Mitochondrial diseases are a heterogeneous group of disorders in which a primary mitochondrial dysfunction is proven by morphological, biochemical, and genetic examinations. The mitral valve has important function in the regulation of blood flow from one chamber to another. Often, the mitral valve becomes abnormal with age, in Rheumatic fever or it is abnormal from birth (Congenital) or it can be destroyed by infection i.e. bacterial endocarditis and needs replacement. Myocardial function depends on energy produced by mitochondria and in any of these disease conditions, mitochondrial functions and enzyme activities may be impaired. With this in view, we analyzed the relationship between preoperative clinical conditions (as per New York heart Association) and extent of mitochondrial enzyme activities in damaged Human mitral valve in two types of heart disease such as Rheumatic Heart Disease (RHD) and Bacterial Endocarditis (BE). Thirty nine Patients undergoing cardiopulmonary bypass (CPB) for routine valvular heart surgery were included in the study. Controls included 11 normal porcine mitral valve samples without any evidence of heart disease. Mitochondrial enzymes like cytochrome oxidase (COX), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), citrate synthase (CS) and ATPase were determined. Mitochondrial COX, SDH, CS and Total ATPase activities were significantly decreased in disease condition like BE and RHD when compared with control (P<0.001). On the other hand as per New York Heart Association (NYHA) preoperative clinical classification, all the mitochondrial enzymes were significantly (p<0.05) impaired in class IV as compared with NYHA class I, II and III. Present study shows that impairment in the mitochondrial enzymes activities are more pronounced in bacterial endocarditis (BE). It also indicates that damage to mitochondrial enzymes are most pronounced in NYHA class IV.     

A study of thyroid status in hyperemesis gravidarum

Study was undertaken to assess thyroid status in hyperemesis gravidarum. 150 women pregnant with <20 weeks of gestation were selected randomly and out of these 100 women presenting with hyperemesis formed study group while 50 normal pregnant women served as controls. 53% of hyperemetic pregnant women were primigravidae and 82% of pregnant women presented with vomiting at less than 12 weeks of gestation. Statistically significant, 22% of hyperemetic women had increased serum T₃ levels while T₄ levels were increased in 67% of women in study group as compared to 8% and 16% respectively in control group. TSH levels were decreased in 18% of hyperemetic women as compared to 8% in control group with decrease in mean TSH level statistically significant. 22% of hyperemetic women had electrolyte disturbances and 7% were ketonuric. In clinically euthyroid women, biochemically altered thyroid function can attribute to vomiting and its prolongation to second trimester     

Isolation ofMycobacterium Tuberculosis 31 kDa antigen protein of diagnostic interest from culture filtrate using anti-ES-31 antibody by affinity chromatography

Proteins secreted into the culture medium byMycobacterium tuberculosis (M. tb) are shown to be source of antigens of immunodiagnostic interest. Anin vitro released 31 kDa antigen ESAS-7F isolated fromM.tb H₃₇Ra culture filtrate by salt precipitation, SDS-PAGE and cation exchange fast protein liquid chromatography (FPLC) was shown earlier to be a diagnostically important antigen fraction. In this report, we describe the isolation of ESAS-7F antigen using monospecific antibody coupled to sepharose CL-4B column. The percentage recovery of ESAS-7F antigen using affinity chromatography was approximately 8% of the total ES antigen proteins compared to 0.05% obtained by conventional purification steps using salt precipitation, SDS-PAGE and FPLC. Similar seroreactivity was observed by the antigen isolated by both the methods in indirect ELISA. Affinity chromatography helped in an increased recovery of ESAS-7F antigen and obviates the need for time consuming conventional purification steps.     

Bilirubin Interference in Serum Creatinine Estimation by Jaffe’s kinetic Method and Its Rectification in Three Different Kits

The negative interference by bilirubin in serum creatinine estimation by Jaffe’s kinetic method is well known. Several approaches have been suggested to overcome this interference. In this article three different creatinine kits (Jaffe’s kinetic method) have been tested for bilirubin interference and its rectification using two simple approaches. The performance of three kits (A, B and C) supplied by three different manufacturers was tested using IQC and EQAS sera and pooled serum with added bilirubin. To overcome the bilirubin interference two approaches viz. NaOH preincubation and TCA precipitation were used. Bilirubin did interfere in creatinine estimation after a certain level (2.3 mg/dl). However, both NaOH preincubation and TCA precipitation approach rectified this interference. The performance of kit A was better than kit B and C. All the three kits showed bilirubin interference upon increasing the bilirubin concentration but kit A performed better than kit B and C. However, NaOH incubation and TCA precipitation methods overcame this interference to a great extent.     

