A polymorphism in the agouti signalling protein (ASIP) is associated with decreased levels of mRNA

To date, a role for agouti signalling protein (ASIP) in human pigmentation has not been well characterized. It is known that agouti plays a pivotal role in the pigment switch from the dark eumelanin to the light pheomelanin in the mouse. However, because humans do not have an agouti banded hair pattern, its role in human pigmentation has been questioned. We previously identified a single polymorphism in the 3'-untranslated region (UTR) of ASIP that was found at a higher frequency in African-Americans compared with other population groups. To compare allele frequencies between European-Australians and indigenous Australians, the g.8818A -> G polymorphism was genotyped. Significant differences were seen in allele frequencies between these groups (P < 0.0001) with carriage of the G allele highest in Australian Aborigines. In the Caucasian sample set a strong association was observed between the G allele and dark hair colour (P = 0.004) (odds ratio 4.6; 95% CI 1.4-15.27). The functional consequences of this polymorphism are not known but it was postulated that it might result in message instability and premature degradation of the transcript. To test this hypothesis, ASIP mRNA levels were quantified in melanocytes carrying the variant and non-variant alleles. Using quantitative real-time polymerase chain reaction the mean ASIP mRNA ratio of the AA genotype to the AG genotype was 12 (P < 0.05). This study suggests that the 3'-UTR polymorphism results in decreased levels of ASIP and therefore less pheomelanin production.

Haemoglobin adducts of acrylonitrile and ethylene oxide in acrylonitrile workers, dependent on polymorphisms of the glutathione transferases GSTT1 and GSTM1

Fifty-nine persons with industrial handling of low levels of acrylonitrile (AN) were studied. As part of a medical surveillance programme an extended haemoglobin adduct monitoring [N-(cyanoethyl)valine, CEV; N- (methyl)valine, MV; N-(hydroxyethyl)valine, HEV] was performed. Moreover, the genetic states of the polymorphic glutathione transferases GSTM1 and GSTT1 were assayed by polymerase chain reaction (PCR). Repetitive analyses of CEV and MV in subsequent years resulted in comparable values (means, 59.8 and 70.3 μg CEV/1 blood; 6.7 and 6.7 μg MV/1 blood). Hence, the industrial AN exposures were well below current official standards. Monitoring the haemoglobin adduct CEV appears as a suitable means of biomonitoring and medical surveillance under such exposure conditions. There was also no apparent correlation between the CEV and HEV or CEV and MV adduct levels. The MV and HEV values observed represented background levels, which apparently are not related to any occupational chemical exposure. There was no consistent effect of the genetic GSTM1 or GSTT1 state on CEV adduct levels induced by acrylonitrile exposure. Therefore, neither GSTM1 nor GSTT1 appears as a major AN metabolizing isoenzyme in humans. The low and physiological background levels of MV were also not influenced by the genetic GSTM1 state, but the MV adduct levels tended to be higher in GSTT1- individuals compared to GSTT1 + persons. With respect to the background levels of HEV adducts observed, there was no major influence of the GSTM1 state, but GST- individuals displayed adduct levels that were about 1/3 higher than those of GSTT1+ individuals. The coincidence with known differences in rates of background sister chromatid exchange between GSTT1- and GSTT1 + persons suggests that the lower ethylene oxide (EO) detoxification rate in GSTT1- persons, indicated by elevated blood protein hydroxyethyl adduct levels, leads to an increased genotoxic effect of the physiological EO background.

云南中甸尼西鸡的血液蛋白多态性研究

　采用水平式淀粉胶凝胶电泳技术和垂直式聚丙烯酰胺凝胶电泳技术检测了云南中 甸尼西鸡的血液蛋白多态，对33个样本的37个基因座位进行了分析，共有9个座位出现多态，分 别为ES-1、ES-2、AKP-1、AKP-2、LAP、PGM、CK、Tf及6PGD。多态位点百分比P=0.2432,平 均杂合度=0.1015,每个座位等位基因的平均数A=1.3784,结果表明尼西鸡的血液蛋白多态程度较 高，在蛋白质水平上的遗传多样性较为丰富。同时发现Amy-1座位只出现杂合子AB型。

