ß-Alanine does not act through branched-chain amino acid catabolism in carp, a species with low muscular carnosine storage.

This study was executed to investigate the effect of dietary ß-alanine (BA) on amino acid (AA) metabolism and voluntary feed intake in carp (Cyprinus carpio) at mildly elevated temperature to exert AA catabolism. Twenty-four fish in 12 aquaria were randomly assigned to either a control diet or the same diet with 500 mg BA/kg. A 14-day period at an ideal temperature (23 °C) was followed by 15 days at chronic mildly elevated temperature (27 °C). After the 15 days, all fish were euthanised for muscle analysis on histidine-containing dipeptides (HCD), whole blood on free AA and carnitine esters. The carnosine and anserine analysis indicated that all analyses were below the detection limit of 5 µmol/L, confirming that carp belongs to a species that does not store HCD. The increases in free AA concentrations due to BA supplementation failed to reach the level of significance. The effects of dietary BA on selected whole blood carnitine esters and their ratios were also not significant. The supplementation of BA tended to increase body weight gain (P = 0.081) and feed intake (P = 0.092). The lack of differences in the selected nutrient metabolites in combination with tendencies of improved growth performance warrants further investigation to unravel the mechanism of BA affecting feed intake. This first trial on the effect of BA supplementation on AA catabolism showed that its metabolic effect in carp at chronic mildly elevated temperature was very limited. Further studies need to evaluate which conditions are able to exert an effect of BA on AA metabolism.

Effect of peripartal feeding strategy on colostrum yield and composition in sows.

Research showed a positive association between back fat (BF) change the week before farrowing and colostrum yield (CY). This study tested the causality of this association, hence to optimize CY by altering the sows' peripartal feeding strategy. Sows were randomly divided into 2 treatment groups at d 108 of gestation. The first group (L, n = 28) received 1.5 kg feed·d(-1), the second group (H, n = 22) received 3 times 1.5 kg feed·d(-1) until farrowing. Daily feed intake and CY were measured. Colostrum was analyzed for nutrient composition, AA and fatty acids, IgG and IgA. Sow serum was obtained at d 108 of gestation and d 1 of lactation after overnight fasting and analyzed for NEFA, (iso)butyrylcarnitine (C4), creatinine, urea, 3-OH-butyrylcarnitine (3-OH-C4), IgG, and IgA. Based on BF at d 108, sows were divided into body condition (BC) groups: skinny (<17 mm, n = 15), moderate (17 to 23 mm, n = 21), fat (>23 mm, n = 14). We performed ANOVA with treatment and BC as fixed factors and Scheffé post-hoc test. The week before farrowing, the L group had the lowest daily feed intake (DFI; 1.5 kg), and within the H group, fat sows (3.8 kg) had a lower DFI than skinny sows (4.3 kg; p = 0.006). The H group tended to have a greater total CY (P = 0.074) and had a greater CY/kg liveborn piglet (P = 0.018) than the L group. Compared with sows in moderate BC, fat sows had a lower total CY (P = 0.044) and a lower CY/kg liveborn piglet (P = 0.005). The H group had a greater concentration of lactose (p = 0.009) and n-3 PUFA (p < 0.001) but a lower concentration of protein (p = 0.040) in colostrum than the L group. The concentration of IgG and IgA did not differ between treatment and BC groups. Serum parameters at d 108 were similar between the treatment groups and BC groups. At d 1, the H group mobilized less body fat (NEFA: p = 0.002) and protein (creatinine: p < 0.001, C4: p = 0.016) reserves but had a greater ratio urea:NEFA (p < 0.001) and less ketone bodies (3-OH-C4: p < 0.001) compared with the L group. This indicates a more balanced entry of metabolites in the citric acid cycle and thus a better support of the maternal peripartal metabolism in the H group. Serum parameters did not differ between BC groups. Both CY and composition can be influenced by the peripartal feeding strategy and BC. The highest CY and most beneficial colostrum composition were obtained when sows entered the farrowing unit in a moderate BC and were provided a high peripartal feeding strategy.

