Yersinia pseudotuberculosis serotype O:1 infection in a captive Seba's short tailed-fruit bat (Carollia perspicillata) colony in Switzerland.

BACKGROUND: Between February and April 2016, a slight increase in mortality was observed in a colony consisting of 400 captive Seba's short-tailed bats (Carollia perspicillata). These animals cohabited with other nocturnal animal species in a dome of a private zoo in Switzerland. RESULTS: Gross and histological analysis of two (14.3%) out of the 13 animals submitted for necropsy within this period revealed a necrosuppurative pneumonia, hepatitis, splenitis, enterocolitis, and endometritis, with abundant intralesional colonies of Gram-negative rods. Yersinia (Y.) pseudotuberculosis serotype O:1 and biotype 1 belonging to the sequence type ST90 was isolated from the affected organs in both animals. Following this diagnosis, » of the colony (99 animals) was culled and submitted for gross and histopathological analysis, and a bacterial culture selective for Yersinia spp. of lung, liver, and spleen was performed. From these 99 animals, one gravid female was tested and found to be positive for Y. pseudotuberculosis in the absence of clinical symptoms and histopathological lesions. PCR analysis of altogether three bacterial isolates for virulence factors revealed the presence of the ail gene, and one isolate was also positive for the virF and yadA plasmid genes. CONCLUSIONS: These findings suggest that Carollia perspicillata are susceptible to lethal yersiniosis but do not represent a regular reservoir for Y. pseudotuberculosis. Culling of » of the population was sufficient to limit the spread of this infection among the colony. Moreover, no infections were detected in cohabitant nocturnal animals and caretakers, indicating that the zoonotic risk in this case was low.

The role of targeted BRCA1/BRCA2 mutation analysis in hereditary breast/ovarian cancer families of Portuguese ancestry.

We report the analysis of altogether 1050 suspected hereditary breast/ovarian cancer (HBOC) families, 524 fully screened for BRCA1/BRCA2 mutations and 526 tested only for the most common mutations. Of the 119 families with pathogenic mutations, 40 (33.6%) had the BRCA2 c.156_157insAlu rearrangement and 15 (12.6%) the BRCA1 c.3331_3334del mutation, the former being specific of Portuguese ancestry and the latter showing a founder effect in Portugal. Interestingly, the two most common mutations were found in a significant proportion of the HBOC families with an a priori BRCAPRO mutation probability <10%. We recommend that all suspected HBOC families from Portugal or with Portuguese ancestry, even those fulfilling moderately stringent clinical-criteria for genetic testing, should be specifically analyzed for the two most common BRCA1/BRCA2 founder mutations, and we here present a simple method for this first tier test. Screening of the entire coding regions of BRCA1 and BRCA2 should subsequently be offered to those families with a mutation probability ≥10% if none of those founder mutations are found.

The MSH2 c.388_389del mutation shows a founder effect in Portuguese Lynch syndrome families.

The MSH2 c.388_389del mutation has occasionally been described in Lynch families worldwide. At the Portuguese Oncology Institute in Porto, Portugal, we have identified 16 seemingly unrelated families with this germline mutation. To evaluate if this alteration is a founder or a recurrent mutation we performed haplotype analysis in the 16 Portuguese index cases and 55 relatives, as well as in four index cases and 13 relatives reported from Germany, Scotland, England, and Argentina. In the Portuguese families we observed a shared haplotype of approximately 10 Mb and all were originated from the north of Portugal. These results suggest that this alteration is a founder mutation in Portugal with a relatively recent origin. In the reported families outside Portugal with this mutation different haplotype backgrounds were observed, supporting the hypothesis that it occurred de novo on multiple occasions. We also conclude that the high proportion of families with the MSH2 c.388_389del mutation indicates that screening for this alteration as a first step may be cost-effective in the genetic testing of Lynch syndrome suspects of Portuguese ancestry, especially those originating from the north of Portugal.

Discordance in treatment of chronic obstructive pulmonary disease following GesEPOC guideline vs. GOLD.