GENETIC DIVERSITY IN THE CHINESE PANGOLIN (MANIS-PENTADACTYLA) INFERRED FROM PROTEIN ELECTROPHORESIS

We examined protein polymorphism of Chinese pangolins (Manis pentadactyla) from Yunnan Province of China, including two forms of three brown and nine dusky Chinese pangolins. Sixty-two genetic loci were screened; 12 loci were found to be polymorphic. The percentage of polymorphic loci (P) is 0.194, the mean individual heterozygosity (H) is 0.078, and the mean number of alleles (A) is 1.258. Furthermore, we calculated the genetic distance (D) between the two forms and found a low level of genetic divergence (D = 0.0206) between them, which indicates an almost-indistinguishable divergence at the level of proteins.

Genetic diversity of Chinese native pigs inferred from protein electrophoresis

We examined protein polymorphism of 20 native pig breeds in China and 3 introduced pig breeds. Thirty loci have been investigated, among which six loci were found to be polymorphic. Especially, the polymorphism of malate dehydrogenase (MDH), adenylate kinase (AK), and two new alleles of adenosine deaminase (ADA) had not been reported in domestic pigs and wild pigs. The percentage of polymorphic loci (P), the mean heterozygosity (H), and the mean number of alleles (A) are 0.200, 0.065, and 1.300, respectively. The degree of genetic variability of Chinese pigs as a whole was higher than that of goats, lower than that of cattle and horses, and similar to that of sheep. Using the gene frequencies of the 30 loci, Nei's genetic distance among the 20 native breeds in China and 3 introduced pig breeds was calculated by the formula of Nei. The program NEIGHBOR in PHYLIP 3.5c was chosen to construct an UPGMA tree and a NJ tree. Our results show that, of the total genetic variation found in the native pig breeds in China, 31% (0.31) is ascribable to genetic differences among breeds. About 69% of the total genetic variation is found within breeds. Most breeds are in linkage disequilibrium. The patterns of genetic similarities between the Chinese native pig breeds were not in agreement with the proposed pig type classification.

Application of Fluorescence Spectroscopy to Quantify Shear-Induced Protein Conformation Change

Rapid and robust methods are required to quantify the effect of hydrodynamic shear on protein conformation change. We evaluated such strategies in this work and found that the binding of the fluorescent probe 4,4'-dianilino-1, 1'-binaphthyl-5,5'-disulfonic acid (bis-ANS) to hydrophobic pockets in the blood protein von Willebrand factor (VWF) is enhanced upon the application of fluid shear to the isolated protein. Significant structural changes were observed when the protein was sheared at shear rates >= 6000/s for similar to 3.5 min. The binding of bis-ANS to multimeric VWF, but not dimeric VWF or control protein bovine serum albumin, was enhanced upon fluid shear application. Thus, high-molecular-weight VWF is more susceptible to conformation change upon tensile loading. Although bis-ANS itself did not alter the conformation of VWF, it stabilized protein conformation once it bound the sheared molecule. Bis-ANS binding to VWF was reduced when the sheared protein was allowed to relax before dye addition. Taken together with functional data in the literature, our results suggest that shear-induced conformation changes in VWF reported by bis-ANS correlate well with the normal function of the protein under physiological/pathological fluid flow conditions. Further, this study introduces the fluorescent dye bis-ANS as a tool that may be useful in studies of shear-induced protein conformation change.

