Experience from an immunohematology reference testing center.

Blood Centres in India lack infrastructure to investigate immunohematology problems. Reference Testing Center (RTC) was established in 2014 to investigate Immunohematological problem as it is not possible for small blood centers to go for complete immunohematology work up due to lack of financial and technical resources in remote and rural areas. Objective of this study is to share our experience as RTC of past 6 years so that more RTC are established across Indian subcontinent. 1456 Discrepant samples received from various hospitals of South India for Immunohematology problems were analysed in 6 years. Maximum requisitions obtained in 2014 were more than 40 years of age and then 21-40 years of age group in 2015 and same was observed till 2020.75% of total samples received were for antibody identification followed by blood group discrepancy resolution, investigation of positive DAT, red cell phenotype and pre-natal evaluation & antibody titration. Single allo-antibodies were identified in 773 cases whereas multiple allo-antibodies were found in 118 cases. Most common single and multiple antibody found was anti D and Anti-D+C. Weak D subgroup was the most common blood group discrepancy.22 cases & 4 cases of Bombay and para-bombay were also investigated.

Study of prevalence of endocrine abnormalities in primary empty sella.

INTRODUCTION: Empty sella is usually an incidental finding. Empty sella is an anatomical condition characterized by the presence of cerebrospinal fluid within the sella with a small pituitary gland compressed above the pituitary floor causing endocrine abnormalities. AIM: The aim of this study is to evaluate hormonal abnormalities associated with empty sella in tertiary care center. MATERIALS AND METHODS: This ongoing study was carried out in patients attending to endocrine out-patient departments from August 2012 to July 2013. A detailed history and examination was done. Hormonal evaluation including free thyroid hormones, thyroid stimulating hormone, growth hormone (GH), follicular stimulating hormone, luteinizing hormone, cortisol and prolactin was done. RESULTS: A total of 16 patients diagnosed clinically and biochemically of hormonal abnormalities were found to have empty sella on magnetic resonance imaging. Hypocortisolemia in 62.5% of cases, hypothyroidism in 50% of cases, in 18.75% hypogonadism, hyper prolactinemia in 18.7.5%, GH deficiency in 12.5% of cases and in 12.5% cases posterior pituitary involvement is seen. CONCLUSION: The high incidence of endocrine abnormalities associated with empty sella necessitates the need for prompt evaluation and early replacement of hormones for better quality-of-life.

Seizure as a presenting manifestation of vitamin D dependent rickets type 1.

There are two types of vitamin D dependent rickets (VDDR) that cause rickets in children. VDDR type 1 (VDDR-I) is caused by an inborn error of vitamin D metabolism, which interferes with renal conversion of calcidiol (25OHD) to calcitriol (1,25(OH)2D) by the enzyme 1-α-hydroxylase. Patients with VDDR-I have mutations of chromosome 12 that affect the gene for the enzyme 1-α-hydroxylase, resulting in decreased levels of 1,25(OH) vitamin D. Clinical features include growth failure, hypotonia, weakness, rachitic rosary, convulsions, tetany, open fontanels and pathologic fractures. We report a case of VDDR-I in 14-month-old male child. Establishing an early diagnosis of these genetic forms of rickets is challenging, especially in developing countries where nutritional rickets is the most common variety of the disease where genetic diagnosis is not always possible because of financial constraints. A prompt diagnosis is necessary to initiate adequate treatment, resolve biochemical features and prevent complications, such as severe deformities that may require surgical intervention.

Studies on salts of 3-nitro-1,2,4-triazol-5-one (NTO) and 2,4,6-trinitroanilino benzoic acid (TABA): potential energetic ballistic modifiers.

The Co/Cu/Ni/Fe salts of 3-nitro-1,2,4-triazol-5-one (NTO) and 2,4,6-trinitroanilino benzoic acid (TABA) were prepared and characterized during this work. All the salts exhibited exothermic decomposition in DSC. The FT-IR spectra of the gaseous products evolved during TGA of NTO salts indicated the release of NO2 and cleavage of NTO ring during the course of decomposition. Thermal decomposition of TABA salts also produced NO2 on decomposition. The transition metal salts enhanced the burning rates of AP-HTPB composite propellant evaluated during this work. The best catalytic effect was obtained with Fe-NTO salt which increased the burning rate to the extent of approximately 80% as well as brought down the pressure index (n) to 0.18 (2-9MPa).

