Alkaptonuric ochronosis: Report of a case and brief review

Alkaptonuria, a metabolic disorder characterized by a triad of homogentisic aciduria, arthritis and ochronosis is one of the first conditions in the charter of group of inborn errors of metabolism proposed to have Mendelian recessive inheritance. It is due to the deficiency of the enzyme homogentisic acid oxidase which catalyzes the conversion of homogentisic acid to maleylacetoacetic acid in the catabolism of tyrosine. Homogentisic acid thus accumulates in cells and body fluids and its oxidized polymers bind to collagen, leading to progressive deposition of grey to bluish black pigment resulting in degenerative changes in cartilage, intervertebral disc and other connective tissues, leading to arthritis which is the only disabling effect in an affected older individual. However the diagnosis can be made in neonates when blackish stain is noticed in an unwashed diaper. Alkaptonuria is treated symptomatically, surgical intervention necessitates in advanced stages, treatment with ascorbic acid (Vitamin C) and dietary restrictions of food containing phenylalanine and tyrosine have proved to be successful in alleviating the symptoms.     

Ochronosis of the Hip Joint: Differential Diagnosis of Inflammatory Arthropathy with Lytic Lesion

Alkaptonuria or ochronosis is a rare inborn disorder of metabolism which is characterized by deficiency of homogentisic acid oxidase. There is accumulation of homogentisic acid in the connective tissues causing brownish black pigmentation and skeletal damage. The most serious complication of this disease is crippling degenerative arthropathy which presents in late years of life. There is no definitive treatment for the condition. Symptomatic management is the main stay. Surgical management such as arthroplasty is done for severe cases of ochronosis. We report a case of ochronosis of the hip joint presenting with inflammation and lytic lesion which can create confusion and lead to error in diagnosis.     

Alkaptonuria: A Case of Familial Inheritance from Hangarki Village in Dharwad District of Karnataka

Alkaptonuria is a genetic disorder, unknown to the general public and ignored by general physicians due to lack of awareness of its high prevalence. Increasing incidences of familial inheritance are recorded. A 41 years 8 months male with the swollen knee, chronic mechanical pain, restricted limping walk and tingling sensation in the limb. Also, has a complaint of the stiffness of back in the morning, which gets relieved by movement. Mild hyper pigmentation of pinna and sclera of eye. X-ray report reveals osteoarthritis of the knee and thoracic kyphosis. After considering clinical finding, report of radiological investigation and Biochemical test results, patient was diagnosed as a case of Alkaptonuria. Screening of entire family revealed a typical case of familial inheritance. Hidden familial inheritance can be disclosed by mass screening of families and medical awareness.     

Maple syrup urine disease: An uncommon cause for neonatal metabolic distress

Maple Syrup Urine Disease is an autosomal recessive disorder caused by a deficiency in the activity of the branched-chain α-ketoacid dehydrogenase complex. This rare disorder represents one of the causes of acute neonatal illness which results in devastating disturbances of neurological development. On investigation of 1750 infants with neurological impairment for inborn errors of amino acid metabolism, 4 neonates with classical maple syrup urine disease were detected. These otherwise normal neonates presented in the first week after birth with seizures, lethargy and refusal of feeds, hypoglycemia and metabolic acidosis. The plasma and urine concentrations of the branched-chain amino acids were increased and there was ketoaciduria. Two of these neonates expired before specific treatment could be instituted. Routine biochemical screening of neonates with acute illness could unearth many cases of this rare inherited metabolic disease.     

