Acid Excretion Rate Research Articles

Effects of insulin resistance and hepatic lipid accumulation on hepatic mRNA expression levels of apoB, MTP and L-FABP in non-alcoholic fatty liver disease

Non-alcoholic fatty liver disease (NAFLD) is considered a hepatic manifestation of metabolic syndrome, which is known to be associated with insulin resistance (IR). NAFLD occurs when the rate of hepatic fatty acid uptake from plasma and de novo fatty acid synthesis is greater than the rate of fatty acid oxidation and excretion as very low-density lipoprotein (VLDL). To estimate the effects of IR on hepatic lipid excretion, mRNA expression levels of genes involved in VLDL assembly were analyzed in NAFLD liver. Twenty-two histologically proven NAFLD patients and 10 healthy control subjects were enrolled in this study. mRNA was extracted from liver biopsy samples and real-time PCR was performed to quantify the expression levels of apolipoprotein B (apoB), microsomal triglyceride transfer protein (MTP) and liver fatty-acid binding protein (L-FABP). Hepatic expression levels of the genes were compared between NAFLD patients and control subjects. In NAFLD patients, we also examined correlations between expression levels of the genes and metabolic factors, including IR, and the extent of obesity and hepatic lipid accumulation. Hepatic expression levels of apoB, MTP and L-FABP were significantly up-regulated in NAFLD patients compared to control subjects. The expression levels of MTP were correlated with those of apoB, but not with those of L-FABP. In the NAFLD liver, the expression levels of MTP were significantly reduced in patients with HOMA-IR >2.5. In addition, a significant reduction in MTP expression was observed in livers with advanced steatosis. Enhanced expression of genes involved in VLDL assembly may be promoted to release excess lipid from NAFLD livers. However, the progression of IR and hepatic steatosis may attenuate this compensatory process.

Effects of Kurozu concentrated liquid on adipocyte size in rats

BackgroundKurozu concentrated liquid (KCL) is used as a health-promoting supplement for the treatment of disorders such as cancer, hyperlipidemia, and hypertension in Japan. We investigated the possible anti-obesity effects of KCL in rats.MethodsMale Sprague Dawley rats were fed American Institute of Nutrition 76 formula diet and were orally administrated KCL or acetic acid at a dose of 100 mg/kg body weight or deionized water for 4 weeks. Adipocyte size, DNA content in subcutaneous adipose tissue, lipid levels in the serum and liver, and the rate of fatty acid excretion were determined. Effects of KCL on pancreatic lipase activity and 3T3-L1 preadipocyte differentiation were investigated in vitro.ResultsIn the KCL group, the average adipocyte size in subcutaneous and perirenal adipose tissues was significantly reduced. The KCL-administered rats displayed greater numbers of small adipocytes in the subcutaneous, perirenal and mesenteric adipose tissues than did rats from the other groups. In the KCL group, the DNA content in subcutaneous adipose tissue was significantly increased. The rate of fatty acid excretion was significantly increased in the KCL group. Furthermore, KCL significantly inhibited pancreatic lipase activity in vitro, and also significantly inhibited fat accumulation and mRNA expression of fatty acid binding protein 2 (aP2) and peroxisome proliferator-activated γ (PPARγ) in 3T3-L1 preadipocyte. The levels of serum and liver lipids, the concentration of serum glucose, and the levels of adiponectin were similar among the 3 groups.ConclusionOral administration of KCL decreases the adipocyte size via inhibition of dietary fat absorption and reductions of PPARγ and aP2 mRNA expression levels in adipocytes.

