Genetic diversity within and among four South European native horse breeds based on microsatellite DNA analysis: implications for conservation.

In the present study, genetic analyses of diversity and differentiation were performed on four Basque-Navarrese semiferal native horse breeds. In total, 417 animals were genotyped for 12 microsatellite markers. Mean heterozygosity was higher than in other horse breeds, surely as a consequence of management. Although the population size of some of these breeds has declined appreciably in the past century, no genetic bottleneck was detected in any of the breeds, possibly because it was not narrow enough to be detectable. In the phylogenetic tree, the Jaca Navarra breed was very similar to the Pottoka, but appeared to stand in an intermediate position between this and the meat breeds. Assuming that Pottoka is the breed less affected by admixture, the others gradually distanced themselves from it through varying influences from outside breeds, among other factors. In a comparative study with other breeds, the French breeds Ardanais, Comtois, and Breton were the closest to the four native breeds. Three different approaches for evaluating the distribution of genetic diversity were applied. The high intrabreed variability of Euskal Herriko Mendiko Zaldia (EHMZ) was pointed out in these analyses. In our opinion, cultural, economic, and scientific factors should also be considered in the management of these horse breeds.

Application of two SH-based methods for metallothionein determination in mussels and intercalibration of the spectrophotometric method: laboratory and field studies in the Mediterranean Sea.

Metallothionein (MT) induction is widely used as a biomarker of exposure to metals in mussels. The aims of the present work were first to compare the suitability of spectrophotometry and differential pulse polarography (DPP) for MT detection in mussels exposed to 200 ppb cadmium for 9 days in a laboratory experiment and in mussels sampled in different seasons from expected pollution gradients along the Mediterranean Sea; second, to intercalibrate the widely used spectrophotometric method using mussels from Saronikos Gulf. In the intercalibration of the spectrophotometric method, similar results (p>0.05) were obtained by two different research teams indicating a good reproducibility of the technique. However, polarographic and spectrophotometric methods gave significantly (p<0.05) different results in laboratory and field studies. In the laboratory experiment, MT values detected with DPP were nine times higher than with spectrophotometry. The results obtained by the two methods were significantly correlated. Both methods could discriminate between control and exposed mussels. In field studies, MT values obtained by DPP were 34-38-fold higher than with spectrophotometry, and MT concentrations measured by both methods were not correlated. This discrepancy could be due to several factors, including the low levels of bioavailable metals in the studied areas and the possibility that the different methods can measure MT isoforms differentially. Further work is needed to decipher the functions of MT isoforms in mussels. This information is relevant for the application of MT as a biomarker in biomonitoring programmes.

Detection of polymorphisms in the tumour necrosis factor alpha candidate gene in sheep.

Primers based on GenBank sequences of the ovine tumour necrosis factor (TNF)-alpha gene were designed to amplify a 273-bp fragment comprising part of the fourth exon and the 3' untranslated region (UTR) of the ovine TNF-alpha gene. Five different single-strand conformational polymorphism (SSCP) patterns were detected in a number of unrelated animals and three different alleles were identified and sequenced. These alleles differed in one deletion and one single nucleotide polymorphism (SNP) and were named TNF*01, TNF*02 and TNF*03. These alleles corresponded to three sequences previously characterized by other groups. In the population analysis, no significant differences were found in the frequencies of the Latxa and Rasa breeds. This is the first description of allelic variation in the ovine TNF-alpha gene.

Analysis of the genetic structure of endangered bovine breeds from the Western Pyrenees using DNA microsatellite markers.

In the Western Pyrenees, three out of four native cattle breeds are in grave danger of extinction. Genetic variation of all four breeds was assessed by analyzing 478 animals using 11 microsatellite markers. A moderate/high within-breed variability was found, a favorable factor to consider when planning conservation and improvement programs. Interestingly, the only selected commercial breed, the Pirenaica, showed depressed heterozygosity levels and a low average number of alleles, perhaps explainable by intensive human selection exacerbated by a bottleneck effect. The Pirenaica also exhibited pronounced genetic differences and was the largest contributor of diversity among the breeds from the Western Pyrenees. Among endangered cattle breeds from this region, our results highlight the singularity of the Betizu. Geographic isolation among herds may be responsible for the large F(IS) value found in the Betizu breed. Lastly, our study suggests that the use of highly selected breeds may be one of the causes of distortion in phylogenetic analyses.

