Neurobiological alterations in the human alcoholic brain: effect of genotype

Long-term alcohol abuse by human subjects leads to selective brain damage that is restricted in extent and variable in severity. Within the cerebral cortex, neuronal loss is most marked in the superior frontal cortex and relatively mild in motor cortex. Cirrhotic alcoholics and subjects with alcohol-related Wernicke-Korsakoff syndrome show more severe and more extensive damage than do uncomplicated cases. Accumulating evidence suggests that the likelihood of developing alcohol dependency is associated with one or more genetic markers. In previous work we showed that GABAA receptor functionality, and the subunit isoform expression that underlies this, differed in region- and disease-specific ways between alcoholics and controls. By contrast, glutamate receptor (NMDA, KA, AMPA) differences were muted or absent. Here we asked if genotype differentiated the form, pharmacology, or expression of glutamate and GABA receptors in pathologically vulnerable and spared cortical regions, with a view to determining whether such subject factors might influence the severity of alcohol-induced brain damage. Cerebrocortical tissue was obtained at autopsy under informed, written consent from uncomplicated and alcoholic-cirrhotic Caucasian (predominantly Anglo-Celtic) cases, together with matched controls and cases with cirrhosis of non-alcoholic origin. All subjects had pathological confirmation of liver and brain diagnosis; none had been polydrug abusers. Samples were processed for synaptic membrane receptor binding, mRNA analysis by quantitative RT-PCR, and protein analysis by Western blot. Genotyping was performed by PCR methods, in the main using published primers. Several genetic markers differentiated between our alcoholic and control subjects, including the GABAA receptor 2 subunit (GABB2) gene ( 2 (3) 10.329, P 0.01), the dopamine D2 receptor B1 (DRD2B) allele ( 2 (3) 10.109, P 0.01) and a subset of the alcohol dehydrogenase-3 (ADH3) alleles ( 2 (2) 4.730, P 0.05). Although neither the type-2 glutamate transporter (EAAT2) nor the serotonin transporter (5HTT) genes were significantly associated with alcoholism, only EAAT2 heterozygotes showed a significant association between ADH3 genotype and alcoholism ( 2 (3) 7.475, P 0.05). Other interactions between genotypes were also observed. DRD2A, DRD2B, GABB2, EAAT2 and 5HTT genotypes did not divide alcoholic cases and controls on NMDA receptor parameters, although in combined subjects there was a significant DRD2B X Area Interaction with glutamateNMDA receptor efficacy (F(1,57) 4.67; P 0.05), measured as the extent of glutamate-enhanced MK801 binding. In contrast, there was a significant Case-group X ADH3 X Area Interaction with glutamateNMDA receptor efficacy (F(3,57) 2.97; P 0.05). When GABAA receptor subunit isoform expression was examined, significant Case-group X Genotype X Area X Isoform interactions were found for EAAT2 with subunit mRNA (F(1,37) 4.22; P0.05), for GABB2 with isoform protein (F(1,37) 5.69; P 0.05), and for DRD2B with isoform protein (F(2,34)5.69; P0.05). The results suggest that subjects’ genetic makeup may modulate the effectiveness of amino acid-mediated transmission in different cortical regions, and thereby influence neuronal vulnerability to excitotoxicity.

Interactions between serotonergic genotype and amino acid transmitter receptor expression in human alcoholics

Serotonin can modulate the activity of neural reward pathways that are strongly implicated in mediating the effects of chronic alcohol misuse, and its treatment, in human subjects. In previous work and as discussed elsewhere at this meeting, we and others have found consistent differences in the parameters of GABA and glutamate receptors, and the expression of their component subunit transcripts and proteins, in areas of the alcoholic brain that are altered by alcoholism. We did not fi nd clear changes in GABA and glutamate transport function in such samples, but a series of microarray analyses showed consistent upregulation of the presynaptic GABA/betaine transporter SLC6A12. Microarray studies showed no signifi cant differences in the expression of transcripts associated with 5HT transmission; however, only a small number of such elements were present on the arrays. Here we partitioned GABAA and NMDA pharmacology, and subunit mRNA and protein expression, measured in samples of frontal and motor cortex obtained at autopsy from alcoholics without comorbid disease, alcoholics with liver cirrhosis, and controls, according to 5HTTLPR (SLC6A4) and 5HT1B (HTR1B) polymorphisms. We found no effect of these genotypes on the expression of GABAA receptor gene products, but there was a signifi cant mRNA Transcript X Area X Group X 5HTTLPR Interaction with NMDA subunit isoform expression measured by Real Time PCR with GAPDH normalization. Further analysis showed the effect to be selective for alcoholics with cirrhosis, to be most marked in the pathologically vulnerable frontal cortex, and to vary with subunit transcript (F2,76 = 6.545, P = 0.002). NR1 expression was most affected, followed by NR2A, with NR2B expression least altered. Pilot data suggest 5HT1B genotype may also modulate NMDA subunit expression. Interactions between amino acid and serotonin transmission may infl uence susceptibility to alcohol dependence or pathogenesis