Enterobacteria Modulate Intestinal Bile Acid Transport and Homeostasis through Apical Sodium-Dependent Bile Acid Transporter (SLC10A2) Expression

In our study, ampicillin (AMP)-mediated decrease of enterobacteria caused increases in hepatic bile acid concentration through (at least in part) elevation of bile acid synthesis in C57BL/6N mice. We investigated the involvement of enterobacteria on intestinal bile acid absorption in AMP-treated mice in the present study. Fecal enterobacterial levels and fecal bile acid excretion rates were markedly decreased in mice treated with AMP (100 mg/kg) for 3 days, whereas bile acid concentrations in portal blood were significantly increased compared with those in mice treated with a vehicle. Ileal apical sodium-dependent bile acid transporter (SLC10A2) mRNA levels and ileal SLC10A2 protein levels in brush-border membranes were significantly increased compared with those in mice treated with the vehicle. In AMP-treated mice, total bile acid levels were increased, whereas levels of enterobacteria-biotransformed bile acid, taurodeoxycholic acid, and cholic acid were decreased in intestinal lumen. These phenomena were also observed in farnesoid X receptor-null mice treated with AMP for 3 days. Discontinuation of AMP administration after 3 days (vehicle administration for 4 days) increased levels of fecal enterobacteria, fecal bile acid excretion, and taurodeoxycholic acid and cholic acid in the intestinal lumen, whereas the discontinuation decreased ileal SLC10A2 expression and bile acid concentrations in the portal blood. Coadministration of taurodeoxycholic acid or cholic acid decreased ileal SLC10A2 expression in mice treated with AMP. These results suggest that enterobacteria-mediated bile acid biotransformation modulates intestinal bile acid transport and homeostasis through down-regulation of ileal SLC10A2 expression.

Enhancing itaconic acid production by Aspergillus terreus

Aspergillus terreus is successfully used for industrial production of itaconic acid. The acid is formed from cis-aconitate, an intermediate of the tricarboxylic (TCA) cycle, by catalytic action of cis-aconitate decarboxylase. It could be assumed that strong anaplerotic reactions that replenish the pool of the TCA cycle intermediates would enhance the synthesis and excretion rate of itaconic acid. In the phylogenetic close relative Aspergillus niger, upregulated metabolic flux through glycolysis has been described that acted as a strong anaplerotic reaction. Deregulated glycolytic flux was caused by posttranslational modification of 6-phosphofructo-1-kinase (PFK1) that resulted in formation of a highly active, citrate inhibition-resistant shorter form of the enzyme. In order to avoid complex posttranslational modification, the native A. niger pfkA gene has been modified to encode for an active shorter PFK1 fragment. By the insertion of the modified A. niger pfkA genes into the A. terreus strain, increased specific productivities of itaconic acid and final yields were documented by transformants in respect to the parental strain. On the other hand, growth rate of all transformants remained suppressed which is due to the low initial pH value of the medium, one of the prerequisites for the accumulation of itaconic acid by A. terreus mycelium.

Effects of serotonergic agents on survival and hemolymph composition of the larval mosquitoAedes aegypti(Diptera: Culicidae, L.)in vivo: does serotonin regulate hemolymph acid–base homeostasis?

The role of serotonin in the regulation of larval Aedes aegypti hemolymph composition was investigated in vivo using two reuptake inhibitors (SSRIs), alaproclate HCl and 6-nitroquipazine maleate, and the receptor antagonist methiothepin mesylate. Larvae were placed in media differing in pH and salinity in the presence and absence of serotonergic agents. The toxicity of each agent was strongly influenced by ambient pH. For each agent, toxicity was negligible in acidic media, intermediate in neutral media and greatest in alkaline media. By contrast, toxicity of all agents was independent of salinity. No effects on mass-specific body water or hemolymph volume were observed whereas hemolymph osmotic pressure, Na(+) concentrations and pH differed significantly among treatments. 6-nitroquipazine caused a decrease in Na(+) from 115+/-1.7 to 103+/-0.9 mmol l(-1), and alaproclate caused alkalosis of the hemolymph from pH 7.55+/-0.026 to pH 7.72+/-0.044. Methiothepin decreased hemolymph osmotic pressure from 329+/-9.9 to 304+/-8.8 and showed the greatest overall toxicity. Control larvae excreted net base in pH 4 media (1.4 micromol g(-1) h(-1)) and net acid in pH 7 (1.2 micromol g(-1) h(-1)) and pH 11 (5.1 micromol g(-1) h(-1)) media. In pH 4 media, alaproclate and methiothepin caused a shift to net H(+) excretion (1.1 and 1.5 micromol g(-1) h(1), respectively) whereas these agents did not influence acid excretion rates in pH 7 or pH 11 media. The hypothesis that serotonin is involved in hemolymph acid-base balance is discussed.