Genetic variability in Larus argentatus and Sterna hirundo

Blood serum and egg-white protein samples from individuals representing seven colonies of Larusargentatus, and four colonies of Sterna hirundo were electrophoretically analysed to determine levels of genetic variability and to assess the utility of polymorphic loci as genetic markers. Variability occurred at five co-dominant autosomal loci. S. hirundo protein polymorphism occurred at the Est-5 and the Oest-l loci, while nineteen loci were monomorphic. L. argentatus samples were monomorphic at seventeen loci and polymorphic at the Ldh-A and the Alb loci. Intergeneric differences existed at the Oalb and the Ldh-A loci. Although LDH-A100 from both species possessed identical electrophoretLc mobilities, the intergeneric differences were expressed as a difference in enzyme the'ITIlostabilities. Geographical distribution of alleles and genetic divergence estimates suggest ~ hirundo population panmixis,at least at the sampled locations. The h argentatus gene pool appears relatively heterogeneous with a discreet Atlantic Coast population and a Great Lakes demic population. These observed population structures may be maintained by the relative amount of gene flow occurring within and among populations. Mass ringing data coupled to reproductive success information and analysis of dispersal trends appear to validate this assumption. Similar results may be generated by either selection or both small organism and low locus sample sizes. To clarify these results and to detect the major factor(s) affecting the surveyed portions of the genome, larger sample sizes in conjunction with precise eco-demographic data are required.

AN IMPROVED ELECTROPHORETIC METHOD FOR THE DETERMINATION OF SERUM MILK PROTEIN VARIANTS IN GYR-HOLSTEIN COWS

Milk serum proteins such as alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG) present biochemical polymorphism which is under the control of codominant autosomal alleles. In the present report, we propose modifications of traditional electrophoretic techniques such as increasing the running gel concentration from 5 to 10% and the addition of 5 M urea to the stacking gel, which permitted the detection of two variants (A and B) at the ALA and BLG loci. About 8 mul of milk serum (6 mg/ml protein) and 10 pl of total fresh milk were applied. Bovine serum albumin (BSA) and immunolactoglobulins (ILG) could also be discriminated. Total fresh milk was as useful as the purified serum milk proteins for the discrimination of ALA and BLG serum milk protein polymorphism by alkaline vertical slab polyacrylamide gel electrophoresis. However, BSA and ILG ran with caseins, which prevented their characterization in this system.

Genetic structure of Blastocerus dichotomus populations in the Parana River basin (Brazil) based on protein variability

The population structure of 147 marsh deer (Blastocerus dichotomus) from three areas in the Parana River basin, Brazil, was studied by observing protein polymorphism at 17 loci. Six loci were polymorphic and 11 monomorphic. The proportion of polymorphic loci (P) was 35.29% and the average heterozygosity (H) was 6.31%. Wright's F-ST indicated that only 4.9% of the total variation in allelic frequencies was due to genetic differences between the three groups. The high value of F-IS (0.246) indicated inbreeding in the marsh deer. Genetic distance values (D = 0.014-0.051) showed little divergence between the three areas. We suggest that probable mechanisms accounting for the genetic structure are female phylopatry and polygyny and also that inbreeding has resulted from decreasing areas of wetland leading to isolation, overhunting, and diseases transmitted by cattle.

Serum protein profiles of juvenile ring-necked pheasants vaccinated or not against newcastle disease

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Role of NOD2 variants in spondylarthritis

Objective. To investigate the role of the gene NOD2 in susceptibility to, and clinical manifestations of, ankylosing spondylitis (AS). Methods. A case-control study of NOD2 polymorphisms known to be associated with Crohn's disease (CD) (Pro268 Ser, Arg702 Trp, GlY908 Arg, and Len1007fsinsC) was performed in 229 cases of primary AS with no diagnosed inflammatory bowel disease (IBD), 197 cases of AS associated with IBD (referred to as colitic spondylarthritis; comprising 78 with CD and 119 with ulcerative colitis [UC]), and 229 ethnically matched, healthy controls. Associations between NOD2 polymorphisms and several clinical features of AS, including disease severity assessed by questionnaire and age at spondylarthritis onset, were also investigated. Exclusion linkage mapping of chromosome 16 was performed in a separate group of 185 multicase families with AS. Results. An association was identified between Gly908 Arg and UC spondylarthritis (P = 0.016, odds ratio [OR] 4.6, 95% confidence interval [95% CI] 1.316), and a nonsignificant trend with a similar magnitude was observed in association with CD spondylarthritis (P = 0.08, OR 3.9, 95% CI 0.8-18). The Pro268Ser variant was inversely associated with UC spondylarthritis (P = 0.003, OR 0.55, 95% CI 0.37-0.82), but not with CD spondylarthritis. No association was demonstrated between NOD2 variants and primary AS, or between other variants of NOD2 and either UC or CD spondylarthritis. Carriage of the Pro268 Ser polymorphism was associated with greater disease activity as measured by the Bath Ankylosing Spondylitis Disease Activity Index (P = 0.002). Although patients with CD had a younger age at spondylarthritis onset than did those with UC (22.4 years versus 26.4 years; P = 0.01), no association was noted between the NOD2 variants linked with CD and age at spondylarthritis onset. In primary AS, the presence of a gene with a magnitude of association >2.0 was excluded (exclusion logarithm of odds score less than -2.0), and no association was observed with the microsatellite D16S3136. Conclusion. NOD2 variants do not significantly affect the risk of developing primary AS, but may influence susceptibility to, and clinical manifestations of, colitic spondylarthritis.

