How does temperature influences the development of lactococcosis? Transcriptomic and immunoproteomic in vitro approaches.

Lactococcus garvieae is the aetiological agent of lactococcosis, a haemorrhagic septicaemia that affects marine and freshwater fish, with special incidence and economic relevance in farmed rainbow trout. Water temperature is one of the most important predisposing factors in the development of lactococcosis outbreaks. Lactococcosis in trout usually occur when water temperatures rise to about 18 °C, while fish carriers remain asymptomatic at temperatures below 13 °C. The aim of this work was to analyse the differences in the complete transcriptome response of L. garvieae grown at 18 °C and at 13 °C and to identify the immunogenic proteins expressed by this bacterium at 18 °C. Our results show that water temperature influences the expression of L. garvieae genes involved in the lysis of part of the bacterial cell population and in the cold response bacterial adaptation. Moreover, the surface immunogenic protein profile at 18 °C suggests an important role of the lysozyme-like enzyme, WxL surface proteins and some putative moonlighting proteins (proteins with more than one function, usually associated with different cellular locations) as virulence factors in L. garvieae. The results of this study could provide insights into the understanding of the virulence mechanisms of L. garvieae in fish.

Pilocarpine-induced status epilepticus increases Homer1a and changes mGluR5 expression.

Homer1a regulates expression of group I metabotropic glutamate receptors type I (mGluR1 and mGluR5) and is involved in neuronal plasticity. It has been reported that Homer1a expression is upregulated in the kindling model and hypothesized to act as an anticonvulsant. In the present work, we investigated whether pilocarpine-induced status epilepticus (SE) would alter Homer1a and mGluR5 expression in hippocampus. Adult rats were subjected to pilocarpine-model and analyzed at 2h, 8h, 24h and 7 d following SE. mRNA analysis showed the highest expression of Homer1a at 8h after SE onset, while immunohistochemistry demonstrated that Homer1a protein expression was significantly increased in hippocampus, amygdala and piriform and entorhinal cortices at 24h after SE onset when compared to control animals. The increased Homer1a expression coincided with a significant decrease of mGluR5 protein expression in amygdala and piriform and entorhinal cortices. The data suggest that during the critical periods of epileptogenesis, overexpression of Homer1a occurs to counteract hyperexcitability and thus Homer1a may be a molecular target in the treatment of epilepsy.

Ultrasound diagnosis of three cases of Mondor's disease.

OBJECTIVE: To describe clinical features and ultrasound findings of three cases of a little-known and relatively infrequent entity in daily clinical activity, which is often unnoticed and under-reported: penile Mondor's disease or superficial penile veins thrombophlebitis. METHODS: We are reporting the cases of three patients aged 33, 25 and 39 years who were referred to our department, the first case with suspicion of inguinal hernia, the second one to rule out testicular pathology because of pubic and perineal discomfort, and the third one for painful induration of the dorsal region of the penis. The three patients underwent Doppler-ultrasound examination (Toshiba®, using a 13-18MHz linear transducer) to establish definitive diagnosis, and had a favorable evolution with conservative management. RESULTS: Ultrasound examination revealed: Case 1. Penile superficial dorsal vein and lateral superficial veins thrombosis. Case 2. Thrombosis of the right branch of the superficial dorsal vein and its perineal distal connections. Case 3. Penile superficial dorsal vein thrombosis. Definitive diagnosis of the three cases was Mondor's disease. CONCLUSIONS: Mondor's disease is an often under-reported entity in daily clinical activity. Doppler-ultrasound findings (echogenic material within veins, lack of any response after compression by the transducer and absence of color flow) confirm de diagnosis. This disease has a favorable evolution and functional prognosis. Knowledge of Mondor's disease by echographists is basic to avoid false-negative results in radiologic examination.

