Genetic variation of physiological response to aflatoxin in Gallus domesticus

Summary A pedigreed, commercial broiler population of 31 sire families was administered dietary aflatoxin at levels of either 0.0 or 5.0 g of aflatoxin per g of diet from 7 to 21 days of age and their response assessed by various physiological parameters.Body weight, gain, packed red blood cell volume (PCV). plasma albumin, plasma protein and cholesterol responses were significantly reduced from control values by the 5.0 g/g aflatoxin diet. Males had greater body weights and gains in both dietary regimes than females. Females had significantly higher PCV, protein, albumin and cholesterol values in the 5.0 g/g aflatoxin group than their male counterparts. These differences resulted in significant sex × aflatoxin level interactions for these parameters. Coefficients of variation were increased for all parameters measured in the 5.0 g/g aflatoxin treatment compared to values for the control group. This increase was greatest for plasma protein, albumin, and cholesterol responses. Heritabilities were calculated for all responses within both treatment groups and were found to be increased in all cases by the 5.0 g/g aflatoxin diet. Highly significant phenotypic correlations were determined between body weight and gain and between plasma albumin and total plasma protein in both treatment groups. High phenotypic correlations among PCV, plasma cholesterol, plasma protein, and plasma albumin were noted in the 5.0 g/g aflatoxin group. Significant genetic correlations were determined between body weight and gain and between plasma albumin and plasma protein in the control group. Body weight and gain and plasma protein, albumin, cholesterol and PCV were genetically correlated in the 5.0 g/g aflatoxin group. Genetic correlations calculated across environments for the same traits were high for PCV, body weight and gain and much lower for plasma albumin, plasma protein, and plasma cholesterol.The results of this study demonstrate that genetic variability for resistance to aflatoxin exists in commercial broiler populations. Strong genetic and phenotypic relationships, and high heritabilities associated with plasma albumin and protein suggest their applicability as selection criteria for aflatoxin resistance. Genetic correlation for these traits across dietary environments indicate that responses for aflatoxin resistance should be measured during aflatoxin challenge and suggest that selection for growth and selection for aflatoxin resistance are not antagonistic.     

Genetic Parameters of Serum Cholesterol Levels, Activity and Growth in Mice

Heritabilities (h²) of, and phenotypic and genetic correlations among, serum cholesterol (SC), levels of physical activity, growth, and body moisture content were estimated from data on 466 sire-son pairs of random-bred, ICR albino mice. Heritability estimates of SC and body moisture content were 0.31 ± 0.07 and 0.29 ± 0.11, respectively. Estimates of h² for activity measured by standard exercise wheels at 28, 49 and 70 days of age were 0.31 ± 0.17, 0.50 ± 0.11 and 0.27 ± 0.14 and for weight at 21, 44 and 67 days of age, were 0.11 ± 0.09, 0.33 ± 0.09 and 0.42 ± 0.08, respectively. The estimates of h² for weight gain ranged from 0.22 ± 0.07 to 0.27 ± 0.08. Significant negative phenotypic correlations occurred between activity and rate of body weight gain. A negative genetic correlation of -0.70 ± 0.11 between SC and body moisture implied that SC and percent body fat are positively correlated. Genetic correlations among SC values and activity scores and between SC values and body weight gains were near zero.     