Production and evaluation of alloantibodies against sheep MHC Class I antigens.

In this work more than 600 sera obtained from three different sources (parous ewes and directed immunizations with whole blood or leucocytes) were tested for cytotoxic antilymphocyte antibodies. A high incidence of cytotoxic antibodies in pregnancy-stimulated animals was confirmed. For the sake of comparison, a comprehensive review of other studies in sheep and other species was performed. There are several different reasons that could explain the differences found between these studies, among them the time of sampling and the cytotoxic assay procedures. Moreover, antibodies were also found in non-pregnant females which may imply that pregnancy is not the only stimulus for antibody production and environmental factors, such as molecular mimicry between infectious agents and lymphocyte antigens, could be the reason for their appearance. In the case of alloimmunizations with leucocytes or whole blood the results were very close to those obtained in goats. Taking into account the Strength Index, the immunization sera were of higher quality. Our results support the methodology of Nesse and Larsen since one injection of whole blood of the lamb is an easy procedure that produces alloantisera with a high value for major histocompatibility complex Class I antigens typing.

Structuring the genetic heterogeneity of the Basque population: a view from classical polymorphisms.

In this study we analyze 18 classical polymorphisms (ABO, Rh, MNSs, Lewis, P, Duffy, Kell, ADA, ESD, PGM1, PGD, AK1, ACP1, GLO1, HP, GC, TF, and PI) in over 2000 autochthonous individuals from 14 natural districts in three provinces of the Basque Country (Alava, Guipuzcoa, and Biscay). Heterogeneity analysis via the chi2 test and a calculation of F(ST) indicate that there is significant genetic heterogeneity between the Basque districts. The R matrix informs us that this heterogeneity is not significantly concentrated in a single district or in the districts of a single province, but is rather distributed among several districts belonging to the three provinces analyzed. We undertake to assess the influence of various historical, geographical, and cultural factors on the genetic structure of the Basque population. Analysis suggests that allele distribution is geographically patterned in the Basque Country. The gradient distributions observed in the case of some alleles (ABO*O, RH*cDE, RH*cde, MNS*MS, and ACP1*C) on the basis of Moran's autocorrelation coefficient I, along with the influence of the two main travel routes through the Basque Country (western route through Bilbao and eastern route through Vitoria), suggest that the gene flow tends toward the coast. As regards other factors considered (administrative division, repopulation processes, linguistic heterogeneity, and north vs. south cultural heterogeneity), we detected only a certain influence exerted by an old tribal differentiation (2000 B.P.), which would diminish with the passage of time.

[Strengthening the interdisciplinary process in student's research programs]. / Fecundando o processo da interdisciplinaridade na iniciação científica.

The modern disciplinary focus is based on specific processes and presents a tendency to reduce knowledge by describing it as a consequence of a highly rational scientific posture that has promoted excessive specialization and fragmentation in detriment of the human being. With the development of societies and the perspective of knowledge interconnection which searches for institutions with global and broad dimensions in the spheres of knowing and doing, disciplinarity will be transcended. In this sense, the purpose of this work is to report the experience of an undergraduate students' research program in the view of interdisciplinarity. Playing the roles of advisor and student from different areas--Nursing and Administration respectively--we are experiencing an interdisciplinary process through the exchange of ideas, actions and the investigation on human resources and their management in the Nursing area. We believe that the dissemination of this experience can contribute to strengthen interdisciplinarity in the context of research in undergraduate programs.

[Recruitment and selection of human resources in a psychiatric hospital at a municipality of São Paulo]. / Recrutamento e seleção de recursos humanos em um hospital psiquiátrico de um município paulista.

This paper aims at disseminating the experience of recruiting and selecting human resources in a psychiatric hospital in the city of Ribeirão Preto, a philanthropic institution with one hundred and four beds that assists pharmaco-dependent patients with mental problems. It presently has eighty-four employees and a high staff turnover in different sectors. As trainees, we realized that the high turnover impaieds the development of activities at the organization as well as prevented a better care delivery to clients. Therefore, we were invited to integrate a team that was made responsible for the recruitment and selection of human resources for this institution. After these procedures and the respective follow-up by those in charge of different sectors, our purpose is to reduce the turnover, implement larger institutional engagement and more synchrony among employees, reduce expenses and bureaucratic activities related to hiring and laying off personnel, reduce operational work and implementing more assisting activities in terms of planning, orientation, execution and evaluation.