Diagnóstico ecográfico de tres caso de enfermedad de Mondor del pene / Ultrasound diagnosis of three cases of Mondor`s disease

OBJETIVO: Describir las características clínicas y los hallazgos ecográficos de tres casos de una entidad poco conocida y relativamente infrecuente en la actividad clínica diaria, lo que ocasiona que sea infradiagnosticada: la enfermedad de Mondor del pene o tromboflebitis aislada de las venas superficiales del pene. MÉTODOS: Los casos corresponden a pacientes de 33, 25 y 39 años remitidos el primero por sospecha de hernia inguinal, el segundo para descartar patología testicular por molestias púbicas y perineales, y el tercero por induración dolorosa del dorso del pene. Se realizó ecografía-Doppler color de alta resolución (Toshiba®, con transductor lineal de 13-18 MHz) y tras el diagnóstico se instauró tratamiento conservador, evolucionando los tres casos de forma favorable. RESULTADOS: En el estudio ecográfico se apreció: Caso 1. Trombosis de la vena dorsal superficial y venas laterales superficiales. Caso 2. Trombosis de la rama derecha de la vena dorsal superficial y de sus conexiones distales perineales. Caso 3. Trombosis de la vena superficial dorsal del pene. Los tres casos fueron diagnosticados como enfermedad de Mondor. CONCLUSIONES: La enfermedad de Mondor es una entidad frecuentemente infradiagnosticada en la práctica diaria. Los hallazgos ecográficos-Doppler (contenido ecogénico en el interior de la vena, falta de respuesta a la compresión y ausencia de flujo) son diagnósticos. Se trata de una entidad clínica con buena evolución y pronóstico funcional. Es fundamental el conocimiento de esta entidad por parte del ecografista para evitar falsos negativos en el estudio radiológico(AU)

OBJECTIVE: To describe clinical features and ultrasound findings of three cases of a little-known and relatively infrequent entity in daily clinical activity, which is often unnoticed and under-reported: penile Mondor’s disease or superficial penile veins thrombophlebitis. METHODS: We are reporting the cases of three patients aged 33, 25 and 39 years who were referred to our department, the first case with suspicion of inguinal hernia, the second one to rule out testicular pathology because of pubic and perineal discomfort, and the third one for painful induration of the dorsal region of the penis. The three patients underwent Doppler-ultrasound examination (Toshiba®, using a 13-18MHz linear transducer) to establish definitive diagnosis, and had a favorable evolution with conservative management. RESULTS: Ultrasound examination revealed: Case 1. Penile superficial dorsal vein and lateral superficial veins thrombosis. Case 2. Thrombosis of the right branch of the superficial dorsal vein and its perineal distal connections. Case 3. Penile superficial dorsal vein thrombosis. Definitive diagnosis of the three cases was Mondor’s disease. CONCLUSIONS: Mondor’s disease is an often under-reported entity in daily clinical activity. Doppler-ultrasound findings (echogenic material within veins, lack of any response after compression by the transducer and absence of color flow) confirm de diagnosis. This disease has a favorable evolution and functional prognosis. Knowledge of Mondor’s disease by echographists is basic to avoid false-negative results in radiologic examination(AU)

Modeling epileptogenesis and temporal lobe epilepsy in a non-human primate.

Here we describe a new non-human primate model of temporal lobe epilepsy (TLE) to better investigate the cause/effect relationships of human TLE. Status epilepticus (SE) was induced in adult marmosets by pilocarpine injection (250mg/kg; i.p.). The animals were divided in 2 groups: acute (8h post-SE) and chronic (3 and 5 months post-SE). To manage the severity of SE, animals received diazepam 5min after the SE onset (acute group: 2.5 or 1.25mg/kg; i.p.; chronic group/; 1.25mg/kg; i.p). All animals were monitored by video and electrocorticography to assess SE and subsequent spontaneous recurrent seizures (SRS). To evaluate brain injury produced by SE or SRS we used argyrophil III, Nissl and neo-Timm staining techniques. Magnetic resonance image was also performed in the chronic group. We observed that pilocarpine was able to induce SE followed by SRS after a variable period of time. Prolonged SE episodes were associated with brain damage, mostly confined to the hippocampus and limbic structures. Similar to human TLE, anatomical disruption of dentate gyrus was observed after SRS. Our data suggest that pilocarpine marmoset model of epilepsy has great resemblance to human TLE, and could provide new tools to further evaluate the subtle changes associated with human epilepsy.