Domestic cooking fuel and lung functions in healthy non-smoking women.

OBJECTIVE: The objective of this study was to compare the pulmonary functions in healthy non-smoking women who used either biomass or liquified petroleum gas (LPG) as their sole cooking fuel. The effects of passive smoking, ventilation, over crowding and cooking index were also taken into account. METHODOLOGY: The study was conducted over a period of two years from January 1994. One hundred healthy non-smoking women were included 50 cooked solely with biomass and 50 cooked with LPG. A standardised respiratory symptoms questionnaire was administered to all the subjects and spirometry was carried out. RESULTS: Passive smoking showed no significant difference between the two groups. No statistically significant differences was found in lung functions in the two groups except for the PEFR, which was significantly lower (P < 0.01) in women using biomass. No correlation was observed between different variables and pulmonary functions. The step-wise multivariate linear regression analysis showed no correlation between cooking fuel and the pulmonary functions. CONCLUSION: The absence of the expected adverse effects of biomass on pulmonary functions was possibly due to better ventilation in the kitchens of subjects in the biomass group compared to previous studies.

Sleep disordered breathing in chronic obstructive pulmonary disease.

We prospectively studied sleep disordered breathing in 50 consecutive patients (39 males) with chronic obstructive pulmonary disease (COPD) with chronic respiratory failure (CRF) (n=33) and without CRF (n=17) by performing polysomnography. Patients with CRF had a lower mean nocturnal oxygen saturation (SaO2 %) (88.6+/-6.7 vs. 96.3+/-0.8; p=0.0001) and a lower minimal nocturnal SaO2 (73.6+/-12.0 vs. 84.3+/-7.3; p=0.002) compared to those without CRF, suggesting that patients with CRF tend to have more severe drops in nocturnal SaO2. Patients with CRF also had a lower FEV1 (% predicted) (p=0.01) and PEFR (% predicted) (p=0.031) compared to those without CRF suggesting an indirect relation to the oxygen saturation. Other pulmonary functions were comparable between both the groups. Among patients with and without CRF, the total sleep time (minutes); the rapid eye movement (REM) stage (% of total sleep time); the non-rapid eye movement (NREM) stage (% of total sleep time) were comparable (p=NS). Only three of the 50 patients with COPD had a significant (>5) apnea-hypopnea index (AHI) (total no. of apneas + total no. of hypopneas/ total sleep time [(hours) = AHI] and these three patients had a mean BMI = 27.7 which was higher than the mean BMI of the whole group (21.1). The AHI was comparable in patients with and without respiratory failure. Multiple regression analysis revealed a positive correlation between AHI and the neck circumference (r=0.41; p=0.005) and BMI (r=0.31; p=NS). There was a small but statistically insignificant negative correlation between AHI and neck length (r= -0.28; p=NS). We conclude that, BMI per se contributes to the AHI and nocturnal desaturation in patients with COPD.

Primary meningococcal pneumonia in elderly patients.

Neisseria meningitidis infection in humans usually manifests as meningitis and septicemia with skin manifestations. Infections of the respiratory tract with N meningitidis have been documented in the past, but often this organism is not routinely considered in the differential diagnosis of pneumonia. The pathogenic role of N meningitidis in lower respiratory tract infections may be underestimated because its isolation is difficult, particularly when oropharyngeal flora are present. We profile 2 elderly patients with primary meningococcal pneumonia to show the importance of Gram stain and culture in early diagnosis. These modalities helped guide treatment and prophylactic measures.

Distribution and localization of monophosphoryl lipid A in selected tissues of the rat.

The distribution and cellular localization of Monophosphoryl Lipid A in the kidney, liver, lung, spleen, and stomach of rat were investigated by immunohistochemistry on paraffin embedded tissue sections. Rats were sacrificed 24h or 48h following intraperitoneal administration of MPL at a dose of 5mg/Kg. The presence of MPL in selected tissues was indicated by a positive reaction to MPL antibody. In the kidneys, MPL was found in collecting tubules and distal convoluted tubules in the medulla, whereas the glomerulus was essentially free of it. Regarding liver, MPL was found to be abundant in hepatocytes but only occasionally present in Kupffer cells. In lungs, both alveolar and bronchiolar macrophages were positive, indicating the lung can also serve as a possible site for the elimination of MPL. In spleen, endothelial cells and macrophages were positive for MPL. In stomach, MPL was detected in the gastric mucosa and vascular endothelial cells. In all the tissues studied the intensity of the peroxidase reaction was significantly weaker in 48h samples as compared to corresponding 24h samples indicating possible elimination of MPL from these tissues.

