Efficacy of Salmonella Bacteriophage S1 Delivered and Released by Alginate Beads in a Chicken Model of Infection.

Modern bacteriophage encapsulation methods based on polymers such as alginate have been developed recently for their use in phage therapy for veterinary purposes. In birds, it has been proven that using this delivery system allows the release of the bacteriophage in the small intestine, the site of infection by Salmonella spp. This work designed an approach for phage therapy using encapsulation by ionotropic gelation of the lytic bacteriophage S1 for Salmonella enterica in 2% w/v alginate beads using 2% w/v calcium chloride as crosslinking agent. This formulation resulted in beads with an average size of 3.73 ± 0.04 mm and an encapsulation efficiency of 70%. In vitro, the beads protected the bacteriophages from pH 3 and released them at higher pH. To confirm that this would protect the bacteriophages from gastrointestinal pH changes, we tested the phage infectivity in vivo assay. Using a model chicken (Gallus gallus domesticus) infected with Salmonella Enteritidis, we confirmed that after 3 h of the beads delivery, infective phages were present in the chicken's duodenal and caecal sections. This study demonstrates that our phage formulation is an effective system for release and delivery of bacteriophage S1 against Salmonella Enteritidis with potential use in the poultry sector.

Punciones esplénicas guiadas por ecoendoscopia: ¿eficaces, seguras, necesarias? / No disponible

INTRODUCCIÓN: la esplenomegalia y/o las lesiones ocupantes de espacio (LOE) esplénicas, han tenido limitaciones en su estudio histopatológico por el riesgo que suponen las punciones esplénicas. Las biopsias percutáneas con aguja fina, son difíciles en especial por interposición de gases, ascitis, obesidad o por antecedentes de cirugía abdominal. En cambio, el ultrasonido endoscópico (USE) aprovecha la proximidad de la pared gástrica con el bazo, para puncionar con visualización de la aguja y de sus movimientos en tiempo real. OBJETIVO: describir la experiencia inicial y resultados obtenidos con USE-PAAF en pacientes con esplenomegalia o LOE esplénica. MATERIALES Y MÉTODOS: estudio observacional descriptivo. Se realizó USE-PAAF del bazo, con técnica de slow-pull, evitando fanning, con 3 pases de aguja en promedio. Las biopsias se enviaron en solución Cytorich RedTM para análisis mediante citología y bloque celular. RESULTADOS: se realizaron punciones en 15 pacientes (9 mujeres), con mediana de 67 años (rango 44-86) en pacientes estudiados por esplenomegalia o LOE esplénica, en contexto de fiebre de origen desconocido, adenopatías, perdida anormal de peso. Se logró un diagnóstico concluyente mediante USE-PAAF en 10 pacientes (66,7 %), de los cuales 4 fueron linfomas no Hodgkin tipo B de célula grande y un linfoma de Hodgkin. No se presentaron complicaciones inmediatas, ni diferidas relacionadas al procedimiento. CONCLUSIONES: las punciones esplénicas USE-guiadas parecen ser seguras, eficaces y podrían ser necesarias en algunos escenarios clínicos para completar la filiación etiológica de esplenomegalia de origen incierto o LOE esplénica y descartar malignidad

No disponible

Endoscopic ultrasound-guided fine-needle aspiration for splenomegaly and focal splenic lesion: is it safe, effective and necessary?

INTRODUCTION: splenomegaly and/or focal splenic lesions (FSL) have limited histopathologic studies due to the risk posed by splenic punctures. Percutaneous biopsies with a fine needle are difficult, especially due to interposition of gases, ascites, obesity or a history of abdominal surgery. On the other hand, endoscopic ultrasound (EUS) takes advantage of the proximity of the gastric wall to the spleen in order to puncture and visualize the needle and its movements in real time. OBJECTIVE: to describe the initial experience and results obtained with EUS-FNA in patients with splenomegaly or FSL. MATERIALS AND METHODS: this was a descriptive observational study. EUS-FNA of the spleen was performed with a slow-pull technique, which avoided fanning with an average of 3 needle passes. Biopsies were sent in Cytorich RedTM solution for analysis by cytology and cell block. RESULTS: punctures were performed in 15 patients (9 females) and the median age was 67 years (range 44-86). Patients studied due to an enlarged spleen or splenic FSL, in the context of fever of an unknown origin, adenopathies and abnormal weight loss were included. A conclusive diagnosis was achieved by EUS-FNA in 10 patients (66.7 %), 4 were large cell type B non-Hodgkin's lymphoma and one Hodgkin's lymphoma. There were no immediate or delayed complications related to the procedure. CONCLUSIONS: EUS-guided splenic punctures appear to be safe, effective and may be necessary in some clinical settings in order to complete the etiologic filiation of splenomegaly of an uncertain origin or FSL and to rule out malignancy.