Can urinary excretion rate of malondialdehyde, uric acid and protein predict the severity and impending death in perinatal asphyxia?

Background Perinatal asphyxia (PA) associated with multi-organ damage is a leading cause of neonatal mortality and morbidity. We evaluated if urinary malondialdehyde:creatinine (UMDA:Cr), uric acid:creatinine (UUA:Cr) and protein:creatinine (UP:Cr) vary with the severity of PA and if these parameters can predict the impending death in PA. Methods Study included 20 asphyxiated and 20 healthy newborn males. Hypoxic-ischemic encephalopathy (HIE) staging, APGAR (activity, pulse, grimace, appearance and respiration) score and urinary protein, uric acid, creatinine and MDA were evaluated. Results UMDA:Cr, UUA:Cr and UP:Cr were significantly higher and correlated with APGAR and HIE in PA. By regression analysis also, urinary parameters were found to have significant association with HIE stage and APGAR in PA. Receiver operating characteristics (ROC) curve of UP:Cr, UUA:Cr and UMDA:Cr showed area under curve of 0.896 ( p = 0.003), 0.859 ( p = 0.008) and 0.849 ( p = 0.010) with cut-off value of 9.04 mg, 2.34 mg and 3.49 µg/mg of creatinine respectively that can optimally predict the impending death in PA. SDS-PAGE of unconcentrated urine detected both high (73 kDa and 68 kDa) and low molecular weight proteins (52 kDa, 47 kDa, 25 kDa and 20 kDa) in PA but not in controls. Conclusion Urinary excretion rate of uric acid, MDA and proteins is higher and has potential to act as biochemical markers for severity evaluation and death prediction in PA.

High-Normal Serum Uric Acid Is Associated with Impaired Glomerular Filtration Rate in Nonproteinuric Patients with Type 1 Diabetes

Early renal function decline begins before the onset of proteinuria in patients with type 1 diabetes. The association of elevated serum uric acid with advanced impaired renal function prompts an examination of its role in early renal function decline in patients before proteinuria develops. Patients with type 1 diabetes and normoalbuminuria or microalbuminuria were recruited to the Second Joslin Kidney Study. A medical history and measurements of BP, hemoglobin A1c, albumin excretion rate, and serum concentrations of uric acid and cystatin C were obtained. Estimated glomerular filtration rate was measured by a cystatin C-based formula. We studied 364 patients with normoalbuminuria and 311 patients with microalbuminuria. Mean glomerular filtration rate in these groups was 119 and 99 ml/min, respectively. Mildly or moderately impaired renal function (<90 ml/min) was present in 10% of those with normoalbuminuria and 36% of those with microalbuminuria. In univariate and multivariate analyses, lower glomerular filtration rate was strongly and independently associated with higher serum uric acid and higher urinary albumin excretion rate, older age, and antihypertensive treatment. Serum uric acid concentration in the high-normal range is associated with impaired renal function in patients with type 1 diabetes. Follow-up studies are needed to confirm that this level of serum uric acid is a risk factor for early renal function decline in type 1 diabetes and to determine whether its reduction would prevent the decline.