Selection for Rate and Efficiency of Lean Gain in the Rat

Full-sib family selection for rate (WP) or efficiency (WP/F) of protein gain in rats from 3 to 9 weeks of age was applied for five generations. Three rats per litter were killed to estimate carcass protein. Standardized response/cumulative selection for WP was.19±.10 for WP,.28±.10 for 3- to 9-week gain,.28±.08 for 9-week weight,.16±.08 for litter size,.22±.12 for skinning loss and -.07±.09 for fraction of protein in the live weight. Response from selection for WP/F was.18±.16 for WP/F,.20±.11 for WP,.21±.11 for weight gain,.16±.11 for 3-week weight,.21±.10 for 9-week weight, but negligible for skinning loss or body protein. Response to WP/F selection was extremely variable among generations, associated with generation differences in weight and composition at 9 weeks. Estimates of heritability from offspring-midparent regression were.20±.12 for WP and.24±.08 for WP/F. Estimates of genotype-generation environment interaction were large for growth, feed intake and skinning loss. Maternal effects were large for weaning weight, fraction of body protein and WP. Sire component genetic correlations were 1.08±.13 for WP with total gain,.92±.08 for WP/F with gross efficiency and.29±.25 for WP with WP/F. A partitional calorimeter was used to evaluate heat production of rats. Lines differed in average heat loss but not in heat loss per unit actual or metabolic weight. Response to selection has been steady for WP but probably could be improved by selecting for WP/F at a constant weight rather than a constant age.     

Variation in Plasma Calcium with Induced Changes in Plasma Specific Gravity,Total Protein,and Albumin

The relationship of plasma calcium levels to changes in plasma specific gravity, total protein, and albumin induced by venous stasis was investigated. Factors were derived for adjusting calcium results to offset the effects of variation in protein concentration and thus to make them of increased discriminatory value to the clinician. The validity of an existing specific gravity correction has been substantiated, but a more exact adjustment of 0·23 mg/100 ml of calcium for every 0·001 change in specific gravity is proposed. We recommend for automated laboratories that the factor based on albumin be used: 0·09 mg/100 ml of calcium should be subtracted from the total calcium value for every increase of 0·1 g in albumin above 4·6 g/100 ml, and a corresponding addition should be made for values of albumin below 4·6 g/100 ml.Using a calcium specific electrode, it has been shown that the ionized calcium concentration does not alter with prolonged venous stasis.     

Repeated Oronasal Exposure to Lipopolysaccharide Induced Mucosal IgA Responses in Periparturient Dairy Cows

This study investigated the effects of repeated oronasal treatment with lipopolysaccharide (LPS) on the humoral immune responses in saliva, vaginal mucus, and the plasma markers of the acute phase response in periparturient dairy cows. One hundred pregnant Holstein cows were administered either 3 increasing doses of LPS (n = 50) as follows: 1) 0.01 µg/kg body weight (BW) on d −28, 2) 0.05 µg/kg BW on d −25, and −21, and 3) 0.1 µg/kg BW on d −18, and −14, or sterile saline solution (controls; n = 50) oronasally for 3 consecutive wk starting at 28 d before parturition. Intensive sampling was conducted on thirty cows (n = 15/group). Multiple saliva, vaginal mucus and blood samples were collected around parturition and analyzed for total immunoglobulin-(Ig)A, plasma serum amyloid A (SAA), lipopolysaccharide-binding protein (LBP), anti-LPS IgA, IgG, IgM, tumour necrosis factor(TNF)-α, and interleukin(IL)-1. Results regarding total secretory IgA (sIgA) antibodies showed greater concentrations in the saliva and an overall tendency for higher total sIgA in the vaginal mucus of the LPS-treated cows. Treatment had no effect on plasma sIgA, IgG, IgM anti-LPS antibodies, haptoglobin, SAA, LBP, TNF-α, and IL-1. Treatments by time interactions were observed for SAA and IL-1 with lowered concentrations of both variables in the plasma of LPS-treated cows after parturition. Overall, repeated oronasal LPS treatment clearly enhanced total sIgA antibodies in the saliva, stimulated their production in vaginal mucus shortly before calving, and lowered plasma IL-1 around parturition, but showed limited effects on markers of the acute phase response in the plasma in dairy cows around parturition.     