Streptomycin hydrazone derivatives: synthesis and molecular recognition in aqueous solution.

Five hydrazone derivatives of streptomycin were synthetized (D0h, D1ph, D2bt, D3dctf, D4ag) and characterized by IR, 1H and 13C NMR spectroscopy, mass spectrometry and elemental analysis. Protonation constants were determined by potentiometry for all derivatives. D1ph and D2bt derivatives were investigated as receptors of dicarboxylates and adenine nucleotides in aqueous solution by potentiometric and 1H NMR titrations. D1ph and D2bt derivatives have the highest affinity with AMP and ATP, respectively, which shows that electrostatic forces are not always the dominant factor in binding of streptomycin derivatives with nucleotides, but the conformational fit between them. Calculated structures at the DFT level of the D1ph derivative bonded with either AMP or ADP showed that the complexes are stabilized by the formation of multiple interactions with the receptors. The antibiotic activity of the derivatives was explored and compared with native streptomycin.

Synthesis and in vitro evaluation of new ethyl and methyl quinoxaline-7-carboxylate 1,4-di-N-oxide against Entamoeba histolytica.

In our search for new antiamoebic agents, a new series of ethyl and methyl quinoxaline-7-carboxylate 1,4-di-N-oxide derivatives have been synthesized using the Beirut reaction. All compounds were characterized by spectroscopic techniques and elemental analysis. Antiamoebic activity was evaluated in vitro against Entamoeba histolytica strain HM1:IMSS by the microdilution method, and the structure-activity relationship was analyzed. We found that eleven quinoxaline derivatives showed greater activity than metronidazole and nitazoxanide with IC50 values in the range 1.99-0.35 µM. Compounds T-001 and T-016 shows IC50 values of 1.41 and 1.47 µM, respectively, with a value of selectivity index >60.

Conducción bajo los efectos de sustancias psicoactivas: correlación de las concentraciones en fluido oral y sangre / Driving under the influence of psychoactive substances: correlation between saliva and blood concentrations

La comunidad científica internacional acepta que el consumo de drogas de abuso o de sustancias psicoactivas por conductores de vehículos a motor supone un incremento del riesgo de sufrir un accidente. Por ello, en los últimos años los distintos países de la Unión Europea han ido modificando su legislación sobre seguridad vial con el objetivo de adaptarla a los nuevos conocimientos existentes. Por regla general, dichas regulaciones establecen que la determinación del consumo de drogas se realizará «in situ» en fluido oral, siendo necesaria la confirmación de dichos resultados. Tras la revisión exhaustiva de la información existente sobre dispositivos de detección, métodos de confirmación y posible correlación entre concentraciones obtenidas en saliva y las existentes en sangre, podemos concluir que los conocimientos actuales no permiten establecer una relación directa entre una determinada concentración en fluido oral y la posibilidad de sufrir un accidente de tráfico, a pesar de que se sabe que el consumo de drogas de abuso afecta a la conducción(AU)

The international scientific community agrees that the use of drugs or psychoactive substances by drivers has an increased risk of suffering a road accident. Therefore, in recent years, various European Union countries have modified their legislation on road safety in order to adapt them to current knowledge. These regulations generally include the provision for determining drug use "in situ" in saliva, with the results being confirmed where necessary. After a comprehensive review of existing information on detection devices, confirmation methods, and possible correlation between saliva and blood concentrations, we can conclude that a relationship between a given salivary fluid concentration and the possibility of having an accident cannot yet be established, even although it is known that drugs consumption impairs driving(AU)

Clinical, genetic, and therapeutic diversity in 2 patients with severe mevalonate kinase deficiency.

Mevalonic aciduria (MA) represents the severest form of mevalonate kinase deficiency due to recessively inherited, loss-of-function MVK mutations. MA is an early-onset disorder characterized by a marked failure to thrive, diverse neurologic symptoms, dysmorphic features, and recurrent febrile episodes. However, significant clinical differences have been reported in the few cases published to date. Here we describe 2 unrelated Spanish patients with MA, emphasizing the clinical heterogeneity observed. One patient presented with the severe classic MA phenotype due to the homozygous p.Ile-268-Thr MVK genotype, with a poor response to conventional treatments. However, the anti-interleukin 1 agent anakinra in this patient resulted in improvement in many clinical and laboratory parameters. The second patient presented with an atypical milder phenotype because of an older age at disease onset, mild neurologic symptoms, absence of febrile episodes and dysmorphic features, and moderate-to-good response to conventional treatments. The novel p.Arg-241-Cys MVK mutation, associated with the already known p.Ser-135-Leu mutation, detected in this patient expands the genetic diversity of mevalonate kinase deficiency. This atypical presentation of MA suggests that it should be included in the differential diagnosis of unclassified patients with psychomotor retardation, failure to thrive or ataxia, even in the absence of febrile episodes.