Environmental oestrogens and sexual development in fish

This brief paper discusses the assumption that watercourses might be harbouring a chemical(s) affecting the sexual development in fish. Male fish was found with the oestrogen-dependent blood protein, vitellogenin, usually found only in maturing females. The author examines a number of man-made chemicals present in the environment have been found to be oestrogenic. The paper concludes that rivers contain environmental oestrogens that are capable of causing disruptions in the sexual development of fish. Whether or not these environmental oestrogens are causing a widespread disruption in reproduction in wild fish, however, has yet to be determined.

滇南小耳猪遗传多样性的血液蛋白电泳研究

文采用蛋白电泳技术研究滇南小耳猪血液蛋白多态性。共分析了滇南小耳猪32个遗传位 点，其中AKP、CAT、ES、PA、6PGD和TF等6个位点检测到多态性，多态位点百分比P=0．1875，平均 杂合度H=0．0712。结果表明滇南小耳猪的血液蛋白多态程度较高，反映在蛋白质水平上的遗传多样 性较为丰富。

Bioquímica en la vida diaria: el equilibrio químico y la función transportadora de la hemoglobina

La hemoglobina es una proteína sanguínea que puede transportar oxígeno, un gas insoluble en medio acuoso, llevándolo a las diferentes partes del organismo en donde es requerido para su buen funcionamiento, así como productos metabólicos como el CO2 y el hidrógeno, para su excreción. Estos procesos se ven condicionados por factores como el pH, la concentración de BPG, las presiones parciales de O2 y CO2, la cooperatividad de la unión entre la hemoglobina y esos compuestos y los cambios conformacionales que la hemoglobina debe sufrir para captar y soltar eficientemente estas moléculas en el sitio del organismo donde son requeridos. Cambios abruptos en la presión atmosférica ligados a la altura, y la exposición a altas concentraciones de otros gases afines a la hemoglobina como el monóxido de carbono, presente en vehículos o recintos cerrados, pueden comprometer el funcionamiento normal del organismo precisamente porque causan efectos sobre esa función transportadora de la hemoglobina. En este escrito, se explicarán fenómenos de la vida diaria relacionados con el transporte de gases por la hemoglobina, como una demostración de que los conocimientos bioquímicos comienzan a ser útiles desde ahora para entender situaciones cotidianas y a dejarnos la expectativa de su valor para entender muchos de los problemas de salud que tendremos en nuestras manos.

Fitness of transgenic Anopheles stephensi mosquitoes expressing the SM1 peptide under the control of a vitellogenin promoter

Three transgenic Anopheles stephensi lines were established that strongly inhibit transmission of the mouse malaria parasite Plasmodium berghei. Fitness of the transgenic mosquitoes was assessed based on life table analysis and competition experiments between transgenic and wild-type mosquitoes. Life table analysis indicated low fitness load for the 2 single-insertion transgenic mosquito lines VD35 and VD26 and no load for the double-insertion transgenic mosquito line VD9. However, in cage experiments, where each of the 3 homozygous transgenic mosquitoes was mixed with nontransgenic mosquitoes, transgene frequency of all 3 lines decreased with time. Further experiments suggested that reduction of transgene frequency is a consequence of reduced mating success, reduced reproductive capacity, and/or insertional mutagenesis, rather than expression of the transgene itself. Thus, for transgenic mosquitoes released in the field to be effective in reducing malaria transmission, a driving mechanism will be required.