Genetic parameters and responses of performance and body composition traits in pigs selected for high and low growth rate on a fixed ration over a set time

Two lines of Large White pigs of common genetic origin were divergently selected over four years for high and low growth rate during a 6 week post-weaning test period in which all pigs were fed the same total amount of food (80% of estimated ad libitum intake). Genetic parameters and direct and correlated responses in performance and carcass traits were estimated on 2884 pigs with pedigrees comprising a total of 5324 animals, with restricted maximum likelihood and best linear unbiased prediction methods applied to a multi-trait animal model. Estimates of heritability (± SE) were 0.19 ± 0.04 for lifetime daily gain (LDG), 0.16 ± 0.03 for test daily gain (TDG), 0.25 ± 0.04 for ultrasound P2 backfat (UBF) and 0.16 ± 0.03 for food conversion ratio during test (TFC), and 0.15 ± 0.04 for daily carcass weight gain (CDG), 0.43 ± 0.06 for carcass backfat (CFT) and 0.40 ± 0.06 for carcass lean percentage (LEAN). Common litter effects for TDG, UBF and TFC were less than 5% and for LDG, 17% of total phenotypic variance. Genetic correlations between performance and carcass traits were moderately to highly favourable. After four years of divergent selection for growth rate, the selection responses in estimated breeding value (EBV) for TDG were 40.14 and -41.11 g (SED 2.95) for the high and low growth lines, respectively. The regressions of EBV on year of birth, indicate that the annual genetic trend for TDG, was 8.73 g/yr in the high and -8.48 g/yr in the low lines (P < 0.001). Correlated genetic responses in the high and low lines respectively were 5.28 g and -12.40 g (SED 2.09) in LDG, -0.35 mm and 0.56 mm (SED 0.009) in UBF, -0.145 units and 0.185 units (SED 0.012) in TFC, 3.17 g and -10.97 g (SED 1.53) in CDG, -1.13 mm and 1.01 mm (SED 0.155) in CFT and 1.24% and -1.27% (SED 0.150) in LEAN. It was concluded that selection for increased post-weaning daily gain on a ration of fixed amount reduces the age at slaughter and the level of backfat and increases the efficiency of food utilisation, weight and leanness of pig carcasses.     

Myocardial and Skeletal Muscle Concentrations of Digoxin in Patients on Long-term Therapy

The digoxin content was measured in samples of left ventricular papillary muscle, skeletal muscle, and plasma obtained during mitral valve replacement from eight patients on maintenance treatment with the drug. The content in papillary muscle ranged from 15·5 to 132 ng/g (mean 77·7) and in skeletal muscle from 7·5 to 23 ng/g (mean 11·3). The ratio of myocardial digoxin concentration to plasma concentration varied between patients from 39:1 to 155:1. No simple relationship exists between plasma levels of digoxin and its concentration in the heart muscle, but total myocardial concentration may not accurately reflect therapeutic activity.     

Unpredictable Feeding Impairs Glucose Tolerance in Growing Lambs

Irregular eating is associated with insulin resistance and metabolic disease in adults but may affect young, growing children differently. We investigated the metabolic effects of unpredictable feeding in female juvenile lambs randomly assigned to receive, for six weeks, maintenance feed given twice daily in equal portions (Control Group, C; n = 24) or the same weekly feed amount in aliquots of variable size at unpredictable times (Unpredictable Group, U; n = 21). Intravenous glucose tolerance tests (IVGTT), insulin tolerance tests (ITT), and measurement of diurnal plasma cortisol concentrations were performed pre and post the dietary intervention. Groups were compared using t test and RM ANOVA. Weight gain was similar in both groups (C 18±2%; U 16±2% of initial body weight). Glucose area under the curve (AUC) was unchanged in C (AUC pre 818±34, post 801±33 mmol.min.l⁻¹), but increased by 20% in U (pre 830±25, post 1010±19 mmol.min.l⁻¹; p<0.0001), with an inadequate insulin response to glucose load (log(AUC insulin first 40 minutes) post intervention C 1.49±0.04 vs U 1.36±0.04 ng.min.ml⁻¹; p = 0.03). Insulin tolerance and diurnal variation of plasma cortisol concentrations were not different between groups. Unpredictable feeding impairs insulin response to glucose in growing lambs despite high quality food and normal weight gain. Irregular eating warrants investigation as a potentially remediable risk factor for disordered glucose metabolism.     

