Ornithine Carbamoyltransferase Activity Research Articles

Fermentation in Pineapple Juice Significantly Enhances Ornithine and Citrulline Production in Lactococcus lactis MSC-3G Isolated from Sugarcane.

Lactic acid bacterial (LAB) fermentation of functional amino acids using fruit juices as a cultivation medium is not well-documented. In the present study, we successfully isolated a high ornithine- and citrulline-producing Lactococcus lactis strain, designated MSC-3G, from sugarcane and investigated the ornithine and citrulline production profile using various fruit juices as a cultivation medium. Among fruit juices, pineapple juice exhibited the highest potentiality to initiate ornithine production (56 mM), while the highest citrulline yield was obtained during lime juice cultivation (34.5 mM). Under the optimal cultivation condition, the highest yield of ornithine and citrulline in pineapple juice reached 98.9 ± 2.2 mM and 211.1 ± 35.7 mM, respectively, both of which were significantly higher than that in the well-known industrial strain of Corynebacterium (C.) glutamicum. Additionally, citrulline production was dependent on oxygen supplementation and increased twofold when grown aerobically. Whole genome sequencing showed that the MSC-3G genome possesses the arginine deiminase (ADI) gene cluster arcABD1C1C2TD2. The results of the ADI pathway enzyme activities of MSC-3G showed a significant increase in arginine deiminase activity, while ornithine carbamoyl transferase activity was decreased, which in turn indicates the high citrulline-accumulation ability of MSC-3G when cultivated in pineapple juice.

Catabolic Ornithine Carbamoyltransferase Activity Facilitates Growth of Staphylococcus aureus in Defined Medium Lacking Glucose and Arginine.

ABSTRACTPrevious studies have found that arginine biosynthesis in Staphylococcus aureus is repressed via carbon catabolite repression (CcpA), and proline is used as a precursor. Unexpectedly, however, robust growth of S. aureus is not observed in complete defined medium lacking both glucose and arginine (CDM-R). Mutants able to grow on agar-containing defined medium lacking arginine (CDM-R) were selected and found to contain mutations within ahrC, encoding the canonical arginine biosynthesis pathway repressor (AhrC), or single nucleotide polymorphisms (SNPs) upstream of the native arginine deiminase (ADI) operon arcA1B1D1C1. Reverse transcription-PCR (RT-PCR) studies found that mutations within ccpA or ahrC or SNPs identified upstream of arcA1B1D1C1 increased the transcription of both arcB1 and argGH, encoding ornithine carbamoyltransferase and argininosuccinate synthase/lyase, respectively, facilitating arginine biosynthesis. Furthermore, mutations within the AhrC homologue argR2 facilitated robust growth within CDM-R. Complementation with arcB1 or arcA1B1D1C1, but not argGH, rescued growth in CDM-R. Finally, supplementation of CDM-R with ornithine stimulated growth, as did mutations in genes (proC and rocA) that presumably increased the pyrroline-5-carboxylate and ornithine pools. Collectively, these data suggest that the transcriptional regulation of ornithine carbamoyltransferase and, in addition, the availability of intracellular ornithine pools regulate arginine biosynthesis in S. aureus in the absence of glucose. Surprisingly, ~50% of clinical S. aureus isolates were able to grow in CDM-R. These data suggest that S. aureus is selected to repress arginine biosynthesis in environments with or without glucose; however, mutants may be readily selected that facilitate arginine biosynthesis and growth in specific environments lacking arginine.