Rapid and sensitive spectrophotometric determination of cerium(IV) with 2,4-dihydroxy benzophenone benzoic hydrazone.

A simple and sensitive spectrophotometric method for the determination of cerium(IV) in an aqueous medium is reported. The metal ion formed a 1:1 orange-red coloured complex with 2,4-dihydroxy benzophenone benzoic hydrazone (DHBPBH) at pH 10.0 showing an absorption maximum at 400 nm. The molar absorptivity and Sandell's sensitivity of the method are found to be 2.0 x 10(4)1/mol/cm and 0.007 mug/cm(2), respectively. Beer's law is obeyed in the range 0.7-7.0 mug/ml. Titanium, vanadium and molybdenum do not interfere. The extent of interferences by other ions is presented. The method is applied for the determination of cerium in simulated rock samples.

Rapid and selective spectrophotometric determination of manganese in steels and alloys using resacetophenone oxime.

A sensitive spectrophotometric method is developed for the determination of manganese in aqueous medium. The metal ion forms a yellowish brown coloured complex with resacetophenone oxime (RPO) in ammonium chloride and ammonium hydroxide buffer of pH 10.5. The 1:1 complex shows maximum absorbance at 380 nm with a Beer's law range of 0.09-1.7 ppm. The molar absorptivity and the Sandell sensitivity are found as 2.5 x 10(4) l.mole(-1).cm(-1) and 0.002 mug/cm(2), respectively. The stability constant of the complex calculated by Job's method is 7.5 x 10(5). The interfering effects of various cations and anions are studied. The present method is applied to the determination of manganese in some steel and alloy samples.

o-Hydroxyacetophenone thiosemicarbazone as a reagent for the rapid spectrophotometric determination of palladium.

o-Hydroxyacetophenone thiosemicarbazone has been synthesized and employed as a new reagent for the spectrophotometric determination of palladium(II), which forms two complex species with it in aqueous dimethylformamide at pH 6.0, these having 1:1 and 1:2 metal-ligand ratios. The Job and molar-ratio plots have an unusual shape that is due to the stepwise conversion of the 1:1 complex into the 1:2 species. The molar absorptivity at 370 nm is 9 x 10(3) l.mole(-1).cm(-1). Beer's law is obeyed over the range 0.42-10.6 mug/ml palladium.

Endothelial cell damage in human and rabbit corneas stored in K-Sol without antioxidants.

Human and rabbit corneas were stored at 4 degrees C in K-Sol with and without antioxidants (ascorbic acid, reduced glutathione, alpha-tocopherol, and retinol acetate) for two to three weeks. All the corneas were then examined visually and by scanning electron microscopy. They appeared clear and slightly oedematous. Scanning electron micrographs were used to grade corneal endothelial cell morphology in a masked manner in terms of cell shape, cell borders, cell swelling, and apical holes. Corneas stored in K-Sol without antioxidants showed changes in cell shape, cell borders, and apical holes. Human corneas showed more morphological changes than rabbit corneas. The results suggest that antioxidants in K-Sol have an important role in the preservation of endothelial cell morphology.

Acylation of disc membrane rhodopsin may be nonenzymatic.