INTRODUCTION: Pharmacological treatment of chronic obstructive pulmonary disease in Spain is usually chosen according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) or Spanish guidelines for chronic obstructive pulmonary disease (GesEPOC). The main objective of this study was to evaluate the degree of concordance between treatment for newly diagnosed chronic obstructive pulmonary disease patients according to GOLD and GesEPOC. MATERIAL AND METHODS: Simulation study. The following variables were used: FEV1%, exacerbations, dyspnoea at first evaluation, blood eosinophilia, personal history of asthma, and degree of bronchodilator reversibility. Four investigators classified and assigned a treatment to each patient (2 using GOLD criteria and the other 2 using GesEPOC). Global Kappa index was calculated. RESULTS: The database included 467 patients. Agreement between treatment decided using GOLD and GesEPOC was poor (Kappa: 0.17, 95% CI: 0.12-0.23). CONCLUSION: There is a poor agreement between GOLD and GesEPOC recommendations for initial chronic obstructive pulmonary disease treatment.

Canine sterile neutrophilic dermatosis (resembling Sweet's syndrome) with severe extracutaneous manifestations.

INTRODUCTION: Sterile neutrophilic dermatosis is a rare disease in dogs, similar to Sweet's syndrome in humans. This case report describes the treatment of a 2-year old Bearded Collie that was presented with a 3-week history of fever, hind-limb weakness, peripheral lymphadenomegaly and leucocytosis. Blood tests revealed severe leukocytosis, renal azotaemia, elevated liver enzymes and bilirubinaemia. Skin lesions started to appear in week four. Histology revealed a sterile neutrophilic dermatitis resembling Sweet's syndrome. The dog displayed extracutaneous manifestations, including fever, polyarthritis, a severe leukemoid reaction, anaemia, hepatopathy and nephropathy. Issues regarding the use of criteria for the diagnosis of Sweet's syndrome in humans that are used for dogs with sterile neutrophilic dermatosis, are discussed in this case report. The condition resolved with dexamethasone and mycophenolate mofetil as a novel steroid-sparing therapy. Three months later the dog relapsed, which rapidly responded to short-term dexamethasone treatment and temporarily increased mycophenolate mofetil dosage.

INTRODUCTION: La dermatose neutrophilique stérile est une maladie rare chez le chien, semblable au syndrome de Sweet chez l'homme. Ce rapport de cas décrit le traitement d'un Bearded Collie de 2 ans présentant des antécédents de fièvre pendant 3 semaines, une faiblesse des membres postérieurs, une lymphadénomégalie périphérique et une leucocytose. Les analyses de sang ont révélé une leucocytose grave, une azotémie rénale, une élévation des enzymes hépatiques et une bilirubinémie. Des lésions cutanées ont commencé à apparaître à la quatrième semaine. L'histologie a révélé une dermatite neutrophilique stérile ressemblant au syndrome de Sweet. Le chien présentait des manifestations extracutanées telles que fièvre, polyarthrite, réaction leucémoïde sévère, anémie, hépatopathie et néphropathie. Les questions relatives à l'utilisation des critères de diagnostic du syndrome de Sweet chez l'homme chez les chiens atteints de dermatose neutrophilique stérile sont abordées dans le présent rapport de cas. La maladie a été traitée avec la dexaméthasone et le mycophénolate mofétil en tant que thérapie innovante permettant d'économiser des stéroïdes. Trois mois plus tard, le chien a rechuté mais a rapidement répondu à un traitement de courte durée à la dexaméthasone et à une augmentation temporairement la dose de mycophénolate mofétil.

Congenital Cutaneous Panadnexal Papillomatous Hamartomas in a Calf.

Several cauliflower-like alopecic masses arose on the head of an otherwise healthy, full-term newborn Jersey × Belgian blue heifer, which was humanely destroyed shortly after birth due to the severity of the skin lesions. Microscopically, the masses were composed of multiple papillary projections displaying well-differentiated sebaceous glands surrounded by a moderate number of well-developed sweat glands, as well as embryonic and fully developed, but dysplastic hair follicles. Thick branching connective tissue stalks supported these adnexal components. The papillated surface, the predominance of sebaceous glands, the presence of embryonic hair follicles and the well-differentiated sweat glands were compatible with nevus sebaceous, a rare form of cutaneous hamartoma described in man, dogs, cats and cattle. However, the cauliflower-like growth pattern, the presence of supportive thick branching connective tissue stalks and the relative abundance of dysplastic hair follicles in association with nevus sebaceous has not been described in the human or veterinary literature. A diagnosis of panadnexal papillomatous hamartoma was made in this case.

Four novel MSH2 / MLH1 gene mutations in portuguese HNPCC families.