Regulation of citrulline synthesis in human enterocytes: Role of hypoxia and inflammation

AbstractIntensive care unit patients and chronic airway inflammatory disease are characterized by chronic systemic hypoxia and inflammation inducing a decrease in nitric oxide release due to impaired l‐arginine (ARG) homeostasis. As ARG is synthesized from circulating l‐citrulline (CIT), an alteration of CIT production in small intestine by ornithine carbamoyltransferase could be involved. Here, we posit that hypoxia and/or inflammation has effects on ornithine carbamoyltransferase regulation in enterocytes. A duodenal explant incubation model was used. Biopsy specimens taken from 25 selected patients were incubated for 6 h in 4 groups: control, inflammation, hypoxia, and hypoxia + inflammation. At the end of the incubation period, we measured CIT concentration in culture media, ornithine carbamoyltransferase activity, ornithine carbamoyltransferase protein and gene expression, protein expression of enzymes involved in the CIT production pathway, and expression of energy status proteins. Inflammation and/or hypoxia conditions did not affect CIT production. Ornithine carbamoyltransferase activity was increased in hypoxia conditions (p = 0.023). Expression of enzymes implicated in the CIT crossroads pathway and enzymes reflecting energy status variation was not affected by inflammation and hypoxia. Data sets were pooled to evaluate the variability of the four quartiles for each parameter. CIT production was found to increase over the quartiles whereas other parameters remained stable. Our results showed that intestinal CIT production is preserved during inflammation and/or hypoxia, thus confirming the significance of this metabolic pathway. This suggests that the CIT deficiency observed in clinical hypercatabolic states could be a consequence of high utilization for ARG synthesis.

Liver involvement in the process of acute respiratory infections in pediatric patients

We aimed to investigate the prevalence of liver involvement in pediatric patients with ARI using both routine tests of hepatic panel, and ornithine carbamoyltransferase (OCT) to identify the most sensitive indicators of early hepatic injury. A prospective cohort study of 84 armenian children with ARI was conducted to evaluate the associated liver involvement. The diagnostic variables of interest were the signs of clinical disease severity, and enzymatic profile of the patients. Serum levels of OCT were increased in 94% of patients versus routine tests of hepatic panel (AST in 41.7%, ALT in 15.5%, etc). Variance analysis by severity groups showed the serum levels of OCT (p < 0.001), ammonia (p < 0.001), phospholipides (p = 0.05), glucose (p = 0.01), TNF-α (p = 0.01), IL-8 (p < 0.001), AST (p < 0.001), and ALP (p < 0.001) were associated with the severity of underlying disease. Moreover, regression analysis revealed the serum activity of OCT (p value < 0.001, OR = 1.27) and ammonia (p value 0.002, OR = 1.1) significantly predict the severity of the disease. Using more sensitive marker of liver damage can detect more cases of ARI with hepatic manifestations. For evaluation of the liver involvement we are suggesting the testing of serum OCT levels as a more sensitive and specific marker. Pediatric patients with ARI and with higher serum OCT levels have 27% more chance to experience increased disease severity, which can affect on liver state and prolong hospitalization time and cost.

Activity of the liver enzyme ornithine carbamoyltransferase (OTC) in blood: LC-MS/MS assay for non-invasive diagnosis of ornithine carbamoyltransferase deficiency.

Liver enzymes are released from hepatocytes into circulation and their activity can be measured in the blood. We examined whether the plasma activity of the liver enzyme ornithine carbamoyltransferase, determined by a novel liquid chromatography-mass spectrometry (LC-MS/MS) assay, could be utilized for the detection of OTC deficiency (OTCD), an X-linked inborn error of the urea cycle. The plasma ornithine carbamoyltransferase (OTC) activity was assayed in the reverse reaction using isotopically labeled citrulline-d4 as a substrate and by determination of the product, ornithine-d4, by LC-MS/MS analysis. The plasma OTC activity in the controls was in the range of 111-658 pkat/L (n=49, median 272 pkat/L), and the activity increased linearly with serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in patients with hepatopathy. The OTC activity was subsequently determined in 32 individuals carrying mutations in the OTC gene, and OTC/ALT and OTC/AST ratios were calculated to account for the degree of hepatopathy, which is a common finding in OTCD. The OTC/ALT ratio enabled clear differentiation of OTCD hemizygotes (n=11, range 0-69×10-6) from controls (504-3440×10-6). This ratio also enabled the detection of 11 of 12 symptomatic heterozygotes (range 38-794×10-6), while this marker did not allow for reliable differentiation of asymptomatic heterozygotes (n=9) from controls. LC-MS/MS assay of plasma OTC activity enabled the detection of all hemizygous and the majority of symptomatic heterozygous OTCD patients in the tested cohort. This study demonstrates that non-invasive assay of enzymes expressed predominantly in the liver could be used as an alternative approach for diagnosing inborn errors of metabolism.