Correlated responses of growth traits to selection for high and low plasma alkaline phosphatase in mice

Summary The effectiveness of two way selection for plasma alkaline phosphatase (ALP) was investigated in order to determine its influences on growth traits through thirteen generations. The responses of the two lines selected for high (HP) and low (LP) ALP at 45 days of age were compared to that of the mice selected for large (L) and small (SM) body size. The selection responses of plasma ALP were very effective for both HP and LP lines, with average responses per generation calculated from linear regressions of 0.227±0.037 and –0.088±0.022 respectively. The final levels of ALP in HP and LP were 5.54±0.71 and 1.27±0.20 in the thirtheenth generation, while the SM, L and base population had levels of 3.49±0.08, 0.86±0.55 and 2.77±0.56 respectively. The body weight at 45 days of age in LP (31.4±1.4 g) as a correlated response was significantly higher than HP (23.4±1.8 g) at generation 10. The correlated response of milk yield, measured by weight gain up to 12 days of age, was significantly greater in the LP line than in HP, but the correlated response of gains after weaning was not so different as the response of milk yield. The response of litter size and weight in LP showed significant higher levels than that of HP, but pups' birth weight did not differ between LP and HP. It is suggested that the correlated response of milk yield contributed more to the divergence of body size between HP and LP than the gain after weaning.Realized heritabilities of ALP were 0.335±0.059 (HP) and 0.279±0.051 (LP). Realized genetic correlations between ALP and 45 days' body weight were –0.27±0.13 (HP with SM) and –0.52±0.19 (LP with L). Realized genetic correlations between ALP and milk yield were –0.95±0.03 (HP) and –0.37±0.29 (LP). Correlations between ALP and postweaning gains were fairly low.     

THE INFLUENCE OF ENVIRONMENTAL TEMPERATURE ON THE UTILIZATION OF FOOD ENERGY IN BABY CHICKS

1. An optimum of environmental temperature is to be expected for the utilization of food energy in warm blooded animals if their food intake is determined by their appetite. 2. Baby chicks were kept in groups of five chicks in a climatic cabinet at environmental temperatures of 21°, 27°, 32°, 38°, and 40°C. during the period of 6 to 15 days of age. The intake of qualitatively complete food was determined by their appetite. Food intake, excretion, and respiratory exchange were measured. Control chicks from the same hatch as the experimental groups were raised in a brooder and were given the same food as the experimental chicks. The basal metabolism of each experimental group was determined from 24 to 36 hours without food at the age of 16 days. 3. The daily rate of growth increased with decreasing environmental temperature from 2.74 gm. at 40°C. to 4.88 gm. at 21°C. This was 4.2 to 6.5 per cent of their body weight. 4. The amount of food consumed increased in proportion to the decrease in temperature. 5. The availability of the food, used for birds instead of the digestibility and defined as See PDF for Structure showed an optimum at 38°C. 6. The CO₂ production increased from 2.95 liters CO₂ per day per chick at 40°C. to 6.25 liters at 21°C. Per unit of the 3/4 power of the body weight, 23.0 liters CO₂ per kilo^3/4 was produced at 40°C. and 43.4 liters per kilo^3/4 at 21°C. The CO₂ production per unit of 3/4 power of the weight increased at an average rate of approximately 1 per cent per day increase in age. The R.Q. was, on the average, 1.04 during the day and 0.92 during the night. 7. The net energy is calculated on the basis of C and N balances. A maximum of 11.8 Cal. net energy per chick per day was found at 32°C. At 21°C. only 6.9 Cal. net per day per chick was produced and at 40°C. an average of 6.7 Cal. 8. The composition of the gained body substance changed according to the environmental temperature. The protein stored per gram increase in body weight varied from 0.217 to 0.266 gm. protein and seemed unrelated to the temperature. The amount of fat per gram gain in weight dropped from a maximum of 0.153 gm. at 32°C. to 0.012 gm. at 21°C. and an average of 0.107 gm. at 40°C. The energy content per gram of gain in weight had its maximum of 2.95 Cal. per gm. at 38°C. and its minimum of 1.41 Cal. per gm. at 21°C. at which temperature the largest amount of water (0.763 gm. per gm. increase in body weight) was stored. 9. The basal metabolism increased from an average of 60 Cal. per kilo^3/4 at an environmental temperature of 40°C. to 128 Cal. per kilo^3/4 at 21°C. No indication of a critical temperature was found. 10. The partial efficiency, i.e. the increase in net energy per unit of the corresponding increase in food energy, seemed dependent on the environmental temperature, reaching a maximum of 72 per cent of the available energy at 38°C. and decreasing to 57 per cent at 21°C. and to an average of 60 per cent at 40°C. 11. The total efficiency, i.e. the total net energy produced per unit of food energy taken in, was maximum (34 per cent of the available energy) at 32°C., dropped to 16 per cent at 21°C., and to an average of 29 per cent at 40°C.     