Hereditary non-polyposis colorectal cancer (HNPCC) is considered to be determined by germline mutations in the mismatch repair (MMR) genes, especially MSH2 and MLH1. While screening for mutations in these two genes in HNPCC portuguese families, 3 previously unreported MSH2 and 1 MLH1 mutations have been identified in families meeting strict Amsterdam criteria. Hum Mutat 15:116, 2000.

p53 alterations in gastric carcinoma: a study of 56 primary tumors and 204 nodal metastases.

We have looked for LOH, mutation, and expression of p53 in a series of 56 primary gastric carcinomas, using Southern blotting, constant denaturant gel electrophoresis (CDGE), direct sequencing, and immunohistochemistry. Two hundred-four lymph node metastases from 36 of these cases were also studied by immunohistochemistry. Loss of constitutional heterozygosity for the p53 locus was detected in five of 16 informative cases; in four of these cases there was a concurrent point mutation at the retained allele. Immunoreactivity for p53 product and mutation of p53 were observed in nine and 10 of the 56 tumors, respectively. There was no significant relationship between p53 immunoreactivity or mutation and pathologic features. The overall prevalence of p53 mutations, as detected by CDGE, was 18%. Direct sequencing confirmed p53 mutations (all G:C-A:T transitions) in seven cases, six of which were missense mutations; the remaining case was a silent mutation. In three of the cases with missense mutations, these occurred at CpG dinucleotides (codons 175, 273, and 282). Three cases with immunoreactivity for p53 product in the primary tumor also showed positive staining in the metastases. In two of these cases (with six and nine metastases, respectively) a positive staining was observed in all lymph node metastases, whereas in the remaining case only two of four nodal metastases were positively stained. In no single case was a positive staining observed in a metastasis where the primary tumor gave a negative staining result. Five of the six cases with p53 mutations and fresh material available for cell culture were successfully grown in vitro and all had abnormal karyotypes.

Increasing levels of MYC and MET co-amplification during tumor progression of a case of gastric cancer.

The cytogenetic study of a nodal metastasis from a gastric carcinoma, after two passages in nude mice, revealed a large number of double minutes. Comparative genomic in situ hybridization (CGH) analysis using DNA extracted from this xenograft revealed the existence of three clear amplification units that originated from the chromosomal subregions 6q24-25, 7q31-32, and 8q24 in the xenograft DNA. Similar, though less prominent, CGH results were found with DNAs extracted from the primary tumor and its metastasis, implying that the same amplicons were also present, albeit less abundantly, in the DNAs of these neoplastic tissues. Southern analysis of the second-passage xenograft detected 18- and 10-fold amplification of MET (located at 7q31) and MYC (located at 8q24), respectively. The retrospective study of the first passage of the xenograft, as well as of the metastatic and primary tumors before xenografting, showed amplification levels of MET of, respectively, 12-, 9-, and 5-fold and MYC of, respectively, 8-, 7-, and 5-fold. Our results suggest that increased levels of co-amplification of MYC and MET correlate with enhanced growth potential in this case of gastric carcinoma.

The interpretation of indicative and subjunctive concessives.

Using a priming paradigm in the context of a reading comprehension task, the possibilities that people keep in mind in order to understand indicative and subjunctive concessive sentences were examined and compared to those from factual and counterfactual 'if A, not-B' conditionals. The length of time it took people to read conjunctive descriptions (i.e., A and B, A and not-B, not-A and B, not-A and not-B) after they had been primed by the different types of linguistic form was measured. The results suggest that, whereas indicative 'even though' concessives and 'if, not' conditionals are understood by keeping in mind just a single possibility ('A and B' and 'A and not-B', respectively), the initial representations of subjunctive 'even if' concessive-conditionals and 'if, not' counterfactuals are compatible with a multiple-model representation. The implications of these results are discussed within the mental models framework.

Genetic diversity and phylogenetic analysis of Feline Coronavirus sequences from Portugal.