The benefits of polyandry in the free-spawning polychaete Galeolaria caespitosa

In many species, females are thought to benefit from polyandry due to the reduced risks of fertilization by genetically incompatible sperm. However, few studies that have reported such benefits have directly attributed variation in female reproductive success to the interacting effects of males and females at fertilization. In this paper, we determine whether male x female interactions influence fertilization in vitro in the free-spawning, sessile polychaete Galeolaria caespitosa. Furthermore, we determined whether polyandry results in direct fertilization benefits for females by experimentally manipulating the number of males contributing towards staged spawning events. To test for male x female interaction effects we performed an initial experiment that crossed seven males with six females (in all 42 combinations), enabling us to assess fertilization rates for each specific male-female pairing and attribute variation in fertilization success to males, females and their interaction. This initial experiment revealed a strong interaction between males and females at fertilization, confirming that certain male-female combinations were more compatible than others. A second experiment tested the hypothesis that polyandry enhances female reproductive success by exposing each female's eggs to either a single male's sperm (monandry) or the sperm from three males simultaneously (polyandry). We performed this second experiment at two ecologically relevant sperm concentrations. This latter experiment revealed a strong fertilization benefit of polyandry, independent of the effects of sperm concentration (which were also significant). We suggest that these direct fertilization gains arising from polyandry will constitute an important source of selection on females to mate multiply in nature.

Genotype effects on body temperature in dairy cows under grazing conditions in a hot climate including evidence for heterosis

We compared diurnal patterns of vaginal temperature in lactating cows under grazing conditions to evaluate genotype effects on body temperature regulation. Genotypes evaluated were Holstein, Jersey, Jersey x Holstein and Swedish Red x Holstein. The comparison of Holstein and Jersey versus Jersey x Holstein provided a test of whether heterosis effects body temperature regulation. Cows were fitted with intravaginal temperature recording devices that measured vaginal temperature every 15 min for 7 days. Vaginal temperature was affected by time of day (P < 0.0001) and genotype x time (P < 0.0001) regardless of whether days in milk and milk yield were used as covariates. Additional analyses indicated that the Swedish Red x Holstein had a different pattern of vaginal temperatures than the other three genotypes (Swedish Red x Holstein vs others x time; P < 0.0001) and that Holstein and Jersey had a different pattern than Jersey x Holstein [(Holstein + Jersey vs Jersey x Holstein) x time, P < 0.0001]. However, Holstein had a similar pattern to Jersey [(Holstein vs Jersey) x time, P > 0.10]. These genotype x time interactions reflect two effects. First, Swedish Red x Holstein had higher vaginal temperatures than the other genotypes in the late morning and afternoon but not after the evening milking. Secondly, Jersey x Holstein had lower vaginal temperatures than other genotypes in the late morning and afternoon and again in the late night and early morning. Results point out that there are effects of specific genotypes and evidence for heterosis on regulation of body temperature of lactating cows maintained under grazing conditions and suggest that genetic improvement for thermotolerance through breed choice or genetic selection is possible.

Simulating growth, development, and yield of tillering pearl millet. III. Biomass accumulation and partitioning