Changes in back fat thickness during late gestation predict colostrum yield in sows.

Directing protein and energy sources towards lactation is crucial to optimise milk production in sows but how this influences colostrum yield (CY) remains unknown. The aim of this study was to identify associations between CY and the sow's use of nutrient resources. We included 37 sows in the study that were all housed, fed and managed similarly. Parity, back fat change (ΔBF), CY and performance parameters were measured. We obtained sow serum samples 3 to 4 days before farrowing and at D1 of lactation following overnight fasting. These were analysed for non-esterified fatty acids (NEFA), urea, creatinine, (iso)butyrylcarnitine (C4) and immunoglobulins G (IgG) and A (IgA). The colostrum samples collected 3, 6 and 24 h after the birth of the first piglet were analysed for their nutrient and immunoglobulins content. The technical parameters associated with CY were parity group (a; parities 1 to 3=value 0 v. parities 4 to 7=value 1) and ΔBF D85-D109 of gestation (mm) (b): CY (g)=4290-842a-113b. (R 2=0.41, P<0.001). The gestation length (P<0.001) and the ΔBF between D109 and D1 of lactation (P=0.050) were identified as possible underlying factors of the parity group. The metabolic parameters associated with CY were C4 at 3 to 4 days before farrowing (a), and 10logC4 (b) and 10logNEFA (c) at D1 of lactation: CY (g)=3582-1604a+1007b-922c (R 2=0.39, P=0.001). The colostrum composition was independent of CY. The negative association between CY and ΔBF D85-D109 of gestation could not be further explained based on our data. Sows that were catabolic 1 week prior to farrowing seemed unable to produce colostrum to their full potential. This was especially the case for sows with parities 4 to 7, although they had a similar feed intake, litter birth weight and colostrum composition compared with parities 1 to 3 sows. In conclusion, this study showed that parity and the use of body fat and protein reserves during late gestation were associated with CY, indicating that proper management of the sow's body condition during late gestation could optimise the intrinsic capacity of the sow's CY.

Questioning the Pathogenic Role of the GLA p.Ala143Thr "Mutation" in Fabry Disease: Implications for Screening Studies and ERT.

Fabry disease is an X-linked inborn error of glycosphingolipid metabolism caused by quantitative or qualitative defects in the lysosomal enzyme alfa-Galactosidase A (aGAL A), ultimately resulting in vital organ dysfunction. Mainly the kidneys, the heart, and the central nervous system are involved. While the classical phenotype of Fabry disease is readily recognizable, screening studies have identified clinical variants. Here, we report the phenotype associated with the GLA p.Ala143Thr (c.427G>A) mutation in 12 patients aged 42-83 years. None of the patients had classical Fabry signs or symptoms as angiokeratoma, hypohidrosis, acroparesthesia, or cornea verticillata. Possible Fabry manifestations were renal failure (5/12), stroke (7/12), and left ventricular hypertrophy (5/12), but these were not necessarily attributable to the p.Ala143Thr mutation, as a cardiac biopsy in one female and left ventricular hypertrophy and kidney biopsies in two males with renal failure and microalbuminuria lacked Gb-3 deposits. The literature data on this mutation as well as data collected in the Fabry Outcome Survey (FOS) database confirm these findings. The association of renal failure, stroke, and left ventricular hypertrophy with this mutation could be the result of selection bias, as most patients were detected in screening studies.We conclude that care should be taken with attribution of vital organ dysfunction to GLA sequence alterations. In case of the p.Ala143Thr mutation, and possibly also other mutations associated with an attenuated phenotype, diagnostic tools such as biopsy and imaging should critically evaluate the relation of end-organ failure with Fabry disease, as this has important consequences for enzyme replacement therapy.

Propionate absorbed from the colon acts as gluconeogenic substrate in a strict carnivore, the domestic cat (Felis catus).