Protein Intake and Plasma Amino-acids of Infants of Low Birth Weight

The plasma amino-acid levels in infants of low birth weight fed on expressed human milk and on a proprietary breast-milk substitute, S26, with a protein intake of not more than 4·5 g/kg/day were compared with those in infants fed on an evaporated milk formula whose protein intake ranged from 6·15 to 12·3 g/kg/day, as well as with normal infants on normal feeds and protein intake. In general, there was little difference between the levels in infants of low birth weight and in normal infants on the same protein intake. The five infants of low birth weight on high protein intake had generally higher levels of plasma amino-acids compared with the group on the lower protein intake, and in particular the levels of tyrosine, phenylalanine, methionine, and cystathionine could be extremely high. Apart from methionine these high levels may be the result both of a reduction in activity of the enzymes involved in the metabolism of these amino-acids, due to the immaturity of the infant, and of the increased stress of a high protein intake. In view of a possible long-term effect of abnormally high plasma amino-acid levels it is suggested that the protein intake of infants of low birth weight should not exceed 6 g/kg/day.     

Identification of Altered Plasma Proteins by Proteomic Study in Valvular Heart Diseases and the Potential Clinical Significance

Background

Little is known about genetic basis and proteomics in valvular heart disease (VHD) including rheumatic (RVD) and degenerative (DVD) valvular disease. The present proteomic study examined the hypothesis that certain proteins may be associated with the pathological changes in the plasma of VHD patients.

Methods and Results

Differential protein analysis in the plasma identified 18 differentially expressed protein spots and 14 corresponding proteins or polypeptides by two-dimensional electrophoresis and mass spectrometry in 120 subjects. Two up-regulated (complement C4A and carbonic anhydrase 1) and three down-regulated proteins (serotransferrin, alpha-1-antichymotrypsin, and vitronectin) were validated by ELISA in enlarging samples. The plasma levels (n = 40 for each) of complement C4A in RVD (715.8±35.6 vs. 594.7±28.2 ng/ml, P = 0.009) and carbonic anhydrase 1 (237.70±15.7 vs. 184.7±10.8 U/L, P = 0.007) in DVD patients were significantly higher and that of serotransferrin (2.36±0.20 vs. 2.93±0.16 mg/ml, P = 0.025) and alpha-1-antichymotrypsin (370.0±13.7 vs. 413.0±11.6 µg/ml, P = 0.019) in RVD patients were significantly lower than those in controls. The plasma vitronectin level in both RVD (281.3±11.0 vs. 323.2±10.0 µg/ml, P = 0.006) and DVD (283.6±11.4 vs. 323.2±10.0 µg/ml, P = 0.011) was significantly lower than those in normal controls.

Conclusions

We have for the first time identified alterations of 14 differential proteins or polypeptides in the plasma of patients with various VHD. The elevation of plasma complement C4A in RVD and carbonic anhydrase 1 in DVD and the decrease of serotransferrin and alpha-1-antichymotrypsin in RVD patients may be useful biomarkers for these valvular diseases. The decreased plasma level of vitronectin – a protein related to the formation of valvular structure – in both RVD and DVD patients might indicate the possible genetic deficiency in these patients.     