Autoimmune thyroid disorders—An update

Background: Autoimmune thyroid disease (AITD), a common organ specific autoimmune disorder is seen mostly in women between 30–50 yrs of age. Thyroid autoimmunity can cause several forms of thyroiditis ranging from hypothyroidism (Hashimoto’s thyroiditis) to hyperthyroidism (Graves’Disease). Prevalence rate of autoimmune mediated hypothyroidism is about 0.8 per 100 and 95% among them are women. Graves’ disease is about one tenth as common as hypothyroidism and tends to occur more in younger individuals. Both these disorders share many immunologic features and the disease may progress from one state to other as the autoimmune process changes. Genetic, environmental and endogenous factors are responsible for initiation of thyroid autoimmunity. At present the only confirmed genetic factor lies in HLA complex (HLA DR-3) and the T cell regulatory gene (CTLA 4). A number of environmental factors like viral infection, smoking, stress & iodine intake are associated with the disease progression. The development of antibodies to thyroid peroxidase (TPO) thyroglobulin (TG) and Thyroid stimulating hormone receptor (TSH R) is the main hallmark of AITD. Circulating T Lymphocytes are increased in AITD and thyroid gland is infiltrated with CD4+ and CD8+ T Cells. Wide varieties of cytokines are produced by infiltrated immune cells, which mediate cytotoxicity leading to thyroid cell destruction. Circulating antibodies to TPO and TG are measured by immunofluorescense, hemagglutination, ELISA & RIA. TSHR antibodies of Graves’ disease can be measured in bioassays or indirectly in assays that detect antibody binding to the receptor.     

Standardization of a colorimetric method for the determination of fructose using o-cresol: Sulphuric acid reagent

Fructose developed a pinkish orange chromogen on treatment with o-cresol: 70% sulphuric acid at 32°C for 15 minutes with a λ max of 500nm. Fructose was 185 times more chromogenic than glucose. Total carbohydrate and fructose values in protein-free filtrate of normal serum samples were in the range, 55.4–86.3 mg/dl and 1.55–3.29 mg/dl, respectively. In diabetes, the observed values were 197–354 mg/dl and 2.91–6.81 mg/dl, respectively.     

Biochemical markers for alcohol consumption

A variety of laboratory tests are available to assist in the diagnosis of alcohol consumption and related disorders. The levels of intake at which laboratory results become abnormal vary from person to person. Laboratory tests are particularly useful in settings where cooperativeness is suspected or when a history is not available. Several biochemical and hematological tests, such as γ-glutamyltransferase (GGT) activity, aspartate aminotransferase (AST) activity, high-density lipoprotein cholesterol (HDL-C) content of serum, and erythrocyte mean corpuscular volume (MCV) are established markers of alcohol intake. Their validity as markers is based largely on correlations with recent intake at a single time point and on decreases in elevated values when heavy drinkers abstain from alcohol. These readily available laboratory tests provide important prognostic information and should be integral part of the assessment of persons with hazardous alcohol consumption. There are several other markers with considerable potential for more accurate reflection of recent alcohol intake. These include carbohydrate deficient transferrin, β-hexosaminidase, acetaldehyde adducts and the urinary ratio of serotonin metabolites, 5-hydroxytryptophol and 5-hydroxyindoleacetic acid. These markers provide hope for more sensitive and specific aids to diagnosis and improved monitoring for intake.     

Should we use carbohydrate deficient transferrin as a marker for alcohol abusers?

Carbohydrate deficient transferrin (CDT) is one of the conventional markers for chronic alcohol consumption, is used by researchers and clinicians. A number of enzymes are affected by ethanol intake. The induction or inhibition of sialyl transferase and plasma sialidase may be involved in the CDT level elevation. An alteration of protein transport during post-translational modification could be a primary mechanism in the impairment of protein metabolism associated with chronic alcohol abuse. Transferrin being a steroid responsive protein, sex-based hormonal variations might contribute to the lower sensitivity of CDT. Varying hormonal statuses such as pregnancy, use of contraceptives, menopause/ menstrual cycle can alter iron homeostasis in women. CDT levels are markedly affected by iron homeostasis. Several CDT assay methods appeared promising, but it is not readily apparent which technique is the most accurate. Moreover, false-positive results of CDT have been reported in non-alcohol related hepatic failure and in rare conditions. Therefore clinical interpretation of CDT needs careful assessment in patients with alcohol-related or non-alcohol-related health disorders.     

Role of ethanol on aluminum induced biochemical changes on rat brain

Aluminum and alcohol, both are well-accepted neurotoxin. The plausible mechanisms for their neurotoxicity are also common. Therefore, the effect of ethanol on aluminum induced biochemical changes in rat brain is being studied. In the present study, ethanol exposure significantly affected the aluminum and protein content of brain. The activities of acid phosphatase and alkaline phosphatase were also changed. Aluminum exposure, on the other hand, contributed significantly in the alterations of aluminum content, acid phosphatase acivity and aspartate aminotransferase activity. Though ethanol co-exposure significantly influenced the aluminum load of brain, the interactions of these two neurotoxins were found to be significant only in case of acid phosphatase activity of brain. Therefore, it can be suggested that general neurotoxicity produced by aluminum is not modified by ethanol. However, the aluminum load caused by aluminum exposure, may be influenced by ethanol co-exposure.