Investigation of hyperuricemia during pegylated‐interferon‐α2b plus ribavirin combination therapy in patients with chronic hepatitis C

Hyperuricemia has been reported as being an adverse effect of pegylated-interferon-alpha2b (Peg-IFNalpha2b) and ribavirin combination therapy for chronic hepatitis C and hyperuricemic changes occur in some patients during the therapy. However, detailed investigation of the elevation of uric acid has not been carried out previously. The incidence and mechanism of hyperuricemia were investigated in this study. The data of 50 patients with chronic hepatitis C who had been treated with Peg-IFNalpha2b and ribavirin combination therapy or pegylated-interferon-alpha2a monotherapy for more than 24 weeks were analyzed. The effects of these treatments were evaluated clinically by the serum uric acid level and its urinary excretion rates. In patients with pegylated-interferon-alpha2a monotherapy, no significant elevation was shown either in serum uric acid concentrations or excretion rates. On the other hand, serum uric acid concentrations were significantly elevated during the combination therapy, reaching > or =7.0 mg/L in men and > or =6.5 mg/L in women in 15% of patients. The urinary uric acid excretion rate was also elevated significantly. Peg-IFNalpha2b plus ribavirin combination therapy induced an elevation of uric acid concentration, although the elevated levels were still within normal limit in many cases. It may be that ribavirin plays a leading role in its elevation and other factors may also be involved.

Hematologic and Urinary Excretion Anomalies in Patients with Chronic Fatigue Syndrome

Patients with chronic fatigue syndrome (CFS) have a broad and variable spectrum of signs and symptoms with variable onsets. This report outlines the results of a single-blind, cross-sectional research project that extensively investigated a large cohort of 100 CFS patients and 82 non fatigued control subjects with the aim of performing a case-control evaluation of alterations in standard blood parameters and urinary amino and organic acid excretion profiles. Blood biochemistry and full blood counts were unremarkable and fell within normal laboratory ranges. However, the case-control comparison of the blood cell data revealed that CFS patients had a significant decrease in red cell distribution width and increases in mean platelet volume, neutrophil counts, and the neutrophil-lymphocyte ratio. Evaluation of the urine excretion parameters also revealed a number of anomalies. The overnight urine output and rate of amino acid excretion were both reduced in the CFS group (P < 0.01). Significant decreases in the urinary excretion of asparagine (P < 0.0001), phenylalanine (P < 0.003), the branch chain amino acids (P < 0.005), and succinic acid (P < 0.0001), as well as increases in 3-methylhistidine (P < 0.05) and tyrosine (P < 0.05) were observed. It was concluded that the urinary excretion and blood parameters data supported the hypothesis that alterations in physiologic homeostasis exist in CFS patients.

Dietary sodium chloride intake independently predicts the degree of hyperchloremic metabolic acidosis in healthy humans consuming a net acid-producing diet

We previously demonstrated that typical American net acid-producing diets predict a low-grade metabolic acidosis of severity proportional to the diet net acid load as indexed by the steady-state renal net acid excretion rate (NAE). We now investigate whether a sodium (Na) chloride (Cl) containing diet likewise associates with a low-grade metabolic acidosis of severity proportional to the sodium chloride content of the diet as indexed by the steady-state Na and Cl excretion rates. In the steady-state preintervention periods of our previously reported studies comprising 77 healthy subjects, we averaged in each subject three to six values of blood hydrogen ion concentration ([H]b), plasma bicarbonate concentration ([HCO(3)(-)]p), the partial pressure of carbon dioxide (Pco(2)), the urinary excretion rates of Na, Cl, NAE, and renal function as measured by creatinine clearance (CrCl), and performed multivariate analyses. Dietary Cl strongly correlated positively with dietary Na (P < 0.001) and was an independent negative predictor of [HCO(3)(-)]p after adjustment for diet net acid load, Pco(2) and CrCl, and positive and negative predictors, respectively, of [H]b and [HCO(3)(-)]p after adjustment for diet acid load and Pco(2). These data provide the first evidence that, in healthy humans, the diet loads of NaCl and net acid independently predict systemic acid-base status, with increasing degrees of low-grade hyperchloremic metabolic acidosis as the loads increase. Assuming a causal relationship, over their respective ranges of variation, NaCl has approximately 50-100% of the acidosis-producing effect of the diet net acid load.