Bovine retinal rod outer segments (ROS) support the incorporation of [3H]palmitate into rhodopsin. [14C] Palmitoyl-CoA serves as the donor with an apparent Km of 40 microM. Solubilization of ROS in the detergent, Emulphogene, results in increased incorporation of label into rhodopsin. A further increase is found when ConA-Sepharose-purified rhodopsin is used as the source of both "enzyme" and acceptor. Failure to separate enzyme from acceptor suggested the possibility of a nonenzymatic reaction. This was confirmed when boiled rhodopsin was found to support the reaction. However, the acylation of rhodopsin is not an artifact since analysis of purified native rhodopsin reveals the presence of covalently bound palmitate and we showed that whole bovine retinas incubated with [3H] palmitate incorporated the fatty acid into rhodopsin (O'Brien, P.J., and Zatz, M. (1984) J. Biol. Chem. 259, 5054-5057). Furthermore, in vivo experiments with rat retinas have revealed that opsin is acylated both in the rod inner and outer segments (St. Jules, R. S., and O'Brien, P.J. (1986) Exp. Eye Res. 43, 929-940). Incubation of labeled rhodopsin with mercaptoethanol resulted in release of the labeled palmitate indicating the presence of a thioester bond. This also illustrates the ease with which a thioester, such as palmitoyl cysteine or palmitoyl-CoA, can transfer the fatty acyl group to a free thiol, such as cysteine or mercaptoethanol.

Arachidonic acid, stearic acid, and diacylglycerol accumulation correlates with the loss of phosphatidylinositol 4,5-bisphosphate in cerebrum 2 seconds after electroconvulsive shock: complete reversion of changes 5 minutes after stimulation.

The effects of electroconvulsive shock (750 msec, 130 V, 150 pps) on the endogenous content of rat cerebral lipids were studied 2, 5, 10, 20, 30, 60, and 300 sec after stimulation. Rapid enzyme inactivation in situ was attained by high-power head-focused microwave irradiation (6.5 kW, 2450 MHz). At 10 sec, phosphatidylinositol 4,5-bisphosphate (PIP2) mass had decreased by 249 nmol per g wet wt, mainly due to loss of arachidonate and stearate. At the same time, the stearoyl-arachidonoyl glycerol accumulated, although to a lesser extent than the loss exhibited in PIP2. Changes in phosphatidylinositol and in phosphatidylinositol 4-phosphate mass were not statistically significant. Free fatty acids and diacylglycerols accumulated to 395 nmol per g wet wt; arachidonic and stearic acids composed 322 nmol of these lipids. Hence, the reduction in content of PIP2 is sufficient to account for 80% of the increases in free fatty acid and diacylglycerol mass. Thirty-three and 12 nmol of accumulated free palmitic and docosahexaenoic acids, respectively, are not accounted for by the loss of PIP2. Sixty seconds after stimulation, PIP2 content returned to 90% of control levels, while diacylglycerol tended to remain below control levels. Free fatty acids had not returned to control levels by 60 sec, with the exception of docosahexaenoic acid. At 300 sec, PIP2, diacylglycerol, and free fatty acids had all returned to control levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Docosahexaenoate metabolism and fatty-acid composition in developing retinas of normal and rd mutant mice.

The fatty-acid composition of retinal lipids in developing control and rd mice (C57BL/6J) was determined. In addition, fatty-acid composition in brain and retina of normal and rd adult animals was compared. At 11 days of postnatal age, rd retinas contained proportionally less docosahexaenoate than controls, whereas the reverse relationship held for oleate at 20 days of age. In contrast, no differences in the fatty-acid composition of brain lipids were observed between rd and control animals. We also examined docosahexaenoate metabolism in rd and control retinas of different postnatal ages in vitro. These studies of [1-14C]docosahexaenoic-acid incorporation into retinal phospholipids and neutral lipids demonstrated significantly higher incorporation into triacylglycerols of the rd retina, beginning at 14-15 days of postnatal age. Incorporation into diacylglycerols and phospholipids was also higher in rd retinas than in controls at 14 days of age and older. Moreover, the concentration of ganglioside (a glycolipid class probably enriched within inner retinal layers) was higher in adult rd retinas. Degeneration of the rd retina can be detected histologically as early as 8-9 days. Therefore, the alterations of fatty-acid composition and docosahexaenoate metabolism described here are probably an effect, rather than a cause, of retinal degeneration in the rd mouse.

Decreased content of docosahexaenoate and arachidonate in plasma phospholipids in Usher's syndrome.

Docosahexaenoate and arachidonate were found to be significantly decreased in plasma phospholipids from Usher's syndrome patients. The fatty acid content of plasma triacylglycerols was not changed in these patients. Usher's syndrome, an autosomal recessive disorder, involves an inherited visual cell degeneration. Photoreceptor membranes are richly endowed with docosahexaenoate and arachidonate, and a metabolic defect affecting these polyunsaturated fatty acids may occur. Moreover, blindness may be due, at least partially, to an alteration in the unsaturated phospholipids of photoreceptor membranes.