Serum Ornithine Carbamoyltransferase Activity and Correlation with Fatty Liver in Dairy Cows with Displaced Abomasum

Ornithine Carbamoyltransferase (OCT) is a hepatocyte-specific enzyme; elevated serum activity indicates liver damage. Displaced abomasum frequently occurs around the time of parturition in dairy cattle and is associated with fatty liver. We analyzed serum OCT activities to investigate the relationship between displaced abomasum and fatty liver. Lactating Holstein cows, 75 clinically healthy and 22 with displaced abomasum, were used. Serum OCT activities were compared between healthy cows and those with displaced abomasum. Liver biopsies of cows with displaced abomasum were classified by the severity of fat accumulation. Blood samples were collected preoperatively and at 4 and 8 days after surgery and analyzed for serum OCT, aspartate aminotransferase, alkaline phosphatase, γ-glutamyltransferase, urea nitrogen, total protein and hematocrit. Serum OCT activity was significantly higher in cows with displaced abomasumthan in healthy cows. The degree of fatty liver in cows with displaced abomasum was classified as mild (13 cows), moderate (5 cows), or severe (4 cows). Statistical analysis revealed no significant differences in blood test results among cows with different degrees of fatty liver. However, our results indicated significant differences in serum OCT, aspartate aminotransferase, urea nitrogen, total protein and hematocrit when stratified by the number of days after surgical treatment for displaced abomasum. Serum OCT activity is increased in dairy cows with displaced abomasum and the influence of the degree of fatty liver on serum OCT activity is unclear. Therefore, OCT should be reconsidered as a marker of fatty liver in cows around the time of parturition, when displaced abomasum frequently occurs. Fatty liver should be evaluated by various clinical parameters, not by one hepatic enzyme.

Expression of the Gene for Resistance to Phaseolotoxin (argK) Depends on the Activity of Genes phtABC in Pseudomonas syringae pv. phaseolicola

The bacterium Pseudomonas syringae pv. phaseolicola produces phaseolotoxin in a temperature dependent manner, being optimally produced between 18°C and 20°C, while no detectable amounts are present above 28°C. Phaseolotoxin is an effective inhibitor of ornithine carbamoyltransferase (OCTase) activity from plant, mammalian and bacterial sources and causes a phenotypic requirement for arginine. To protect the cell from its own toxin, P. syringae pv. phaseolicola synthesizes a phaseolotoxin-resistant OCTase (ROCT). The ROCT is the product of the argK gene and is synthesized only under conditions leading to phaseolotoxin synthesis. The argK gene is included in a chromosomal fragment named Pht cluster, which contains genes involved in the synthesis of phaseolotoxin. The aim of the present work was to investigate the possible involvement of other genes included in the Pht cluster in the regulation of gene argK. We conducted transcriptional analyses of argK in several mutants unable to produce phaseolotoxin, transcriptional fusions and electrophoretic mobility shift assays, which allowed us to determine that genes phtABC, located within the Pht cluster, participate in the transcriptional repression of gene argK at temperatures not permissive for phaseolotoxin biosynthesis. This repression is mediated by a protein present in both toxigenic and nontoxigenic strains of P. syringae and in E. coli, and requires the coordinated participation of phtA, phtB and phtC products in order to carry out an efficient argK repression.