Evaluation of urinary abnormalities in nephrolithiasis patients from Marathwada region

Urinary abnormalities were evaluated in 100 renal stone patients with first episode of renal stone having age 22 to 45 years from both sex and compared to 100 normal healthy control group having same age group from both sex. Twenty-four hours urinary oxalate, calcium, uric acid, sodium, magnesium, phosphorus and citrate were estimated. The urinary pH was also determined. In stone formers urinary oxalate, calcium, sodium and uric acid excretions were significantly higher when compared with control group. Whereas citrate, phosphate and magnesium excretion were significantly lower in stone formers when compared with control. The pH of urine in stone formers was lower than the controls. High dietary intake of purine rich diet causes elevated excretion of uric acid, which leads to calcium oxalate crystal formation and precipitation. Other risk factors such as urinary oxalate, calcium also related to formation of renal calculi. Hypocitraturia is the main cause of renal calculi along with hypomagnesiuria and hypophosphaturia in the patient of Marathwada region. On the basis of urinary abnormalities further stone formation in the patient can be prevented by dietary modifications.     

Assessment of renal acidification capacity of stone formers by acid challenge test

Acid challenge test was carried out on 87 subjects (37 normal and 50 radiologically proven stone formers) to assess their renal acidification capacity. NS were given three doses viz. 75, 100 and 150 mg NH₄Cl/Kg body weight while the SF were tested with only 150 mg dose which was found to be an adequate dose. Ammonium chloride was given in gelatin capsules with breakfast and hourly urine samples were collected for next 7 hours. pH, creatinine, calcium, inorganic phosphorus, citrate, titrable acid and ammonium were analysed in all the samples. The incidence of renal tubular acidosis (RTA) in NS, tested with 75 mg and 100 mg/kg doses of ammonium chloride, was 50% and 10% respectively while it was nil and 24% respectively in NS and SF tested with 150mg/kg dose. Complete RTA was detected in one SF. No significant difference was observed in urinary profile of NS and SF except that the hourly titrable acid excretion was lower in later. A comparison between with and without RTA SF revealed that titrable acid and phosphate excretion decreased in RTA defect. Overall data indicate that (a) RTA was an etiologic factor in one fourth of the SF; (b) an oral acid challenge test is a good adjunct to detect this condition and (c) acid challenge increases calcium, phosphate and titrable acid excretion and decreases citrate.     

Serum markers for early diagnosis of non-‘Q’ wave and ‘Q’ wave myocardial infarction

The present study conducted on twenty-five uncomplicated cases of acute myocardial infarction diagnosed by clinical and electrocardiographic findings indicated significantly increased level of cardiac Troponin-T and increased activities of the enzymes total creatine kinase, creatine kinase-MB, aspartate transaminase and lactate dehydrogenase as compared to the twenty-five healthy control subjects. The level of cardiac Troponin-T and the activities of the enzymes total creatine kinase, creatine kinase-MB, aspartate transaminase and lactate dehydrogenase was found to be higher in “Q” wave myocardial infarction patients as compared to the non-“Q” wave and the controls. Since cardiac Troponin-T has been shown to increase in unstable angina and renal failure without cardiac disease and creatine kinase-MB activity has been found to be normal in patients with unstable angina and increase very slightly in patients with renal failure, it was concluded that a combination of cardiac Troponin-T and creatine kinase-MB activity was sufficient for the early diagnosis of acute myocardial infarction.     

拉萨河流域健康湿地与退化湿地大型土壤动物群落比较研究

青藏高原是我国重要的高寒湿地分布区,高寒湿地也是我国特有的湿地类型。随着社会经济发展,人口不断的增长及农牧活动的增加,湿地面积、植被和土壤水分条件都发生了明显的变化。大型土壤动物是构成湿地生态系统中不可缺少的重要组成部分,与植物群落间有密切的联系。本文以拉萨河流域健康湿地和退化湿地为研究对象,探讨大型土壤动物群落特征,为高原湿地的监测及保护提供土壤动物学依据。研究结果表明,退化湿地大型土壤动物类群数和个体数均高于健康湿地,退化湿地稀有类群极为丰富,由于退化湿地生境较为干燥,出现了蚁科、隐翅虫科、步甲科等动物类群。显著性检验结果显示,健康湿地和退化湿地大型土壤动物存在明显的差异性,产生这种差异的主要原因是受生境中植被和土壤中含水率的影响。     