[Chemical composition of a typical bread and its energy and protein supply in the population from the northeast of Mexico]. / Composición química del pan tradicional e industrial y su aporte de energía y proteína en la población del Noroeste de México.

Bread from wheat flour is one of the most widely consumed products by the Mexican population. In this study, proximate composition, and mineral content were determined, and cost per/gram of protein and energy of traditional and industrial bread were compared. Seven types of bread were analyzed: bolillo, virginia, white bread and pastries conchita, croissant, breadroll and donuts. Products were analyzed for proximate composition by official methods, and content of Na, K, Ca, Mg, Fe, y Zn by atomic absorption spectrometry. Based on 24 hour recall interviews, the consumption of each type of bread as well as the percent of protein and energy allowances were calculated. Significant differences (p < 0.05) were found with respect to nutrient content between traditional and industrial breads. White bread and pastries from industrial origin showed higher costs (p < 0.05) per gram of protein and energy than traditional products in most cases. Both industrial and traditional breads showed higher fat content than that established by Official Mexican regulations. A high content of Ca was found in bread from the industrial origin and K, Mg, Fe y Zn were higher (p < 0.05) than those from the traditional bakery.

Inhibition of esterases in the marine gastropod Littorina littorea exposed to cadmium.

Electrophoretic analysis of Littorina littorea esterases indicated the presence of a cathodal migration isoenzyme (EST-C), which was identified in extracts obtained from the gonad-digestive gland complex. A loss of EST-C enzyme activity was observed in individuals exposed to Cd2+. This loss of activity was complete in 86% of individuals dead by the action of Cd2+ and in 68% of those individuals that survived Cd2+ exposure. This difference was statistically significant. In vitro inhibition of esterases by different concentrations of Cd2+ and Cu2+ was studied in individuals not experimentally exposed to heavy metals, to determine whether EST-C inhibition was caused by direct binding of Cd2+ to the esterase molecule or by Cd2+ displacement of a metal ion from essential Cu2+- or Zn2+-containing proteins, which may then be responsible for the inhibition of esterase. Contrary to what happened when individuals were exposed to Cd2+, in the in vitro experiment with Cd2+, inhibition of anodal but not cathodal systems was observed. At the same time, when Cu2+ was used in the in vitro experiment, both anodal and cathodal systems were inhibited. These results suggest that the inhibition of EST-C activity by Cd2+ takes place in vivo and seems not to be due to the direct action of Cd2+ on the molecule, but rather is a process in which transcriptional, posttranscriptional, and other inhibitory processes may be involved.

Alpha-2-HS-glycoprotein gene frequencies in Basques.

The genetic polymorphism of human alpha 2-HS-glycoprotein (AHSG) was analyzed in a sample of 736 native individuals from the three provinces of the Basque Autonomous Community (Guipúzcoa, Vizcaya and Alava). The Basque population shows the highest frequency of the allele AHSG*3 described to date in European populations. The same is not true for the frequency of allele AHSG*2, which fits well into the genocline described in Europe, in accordance with the adaptive hypothesis.

An improved method for typing of plasma alpha 1-protease inhibitor (PI1) variants in sheep.

Sheep plasma alpha 1-protease inhibitor (PI1) variants were typed by analysing neuraminidase treated samples by isoelectric focusing in ultrathin polyacrylamide gels followed by staining for trypsin inhibition. The 10 different PI1 phenotypes observed were shown to be controlled by four codominant alleles. The probability of sire exclusion provided by PI1 polymorphism was about 0.45 in both of the studied milk sheep breeds (Latxa and Karrantzar) of the Basque Country.

Are the Basques a single and a unique population?