Pharmacological PPARα Activation Markedly Alters Plasma Turnover of the Amino Acids Glycine,Serine and Arginine in the Rat

The current study extends previously reported PPARα agonist WY 14,643 (30 µmol/kg/day for 4 weeks) effects on circulating amino acid concentrations in rats fed a 48% saturated fat diet. Steady-state tracer experiments were used to examine in vivo kinetic mechanisms underlying altered plasma serine, glycine and arginine levels. Urinary urea and creatinine excretion were measured to assess whole-body amino acid catabolism. WY 14,643 treated animals demonstrated reduced efficiency to convert food consumed to body weight gain while liver weight was increased compared to controls. WY 14,643 raised total amino acid concentration (38%), largely explained by glycine, serine and threonine increases. ³H-glycine, ¹⁴C-serine and ¹⁴C-arginine tracer studies revealed elevated rates of appearance (R_a) for glycine (45.5±5.8 versus 17.4±2.7 µmol/kg/min) and serine (21.0±1.4 versus 12.0±1.0) in WY 14,643 versus control. Arginine was substantially decreased (−62%) in plasma with estimated R_a reduced from 3.1±0.3 to 1.2±0.2 µmol/kg/min in control versus WY 14,643. Nitrogen excretion over 24 hours was unaltered. Hepatic arginase activity was substantially decreased by WY 14,643 treatment. In conclusion, PPARα agonism potently alters metabolism of several specific amino acids in the rat. The changes in circulating levels of serine, glycine and arginine reflected altered fluxes into the plasma rather than changes in clearance or catabolism. This suggests that PPARα has an important role in modulating serine, glycine and arginine de novo synthesis.     

Temperature-Induced Protein Conformational Changes in Barley Root Plasma Membrane-Enriched Microsomes: II. Intrinsic Protein Fluorescence

The membrane-bound proteins of barley (Hordeum vulgare L. cv Conquest) root plasma membrane-enriched microsomes displayed fluorescence typical of protein-associated trytophan residues. The protein fluorescence intensity was sensitive to variations in sample temperature. The temperature-induced decline in protein fluorescence intensity was nonlinear with slope discontinuities at about 12 and 32°C. Detergents at levels above their critical micelle concentration enhanced protein fluorescence. Glutaraldehyde reduced protein fluorescence. Protein fluorescence polarization increased at temperatures above 30°C. Both the rate of tryptophan photoionization and the fluorescence intensity of the photoionization products suggested alterations in membrane protein conformation between 12 and 32°C. The quenching of the intrinsic protein fluorescence by acrylamide and potassium iodide indicated changes in accessibility of the extrinsic agents to the protein tryptophan residues beginning at about 14°C. The results indicate thermally induced changes in the dynamics of the membrane proteins over the temperature range of 12 to 32°C which could account for the complex temperature dependence of the barley root plasma membrane ATPase.     

The Response of Metallothionein and Malondialdehyde after Exclusive and Combined Cd/Zn Exposure in the Crab Sinopotamon henanense

The purpose of this paper is to show the interactions of Cd and Zn in the freshwater crab Sinopotamon henanense through metallothionein (MT) and malondialdehyde (MDA) level measurements. Laboratory acclimated S.henanense were exposed to Cd (50 µg/L, 100 µg/L, 500 µg/L ), and Zn (100 µg/L, 1000 µg/L) alone and in combined treatments (100 µg/L Zn+50 µg/L Cd, 100 µg/L Zn+100 µg/L Cd, 100 µg/L Zn+500 µg/L Cd, 1000 µg/L Zn+50 µg/L Cd, 1000 µg/L Zn+100 µg/L Cd, 1000 µg/L Zn+500 µg/L Cd) for 7, 14, 21, 28, 35 days. The results demonstrated that the MDA contents increased with exposure time and dose and showed time- and dose-dependence in both gills and hepatopancreas of S.henanense after single Cd exposure, while the changes of MDA levels were not significant with single Zn exposure. The MDA levels decreased when the crabs were exposed to metal mixtures compared to Cd exposure alone, indicating that Zn mediated the cellular toxicity of Cd. MT contents increased after single Cd exposure and also showed a time- and dose-dependence, in a tissue-specific way. Zn showed a limited ability of MT induction both in gills and hepatopancreas of S.henanense. The MT contents represented not a simple addition of single metal exposures but were enhanced at a higher concentration of Zn combined with different Cd concentrations compared to single metal exposure. Whether MT can be used as a biomarker for complex field conditions need to be considered cautiously since different induction patterns of MT were found among single Zn, Cd and combined groups. It is suggested that several biomarkers together as a suite should be used in the monitoring of heavy metal pollution in the aquatic environment.     