The distribution of FCoV Types I and II in a Portuguese cat population was studied by a RT-PCR assay targeting the 3'-end of the viral RNA. For a period of 3 years, 120 samples were collected and 57 were found positive for FCoV RNA. Within the positive samples the presence of FCoV Type I was found in 79%. Type II was only detected in 3.5% in animals with Feline Infectious Peritonitis. The remaining 17.5% could not be differentiated. These viral sequences, comprising a region within gene S were further subjected to a heteroduplex mobility assay (HMA) detecting the presence of viral quasispecies in 17% of the samples. Phylogenetic analysis for FCoV Type I revealed high genetic diversity between the Portuguese sequences and other previously characterized strains, while Type II tree showed a higher genetic homogeneity. This study confirmed the presence of FCoV Types I and II circulating in Portugal and detected high genetic diversity between circulating strains suggesting that the virus persists within the host as mixed viral populations.

Valor nutricional de híbridos de sorgo em diferentes estádios de maturação / Nutritional value of hybrids of sorghum in different maturation stages

Avaliaram-se a composição bromatológica e a digestibilidade in vitro das plantas, folhas, colmos e panículas de três híbridos de sorgo (BRS 610, BR 700 e BRS 655) colhidos em três estádios de maturação (leitoso, pastoso e farináceo). O delineamento utilizado foi o inteiramente casualizado, em arranjo fatorial 3x3 (híbridos x estádios de maturação), sendo as médias comparadas pelo teste SNK (P<0,05). Os teores de matéria seca (MS) das plantas variaram de 25,73% a 43,96% e aumentaram com a maturidade. A concentração de proteína bruta (PB) das plantas manteve-se inalterada (P>0,05) entre as idades de corte para todos os híbridos. Os teores de fibra insolúvel em detergente neutro (FDN) e fibra insolúvel em detergente ácido (FDA) das plantas não foram influenciados pelo estádio de maturação. Os coeficientes de digestibilidade in vitro da MS (DIVMS) das plantas do BRS 610 não variaram com a maturidade, mas para o BR 700 e BRS 655 apresentaram redução entre o estádio leitoso e o farináceo. As variações observadas nas porcentagens de MS, PB, FDN, FDA e DIVMS das frações folha, colmo e panícula ocorreram de forma diferente entre os híbridos com o avanço da maturidade. Os híbridos BR 700 e BRS 655 devem ser ensilados no estádio leitoso, enquanto o BRS 610 pode ser colhido nos três estádios de maturação avaliados.

The nutritional value of the plants, leaves, stems and panicles of three hybrids of sorghum (BRS 610, BR 700 and BRS 655) at three maturation stages (milk, soft dough and flour) were evaluated. A complete randomized design was used in a factorial arrangement 3x3 (hybrids x ages of cut), and the means were compared by SNK (P<0.05). Dry matter contents of the plants varied from 25.73% to 43.96% and increased with maturity. The percentage of crude protein (CP) of the plants remained constant (P>0.05) among cuts for all the hybrids. The values of neutral detergent fiber (NDF) and acid detergent fiber (ADF) of the plants were not affected by maturation stages. The in vitro dry matter digestibility (IVDMD) of the plants of BRS 610 didn't change with maturity, but decreased between milk and floury stages for BR 700 and BRS 655. Differences in the percentages of DM, CP, NDF, ADF and IVDMD of the plant fractions (leaves, steam and panicle) occurred differently among hybrids with the advance of maturity. The hybrids BR 700 and BRS 655 should be ensiled at milk stage, while the BRS 610 can be harvested at the three maturation stages evaluated.

Improving reading comprehension: From metacognitive intervention on strategies to the intervention on working memory executive processes / Mejorando la comprensión lectora: Desde la intervención metacognitiva en estrategias a la intervención en los procesos ejecutivos de la memoria operativa

Many students may read fluently but have difficulties constructing meaning from texts. Difficulties with reading comprehension have many implications at school. In particular, problems understanding texts interfere with studying and learning from text. Reading comprehension has improved in the last 30 years focusing on intervention programs that work with strategies in which metacog-nition plays a crucial role. However, recent years have seen relevant advances in the study of the relationship between working memory (WM), particularly executive processes, and reading comprehension. In this paper, we present how the last 20 years of our research has evolved regarding metacognitive intervention from text comprehension strategies, as the main idea and summarization to the intervention on WM's executive processes during reading. Thus, our more recent empirical data has shown that text comprehension can be improved after specific training on the executive functions of working memory (e.g., focusing, switching, connecting and updating mental representations, and the inhibition of irrelevant information) in Primary school students.