Detection of inherited metabolic diseases in children with mental handicap

Mental Retardation is a condition where complex interactions of intrinsic and extrinsic factors hamper mental and sometimes physical growth of the child during developmental period. This study was carried out to detect cause of Mental Retardation in 2000 cases of developmental delay by a multidisciplinary team comprising of a Pediatrician, Cytogeneticist, Biochemist, Psychologist and Speech and Occupational therapists. The causes for developmental delay are broadly divided into Genetic, Environmental and Idiopathic (no specific cause found) factors. The complete diagnosis was possible in 1192(60%) cases. Genetic factors were found in 477(23.8%) cases and environmental factors covered 692(34.6%) cases as a cause of retardation. The most common genetic cause is chromosomal abnormalities which were found in 355 (17.75%) cases. Another major group comprising of 122 (6.1%) cases was that of the disorders which follow Mendelian inheritance. Autosomal recessive conditions, which covers majority of metabolic disorders, are detected in 48(2.4%) cases. During this screening the most common inherited metabolic condition detected is Mucopolysaccharidosis, a Lysosomal Storage Disorder followed by Amino acid abnormalities like Phenylketonuria, Alkaptonuria and Tyrosinuria.     

Lipid peroxidation and antioxidant status in patients with rheumatoid arthritis

The changes in the erythrocyte lipid peroxidation products (MDA), levels of glutathione (GSH), ascorbic acid and plasma vitamin E (non enzymatic antioxidant parameters) and activities of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GP_X), catalase in erythrocytes and plasma glutathione - S - transferase (GST) activity were estimated in patients with rheumatoid arthritis. This work was undertaken to assess oxidative stress and anti oxidant status in patients with rheumatoid arthritis. It was observed that there was a significant increase in erythrocyte MDA levels, activities of SOD, GP_X, plasma GST and a significant decrease in erythrocyte GSH, ascorbic acid, plasma vitamin E levels and catalse activity in patients with rheumatoid arthritis when compared to controls. The results of our study suggests higher oxygen free radical production, evidenced by increased MDA and decreased GSH, ascorbic acid, vitamin E and Catalase activity, support to the oxidative stress in rheumatoid arthritis. The increased activities of antioxidant enzymes may be a compensatory regulation in response to increased oxidative stress.     

Study of some factors affecting stability of N-acetyl-β-D-glucosaminidase and aminopeptidase N in urine at 37°C

Little is known of conditions which influence the stability of urinary enzymes upon storage in the bladder at 37°C. Using a continuous flow system simulatingin vivo conditions, we studied the influence of the pH of urine on the stability of two renal parenchymal enzymes N—Acetyl—β—D—Glucosaminidase (2—acetamido—2—deoxy—β—D—glucoside acetamidodeoxy glucohydrolase, NAG, EC 3.2.1.30) and L—Alanine aminopeptidase (Aminopeptidase N, AAP, EC 3.4.11.2). This continuous flow model that we have described can be employed to study the influence of pH on the stability of any renal enzyme excreted in urine. We also studied thein vitro effects of varying concentrations of low molecular weight regulatory metabolites such as urea, creatinine and uric acid and of some drugs excreted in urine, on the assay of these two enzymes. Urinary pH, urea content and some antibiotics seem to influence measured urinary NAG and AAP activities and we therefore express the need for caution before diagnostic interpretation of the urinary enzyme activities are made.     

Alcohol induced effects on kidney

After administration ethanol and its metabolites go through kidneys and are excreted into urine, and its content in the urine is higher than that of the blood and the liver. Chronic ethanol administration decreases the renal tubular reabsorption and reduces renal function. Multiple functional abnormalities of renal tubules may be associated with ethanol-induced changes in membrane composition and lipid peroxidation. The vulnerability of the kidney to oxidative damage has been partly attributed to its high content of long-chain polyunsaturated fatty acids. Renal ultra structural abnormalities due to ethanol exposure may be important in the genesis of functional disturbances. Increased oxidative stress and endothelial dysfunction with their complex interrelationships are relevant aspects of atherogenesis in chronic renal failure. Antioxidants, particularly polyphenols are expected to decrease the vulnerability of the kidney to oxidative challenges.     