Piroxicam and meloxicam ameliorate hepatic oxidative stress and protein carbonylation in Kupffer and sinusoidal endothelial cells promoted by ischemia-reperfusion injury

The present study was aimed to assess the effect of protein carbonylation (PC) in hepatic cells and effects of nonsteroidal anti-inflammatory drugs (NSAIDs) on indicators of tissue damage induced by liver ischemia-reperfusion injury (LIRI). Warm ischemia was performed by partial vascular occlusion during 90 min in Wistar rats. In serum, we determined the catalytic activity of Alanine Aminotransferase, Aspartate Aminotransferase, Lacticate Dehydrogenase, and Ornithine Carbamoyltransferase. In liver samples, we studied cellular alterations by means of histologic studies, lipid peroxidation, PC by immunohistochemistry, apoptosis and reactive oxygen species in bile by electron paramagnetic resonance. Based on PC data, sinusoidal endothelial cells (SEC) and Kupffer cells (KC) were the first to exhibit LIRI-associated oxidative damage and prior to parenchymal cells. Administration of piroxicam or meloxicam during the pre-ischemic period produced a highly significant decrease in all studied injury indicators. No significant differences were revealed between the protective action of the two drugs. The data shown here suggest the potential use of NSAIDs such as piroxicam or meloxicam in minimizing ischemic event-caused damage in liver. We also propose that PC may be employed as an adequate tool to assess tissue damage after oxidative stress.

Effects of crowding on ornithine–urea cycle enzyme mRNA expression and activity in gulf toadfish (Opsanus beta)

The gulf toadfish (Opsanus beta) is a facultatively ureotelic fish that excretes primarily urea under conditions of crowding or confinement. To examine the relationship between ammonia production, urea production and the ornithine-urea cycle (O-UC) enzyme activity and mRNA expression, we subjected toadfish to two-day and seven-day crowding regimes. Plasma cortisol levels were measured and liver tissue was assayed for ammonia and urea concentrations. Liver glutamine synthetase (GS), carbamoyl phosphate synthetase III (CPS), ornithine carbamoyl transferase (OCT) and arginase (ARG) activities were also measured. Quantitative PCR was utilized to determine liver GS, CPS, OCT, ARG, argininosuccinate synthetase (ASS) and argininosuccinate lyase (ASL) mRNA expression. Hepatic ammonia concentrations decreased with increased duration of crowding whereas liver urea and circulating cortisol levels increased. An elevation in enzyme activity with increased duration of crowding was observed for all four O-UC enzymes examined. By contrast, mRNA expression was variable for the O-UC enzymes and only CPS and ASS had mRNA expression levels that were elevated in crowded fish. These results suggest that the activities of O-UC enzymes are better predictors for urea production than O-UC enzyme mRNA expression levels.

Lysine 88 Acetylation Negatively Regulates Ornithine Carbamoyltransferase Activity in Response to Nutrient Signals

Ornithine carbamoyltransferase (OTC) is a key enzyme in the urea cycle to detoxify ammonium produced from amino acid catabolism. OTC deficiency is an X-linked genetic disorder ranging from fatal in newborns to hyperammonemia and anorexia in adults. Through affinity purification of acetylated peptides and mass spectrometry, we identified that OTC is acetylated on lysine residues, including Lys88, which is also mutated in OTC-deficient patients. OTC acetylation was confirmed to occur under physiological conditions. Biochemical characterizations revealed that OTC Lys88 acetylation decreases the affinity for carbamoyl phosphate, one of the two OTC substrates, and the maximum velocity, whereas the K(m) for ornithine, the other OTC substrate, is not affected. Furthermore, Lys88 acetylation is regulated by both extracellular glucose and amino acid availability, indicating that OTC activity may be regulated by cellular metabolic status. Our results provide an example of the novel mechanism of regulating metabolic enzyme activity through protein acetylation.

Lysine 88 Acetylation Negatively Regulates Ornithine Carbamoyltransferase Activity in Response to Nutrient Signals

Lysine 88 Acetylation Negatively Regulates Ornithine Carbamoyltransferase Activity in Response to Nutrient Signals

Influences of temperature and threshold effect of NaCl concentration on Alpias vulpinus OCT