Dietary acetic acid reduces serum cholesterol and triacylglycerols in rats fed a cholesterol-rich diet

To investigate the efficacy of the intake of vinegar for prevention of hyperlipidaemia, we examined the effect of dietary acetic acid, the main component of vinegar, on serum lipid values in rats fed a diet containing 1 % (w/w) cholesterol. Animals were allowed free access to a diet containing no cholesterol, a diet containing 1 % cholesterol without acetic acid, or a diet containing 1 % cholesterol with 0.3 % (w/w) acetic acid for 19 d. Then, they were killed after food deprivation for 7 h. Cholesterol feeding increased serum total cholesterol and triacylglycerol levels. Compared with the cholesterol-fed group, the cholesterol and acetic acid-fed group had significantly lower values for serum total cholesterol and triacylglycerols, liver ATP citrate lyase (ATP-CL) activity, and liver 3-hydroxy-3-methylglutaryl-CoA content as well as liver mRNA levels of sterol regulatory element binding protein-1, ATP-CL and fatty acid synthase (P<0.05). Further, the serum secretin level, liver acyl-CoA oxidase expression, and faecal bile acid content were significantly higher in the cholesterol and acetic acid-fed group than in the cholesterol-fed group (P<0.05). However, acetic acid feeding affected neither the mRNA level nor activity of cholesterol 7alpha-hydroxylase. In conclusion, dietary acetic acid reduced serum total cholesterol and triacylglycerol: first due to the inhibition of lipogenesis in liver; second due to the increment in faecal bile acid excretion in rats fed a diet containing cholesterol.

Anthropometrics Provide a Better Estimate of Urinary Organic Acid Anion Excretion than a Dietary Mineral Intake-Based Estimate in Children, Adolescents, and Young Adults

The role of elevated net endogenous acid production (NEAP) in the causation of osteoporosis, muscle wasting, and kidney stones is currently under discussion. The aim of this study was to examine whether urinary organic acid anion excretion, a major component of NEAP, is predicted primarily by anthropometric- (OA(anthro)) or diet- (OA(diet)) based estimates. Dietary intakes, anthropometric data, and 24-h urinary excretion rates of organic acids (24h-OA(urine)) were determined cross sectionally in healthy children (6-7 y; n = 217), adolescents (13-14 y; n = 91), and young adults (18-22 y; n = 82). OA(anthro) was computed from body surface area and OA(diet) calculated using a published algorithm based on dietary intakes of mineral anions and cations. There was a significant increase (P < 0.0001) in 24h-OA(urine) across the age groups that was no longer discernible after correction for body surface area. In almost all sex-stratified subsamples, OA(anthro) had a higher correlation with 24h-OA(urine) than OA(diet). Multiple regression analyses, using energy-corrected diet variables, revealed that OA(anthro) was consistently the primary predictor of 24h-OA(urine) (R(2) varying from 0.15 to 0.39) and dietary fat and protein were sporadic predictors. In accordance with the observed age independency of 24h-OA(urine) after body surface area correction, our findings indicate that OA(anthro) is a better estimate of 24h-OA(urine) in healthy children, adolescents, and young adults than OA(diet). This further confirms that the (principally diet-dependent) NEAP comprises a component, i.e., organic acid anions, that is reasonably predictable by anthropometrics. Consequently, the other component, i.e., the potential renal acid load, appears to be the primary parameter that characterizes the diet-induced acid load.