In six normal-weight and six obese cats, the metabolic effect of propionate absorbed from the colon was assessed. Two colonic infusions were tested in a crossover design with intervals of 4 weeks. The test solution contained 4 mmol sodium propionate per kg ideal body weight in a 0.2% NaCl solution. Normal saline was given as control solution. Solutions were infused into the hindgut over 30 min. Blood samples were obtained prior to and at various time points after starting the infusion. As body condition did not affect evaluated parameters, all data were pooled. Plasma glucose concentrations showed differences neither over time nor during or after infusion with propionate or control. Plasma amino acid concentrations rose over time (p < 0.001), but were similar for both infusions. Plasma propionylcarnitine rose markedly towards the end of the propionate infusion and decreased afterwards (p < 0.001), whereas 3-hydroxy-3-methylglutarylcarnitine was lower 30 (p = 0.005) and 60 min (p = 0.032) after ending propionate infusions and acetylcarnitine tended to fall at the same time points (p = 0.079; p = 0.080), suggesting inhibition of gluconeogenesis from pyruvate and amino acids, but initiation of propionate-induced gluconeogenesis. In conclusion, propionate absorbed from the colon is hypothesized to act as gluconeogenic substrate, regardless of the cat's body condition.

Methods for a prompt and reliable laboratory diagnosis of Pompe disease: report from an international consensus meeting.

Pompe disease is an autosomal recessive disorder of glycogen metabolism caused by a deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA). It presents at any age, with variable rates of progression ranging from a rapidly progressive course, often fatal by one-year of age, to a more slowly, but nevertheless relentlessly progressive course, resulting in significant morbidity and premature mortality. In infants, early initiation of enzyme replacement therapy is needed to gain the maximum therapeutic benefit, underscoring the need for early diagnosis. Several new methods for measuring GAA activity have been developed. The Pompe Disease Diagnostic Working Group met to review data generated using the new methods, and to establish a consensus regarding the application of the methods for the laboratory diagnosis of Pompe disease. Skin fibroblasts and muscle biopsy have traditionally been the samples of choice for measuring GAA activity. However, new methods using blood samples are rapidly becoming adopted because of their speed and convenience. Measuring GAA activity in blood samples should be performed under acidic conditions (pH 3.8-4.0), using up to 2 mM of the synthetic substrate 4-methylumbelliferyl-alpha-D-glucoside or glycogen (50 mg/mL), in the presence of acarbose (3-9 microM) to inhibit the isoenzyme maltase-glucoamylase. The activity of a reference enzyme should also be measured to confirm the quality of the sample. A second test should be done to support the diagnosis of Pompe disease until a program for external quality assurance and proficiency testing of the enzymatic diagnosis in blood is established.

Molecular and clinical characterization of a Moroccan Cog7 deficient patient.

Mutations in the N-linked glycosylation pathway cause rare autosomal recessive defects known as Congenital Disorders of Glycosylation (CDG). A previously reported mutation in the Conserved Oligomeric Golgi complex gene, COG7, defined a new subtype of CDG in a Tunisian family. The mutation disrupted the hetero-octomeric COG complex and altered both N- and O-linked glycosylation. Here we present clinical and biochemical data from a second family with the same mutation.

Determination of carbohydrate-deficient transferrin using capillary zone electrophoresis.