Pearl millet landraces from Rajasthan, India, yield significantly less than improved cultivars under optimum growing conditions, but not under stressed conditions. To successfully develop a simulation model for pearl millet, capable of capturing such genotype x environment (G x E) interactions for grain yield, we need to understand the causes of the observed yield interaction. The aim of this paper is to quantify the key parameters that determine the accumulation and partitioning of biomass: the,light extinction coefficient, radiation use efficiency (RUE), pattern of dry matter allocation to the leaf blades, the determination of grain number, and the rate and duration of dry matter accumulation into individual grains. We used data on improved cultivars and landraces, obtained from both published and unpublished sources collected at ICRISAT, Patancheru, India. Where possible, the effects of cultivar and axis (main shoot vs. tillers) on these parameters were analysed, as previous research suggested that G x E interactions for grain yield are associated with differences in tillering habit. Our results indicated there were no cultivar differences in extinction coefficient, RUE, and biomass partitioning before anthesis, and differences between axes in biomass partitioning were negligible. This indicates there was no basis for cultivar differences in the potential grain yield. Landraces, however, produced consistently less grain yield for a given rate of dry matter accumulation at anthesis than did improved cultivars. This was caused by a combination of low grain number and small grain size. The latter was predominantly due to a lower grain growth rate, as genotypic differences in the duration of grain filling were relatively small. Main shoot and tillers also had a similar duration of grain filling. The low grain yield of the landraces was associated with profuse nodal tillering, supporting the hypothesis that grain yield was below the potential yield that could be supported by assimilate availability. We hypothesise this is a survival strategy, which enhances the prospects to escape the effects of stress around anthesis. (C) 2002 E.J. van Oosterom. Published by Elsevier Science B.V. All rights reserved.

Methods to classify maize cultivars in use efficiency and response to nitrogen

n plant breeding programs that aim to obtain cultivars with nitrogen (N) use efficiency, the focus is on methods of selection and experimental procedures that present low cost, fast response, high repeatability, and can be applied to a large number of cultivars. Thus, the objectives of this study were to classify maize cultivars regarding their use efficiency and response to N in a breeding program, and to validate the methodology with contrasting doses of the nutrient. The experimental design was a randomized block with the treatments arranged in a split-plot scheme with three replicates and five N doses (0, 30, 60, 120 and 200 kg ha-1) in the plots, and six cultivars in subplots. We compared a method examining the efficiency and response (ER) with two contrasting doses of N. After that, the analysis of variance, mean comparison and regression analysis were performed. In conclusion, the method of the use efficiency and response based on two N levels classifies the cultivars in the same way as the regression analysis, and it is appropriate in plant breeding routine. Thus, it is necessary to identify the levels of N required to discriminate maize cultivars in conditions of low and high N availability in plant breeding programs that aim to obtain efficient and responsive cultivars. Moreover, the analysis of the interaction genotype x environment at experiments with contrasting doses is always required, even when the interaction is not significant.

New strategies to detect and understand genotype-by-environment interactions and QTL-by-environment interactions

Dissertação para obtenção do Grau de Doutor em Estatística e Gestão do Risco, especialidade em Estatística

Viral genotype-specific role of PNPLA3, PPARG, MTTP, and IL28B in hepatitis C virus-associated steatosis.

BACKGROUND & AIMS: Steatosis is a prominent feature of hepatitis C, especially in patients infected with genotype 3. The analysis of genetic polymorphisms influencing steatosis in chronic hepatitis C has been limited by the studies' small sample size, and important single nucleotide polymorphisms (SNPs), such as those in the patatin-like phospholipase family 3 protein (PNPLA3), were never evaluated. METHODS: We analyzed the role of SNPs, from 19 systematically selected candidate genes, on steatosis in 626 Caucasian hepatitis C virus (HCV) infected patients. SNPs were extracted from a genome-wide association-generated dataset. Associations of alleles with the presence and/or different severity of steatosis were evaluated by univariate and multivariate logistic regression, accounting for all relevant covariates. RESULTS: The risk of steatosis was increased by carriage of I148M in PNPLA3, but only in patients with HCV genotypes non-3 (odds ratio [OR]=1.9, 95% confidence interval [CI]=1.6-2.3, p<0.001) and similar, albeit weaker associations were found for SNPs in peroxisome proliferator-activated receptor-γ (PPARG) and interleukin-28B (IL28B). Carriage of a SNP in the microsomal triglyceride transfer protein (MTTP) increased the risk of steatosis, but only in patients with HCV genotype 3 (rs1800803, OR=3.4, 95% CI=2.4-4.9, p=0.001). CONCLUSIONS: The rs738409 SNP in PNPLA3 is associated with an increased risk of steatosis in patients infected with HCV genotypes non-3. Host genes affect steatosis depending on the infecting HCV genotype, suggesting their interaction with viral factors.