Polymorphic Variants of SCN1A and EPHX1 Influence Plasma Carbamazepine Concentration,Metabolism and Pharmacoresistance in a Population of Kosovar Albanian Epileptic Patients

Aim

The present study aimed to evaluate the effects of gene variants in key genes influencing pharmacokinetic and pharmacodynamic of carbamazepine (CBZ) on the response in patients with epilepsy.

Materials & Methods

Five SNPs in two candidate genes influencing CBZ transport and metabolism, namely ABCB1 or EPHX1, and CBZ response SCN1A (sodium channel) were genotyped in 145 epileptic patients treated with CBZ as monotherapy and 100 age and sex matched healthy controls. Plasma concentrations of CBZ, carbamazepine-10,11-epoxide (CBZE) and carbamazepine-10,11-trans dihydrodiol (CBZD) were determined by HPLC-UV-DAD and adjusted for CBZ dosage/kg of body weight.

Results

The presence of the SCN1A IVS5-91G>A variant allele is associated with increased epilepsy susceptibility. Furthermore, carriers of the SCN1A IVS5-91G>A variant or of EPHX1 c.337T>C variant presented significantly lower levels of plasma CBZ compared to carriers of the common alleles (0.71±0.28 vs 1.11±0.69 μg/mL per mg/Kg for SCN1A IVS5-91 AA vs GG and 0.76±0.16 vs 0.94±0.49 μg/mL per mg/Kg for EPHX1 c.337 CC vs TT; P<0.05 for both). Carriers of the EPHX1 c.416A>G showed a reduced microsomal epoxide hydrolase activity as reflected by a significantly decreased ratio of CBZD to CBZ (0.13±0.08 to 0.26±0.17, p<0.05) also of CBZD to CBZE (1.74±1.06 to 3.08±2.90; P<0.05) and CDR_CBZD (0.13±0.08 vs 0.24±0.19 μg/mL per mg/Kg; P<0.05). ABCB1 3455C>T SNP and SCN1A 3148A>G variants were not associated with significant changes in CBZ pharmacokinetic. Patients resistant to CBZ treatment showed increased dosage of CBZ (657±285 vs 489±231 mg/day; P<0.001) but also increased plasma levels of CBZ (9.84±4.37 vs 7.41±3.43 μg/mL; P<0.001) compared to patients responsive to CBZ treatment. CBZ resistance was not related to any of the SNPs investigated.

Conclusions

The SCN1A IVS5-91G>A SNP is associated with susceptibility to epilepsy. SNPs in EPHX1 gene are influencing CBZ metabolism and disposition. CBZ plasma levels are not an indicator of resistance to the therapy.     

Reciprocal Recurrent Selection Compared to within-Strain Selection for Increasing Rate of Egg Lay of Tribolium under Optimal and Stress Conditions

A replicated comparison of reciprocal recurrent selection (rrs) based on crossbred performance and within strain-selection (wss) based on purebred performance was made in three diverse environments over ten generations for the improvement of a heterotic trait, 4-day virgin egg lay of Tribolium castaneum. A selection intensity of 20% based on performance in either an optimum (33°), a mild stress (38°), or a severe stress (28°) environment was applied uniformly. Periodically, the performance of each population was measured in all three environments to provide both direct and correlated responses.—Heritability of egg lay in the base population ranged from 0.36 ± 0.03 in optimum to 0.26 ± 0.03 in severe stress. Estimates of dominance effects assumed significant proportions in severe stress only. Genetic correlations for egg lay in diverse environments were large and positive (.6 to.8).—Only in severe stress did the rrs response significantly exceed that for wss. Quadratic adjustments fitted to response curves revealed that small initial genetic gains under rrs were followed by significantly increasing rates of gain in late generations of selection. The reverse was true for wss. This and evidence from realized heritabilities and genetic correlations suggested that rrs had utilized both additive and dominance effects, but wss response was limited to additive effects.—These results agree with selection theory in demonstrating that purebred selection is more efficient than crossbred selection in utilizing additive gene effects. The latter method has merit when non-additive effects assume significant proportions, and this is the more probable case for severe stress conditions.     