Neurobehavioral effect of essential oil of Cymbopogon citratus in mice.

Tea obtained from leaves of Cymbopogon citratus (DC) Stapf is used for its anxiolytic, hypnotic and anticonvulsant properties in Brazilian folk medicine. Essential oil (EO) from fresh leaves was obtained by hydrodistillation and orally administered to Swiss male mice 30 min before experimental procedures. EO at 0.5 or 1.0 g/kg was evaluated for sedative/hypnotic activity through pentobarbital sleeping time, anxiolytic activity by elevated plus maze and light/dark box procedures and anticonvulsant activity through seizures induced by pentylenetetrazole and maximal electroshock. EO was effective in increasing the sleeping time, the percentage of entries and time spent in the open arms of the elevated plus maze as well as the time spent in the light compartment of light/dark box. In addition, EO delayed clonic seizures induced by pentylenetetrazole and blocked tonic extensions induced by maximal electroshock, indicating the elevation of the seizure threshold and/or blockage of seizures spread. These effects were observed in the absence of motor impairment evaluated on the rotarod and open field test. Our results are in accord with the ethnopharmacological use of Cymbopogon citratus, and after complementary toxicological studies it can support investigations assessing their use as anxiolytic, sedative or anticonvulsive agent.

[The role of radiologists in the management of thyroid nodules]. / Papel del radiólogo en el manejo del nódulo tiroideo.

We review the different techniques in the diagnostic of thyroid nodule. The ultrasound and, especially, sonographically guided fine-needle aspiration (US-FNA), are the most accurate diagnostics tests to achieve a correct diagnosis of thyroid nodule, which is only done better by the surgery treatment and the pathology study of all lesion. We review the situations in which US-FNA is necessary and the different diagnostic and therapeutics options. Finally, we propose algorithms for the management of a solitary thyroid nodule, multinodular goiter and casually discovered nodule.

Papel del radiólogo en el manejo del nódulo tiroideo / The role of radiologists in the management of thyroid nodules

Se revisan las diferentes herramientas existentes para el diagnóstico del nódulo tiroideo. Dentro de ellas la ecografía, y sobre todo la punción-aspiración con aguja fina mediante guía ecográfica (ECO-PAAF), constituyen hoy en día las pruebas más fiables para determinar la naturaleza de un nódulo tiroideo, sólo superadas por la extirpación quirúrgica y el estudio anatomopatológico de la pieza completa. Se revisan las situaciones en las que se debe realizar ECO-PAAF, los diagnósticos posibles y las alternativas terapéuticas en función del diagnóstico obtenido. Por último, se proponen algoritmos de manejo diagnóstico y terapéutico para el nódulo solitario, el bocio multinodular y el nódulo asintomático descubierto casualmente

We review the differents techniques in the diagnostic of thyroid nodule. The ultrasound and, especially, sonographically guided fine-needle aspiration (US-FNA), are the most accurate diagnostics tests to achieve a correct diagnosis of thyroid nodule, which is only done better by the surgery treatment and the pathology study of all lesion. We review the situations in which US-FNA is necessary and the differents diagnostic andtherapeutics options. Finally, we propose algorithms for the management of a solitary thyroid nodule, multinodular goiter and casually discovered nodule

Sedative and anticonvulsant effects of hydroalcoholic extract of Equisetum arvense.

The hydroalcoholic extract of Equisetum arvense (HAE) tested at the doses of 200 and 400 mg/kg showed a significant activity on the open-field, enhanced the number of falls in the rota-rod reducing the time of permanence in the bar and increased the sleeping time (46% and 74%) in the barbiturate-induced sleeping time. In the pentylenetetrazole-seizure, it increased the first convulsion latency, diminished the severity of convulsions, reduced the percentage of animals which developed convulsion (50% and 25%) and protected animals from death. On the contrary, in the elevated plus maze, the doses 50, 100 and 150 mg/kg did not affect the evaluated parameters. Thus, HAE presented anticonvulsant and sedative effects. Phytochemical analysis detected the presence of tannins, saponins, sterols and flavonoids.