The influence of allopurinol on urinary purine loss after repeated sprint exercise in man

The influence of allopurinol on urinary purine loss was examined in 7 active male subjects (age 24.9 ± 3.0 years, weight 82.8 ± 8.3 kg, V˙ o 2peak 48.1 ± 6.9 mL · kg −1 · min −1). These subjects performed, in random order, a trial with 5 days of prior ingestion of a placebo or allopurinol. Each trial consisted of eight 10-second sprints on an air-braked cycle ergometer and was separated by at least a week. A rest period of 50 seconds separated each repeated sprint. Forearm venous plasma inosine, hypoxanthine (Hx) and uric acid concentrations were measured at rest and during 120 minutes of recovery from exercise. Urinary inosine, Hx, xanthine, and uric acid excretion were also measured before and for 24 hours after exercise. During the first 120 minutes of recovery, plasma Hx concentrations, as well as the urinary Hx and xanthine excretion rates, were higher ( P < .05) with allopurinol compared with the placebo trial. In contrast, plasma uric acid concentration and urinary uric acid excretion rates were lower ( P < .05) with allopurinol. The total urinary excretion of purines (inosine + Hx + xanthine + uric acid) above basal levels was higher in the allopurinol trial compared with placebo. These results indicate that the total urinary purine excretion after intermittent sprint exercise was enhanced with allopurinol treatment. Furthermore, the composition of urinary purines was markedly affected by this drug.

Correlation of uric acid and urinary albumin excretion rate in patients with type 2 diabetes mellitus in Taiwan

Uric acid is detrimental to the kidneys in animal models. However, its role in human diabetic nephropathy has not been extensively studied. This study evaluated the association between serum uric acid and urinary albumin-to-creatinine ratio (ACR) among patients with type 2 diabetes mellitus in Taiwan. A total of 343 patients (144 men and 199 women), aged 62.8 +/- 10.8 years and not using uric acid-lowering agents, diuretics, or alcohol, were recruited. Serum uric acid and urinary ACR were determined. Normoalbuminuria, microalbuminuria, and macroalbuminuria were defined as ACR <30.0, 30.0 to 299.9, and > or =300.0 microg/mg, respectively. The respective uric acid levels for normoalbuminuria (N= 166), microalbuminuria (N= 130), and macroalbuminuria (N= 47) were 5.2 +/- 1.6 mg/dL, 5.6 +/- 1.9 mg/dL, and 6.7 +/- 2.1 mg/dL (P < 0.001). The mean +/- SD (minimum-maximum) values of uric acid for the first to the fourth quartile were 3.4 +/- 0.6 (1.7-4.2), 4.9 +/- 0.4 (4.3-5.4), 6.0 +/- 0.3 (5.5-6.5), and 8.1 +/- 1.2 (6.6-12.2), respectively. Prevalence of abnormal albuminuria (microalbuminuria plus macroalbuminuria) for the respective quartiles were 38.4%, 51.2%, 50.6%, and 66.3% (P trend <0.01). In men, uric acid correlated positively with triglycerides and natural logarithmic [ln (ACR)] (gamma= 0.168, P < 0.05). In women, uric acid correlated positively with triglycerides, ln (ACR) (gamma= 0.277, P < 0.01) and body mass index (borderline significant P < 0.1), but negatively with calculated creatinine clearance. The standardized regression coefficient for ln (ACR) and the odds ratio for abnormal albuminuria for every 1 mg/dL increment of uric acid after adjusting for calculated creatinine clearance and other confounders were 0.138 (P < 0.05) and 1.183 (1.025-1.364), respectively. The results after excluding 127 cases with a history of hypertension were similar. Serum uric acid is an independent correlate of urinary ACR in Taiwanese patients with type 2 diabetes mellitus.