Genetical and environmental analyses of yield in six biparental soybean crosses

The yearly genetic progress obtained by breeding for increased soybean yield has been considered acceptable worldwide. It is common sense, however, that this progress can be improved further if refined breeding techniques, developed from the knowledge of the genetic mechanisms controlling soybean yield, are used. In this paper, data from four cultivars and/or lines and their derived sets of F2, F3, F7, F8, F9 and F10 generations assayed in 17 environments were analyzed to allow an insight of the genetic control of soybean yield under different environmental conditions. The general picture was of a complex polygene system controlling yield in soybeans. Additive genetic effects predominated although dominance was often found to be significant. Complications such as epistasis, linkage and macro and micro genotype x environment (G x E) interactions were also commonly detected. The overall heritability was 0.29. The relative magnitude of the additive effects and the complicating factors allowed the inference that the latter are not a serious problem to the breeder. The low heritability values and the considerable magnitude of G x E interactions for yield, however, indicated that careful evaluation through experiments designed to allow for the presence of these effects is necessary for successful selection.

Repeatability of adaptability and stability parameters of common bean in unpredictable environments

The objective of this work was to estimate the repeatability of adaptability and stability parameters of common bean between years, within each biennium from 2003 to 2012, in Minas Gerais state, Brazil. Grain yield data from trials of value for cultivation and use common bean were analyzed. Grain yield, ecovalence, regression coefficient, and coefficient of determination were estimated considering location and sowing season per year, within each biennium. Subsequently, a analysis of variance these estimates was carried out, and repeatability was estimated in the biennia. Repeatability estimate for grain yield in most of the biennia was relatively high, but for ecovalence and regression coefficient it was null or of small magnitude, which indicates that confidence on identification of common bean lines for recommendation is greater when using means of yield, instead of stability parameters.

Progress in the Spanish National Barley Breeding Program

The Spanish Barley Breeding Program is carried out by four public research organizations, located at the most representative barley growing regions of Spain. The aim of this study is to evaluate the program retrospectively, attending to: i) the progress achieved in grain yield, and ii) the extent and impact of genotype-by-environment interaction of grain yield. Grain yields and flowering dates of 349 advanced lines in generations F8, F9 and F10, plus checks, tested at 163 trials over 11 years were analized. The locations are in the provinces of Albacete, Lleida, Valladolid and Zaragoza. The data are highly unbalanced because the lines stayed at the program for a maximum of three years. Progress was estimated using relative grain yield and mixed models (REML) to homogenize the results among years and locations. There was evident progress in the program over the period studied, with increasing relative yields in each generation, and with advanced lines surpassing the checks in the last two generations, although the rate of progress was uneven across locations. The genetic gain was greater from F8 to F9 than from F9 to F10. The largest non-purely environmental component of variance was genotype-by-location-by-year, meaning that the genotype-by-location pattern was highly unpredictable. The relationship between yield and flowering time overall was weak in the locations under study at this advanced stage of the program. The program can be continued with the same structure, although measures should be taken to explore the causes of slower progress at certain locations.

Genetic interactions involving major genes at the dwarf locus in egg-type chickens

Three egg-type stocks segregating dwarf (dw) and bantam (dwB) genes in female progeny were produced from the same 18 heterozygous (dwB/dw) sires used to inseminate dams of three different genotypes: normal (dw+), dwarf (dw) and bantam (dwB) dams. The heritability of 8-week body weight estimated from full-sibs of the same phenotype of progeny was 0.40, and that estimated from paternal half-sibs of the same phenotype (dwarf or bantam), and from the same genotype of dam was 0.38. Therefore, maternal and non-additive effects within genotypic classes of dam made little contribution to the genetic variance for 8-week body weight among their progeny. The interaction of sires (S) with genotypes (dw+, dw and dwB) of dam (G) was significant at the 5% level. This indicates that the rankings of the sires within each one of the three genotypes of dam were not the same, probably due to non-additive genetic variation among genotypes of dams. The evidence indicated that in general the genes from individual sires combined differently with each type of dam (G). Those genes which combined well with the genes from normal (dw+) dams combined poorly with both the genes from the dwarf (dw) and the genes from the bantam (dwB) dams. The interaction of sires (S) with phenotypes (dwarf and bantam) of progeny (P) was significant at the 10% level. The results indicated a probable gene x genotype interaction for 8-week weight between genes at the dwarf locus (dw and dwB) and the background genotype (single and/or polygenes). The correlation among paternal half-sibs was influenced more by the S x G than by the S x P interaction, but the effects tended to be cumulative

Effect of sex and genotype on cardiovascular biomarker response to fish oils: the FINGEN Study

