New polymorphic microsatellite markers for the limpet Patella rustica and cross-priming testing in four Patella species.

Eighteen microsatellite markers have been characterized from the Patella rustica genome. An average of 10 alleles per locus and an expected heterozygosity ranging from 0.054 to 0.937 were observed in a sample of 32 wild individuals from Viana do Castelo, Portugal. Distinct cross-priming amplification rates were recovered on four additional Patella species. Three monomorphic P. rustica loci were polymorphic in other Patella species. The microsatellites developed herein could be a useful intraspecific genetic tool to undertake fine population studies in the genus Patella.

Genetic characterization of southwestern European bovine breeds: a historical and biogeographical reassessment with a set of 16 microsatellites.

The origin of Iberian cattle has been suggested by some authors to be the product of European and north African cattle entrances during the last few thousands of years. However, these hypotheses were mainly based on morphological similarities. This study analyzed 889 unrelated individuals from 15 representative Iberian breeds and 3 French breeds for 16 microsatellite loci. Statistical tests were used to calculate interpopulation genetic distances (D(A)) and principal components analysis (PCA). To visualize the geographical distribution of the genetic differentiation between Iberian cattle breeds, data from the PCA analysis were used to construct synthetic maps. Genetic similarity among neighboring Iberian breeds is mainly caused by gene flow. However, recent demographic fluctuations and reproductive isolation in Alistana, Mirandesa, and Tudanca has increased genetic drift, which may be the main cause for the relatively high differentiation of these populations. The synthetic maps constructed with the first and second PCs revealed (1) a large differentiation between Northern Iberian breeds rather than between more geographically distant breeds, and (2) a clear east-west gradient that may be related with the model of demic diffusion of agriculture. Finally, we detected no strong evidence for an African genetic influence in the Iberian cattle breeds analyzed in this study.

Hemobilia in hereditary hemorrhagic telangiectasia: an unusual complication of endoscopic retrograde cholangiopancreatography.

Hepatobiliary manifestations of hereditary hemorrhagic telangiectasia (HHT) are rare, but often involve cholestasis. We report here a case of HHT associated with cholestasis due to common bile duct stenosis. Attempted balloon dilation of the stenosis during endoscopic retrograde cholangiopancreatography (ERCP) resulted in hemobilia. Hemostasis was achieved by adjusting the nasobiliary drain. The aim of this report is to highlight the biliary manifestations of HHT and draw attention to an unusual complication of ERCP in this setting.

Spontaneous bacterial peritonitis in patients with hepatic cirrhosis: evaluation of a treatment protocol at specialized units.

INTRODUCTION: Spontaneous bacterial peritonitis is a common and severe complication in patients with cirrhosis and ascitis. Its prognosis clearly depends on its precocious clinical recognition and efficacious therapy. AIM: To optimize a treatment protocol, after auditing clinical efficacy and describe microorganisms implicated at our institution. MATERIAL AND METHODS: Retrospective study of clinical files of patients with hepatic cirrhosis with positive culture of ascitic fluid (AF) and/or an AF polymorphonuclear (PMN) count of more than 250/mm3, treated at our units between 1st January, 2000 and 31st December, 2001 (n = 38). Patients showed a median age of 49 years (30-76), 63% of which were male. Forty-eight percent were classified as belonging to Child-Pugh B class, and 52% to C. RESULTS: First, considering cases with PMN > 250/mm3 (n = 29), antibiotics were given to all patients (cefotaxime and ampiciline). Fifty-two percent had hepatic encephalopathy, 42% had fever, 66% abdominal pain. In 42% a microorganism was isolated. Although 24% of fatal cases (only two related to infection), we noted a 73% clinical and laboratorial response. Five patients (72%) that died, showed renal failure by the time of death. Second, in all cases with positive culture of ascitic fluid (n = 21), 42% of which with PMN > 250/mm3 and 9 monobacterial nonneutrocytic bacterascites' cases, one only agent was found: E. coli in 36%, Streptococci (37%), Staphylococci (14%), and other (14%): Klebsiella oxytoca, n = 1; Salmonella enteritidis, n = 1; Enterococcus faecium, n = 1, Acinectobacter anitratus, n = 1. Only one of the agents, E. faecium (3%) showed in vitro sensitivity exclusively to ampiciline; all other were cefotaxime sensitivite. CONCLUSIONS: Our protocol will be modified, to treat patients with spontaneous bacterial peritonitis with cefotaxime, as monotherapy. Albumin infusion will also be added to the protocol, as, we found renal failure to be an important negative prognosis factor.