Different analyses of genetic polymorphisms performed on the Basque population have suggested a possible heterogeneity of the Basques and a singularity of their genetic characteristics. In this paper, both aspects are analyzed by means of the genetic study of seven polymorphic systems--ACP, ADA, AK, ESD, PGD, GC, and HP--in 854 autochthonous individuals from the province of Vizcaya. The individuals were classified as being from the regions of Arratia, Guernica, Durango, Uribe, Marquina, Lea, and Bilbao, on the basis of the birthplaces of their four grandparents. Analyses for heterogeneity of the gene frequencies distribution suggest that there is a moderate genetic heterogeneity, probably produced by centuries of geographical and administrative isolation of these regions. The comparison with caucasoid populations, performed using the principal components analysis and Cavalli-Sforza and Edwards arc distance, indicates that the subpopulations of the province of Vizcaya have experienced little genetic exchange with other caucasoids and that the distribution of their genetic frequencies differentiates them from other populations.

Acid phosphatase, adenosine deaminase and esterase D polymorphisms in the Spanish Basque population.

The 3 red-cell polymorphic systems acid phosphatase (ACP), adenosine deaminase (ADA) and esterase D (ESD) have been studied in a random sample of 1,112 individuals from the Basque country: The allelic frequencies obtained were ACP*A = 0.275, ACP*B = 0.718 and ACP*C = 0.007; ADA*2 = 0.021, and, ESD*2 = 0.066. The allelic frequencies have been compared with those of other Basque and other European populations. In comparison with Basques, significant differences were detected only for ACP, whereas as regards other Europeans significant differences were obtained with practically all the populations compared for the 3 genetic systems studied. The low values of the less frequent alleles, especially that for the ACP*C allele which is the lowest reported in Europe, are noteworthy.

AK1, PGD, GC and HP frequencies in the Basque population: a review.

A random sample from the Basque population has been studied for 4 polymorphic genetic markers. The gene frequencies are AK1*1 = 0.954, PGD*A = 0.991, GC*1 = 0.663, HP*1 = 0.442. The comparison between the data obtained and other existing studies on Basques shows significant differences. The overall data on the Basque population is heterogeneous for the markers investigated, and the comparison with neighbouring non-Basque populations corroborates this heterogenity.

Some red cell enzymes and haptoglobin gene frequencies in two Basque regions and León.

A study was conducted to determine the distribution of phenotypes and gene frequencies of haptoglobin, phosphoglucomutase 1, esterase D, 6-phosphogluconate dehydrogenase and three monomorphic systems SOD, sMDH and NADH DIA I. Results obtained from two Basque regions and León were compared with those from other Spanish populations. Gene frequencies observed in the León sample were similar to those obtained from the other Spanish samples. The Basque samples differed from the other Spanish populations in the gene frequencies of esterase D.

Some genetic markers in Castillian populations.

Haptoglobin, phosphoglucomutase 1 and 6-phosphogluconate dehydrogenase gene frequencies have been found for some Castillian provinces. The NADH diaphorase I, superoxide dismutase, lactate dehydrogenase, soluble oxaloacetate transaminase, soluble malate dehydrogenase, carbonic anhydrase I and carbonic anhydrase II are non-polymorphic red cell enzymes in Caucasoids and we have verified this in Castillian samples.

Polymorphism of delta-aminolevulinic acid dehydratase in Basque populations.

Human red cell delta-aminolevulinic acid dehydratase (ALADH; EC 4.2.1.24) polymorphism was studied in three population samples of the Basque Country. The frequency of the ALADH2 was around 0.08 and similar to that in other European countries.

A cline in the acid phosphatase1 distribution in the Iberian Peninsula.

Blood samples from autochthonous and unrelated persons were examined for acid phosphatase (ACP1) phenotypes. The subjects were 143 Spanish Basques, 118 people from León and 295 from Castile. The samples were typed using starch gels, with one surface coloured with phenolphthalein diphosphate (Na5 salt) and the other surface revealed with 4-methylumbelliferyl-dihydrogenphosphate. The gene frequencies observed are compared with those obtained by other authors.

Red cell glyoxalase I polymorphism in Basque and Castilian populations.

In samples of Arratia and Gernika populations, who are indigenous Basque ethnic groups, the frequency of the GLOI1 allele was respectively 0.457 +/- 0.042 and 0.459 +/- 0.041. Four samples belonging to Castilian populations were also studied. The results of this work suggested homogeneity between the Spanish and other populations.