脑梗死诊治临床教学体会

随着生活方式的转变,我国脑血管病的发病率逐年上升,脑梗死不仅是神经科的常见病、多发病,而且也是我国致残率和死亡率较高的病种之一。因此,提高脑梗死的诊治水平迫在眉睫,笔者就近年来脑梗死临床教学中的点滴体会,总结如下。     

Quantification of urinary urate with alkylamine glass-bound uricase and arylamine glass-bound peroxidase

Uric acid in urine was analyzed using porcine liver uricase and horseradish peroxidase immobilized on alkylamine and arylamine glass beads (pore diameter 55nm) respectively. The minimum detection limit was 5.0mg/0.1 ml urine. The recovery of added uric acid was 92%. Within and between assay CVS were <1.3% and <5.3% respectively. A good correlation (r=0.93) was found between urinary urate values obtained by a commercial kit method and the present method. The uric acid in 24hr urine of apparently healthy adults and person sufering from various diseases was found to be 450 to 900mg/24hr, 659mg/24hr (range, mean) and 910–1400mg/24hr, 1145 mg/24hr (range, mean) respectively with the present method.     

Antioxidant Status and Oxidative Stress in the Circulation of Younger and Elderly Human Subjects

Extensive research has demonstrated the protective properties of antioxidants, which scavenge reactive oxygen species and their precursors, as well as up-regulate enzymes involved in the repair of cellular damage. Several case–control studies have showed higher blood levels of antioxidants and decreased oxidative stress in younger individuals when compared with older ones. Cell damage caused by free radicals appears to be a major contributor in aging and degenerative diseases of aging such as cancer, cardiovascular disease, cataracts, compromised immune system, rheumatoid arthritis and brain dysfunction. The objective of this study was to determine the variation of Circulating levels of selected antioxidants (enzymic and non enzymic) and oxidative stress marker in younger and older humans. The results showed that a majority of the younger age group participants showed a significant increase in enzymic and nonenzymic antioxidant status and a decrease in oxidative stress when compared with the older age group.     

Sex Dimorphism in Serum Lecithin: Cholesterol Acyltransferase and Lipoprotein Lipase Activities in Adult Sickle Cell Anaemia Patients with Proteinuria

Proteinuria in subjects with sickle cell anaemia (SCA) is an indication of an ongoing renal insufficiency and it’s prevalence varies between sexes. We evaluated sex differences in the activities of Lecithin: cholesterol acyltransferase (LCAT), Lipoprotein lipase (LPL) and the levels of lipoproteins in SCA patients with proteinuria. Fifty SCA patients (30 males aged: 26.4 ± 7.3 years and 20 females, aged 25.4 ± 2.6 years) and 50 age and sex matched control SCA patients were recruited for the study. Random urine specimens were collected and tested for the presence of albumin by urine dipstick technique. A 24 h urinary protein was quantitated using sulphosalicylic acid technique. Fasting serum total cholesterol, triglyceride, urea and creatinine were determined using enzymes catalyzed colorimetric methods. HDL cholesterol was determined in the supernatant after precipitation with manganese chloride–phosphotungstic acid solution. LCAT was measured using the Anasolv LCAT assay with proteoliposome as substrate. LPL was determined by incubating the serum in glyceryl trioleate substrate, the glycerol liberated was measured in an aliquot of the incubating mixture. In male SCA controls there was 18.2 and 6.9% increase in the activities of LPL and LCAT respectively when compared with females but in SCA patients with proteinuria there was 8.4 and 5.2% decreases in the male SCA patients compared with females. The concentration of 24 h urine protein in the SCA male subjects with proteinuria was significantly higher (0.25 g/day; P < 0.001) compared with the SCA female patients with proteinuria (0.09 g/day). There are sex differences in the activities of LCAT and LPL in SCA patients with proteinuria. Metabolism of these lipolytic enzymes may be modulated differently in SCA patients with proteinuria.     