BACKGROUND: Current methods for carbohydrate-deficient transferrin (CDT) often suffer from low precision, complexity, or risk of false positives attributable to genetic variants. In this study, a new capillary zone electrophoresis (CZE) method for CDT was developed. METHODS: CZE was performed on a P/ACE 5000 using fused-silica capillaries [50 microm (i.d.) x 47 cm] and the CEOFIX CDT buffer system with addition of 50 microL of anti-C3c and 10 microL of anti-hemoglobin. Native sera were loaded by high-pressure injection for 3 s, separated at 28 kV over 12 min, and monitored at 214 nm. RESULTS: CDT was completely resolved by differences in migration times (di-trisialotransferrin, 9.86 +/- 0.05 min; monosialotransferrin, 9.72 +/- 0.05 min; asialotransferrin, 9.52 +/- 0.04 min), with a CV of 0.15%. The number of theoretical plates was 312,000 +/- 21,000 for the mono- and 199 000 +/- 6500 for the di-trisialylated transferrin. Genetic CB and CD variants showed prominent peaks with migration times of 10.12 +/- 0.06 and 9.89 +/- 0.03 min, respectively, and the carbohydrate-deficient glycoprotein syndrome could be detected, excluding false-positive results. CZE results (as a percentage; y) correlated with the Axis %CDT TIA (x) values by Deming regression analysis: y = 1.92x - 7.29; r = 0.89. CDT values in 130 healthy nonalcoholics were determined. The 2.5th and 97.5th percentiles were 1.84% and 6.79%. CONCLUSIONS: CZE without sample pretreatment can determine CDT with good precision, allows detection of variants, and correlates with ion-exchange chromatography.

Carbohydrate-deficient transferrin and chronic alcohol ingestion in subjects with transferrin CD-variants.

Carbohydrate-deficient transferrin (CDT) is widely accepted as screening test for excessive alcohol consumption. However, results from subjects with transferrin variants must be interpreted with caution since chromatography-based methods may give false-positive results. Furthermore, due to the co-elution in HPLC or the co-migration in capillary zone electrophoresis (CZE) of the di- and trisialylated C transferrins with the tetrasialylated D peak, exact measurement of CDT is impossible in CD-variants. Therefore, in this study, we tried to offer a different solution, including only the asialo-D, asialo-C, monosialo-D, monosialo-C, disialo-D and trisialo-D transferrins in the CDT calculation and referring to a different cut-off value for CDT in transferrin CD-variants. Comparison of alcohol consumers with teetotalers demonstrated area under the receiver operating characteristic curve of 0.79 and 0.76 for carbohydrate-deficient transferrin, 0.71 and 0.71 for mean corpuscular volume and 0.51 and 0.68 for gamma-glutamyltransferase in 43 subjects with transferrin CD-variants and 225 subjects with CC-phenotypes, respectively. Since false-positive carbohydrate-deficient transferrin results due to a transferrin CD-variant have major social implications, capillary electrophoresis-based or similar methods (HPLC, FPLC) should be preferred in populations carrying a high D-allele frequency.

A new method for fast haptoglobin phenotyping and hemoglobin binding capacity calculation based on capillary zone electrophoresis.

A capillary zone electrophoresis method was developed for haptoglobin (Hp) phenotyping in hemoglobin (Hb) supplemented serum. The method allows a complete resolution of the major haptoglobin phenotypes Hp 1-1, Hp 2-1, and Hp 2-2 based on the difference in charge-to-mass ratio of their Hb-Hp complexes. Identification of these phenotypes was achieved by their significant differences in migration times and their marked difference in electrophoretic pattern. Our method showed full agreement with starch gel electrophoresis. Furthermore, following neuraminidase treatment of the serum, the Hp subtypes Hp1-1FF, FS and SS could be resolved, based on the same criteria as the phenotyping. The new electrophoretic method allowed typing of the rare phenotypes Hp 2-1 modified (Hp 2-1M) and Hp Johnson. The calculated hemoglobin binding capacity of serum correlates well with the nephelometrically determined haptoglobin concentration. The new method for typing haptoglobin gives prospectives for fast haptoglobin typing and Hp 1-1 subtyping.

Transferrin polymorphism influences iron status in blacks.