Analysis of the In Vivo Functions of Mrp3

Multidrug resistance protein 3 (MRP3) is an ATP-binding cassette transporter that is able to confer resistance to anticancer agents such as etoposide and to transport lipophilic anions such as bile acids and glucuronides. These capabilities, along with the induction of the MRP3 protein on hepatocyte sinusoidal membranes in cholestasis and the expression of MRP3 in enterocytes, have led to the hypotheses that MRP3 may function in the body to protect normal tissues from etoposide, to protect cholestatic hepatocytes from endobiotics, and to facilitate bile-acid reclamation from the gut. To elucidate the role of Mrp3 in these processes, the Mrp3 gene (Abcc3) was disrupted by homologous recombination. Homozygous null animals were healthy and physically indistinguishable from wild-type mice. Mrp3(-/-) mice did not exhibit enhanced lethality to etoposide phosphate, although an analysis of transfected human embryonic kidney 293 cells indicated that the potency of murine Mrp3 toward etoposide ( approximately 2.0- to 2.5-fold) is comparable with that of human MRP3. After induction of cholestasis by bile duct ligation, Mrp3(-/-) mice had 1.5-fold higher levels of liver bile acids and 3.1-fold lower levels of serum bilirubin glucuronide compared with ligated wild-type mice, whereas significant differences were not observed between the respective sham-operated mice. Bile acid excretion, pool size, and fractional turnover rates were similar in Mrp3(-/-) and wild-type mice. We conclude that Mrp3 functions as an alternative route for the export of bile acids and glucuronides from cholestatic hepatocytes, that the pump does not play a major role in the enterohepatic circulation of bile acids and that the lack of chemosensitivity is probably attributable to functional redundancy with other pumps.

Delineation of molecular changes in intrahepatic cholesterol metabolism resulting from diminished cholesterol absorption

The absorption of cholesterol by the small intestine is a major route for the net entry of cholesterol into the body and can therefore affect the plasma low density lipoprotein-cholesterol (LDL-C) concentration. These studies used ezetimibe, a potent inhibitor of cholesterol absorption, to delineate the biochemical and molecular changes in intrahepatic metabolism and biliary lipid secretion when there is a major reduction in chylomicron cholesterol delivery to the liver. In female LDL receptor (LDLR)-deficient (LDLR-/-) mice fed a basal diet containing ezetimibe (0-10 mg/day/kg body weight), cholesterol absorption was reduced up to 91%, fecal neutral sterol excretion was increased up to 4.7-fold, and plasma total cholesterol concentrations decreased by up to 18%. Blocking cholesterol absorption prevented the accumulation of very low density lipoproteins and LDL in the circulation of LDLR-/- mice fed a lipid-rich diet. In female LDLR+/+ mice fed the lipid-rich diet with ezetimibe, the relative mRNA level for the LDLR in the liver was 2-fold greater than in matching mice given the lipid-rich diet alone. We conclude that in the mouse the reduction in plasma LDL-C levels induced by blocking cholesterol absorption reflects both a diminished rate of LDL-C production and a modest increase in hepatic LDLR expression.

β-Thalassemia major and its effect on amino acid metabolism and growth in patients in the United Arab Emirates

Background There may be a marked reduction in essential amino acids in the serum of children with thalassemia major and this is related to decreased growth in affected children. Methods One hundred patients with β-thalassemia and 50 control children selected from among those who had presented with minor disorders unrelated to hematological disease were recruited. Urine and heparinized blood were collected from fasting thalassemic patients. After deproteinization and dilution, amino acid concentrations were measured using ion-exchange chromatography. Results Isoleucine ( p<0.0001), phenylalanine ( p<0.05), tyrosine ( p<0.0001), taurine ( p<0.0001) and glutamine ( p<0.01) were significantly decreased in the plasma of thalassemic patients compared to the control group. Whereas glutamate ( p<0.0001), serine ( p<0.05) and proline ( p<0.05) were significantly higher in thalassemic patients, threonine, glycine, alanine, valine, methionine, leucine, ornithine, lysine, histidine and arginine values were not different. The essential amino acids taurine ( p<0.0001), methionine ( p<0.01), valine ( p<0.01), phenylalanine ( p<0.01) and leucine ( p<0.05) were significantly decreased in urine of thalassemic patients vs. controls, but threonine and ornithine were not different. The mean urinary excretion rate of β-aminoisobutyric acid was not different (69±96 in thalassemics vs. 41±52 in controls). However, most plasma and urinary essential amino acids were found to be lower in thalassemics. Thalassemic patients were also found to be significantly growth impaired for age, both in height and weight compared to controls. Conclusion Lower plasma values of essential amino acids and a decrease in urinary amino acids occur in thalassemic patients. Growth impairment both in height and weight also occurs in thalassemic patients compared to a control population.