Serum and platelet sialic acid in acute myocardial infarction

Blood samples from 39 patients with acute myocardial infarction and 15 healthy controls were analysed for serum and platelet sialic acid. Serum sialic acid levels in patients with acute myocardial infarction were significantly higher than controls (mean 2.7±0.46 μmol/ml Vs. 1.91±0.17 μmol/ml respectively). Levels of serum orosomucoid, an acute phase reactant, containing sialic acid, were also higher in these patients, suggesting a possible non-specific mechanism of increase in serum sialic acid concentration. In contrast, platelets contained significantly less sialic acid in patients with acute myocardial infarction than control (26.73±1.57 nmol/mg protein and 31.97±2.68 nmol/mg protein respectively).     

Comparison of Urinary Total Proteins by Four Different Methods

The total proteins in human urine have been compared by sulfosalicylic acid, sulfosalicylic acid with sodium sulphate and trichloroacetic acid methods with pyrogallol red molybdate method as there are no studies found quantifying imprecision and bias components. Fresh urine of 36 patients was analyzed by four methods. Imprecision and inaccuracy were determined by repeated analysis and method comparison studies using correlation plots, Bland and Altman, and Passing and Bablok regression analyses respectively. The coefficient of variation was 5.07 % for pyrogallol red molybdate; 6.84 % for sulfosalicylic acid; 3.97 % for sulfosalicylic acid with sodium sulphate and 5.93 % for trichloroacetic acid methods. Bland and Altman analysis showed a bias of 5.8, 1.7 and ?5.4 for pyrogallol red molybdate versus sulfosalicylic acid, sulfosalicylic acid with sodium sulphate and trichloroacetic acid methods respectively. Passing and Bablok regression revealed a constant bias for pyrogallol red molybdate versus all turbidimetric methods but a proportional bias only with trichloroacetic acid method. Sulfosalicylic acid with sodium sulphate method is preferred to sulfosalicylic acid and trichloroacetic acid methods.     

Uric acid a better scavenger of free radicals than vitamin C in rheumatoid arthritis

Uric acid an endogenous aqueous antioxidant in normal humans is present in much higher concentrations than vitamin C and has been known to cover 2/3^rd of the free radical scavenging capacity in plasma. In the present study average uric acid levels of patients of rheumatoid arthritis were found to be close to the normal individuals. A unique feature was observed after classifying the patients on the basis of the duration of suffering, the patients having longer duration of disease had least uric acid levels as compared to those suffering from relatively lesser period, similar trend was observed in the ascorbic acid estimations. The decline in uric acid values with progression of the disease was much more than what was observed in case of ascorbic acid suggesting the significant role of uric acid in scavenging of free radicals. Too much lowering of Uric Acid should be checked and vitamin C should be supplemented in diet for maintaining the balance between pro oxidant and antioxidant forces to check pro oxidant insult in rheumatoid arthritis.     

Lipid peroxidation and antioxidant status in blood of rheumatoid arthritis patients

The main objective of the study was to assess the oxidative stress in plasma and erythrocytes of rheumatoid arthritis patients by measuring the levels of thiobarbituric acid reactive substances (TBARS), non-enzymatic antioxidants (vitamin E, C and reduced glutathione) and enzymatic antioxidants [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSHPx)]. This study has been conducted on twenty-two adult female rheumatoid arthritis patients and an equal number of healthy subjects. Elevated lipid peroxidation and multidirectional changes in the antioxidant defence system were noticed in patients with rheumatoid arthritis. The enhanced lipid peroxidation accompanied by disturbance in antioxidant status indicates that rheumatoid arthritis patients are more prone to free radical mediated oxidative damage.