Thermodynamic, circular dichroism (CD), and activity measurements have been used to characterize the different conformational states and the effects of NaCl concentrations (0.0–3.0 M) on thermal unfolding of ornithine carbamoyltransferase (OCT) from Alopias vulpinus. Furthermore conformational changes in whole enzyme structure have been monitored by titration of SH-groups. OCT unfolding process follows an irreversible two-state mechanism with a first-order kinetic of denaturation, without breaking-point. NaCl shows very little stabilization effects at low concentration and its action become very important over 1.5 M concentration. The presence of 3.0 M NaCl completely avoids OCT unfolding at 60, 64 and 66 °C. Kinetic and thermodynamic parameters are strongly influenced by the presence of high NaCl concentration. Our experiments showed that NaCl stabilization process involved changes in preferential binding, in electrostatic and van der Waals interactions and exposure of buried site and SH-groups. During thermal denaturation, UV–vis and CD spectroscopy show that high salts concentration preserves OCT activity, avoiding exposure of hydrophobic site and destruction of secondary and tertiary structure elements.

Clinicopathologic Evaluation of Hepatic Lipidosis in Periparturient Dairy Cattle

Background: Fatty change of the liver (FCL) is very common in dairy cattle periparturiently. Many laboratory methods have been implicated in order to assist the diagnosis.Hypothesis: To investigate whether FCL in dairy cattle could be evaluated by assessment of ornithine carbamoyl transferase (OCT) by means of an assay modified for bovine serum, other enzyme activity, serum bile acids (SBA) concentration, or other biochemical constituents.Animals: A total of 187 dairy cattle were included: 106 were suspected to have liver dysfunction and were examined after referral by veterinarians; 70 were clinically healthy with mild FCL; and 11 were clinically healthy without FCL.Methods: Blood and liver biopsy samples were obtained after clinical examination. Histologic examination by light microscopy and classification of samples according to the severity of FCL was done, and total lipid and triglyceride concentration was measured. In serum, OCT, aspartate aminotransferase (AST), alanine aminotransferase (ALT), sorbitol dehydrogenase (SDH), glutamate dehydrogenase (GDH), alkaline phosphatase (ALP), and γ‐glutamyltransferase (γ‐GT) activity as well as SBA, glucose, ketones, total bilirubin (tBIL), and nonesterified fatty acids (NEFA) concentration were measured.Results: OCT and AST activity and tBIL concentration correlate well with the degree of FCL. SBA concentration does not contribute well to FCL diagnosis. The majority of FCL cases appeared within the first 21 days‐in‐milk (DIM). The majority of moderate‐to‐severe and severe FCL cases arose in the first 7 DIM.Conclusions and Clinical Importance: Except for OCT, AST, and tBIL, none of the biochemical tests used, including SBA, had sufficient discriminatory power to differentiate reliably between mild and severe FCL because of poor sensitivity. A weak correlation between clinical signs and the extent of FCL was evident.

Clinicopathologic Evaluation of Hepatic Lipidosis in Periparturient Dairy Cattle

Fatty change of the liver (FCL) is very common in dairy cattle periparturiently. Many laboratory methods have been implicated in order to assist the diagnosis. To investigate whether FCL in dairy cattle could be evaluated by assessment of ornithine carbamoyl transferase (OCT) by means of an assay modified for bovine serum, other enzyme activity, serum bile acids (SBA) concentration, or other biochemical constituents. A total of 187 dairy cattle were included: 106 were suspected to have liver dysfunction and were examined after referral by veterinarians; 70 were clinically healthy with mild FCL; and 11 were clinically healthy without FCL. Blood and liver biopsy samples were obtained after clinical examination. Histologic examination by light microscopy and classification of samples according to the severity of FCL was done, and total lipid and triglyceride concentration was measured. In serum, OCT, aspartate aminotransferase (AST), alanine aminotransferase (ALT), sorbitol dehydrogenase (SDH), glutamate dehydrogenase (GDH), alkaline phosphatase (ALP), and gamma-glutamyltransferase (gamma-GT) activity as well as SBA, glucose, ketones, total bilirubin (tBIL), and nonesterified fatty acids (NEFA) concentration were measured. OCT and AST activity and tBIL concentration correlate well with the degree of FCL. SBA concentration does not contribute well to FCL diagnosis. The majority of FCL cases appeared within the first 21 days-in-milk (DIM). The majority of moderate-to-severe and severe FCL cases arose in the first 7 DIM. Except for OCT, AST, and tBIL, none of the biochemical tests used, including SBA, had sufficient discriminatory power to differentiate reliably between mild and severe FCL because of poor sensitivity. A weak correlation between clinical signs and the extent of FCL was evident.