Rediseño del proceso asistencial de una unidad multidisciplinaria de patología mamaria. Análisis de la repercusión en el consumo de recursos durante un periodo de 5 años / Redesign of the nursing process of a multi-disciplinary breast pathology unit. Analysis of the repercussions in the consumption of resources over a period of five years

El objetivo de este artículo es analizar el impacto en el consumo de recursos sanitarios del rediseño del proceso asistencial diagnóstico en una Unidad de Patología Mamaria. Con la introducción del nuevo sistema de gestión durante un periodo de cinco años, (en el que se han atendido 10.804 pacientes), se han evitado un total de 15.781 consultas, lo que ha supuesto una reducción de costes económicos estimados entre 333.218,58 € y 594.012,18 € en función del tipo de hospital al que se aplique. El modelo de gestión desarrollado es eficiente en cuanto a reducción de consumo de recursos sanitarios en el diagnóstico de patología mamaria (AU)

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Multiplex PCR assay for detection of bacterial pathogens associated with warm-water Streptococcosis in fish.

A multiplex PCR-based method was designed for the simultaneous detection of the main pathogens involved in warm-water streptococcosis in fish (Streptococcus iniae, Streptococcus difficilis, Streptococcus parauberis, and Lactococcus garvieae). Each of the four pairs of oligonucleotide primers exclusively amplified the targeted gene of the specific microorganism. The sensitivity of the multiplex PCR using purified DNA was 25 pg for S. iniae, 12.5 pg for S. difficilis, 50 pg for S. parauberis, and 30 pg for L. garvieae. The multiplex PCR assay was useful for the specific detection of the four species of bacteria not only in pure culture but also in inoculated fish tissue homogenates and naturally infected fish. Therefore, this method could be a useful alternative to the culture-based method for the routine diagnosis of warm-water streptococcal infections in fish.

Lactococcus lactis subsp. lactis infection in waterfowl: first confirmation in animals.

We report the first description, confirmed by bacteriologic and molecular (polymerase chain reaction and pulsed-field gel electrophoresis) analysis, of an infection in animals caused by Lactococcus lactis subsp. lactis, affecting waterfowl.

Molecular typing by pulsed-field gel electrophoresis of Spanish animal and human Listeria monocytogenes isolates.

A total of 153 strains of Listeria monocytogenes isolated from different sources (72 from sheep, 12 from cattle, 18 from feedstuffs, and 51 from humans) in Spain from 1989 to 2000 were characterized by pulsed-field gel electrophoresis. The strains of L. monocytogenes displayed 55 pulsotypes. The 84 animal, 51 human, and 18 feedstuff strains displayed 31, 29, and 7 different pulsotypes, respectively, indicating a great genetic diversity among the Spanish L. monocytogenes isolates studied. L. monocytogenes isolates from clinical samples and feedstuffs consumed by the diseased animals were analyzed in 21 flocks. In most cases, clinical strains from different animals of the same flock had identical pulsotypes, confirming the existence of a listeriosis outbreak. L. monocytogenes strains with pulsotypes identical to those of clinical strains were isolated from silage, potatoes, and maize stalks. This is the first study wherein potatoes and maize stalks are epidemiologically linked with clinical listeriosis.

Phenotypic and genetic characterization of Lactococcus garvieae isolated in Spain from lactococcosis outbreaks and comparison with isolates of other countries and sources.