Background: The lipid-modulatory effects of high intakes of the fish-oil fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are well established and likely to contribute to cardioprotective benefits. Objectives: We aimed to determine the effect of moderate EPA and DHA intakes (< 2 g EPA + DHA/d) on the plasma fatty acid profile, lipid and apolipoprotein concentrations, lipoprotein subclass distribution, and markers of oxidative status. We also aimed to examine the effect of age, sex, and apolipoprotein E (APOE) genotype on the observed responses. Design: Three hundred twelve adults aged 20-70 y, who were prospectively recruited according to age, sex, and APOE genotype, completed a double-blind placebo-controlled crossover study. Participants consumed control oil, 0.7 g EPA + DHA/d (0.7FO), and 1.8 g EPA + DHA/d (1.8FO) capsules in random order, each for an 8-wk intervention period, separated by 12-wk washout periods. Results: In the group as a whole, 8% and 11% lower plasma triacylglycerol concentrations were evident after 0.7FO and 1.8FO, respectively (P < 0.001): significant sex x treatment (P = 0.038) and sex x genotype x treatment (P = 0.032) interactions were observed, and the greatest triacylglycerol-lowering responses (reductions of 15% and 23% after 0.7FO and 1.8FO, respectively) were evident in APOE4 men. Furthermore, lower VLDL-cholesterol (P = 0.026) and higher LDL-cholesterol (P = 0.010), HDL-cholesterol (P < 0.001), and HDL2 (P < 0.001) concentrations were evident after fish-oil intervention. Conclusions: Supplements providing EPA + DHA at doses as low as 0.7 g/d have a significant effect on the plasma lipid profile. The results of the current trial, which used a prospective recruitment approach to examine the responses in population subgroups, are indicative of a greater triacylglycerol-lowering action of long-chain n-3 polyunsaturated fatty acids in males than in females.

Effect of sex and genotype on cardiovascular biomarker response to fish oils: the FINGEN Study

Background: The lipid-modulatory effects of high intakes of the fish-oil fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are well established and likely to contribute to cardioprotective benefits. Objectives: We aimed to determine the effect of moderate EPA and DHA intakes (< 2 g EPA + DHA/d) on the plasma fatty acid profile, lipid and apolipoprotein concentrations, lipoprotein subclass distribution, and markers of oxidative status. We also aimed to examine the effect of age, sex, and apolipoprotein E (APOE) genotype on the observed responses. Design: Three hundred twelve adults aged 20-70 y, who were prospectively recruited according to age, sex, and APOE genotype, completed a double-blind placebo-controlled crossover study. Participants consumed control oil, 0.7 g EPA + DHA/d (0.7FO), and 1.8 g EPA + DHA/d (1.8FO) capsules in random order, each for an 8-wk intervention period, separated by 12-wk washout periods. Results: In the group as a whole, 8% and 11% lower plasma triacylglycerol concentrations were evident after 0.7FO and 1.8FO, respectively (P < 0.001): significant sex x treatment (P = 0.038) and sex x genotype x treatment (P = 0.032) interactions were observed, and the greatest triacylglycerol-lowering responses (reductions of 15% and 23% after 0.7FO and 1.8FO, respectively) were evident in APOE4 men. Furthermore, lower VLDL-cholesterol (P = 0.026) and higher LDL-cholesterol (P = 0.010), HDL-cholesterol (P < 0.001), and HDL2 (P < 0.001) concentrations were evident after fish-oil intervention. Conclusions: Supplements providing EPA + DHA at doses as low as 0.7 g/d have a significant effect on the plasma lipid profile. The results of the current trial, which used a prospective recruitment approach to examine the responses in population subgroups, are indicative of a greater triacylglycerol-lowering action of long-chain n-3 polyunsaturated fatty acids in males than in females.

Location, location, location: development of spatiotemporal sequence learning in infancy

We investigated infants' sensitivity to spatiotemporal structure. In Experiment 1, circles appeared in a statistically defined spatial pattern. At test 11-month-olds, but not 8-month-olds, looked longer at a novel spatial sequence. Experiment 2 presented different color/shape stimuli, but only the location sequence was violated during test; 8-month-olds preferred the novel spatial structure, but 5-month-olds did not. In Experiment 3, the locations but not color/shape pairings were constant at test; 5-month-olds showed a novelty preference. Experiment 4 examined "online learning": We recorded eye movements of 8-month-olds watching a spatiotemporal sequence. Saccade latencies to predictable locations decreased. We argue that temporal order statistics involving informative spatial relations become available to infants during the first year after birth, assisted by multiple cues.