3-Hydroxypropionic Acid and Methylcitric Acid Are Not Reliable Indicators of Marginal Biotin Deficiency in Humans

In two studies comprising 10 and 11 subjects, respectively, marginal biotin deficiency was induced experimentally by an egg-white diet in healthy men and women. The following urinary organic acids were assessed for their usefulness in detecting marginal biotin status: 1) 3-hydroxypropionic acid and methylcitric acid, organic acids that reflect decreased activity of the biotin-dependent enzyme propionyl-CoA carboxylase and 2) methylcrotonylglycine and isovalerylglycine, organic acids that reflect decreased activity of methylcrotonyl-CoA carboxylase. Mean 3-hydroxypropionic acid excretion rates remained normal during biotin depletion in both studies. By the end of the depletion period, 3-hydroxypropionic acid excretion identified only 5 of 21 marginally deficient subjects. Mean methylcitric acid excretion increased (P < 0.0001) in the first study but not in the second. Mean methylcrotonylglycine excretion increased in each study (P < 0.004 and P < 0.05, respectively); methylcrotonylglycine excretion identified 13 of 21 marginally deficient subjects. Mean isovalerylglycine excretion increased only in the first study (P = 0.006) and identified only 6 of 21 deficient subjects. We conclude that none of these organic acids is as sensitive an indicator of marginal biotin deficiency as 3-hydroxyisovaleric acid, which reflects decreased methylcrotonyl-CoA carboxylase.

Assessment of DNA damage in glue sniffers by use of the alkaline comet assay

Toluene is used widely, not only in industry, but also in households where toluene exposure and abuse can occur. To estimate the genotoxic risk of toluene exposure, DNA damage was determined in peripheral lymphocytes of 20 glue sniffers and 20 age-matched controls by use of the alkaline comet assay. Urinary hippuric acid and o-cresol excretion rates, which are used as a marker for toluene exposure, were also measured in sniffers and compared with historical control values. The increase in genetic damage in sniffers was statistically significant as compared to control subjects ( P<0.0001). The mean values of the hippuric acid and o-cresol excretion rate for glue sniffers was 73- and 1582-fold higher, respectively, than in controls and confirms the putative exposure. Education of the general public and efforts to keep adolescents away from volatile solvent-based products, which may lead to a desire of sniffing in the future, would be advisable.

Resistance of SHP-null Mice to Bile Acid-induced Liver Damage

The orphan nuclear hormone receptor SHP (gene designation NROB2) is an important component of a negative regulatory cascade by which high levels of bile acids repress bile acid biosynthesis. Short term studies in SHP null animals confirm this function and also reveal the existence of additional pathways for bile acid negative feedback regulation. We have used long term dietary treatments to test the role of SHP in response to chronic elevation of bile acids, cholesterol, or both. In contrast to the increased sensitivity predicted from the loss of negative feedback regulation, the SHP null mice were relatively resistant to the hepatotoxicity associated with a diet containing 0.5% cholic acid and the much more severe effects of a diet containing both 0.5% cholic acid and 2% cholesterol. This was associated with decreased hepatic accumulation of cholesterol and triglycerides in the SHP null mice. There were also alterations in the expression of a number of genes involved in cholesterol and bile acid homeostasis, notably cholesterol 12alpha-hydroxylase (CYP8B1), which was strongly reexpressed in the SHP null mice, but not the wild type mice fed either bile acid containing diet. This contrasts with the strong repression of CYP8B1 observed with short term bile acid feeding, as well as the effects of long term feeding on other bile acid biosynthetic enzymes such as cholesterol 7alpha-hydroxylase (CYP7A1). CYP8B1 expression could contribute to the decreased toxicity of the chronic bile acid treatment by increasing the hydrophilicity of the bile acid pool. These results identify an unexpected role for SHP in hepatotoxicity and suggest new approaches to modulating effects of chronically elevated bile acids in cholestasis.