The phenotypic and genetic analysis results for 84 isolates of Lactococcus garvieae (including 62 strains from trout with lactococcosis from four different countries, 7 strains from cows and water buffalos with subclinical mastitis, 3 from water, and 10 from human clinical samples) are presented. There was great phenotypic heterogeneity (13 different biotypes) based on the acidification of saccharose, tagatose, mannitol, and cyclodextrin and the presence of the enzymes pyroglutamic acid arylamidase and N-acetyl-beta-glucosaminidase. L. garvieae also exhibited high genetic diversity by pulsed-field gel electrophoresis (PFGE), with 19 different pulsotypes among the isolates of L. garvieae studied. Only epidemiologically related strains, like the Spanish and Italian fish isolates and the cow and water buffalo isolates, displayed a close genetic relationship by PFGE, while the strains isolated from sporadic clinical cases, like the human isolates, were genetically unrelated. Overall, a general correlation between phenotypic and genetic data was observed. Epidemiological analysis of biotype and PFGE results indicated that the trout lactococcosis outbreaks in Spain and Portugal and those in France and Italy were produced by genetically unrelated clones. In Spain, two different clones were detected; the outbreaks diagnosed from 1995 onward were produced by a clone (biotype 2, pulsotype A1) which, although genetically related, was different from the one that was responsible for the outbreaks studied between 1991 and 1994 (biotype 1, pulsotype B). The Portuguese isolate had a biochemical profile identical to that of the Spanish strain isolated from 1995 onward and is also genetically closely related to this strain (pulsotype A2). There was a close relationship between the two pulsotypes (E and F) found in the Italian isolates. The French isolate (biotype 3, pulsotype D) was not genetically related to any other L. garvieae fish isolate. These results suggest the existence of diverse infection sources for the different lactococcosis outbreaks.

Development of a PCR assay for detection of Yersinia ruckeri in tissues of inoculated and naturally infected trout.

A PCR-based method was developed for the specific detection of Yersinia ruckeri in tissues of inoculated trout and naturally infected trout. No amplification products were obtained with other yersiniae, bacterial fish pathogens, or phylogenetically related bacteria (n = 34). The sensitivity of PCR detection was 60 to 65 bacterial cells per PCR tube, which was decreased to 10 to 20 cells by hybridization with a nonradioactive probe. The PCR assay proved to be as reliable as and faster than the conventional culture method for the detection of Y. ruckeri in infected trout tissues.

Taxonomic note: a proposal for reviewing the interpretation of the CAMP reaction between Listeria monocytogenes and Rhodococcus equi.

The discrepancies between the current description of the CAMP test between Listeria monocytogenes and Rhodococcus equi in the latest edition of Bergey's Manual of Determinative Bacteriology (L. monocytogenes is described as CAMP test negative with R. equi) and routine findings (positive reactions are usually described in many laboratories) make it advisable to review the current interpretation of the CAMP test to avoid confusion among people working in microbiological laboratories. Overall, 98.4% of the L. monocytogenes strains examined in this study, regardless of their source or the intensity of their hemolytic activity, displayed a synergic hemolytic reaction (CAMP phenomenon) with R. equi, indicating that L. monocytogenes can generally be considered CAMP positive with R. equi. We propose that L. monocytogenes, together with Listeria ivanovii, should be considered CAMP test positive with R. equi (circular or racket and semicircular or shovel shapes, respectively).

Cholesterol oxidase from Rhodococcus equi is likely the major factor involved in the cooperative lytic process (CAMP reaction) with Listeria monocytogenes.

The CAMP reaction between Listeria monocytogenes and Rhodococcus equi was studied by a diffusion assay. Listeria monocytogenes displayed identical cooperative haemolytic effect with supernatant cultures of R. equi or with commercial cholesterol oxidase (COX). This result, even with enzymes of different sources (commercial COX is obtained from Pseudomonas spp.) suggests that this enzyme secreted by R. equi has a crucial role in the synergistic haemolytic (CAMP) reaction with L. monocytogenes. The mechanism of the cooperative lytic process between L. monocytogenes and R. equi may represent a different and novel mechanism reaction, in which the COX may not act as a conventional second-step factor, and a reaction different to the direct interaction with the cholesterol of the erythrocyte membrane may be involved.

Serological response in rabbits to Listeria monocytogenes after oral or intragastric inoculation.

The serological response in rabbits against Listeria monocytogenes after oral or intragastric inoculation was investigated. Both the number of sero-positive animals and the average serum titres were higher in animals inoculated by the oral route. This difference was especially marked in rabbits inoculated with the lower dose (1 x 10(3) colony-forming units (cfu)), which developed a strong serological response (average serum titre of 1280 after 4 inoculations) in most of the inoculated animals (80%), without any clinical signs. The implication of these results in the epidemiology of listeriosis is discussed.