Effects of short-term continuous subcutaneous insulin infusion on fasting plasma fibroblast growth factor-21 levels in patients with newly diagnosed type 2 diabetes mellitus

Background

To investigate the effects of short-term continuous subcutaneous insulin infusion (CSII) on plasma fibroblast growth factor-21 (FGF-21) levels in patients with newly diagnosed type 2 diabetes mellitus (nT2DM).

Method

Sixty-eight patients with nT2DM (nT2DM group), and 52 gender-, age- and body mass index (BMI) -matched normal glucose tolerance (NGT group) controls participated in the study. 30 nT2DM patients with FBG≥14.0 mmol/L were treated with CSII for 2 weeks, and were underwent a euglycemic–hyperinsulinemic clamp pre- and post-treatment. Plasma FGF-21 concentrations were measured with a commercial ELISA kit. The relationship between plasma FGF-21 levels and metabolic parameters was also analyzed.

Results

Fasting plasma FGF-21 levels were higher in the nT2DM group than in NGT groups (1.60±0.08 vs. 1.13±0.26 µg/L, P<0.01). In nT2DM patients, fasting plasma FGF-21 concentrations were significantly decreased after CSII treatment for 2 weeks (1.60±0.08 vs.1.30±0.05 µg/L, P<0.05), accompanied by a significant increase in the whole body glucose uptake (M value) and blood glucose control. The changes in plasma FGF-21 levels (ΔFGF-21) were positively associated with the amelioration of insulin resistance shown by the changes in M value.

Conclusion

Plasma FGF-21 level is associated with whole body insulin sensitivity and significantly reduced following short-term CSII treatment.     

The Effect of Lutein Supplementation on Blood Plasma Levels of Complement Factor D,C5a and C3d

Lutein is selectively taken up by the primate retina and plays an important role as a filter for harmful blue light and as an antioxidant. Recent studies have shown that lutein has systemic anti-inflammatory properties. Dietary lutein has been associated with reduced circulating levels of inflammatory biomarkers such as CRP and sICAM. Whether lutein also affects activation of the complement system has not yet been addressed and was the purpose of the study described here. Seventy-two subjects with signs of early macular degeneration were randomly assigned to receive either a 10 mg lutein supplement or a placebo during one year. EDTA blood samples were collected at 0, 4, 8 and 12 months. Complement factor D (CFD), a rate limiting component of the alternative pathway of complement activation and the complement activation products C5a and C3d were determined in the plasma samples by ELISA. A significant 0.11 µg/ml monthly decrease in plasma CFD concentration was observed in the lutein group (p<0.001), resulting in a 51% decrease from 2.3 µg/ml at baseline to 1.0 µg/ml at 12 months. The C5a concentration showed a significant 0.063ng/ml monthly decrease in the lutein group (p<0.001) resulting in a 36% decrease from 2.2ng/ml at baseline to 1.6ng/ml at 12 months. The C3d concentration showed a significant 0.19µg/ml monthly decrease in the lutein group (p=0.004) that gave rise to a 9% decrease from 15.4µg/ml at baseline to 14.4µg/ml at 12 months. In the placebo group we found a significant 0.04 µg/ml monthly decrease in plasma CFD concentration, whereas no changes were observed for C5a and C3d. Lutein supplementation markedly decreases circulating levels of the complement factors CFD, C5a and C3d levels, which might allow a simple method to control this inflammatory pathway of the innate immune system.