Evaluation of ornithine carbamoyl transferase and other serum and liver-derived analytes in diagnosis of fatty liver and postsurgical outcome of left-displaced abomasum in dairy cows

To evaluate postsurgical outcome in dairy cows with left-displaced abomasum (LDA) with regard to severity of fatty liver and assess the usefulness of preoperative determination of serum ornithine carbamoyl transferase (OCT) activity, bile acids concentration, and other variables for evaluating liver function during the postsurgical convalescence period. Prospective study. 68 Holstein cows. Blood and liver biopsy specimens were obtained during standing LDA surgery. Liver tissue was examined histologically and classified by severity of fatty change. Serum activities of liver-derived enzymes and concentrations of total lipids, triglycerides, bile acids, glucose, beta-hydroxybutyric acid, bilirubin, and nonesterified fatty acids were determined. Most cows with LDA and cows with severe fatty liver were detected within the first month after calving. Postsurgical outcome was related to severity of fatty liver. All cows that died had severe fatty liver. Serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration were sensitive indicators of fatty liver. Serum bile acids concentration was not an accurate indicator of fatty liver. Postsurgical outcome of cows undergoing surgery to correct LDA was related to fatty liver severity. Assessment of serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration is recommended for diagnosis of fatty liver in dairy cows with LDA, whereas determination of bile acids concentration is not. The strong correlation between OCT activity and degree of hepatocellular damage supports use of this enzyme for assessing severity of fatty liver and predicting postsurgical outcome in cows with LDA.

Suppressive effects of dietary D-penicillamine on the gene expression of hepatic metallothionein-1 and protein level of hepatic metallothionein in Long-Evans Cinnamon rats

We investigated the effects of dietary D-penicillamine on copper and zinc contents in the liver, serum, and urine of Long-Evans Cinnamon (LEC) rats, as well as the gene expression of hepatic metallothionein-1 and hepatic metallothionein protein level in these animals. Young male LEC and Fischer rats were fed a basal or D-penicillamine (5 g D-penicillamine/kg diet) diet for 16 days. The urinary output of copper and zinc was markedly increased by the D-penicillamine diet, and the copper and zinc contents in the liver were significantly decreased in LEC rats. Excess copper accumulation in the liver induced hepatic metallothionein in LEC rats, but the D-penicillamine diet depressed the level of hepatic metallothionein-1 mRNA as well as the hepatic metallothionein protein level. When fed the basal diet, serum ornithine carbamoyltransferase activity was higher in LEC rats than in Fischer rats, but a significant decrease in serum ornithine carbamoyltransferase activity rats was observed in LEC rats on feeding the D-penicillamine diet. These results indicated that dietary D-penicillamine significantly reduced excess copper and zinc in the livers of LEC rats through urinary excretion, and the decrease in hepatic copper and zinc contents caused a decrease in the levels of metallothionein-1 mRNA and metallothionein protein in LEC rats.

乳牛における血漿オルニチンカルバミルトランスフェラーゼ活性の病態的意義

Ornithine Carbamoyltransferase (OCT) is recognized as a liver parenchyma enzyme. To clarify the pathophysiological significance of its activities, γ-glutamyl transferase (CGT) and aspartate transaminase (AST) were evaluated in 16 healthy and 106 sick dairy cows. Liver weight and liver triglyceride (TG) levels were measured in some of the animals, on which autopsies were performed. Although plasma OCT activity was 2-9 IU in healthy cows, it tended to rise in some diseased cows, especially when conditions like liver congestion brought on by cardiovascular disorders and hepatic fibroses resulted in liver enlargement. It did not necessarily increase, however, in cases of fatty liver. Hepatic disorder was more probable when OCT was high than when GGT or AST was high. Examination of plasma OCT activities provides useful information for evaluating liver disturbances in dairy cows.