Peritonitis bacteriana espontánea en pacientes con cirrosis hepática: evaluación de un protocolo de tratamiento en unidades especializadas / Spontaneous bacterial peritonitis in patients with hepatic cirrhosis: evaluation of a treatment protocol at specia lized units

Introducción: la peritonitis bacteriana espontánea es una complicación frecuente y grave en los pacientes con cirrosis y ascitis. Su pronóstico depende claramente de su reconocimiento clínico precoz y de que el tratamiento sea eficaz. Objetivo: optimizar un protocolo de tratamiento, después de auditar su eficacia clínica, y describir los microorganismos implicados en nuestra institución. Material y métodos: estudio retrospectivo de las historias clínicas de los pacientes con cirrosis hepática y cultivo positivo de líquido ascítico (LA), y / o un recuento de polimorfonucleares (PMN) en LA de más de 250/mm3, tratados en nuestras unidades entre el 1 de enero de 2000 y el 31 de diciembre de 2001 (n = 38). Los pacientes presentaban una mediana de edad de 49 años (30-76), siendo varones el 63 por ciento de ellos. El 48 por ciento se clasificaron como pertenecientes a la categoría Child-Pugh B, y el 52 por ciento a la C. Resultados: en primer lugar, considerando los casos con PMN > 250/mm3 (n = 29), se administraron antibióticos a todos los pacientes (cefotaxima y ampicilina). El 52 por ciento tenían encefalopatía hepática, el 42 por ciento fiebre y el 66 por ciento dolor abdominal. En el 42 por ciento se aisló un microorganismo. Aunque el 24 por ciento de los casos fueron fatales (sólo 2 en relación con infecciones), observamos un 73 por ciento de respuestas clínicas y analíticas. Cinco de los pacientes (72 por ciento) que fallecieron presentaban insuficiencia renal en el momento de morir. En segundo lugar, en todos los casos con cultivo positivo de líquido ascítico (n= 21), el 42 por ciento de los cuales presentaban PMN > 250 mm3, con nueve casos de ascitis monobacteriana no neutrocítica, sólo se halló un agente: E. coli en el 36 por ciento, estreptococos (37 por ciento), estafilococos (14 por ciento) y otros, como Klebsiella oxytoca, n = 1; Salmonella enteritidis, n = 1; Enterococcus faecium, n = 1, y Acinectobacter anitratus, n = 1. Sólo uno de los agentes, E. faecium (3 por ciento) mostró sensibilidad exclusivamente a la ampicilina in vitro. Todos los demás fueron sensibles a la cefotaxima. Conclusiones: nuestro protocolo se modificará para tratar con cefotaxima en monoterapia a los pacientes con peritonitis bacteriana espontánea. También se añadirán al protocolo las infusiones de albúmina, ya que encontramos que la insuficiencia renal suponía un importante factor pronóstico negativo. (AU)

Genetic diversity measures of local European beef cattle breeds for conservation purposes.

This study was undertaken to determine the genetic structure, evolutionary relationships, and the genetic diversity among 18 local cattle breeds from Spain, Portugal, and France using 16 microsatellites. Heterozygosities, estimates of Fst, genetic distances, multivariate and diversity analyses, and assignment tests were performed. Heterozygosities ranged from 0.54 in the Pirenaica breed to 0.72 in the Barrosã breed. Seven percent of the total genetic variability can be attributed to differences among breeds (mean F(st) = 0.07; P<0.01). Five different genetic distances were computed and compared with no correlation found to be significantly different from 0 between distances based on the effective size of the population and those which use the size of the alleles. The Weitzman recursive approach and a multivariate analysis were used to measure the contribution of the breeds diversity. The Weitzman approach suggests that the most important breeds to be preserved are those grouped into two clusters: the cluster formed by the Mirandesa and Alistana breeds and that of the Sayaguesa and Tudanca breeds. The hypothetical extinction of one of those clusters represents a 17% loss of diversity. A correspondence analysis not only distinguished four breed groups but also confirmed results of previous studies classifying the important breeds contributing to diversity. In addition, the variation between breeds was sufficiently high so as to allow individuals to be assigned to their breed of origin with a probability of 99% for simulated samples.

A highly polymorphic plasma protein locus in brown trout (Salmo trutta L.) populations from Portugal.

Genetic polymorphism of an unidentified plasma protein (PX) is described for the first time in Salmo trutta (L.) by means of isoelectric focusing. The analysis of 414 individuals from different geographic origins in Portugal allowed the identification of nine alleles. Heterozygosity in natural populations is generally above 0.60, thus giving similar values to those reported for brown trout microsatellite loci. Substructuring of Portuguese brown trout is evident between northern and southern basins. Genetic affinities between the southernmost rivers and the hatchery stock were detected, suggesting the existence of recent stocking influences.

Mitochondrial haplotype diversity among Portuguese brown trout Salmo trutta L. populations: relevance to the post-pleistocene recolonization of northern Europe.

Mitochondrial haplotype diversity in seven Portuguese populations of brown trout, Salmo trutta L., was investigated by sequencing the 5' end of the mitochondrial DNA (mtDNA) control region. Five new haplotypes were described for this species, each two to three mutational steps distant from the common north Atlantic haplotype. Significant population subdivision of mtDNA haplotypes was also apparent. Based on these results, as well as on published data describing the distribution of both mtDNA haplotypes and allozyme alleles throughout Europe, the postglacial recolonization of northern Europe was re-evaluated. It is argued that the available data do not support the contribution of two major glacial refugia (southwest Atlantic and Ponto-Caspian Basin) to this postglacial recolonization, as proposed in a recently published model. The unique genetic architecture of Portuguese brown trout within the Atlantic-basin clade of this species represents a highly valuable genetic resource that should be protected from introgression with nonendemic strains of hatchery fish.