Muchos estudiantes pueden leer de forma fluida pero presentan dificultades para construir significados a partir de los textos. Las dificultades de compresión lectora tienen varias implicaciones en la escuela. En particular, los problemas de comprensión de textos interfieren con el estudio y el aprendizaje desde el texto. La comprensión de lectura se ha mejorado en los últimos 30 años enfocándose en los programas de intervención que trabajan con estrategias en las cuales la metacognición juega un papel crucial. Sin embargo, en años recientes han sido relevantes los avances en el estudio de las relaciones entre la memoria de trabajo (WM), particularmente el proceso ejecutivo, y la comprensión de lectura. En este artículo presentamos la manera como se ha desarrollado nuestra investigación en los últimos 20 años, en relación con intervención metacognitiva desde las estrategias de comprensión de textos, tales como la idea principal y el resumen en la intervención sobre el proceso ejecutivo de WM durante la lectura. Así, nuestros datos empíricos recientes han mostrado que la comprensión de textos puede ser mejorada después del tratamiento específico sobre las funciones ejecutivas de memoria de trabajo (e.g., enfocándose, cambiando, conectando y actualizando las representaciones mentales y la inhibición de información irrelevante) en niños de escuela primaria.

MSH6 germline mutations in early-onset colorectal cancer patients without family history of the disease.

Germline MLH1 and MSH2 mutations are scarce in young colorectal cancer patients with negative family history of the disease. To evaluate the contribution of germline MSH6 mutations to early-onset colorectal cancer, we have analysed peripheral blood of 38 patients diagnosed with this disease before 45 years of age and who presented no family history of hereditary nonpolyposis colorectal cancer-related cancers. Blood samples from 108 healthy volunteers were analysed for those genetic alterations suspected to affect the function of MSH6. Of the seven (18.4%) MSH6 alterations found, we have identified three novel germline mutations, one 8 bp deletion leading to a truncated protein and two missense mutations resulting in the substitution of amino acids belonging to different polarity groups. High-frequency microsatellite instability was found in the patient with the MSH6 deletion, but not in the other 27 carcinomas analysed. No MLH1 promoter methylation was detected in tumour tissue. Our findings suggest that germline MSH6 mutations contribute to a subset of early-onset colorectal cancer. Further studies are warranted to understand the genetic and environmental factors responsible for the variable penetration of MSH6 germline mutations, as well as to identify other causes of early-onset colorectal cancer.

Pathological exon skipping in an HNPCC proband with MLH1 splice acceptor site mutation.

One of the most commonly mutated mismatch repair genes in human nonpolyposis colorectal cancer (HNPCC) is MLH1. We identified a splice site mutation in MLH1 in a colorectal cancer proband (T-to-A at position -11 of intron 1 splice acceptor) and investigated its functional consequences by RT-PCR, using lymphocyte mRNA from the proband, two noncarrying siblings, and one unrelated individual. Subcloning of PCR products followed by sequencing of individual clones revealed increased transcript heterogeneity in the mutation carrier, attributable to the presence of a variety of mRNA forms lacking exon 2, or combinations of exons 2, 4, 6, 9, and 10. The full-length transcript subcloned from the mutation carrier was detected with a much reduced frequency, suggesting that only the wild-type allele produced functional MLH1 mRNA. The three noncarriers expressed some previously described transcripts and several novel variants, but none that lacked exon 2. The results are consistent with the hypothesis that this splice site mutation causes skipping of MLH1 exon 2 in a large proportion of mRNA transcripts derived from the mutated allele. Such an observation strengthens the case for identifying the mutation as pathogenic in this HNPCC family, which is of interest given the rarity of exon skipping defects resulting from splice acceptor site mutations outside the invariant AG dinucleotide.

Detection of prognostic significant translocations in childhood acute lymphoblastic leukaemia by one-step multiplex reverse transcription polymerase chain reaction.

The search for chromosomal translocations in de novo cases of childhood acute lymphoblastic leukaemia (ALL) is crucial for the selection of the appropriate therapeutic protocol. In this work, we describe a new method - one-step multiplex reverse transcription polymerase chain reaction (RT-PCR) - to screen for prognostic significant translocations in childhood ALL. Our approach involves a single PCR reaction for the simultaneous detection of the molecular rearrangements resulting from the t(9;22), t(12;21), t(4;11) and t(1;19), with a turnaround time of less than 24 h. This assay proved to be highly sensitive, specific, reproducible and easy to implement in a routine genetics laboratory.