Fatty acid composition and arachidonic acid metabolism in vitreous lipids from canine and human eyes.

About 55% of the acyl groups of dog and human vitreous are unsaturated fatty acids. The major components are oleate (18:1, n-9) and arachidonate (20:4, n-6) with moderate amounts of linoleate (18:2, n-6) and docosahexaenoate (22:6, n-3). Palmitate (16:0) and stearate (18:0) are the major saturated fatty acids. There are no significant changes between ages 37-82 years in the fatty acyl group content and composition of human vitreous. In vitreous from Irish setters with hereditary rod-cone dysplasia (RCD) the levels of oleate are decreased with a concomitant increase in arachidonate. [1-14C]Arachidonic acid was actively incorporated into canine vitreous glycerolipids both in vitro and in vivo. The incorporation was mainly into phosphatidylinositol, triacylglycerol, phosphatidylcholine and phosphatidylethanolamine. There were some differences in the pattern of incorporation between human and dog and between in vivo and in vitro incubations of canine vitreous. Glycerolipid acylation was significantly increased in phosphatidylinositol and phosphatidylcholine in RCD canine vitreous. The pattern of incorporation of [U-14C]docosahexaenoic acid into vitreous glycerolipids was different from arachidonic acid incorporation. Although vitreous did not produce any measurable enzymatic synthesis of cyclooxygenase and lipoxygenase products from [1-14C]-arachidonic acid in vitro, there was significant generation of autooxidation products. These results suggest an active lipid metabolism in vitreous.

Enhanced synthesis of prostaglandins and hydroxyeicosatetraenoic acids in retina from a canine model of Batten's disease.

The metabolism of [1-14C]arachidonic acid (20:4, n-6) was studied in intact retina and retinal pigment epithelial cells from normal English setters and English setters affected with hereditary canine ceroid lipofuscinosis. Acylation of arachidonic acid into membrane glycerolipids and oxygenation by lipoxygenase and cyclooxygenase to eicosanoids were measured by radiochromatographic techniques. In addition, the histopathology of accumulated ceroid particles in retinal ganglion cells and pigment epithelial cells was studied by electron microscopy. Synthesis of prostaglandins and hydroxyeicosatetraenoic acids was increased in canine ceroid lipofuscinosis retina, but not in retinal pigment epithelium. Prostaglandin D2, the putative neuronal eicosanoid, was increased nearly eightfold, whereas other eicosanoids increased two- to threefold. Ultrastructural studies revealed accumulation of ceroid and deterioration of neuronal and pigment epithelial cell architecture. These experiments demonstrate that, although lipopigment accumulates in both tissues, alterations of eicosanoid synthesis are specific for the retina, a neuronal tissue. The specific increase in prostaglandin D2 and the specificity of changes for the retina indicate that enhanced eicosanoid synthesis may be a result of an impairment of the control of oxygenation of arachidonic acid in neurons.

Effects of neonatal undernutrition of rats on the synthesis of phosphatidylcholine and phosphatidylethanolamine by microsomes from gray matter and white matter.

Undernutrition of rats during the suckling period produces deficits of glycerophospholipids in white matter but not in gray matter. The synthetic enzymes, CDPethanolamine:diacylglycerol phosphoethanolaminetransferase (EC 2.7.8.1) and CDPcholine:diacylglycerol phosphocholinetransferase (EC 2.7.8.2), were assayed in washed microsomal fractions from moderately and severely undernourished rats at 21 days of age. Deficits in brain weight were 14 and 31%, respectively. The transferase activities in white matter were more than 2-fold greater than in gray matter. Moderate undernutrition has less effect on total transferase activities but Km values were increased for CDPethanolamine with phosphoethanolamine transferase and for CDPcholine and dicaprin with phosphocholine transferase. Severe undernutrition produced by feeding the dams a diet with 4% protein gave deficits for phosphoethanolamine transferase of 52 and 55% and for phosphocholine transferase of 40 and 46% for gray matter and white matter, respectively. These results are consistent with the deficits of glycerophospholipids in white matter. Additional factors, such as synthetic rates in vivo and a reduction of turnover rates, must be sought for the sparing of the glycerophospholipid content of gray matter during undernutrition.