Synthesis of citrulline from ornithine by the small intestinal mucosa of cattle

ABSTRACTThree segments of cattle small intestine (duodenum, upper jejuno‐ileum and lower jejuno‐ileum) were examined in an in vitro system for activity of ornithine carbamoyltransferase (OCT; EC 2.1.3.3) which is involved in the synthesis of citrulline (Cit) from ornithine (Orn). The mucosa of the three segments of small intestine was collected from Japanese black cattle, homogenized and then centrifuged. The supernatant fraction was used as the crude OCT enzyme solution. The OCT activity was assayed by the production of Cit from Orn determined directly by HPLC. The optimal pH and temperature for OCT activities in the duodenum, upper jejuno‐ileum and lower jejuno‐ileum of cattle small intestine were 7.47 and 39°C. Little difference was observed between the three segments. The OCT activity in cattle kidney was also examined for comparison, and almost no activity was found. The OCT activities in crude enzyme solutions of the three segments of small intestine were stable for up to one month of storage at −20°C in Tris HCl buffer solution. Finally, the role of the small intestine in supplying Cit as a precursor for arginine synthesis in cattle kidney was discussed.

Degradation of arginine and other amino acids by butyrate-producing asaccharolytic anaerobic Gram-positive rods in periodontal pockets

The use of 20 amino acids by butyrate-producing asaccharolytic anaerobic Gram-positive rods (AAGPRs) in periodontal pockets, i.e. Eubacterium minutum, Filifactor alocis, E. infirmum, E. sulci and E. saphenum, was studied. E. minutum used only arginine and lysine, and produced substantial amounts of butyrate and ammonia as the main metabolic products from arginine, and acetate, butyrate and ammonia from lysine. Fi. alocis used arginine alone and produced butyrate and ammonia. E. infirmum, E. sulci and E. saphenum used lysine alone and produced acetate, butyrate and ammonia. The growth of these bacterial species was supported and enhanced by arginine and/or lysine enriched to culture media, but not by the other amino acids. Arginine deiminase, ornithine carbamoyltransferase and carbamate kinase activity were detected in the cell-free extract of E. minutum, suggesting that arginine was metabolised to citrulline initially, and subsequently to ornithine and carbamoyl phosphate. Ornithine and carbamoyl phosphate were further converted to butyrate, and carbon dioxide and ammonia, respectively. Enzymatic activity of arginine deiminase and ornithine carbamoyltransferase was not detected in Fi. alocis, indicating that Fi. alocis converted arginine to ornithine directly, not via citrulline, and further to butyrate.

Serum Ornithine Carbamoyl Transferase as a Surrogate Marker in Malaria.

Ornithine carbamoyl transferase (OCT) activity and other liver function tests were studied in a total of 50 patients of clinical malaria and 15 controls. They were grouped as group I (positive for malarial parasite on peripheral blood smear, n=18), group II (negative for malarial parasite on peripheral blood smear (PBS) but responded to antimalarials, n=17) and group III (peripheral blood smear negative and did not respond to antimalarial therapy, n=15). The mean OCT levels were significantly raised in group I (6.79 ± 1.84 IU/L, p value = 0.006) and group II (5.0 ± 1.15 IU/L, p value = 0.014) as compared to controls (2.5 ± 1.13 IU/L) and returned to normal after treatment In contrast, group III had normal levels except in a case of kala azar and septicemia where OCT levels were high and increased further on treatment. Taking PBS positivity as a gold standard of diagnostic criteria, OCT had a sensitivity of 83% and specificity of 86% with a high positive predictive value of 88% as compared to ALT which had a lower sensitivity of 55% and specificity of 80%. The clinical response rate in PBS negative cases of fever having high OCT level was 83% as compared to 35% in cases with normal OCT level, making OCT a good surrogate marker of malaria. OCT levels could also be of prognostic significance as 2 cases of cerebral malaria had high OCT levels of 11.1 UAL and 10.7 IU/L, respectively.