Acid phosphatase polymorphism in European shad (Fish: Clupeids).

Genetic polymorphism of acid phosphatases was investigated in 11 populations of the two European Alosa species using isoelectric focusing after sample treatment with neuraminidase. Two distinct loci, ACP1 and ACP2, were detected being ACP2 polymorphic. The observed genetic diversity between the species at the ACP2 locus supports other studies which indicate that A. alosa is the less polymorphic species of the two. This locus shows a higher geographic than interspecific pattern of differentiation and the ACP*2 allele is essentially confined to the Mediterranean.

Hereditary protein C deficiency and portal-vein thrombosis.

Inherited defects of the natural coagulation inhibitors predispose patients to thrombosis. These disorders have similar clinical presentations with a strong family history of thrombosis, episodes of recurrent venous thromboembolism, beginning in early adulthood. We report a case of upper gastrointestinal bleeding in a patient with portal hypertension due to portal-vein thrombosis secondary to hereditary protein C deficiency, an association that has seldom been reported.

[Hereditary protein C deficiency and portal vein thrombosis]. / Deficiência hereditária de proteína C e trombose da veia porta.

Inherited defects of the natural coagulation inhibitors predispose patients to thrombosis. These disorders have similar clinical presentations with a strong family of thrombosis, episodes of recurrent venous thromboembolism, beginning in early adulthood. We report a case of upper gastrointestinal bleeding in a patient with portal hypertension due to portal-vein thrombosis secondary to hereditary protein C deficiency, an association that has seldom been reported. We conclude that protein C deficiency should be investigated in thrombotic states, namely after more frequent causes have been excluded.

Propranolol or endoscopic sclerotherapy in the prevention of recurrence of variceal bleeding. A prospective, randomized controlled trial.

Endoscopic sclerotherapy (ES) and continuous propranolol (P) treatment have both been proposed as useful methods to prevent recurrent esophageal variceal bleeding. We report a prospective randomized trial in 65 patients with a previous history of endoscopically proven esophageal variceal bleeding. Patients were randomized by sealed envelopes stratified for Child's A and B groups to receive either endoscopic sclerotherapy (n = 31) or propranolol (n = 34). The dose of oral propranolol was based on a reduction of the resting pulse rate by 25%. Intravascular ethanolamine oleate was used for the endoscopic sclerotherapy in a 3-week schedule. The follow-up period ranged from 17 to 57 months (median: ES = 31; P = 28 months). There was no difference in the cumulative percentages of patients free of rebleeding from any source: esophageal and gastric varices, acute esophageal and gastric ulcers or erosions (ES = 37%; P = 16%). Also, there was no difference in the cumulative survival (ES = 69%; P = 54%). However, patients in the propranolol group had significantly more variceal rebleeding from the esophagus (n = 21) than did those in the sclerotherapy group (n = 9). The cumulative percentages of patients free of esophageal variceal rebleeding after inclusion were 67% in the endoscopic sclerotherapy group and 25% in the propranolol group (log-rank test, P less than 0.02). These differences indicated that sclerotherapy should be used in 29% of the propranolol patients who rebled. Based on these results we recommend elective sclerotherapy as long-term therapy for preventing rebleeding of esophageal varices.

Controlled trial of vasopressin and balloon tamponade in bleeding esophageal varices.

In a randomized controlled trial, the effect of continuous intravenous administration of vasopressin was compared with Sengstaken-Blakemore balloon tamponade in 37 episodes of bleeding esophageal varices in patients with cirrhosis. The majority were Group A and B of Child's classification. Bleeding was controlled in 11 of 17 (65%) patients on vasopressin and in 14 of 20 (70%) patients on tamponade. The patients who failed to respond initially (6 episodes on vasopressin and 5 on tamponade) were treated successfully with the alternative method. Overall mortality was similar in both groups: 3 patients in the vasopressin group and 4 in the tamponade group died. Only one patient died of uncontrolled bleeding; 4 patients probably died of complications of treatment, 2 of cardiac ischemia after vasopressin and 2 of pulmonary infection after tamponade. The vasopressin group required significantly fewer blood transfusions than did the tamponade group.

Endoscopy in the upper G.I. bleedings.

From 388 patients with upper G.I. bleeding investigated by endoscopy, radiology or emergent surgery, one third bled from duodenal ulcer, one third oesophageal varices, and from the remain the most frequent were gastric ulcer (14%) and gastric cancer (9%). From a sample of 53 patients with liver cirrhosis, 66% bled from varices and 34% from other lesions. The proportion of patients who bled from oesophageal varices is higher under 60 yrs. The mortality was higher after 60 yrs, except when there was associated chronic liver disease or renal or cardio-respiratory failure. In this group of patients, near half in our series, the mortality is the same under and above 60 years.