BACKGROUND: Genetic variants of human transferrin (TF) have been described, but little is known about their functional differences. We studied iron status according to TF phenotype in a healthy Zimbabwean population and in subjects at risk of African iron overload. METHODS: The study population consisted of 483 nondrinkers, 31 drinking spouse pairs, and 5 family pedigrees (n = 88) with index cases of iron overload. TF phenotypes were determined using starch gel electrophoresis. To evaluate iron status, serum iron, total iron-binding capacity (TIBC), ferritin, and soluble TF receptors were measured, and the percentage of saturation and the serum iron:TF ratio were calculated. The binding of the TF variants was studied by equilibrium dialysis. RESULTS: The reference population was characterized by a high TF D allele frequency (0.050) and a complete absence of homozygous TF DD individuals. Similar allele frequencies were observed in subjects at risk of African iron overload. In the reference population, male TF CD heterozygotes had significantly lower (P <0.01) values for serum iron, TIBC, TF saturation, and serum iron:TF ratio than the TF CC homozygotes; in females, only TIBC was significantly different. Overall red blood cell indices did not differ according to TF phenotype. In the population at risk of African iron overload, only serum iron:TF ratio was consistently significantly lower in TF CD phenotypes (P <0.05). After equilibrium dialysis, the amount of iron bound by TF was significantly lower (P <0.01) in TF CD individuals. CONCLUSIONS: The present data demonstrate a functional difference between TF phenotypes in blacks.

Natural killer (NK) cell leukaemia in a patient with a B cell non-Hodgkin's lymphoma.

We describe a 74-year-old woman with the diagnosis of natural killer (NK)-cell leukaemia and autoimmune pathology. Four years previously, a diffuse large B cell non-Hodgkin's lymphoma had been diagnosed and treated effectively. Although NK-cell leukaemia has been thought to be a distinct highly aggressive clinicopathological entity, our case shows no further evolution at the present time. As far as we know, this association has not been previously described in the literature.

Effect of haptoglobin phenotypes on growth of Streptococcus pyogenes.

The haptoglobin (Hp) 2-1 and 2-2 phenotypes have been shown to agglutinate Streptococcus pyogenes carrying the membrane antigen T4. In this study, the growth rate of two strains of Streptococcus pyogenes (T1 and T4) in human serum was compared among haptoglobin phenotypes in vitro. During incubation for 16 hours in serum of different haptoglobin types, only Hp 2-1 and Hp 2-2 sera showed an inhibitory effect on growth, Hp 2-2 being 1.85 times more potent than Hp 2-1. Growth of Streptococcus pyogenes T4 negatively correlated with the serum concentration of Hp 2-1 (r = -0.908) and Hp 2-2 (r = -0.953). Haptoglobin-depleted serum had no inhibitory effect on bacterial growth. Addition of haemoglobin and ferric citrate to the serum accelerated the growth of Streptococcus pyogenes T4 (P <0.05) but not in Hp 2-2 serum. Haptoglobin types 2-1 and 2-2 can be regarded as inhibitors of Streptococcus pyogenes growth in vitro. These data point towards a potential protective role of Hp 2-2 in Streptococcus pyogenes infection in vivo, independently of iron uptake.

Angiotensin I-converting enzyme insertion/deletion polymorphism: clinical implications.

Since the identification of an Insertion/Deletion polymorphism in the ACE gene, numerous studies have evaluated the potential risk of the DD genotype in cardiovascular disease and hypertension. The report of many conflicting publications reveals a strong need for reviewing the most important data. There is evidence of the absence of an association between the ACE polymorphism and hypertension in Caucasians. In blacks a positive association between the D allele and high blood pressure was seen, Japanese studies show discrepant results. Several studies showed no association between the ACE polymorphism and the risk of myocardial infarction. However, in certain subpopulations, such as low risk patients or coronary care unit patients, an increased risk of myocardial infarction in DD type is present, and a meta-analysis supports this proposition. Because of conflicting data, the potential association between the ACE polymorphism and coronary artery disease, cerebrovascular disease, left ventricular hypertrophy, hypertrophic and idiopathic dilated cardiomyopathy, carotid artery disease and diabetic and immunoglobin A nephropathy, remains inconclusive.