What do we know about Q fever in Mexico?

In Mexico, Q fever is considered a rare disease among humans and animals. From March to May of 2008, three patients were referred, from the state of Hidalgo to a tertiary-care center in Mexico City, with an acute febrile illness that was diagnosed as Q fever. We decided to undertake a cross sectional pilot study to identify cases of acute disease in this particular region and to determine the seroprevalence of Coxiella burnetii among healthy individuals with known risk factors for infection with this bacteria. Q fever was defined according to the Centers for Disease Control and Prevention criteria. All subjects were interviewed for signs and symptoms of the disease, demographic and household characteristics and occupational exposure to cattle. Blood samples were taken from hospitalized and outpatients with symptoms suggestive of Q fever, as well as from asymptomatic individuals with direct and daily exposure to cattle (slaughterers, butchers, farmers, shepherds and veterinarians) in the five municipalities. We report the occurrence of 17 cases with positive antibodies against C. burnetii in a rural area of central Mexico; eight cases had clinical criteria of acute Q fever disease. Results from this pilot study underscore the need for active surveillance programs and comprehensive studies to further define the prevalence and risk factors associated with the disease in Mexico, to know more about its clinical presentation and to characterize bacterial factors involved in its pathogenesis.

Primary cutaneous CD4+ small/medium-sized pleomorphic T-cell lymphoma expresses follicular T-cell markers.

Cutaneous CD4 small/medium-sized pleomorphic T-cell lymphoma (CSTCL) is a cutaneous T-cell lymphoma defined by a predominance of small-to-medium-sized CD4 pleomorphic T cells, with a favorable clinical course. Cases are also characterized by the presence of a rich infiltrate of reactive B cells. Recently, it has been reported that follicular helper T cells (TFH cells) display a distinct gene expression profile, positive for PD-1, CXCL13, and BCL-6. We report for the first time the expression of PD-1 and other TFH cell markers in CSTCLs and discuss its biologic significance. Sixteen CSTCLs were included in this study, and also 20 reactive inflammatory conditions, 10 primary cutaneous marginal zone, 10 follicular center lymphomas, and 5 primary CD30 cutaneous lymphomas. They were immunohistochemically analyzed for a large panel of markers. Double immunoperoxidase labeling of paraffin sections was performed for PD-1, OCT-2, and BCL-6. Clonal Ig and T-cell receptor rearrangements and Epstein-Barr virus-encoded RNA expression were also evaluated. Morphologic and clinical data were reviewed. Histologic examination showed a dense polymorphic lymphoid infiltrate throughout the dermis. Atypical large CD4 cells were positive for PD-1, CXCL13, and BCL-6 in all cases, and were attached in small clusters, or formed rosettes around CD30/OCT-2+ B blast cells. Epstein-Barr virus was not apparent in any of the cases. A dominant T-cell clone was identified in 14 cases, whereas polymerase chain reaction IgH gene rearrangement studies showed that all cases were polyclonal. None of the patients had lymphadenopathy or showed any evidence of systemic disease, nor did they have any previous history of mycosis fungoides or drug reactions. FTH cell markers are not exclusive to angioimmunoblastic lymphadenopathy but may also be seen in neoplastic cells of CSTCLs. Moreover, these findings suggest that B-cell stimulation by FTH could also take place in some cutaneous T-cell lymphomas.

Endobronchial instillations of amphotericin B: complementary treatment for pulmonary mucormycosis.

Pulmonary mucormycosis is a rare fungal infection that appears in patients with hematologic diseases who are immunosuppressed by chemotherapy. We report a case of pulmonary mucormycosis in a 21-year-old man with acute lymphoblastic leukaemia. During the period of pancytopenia he developed invasive mucormycosis. His initial symptoms were pleuritic chest pain with fever and unresponsiveness to broad-spectrum antibiotics. The outcome was favorable after long-term systemic and aerosolized amphotericin B, endobronchial instillations of amphotericin B, posaconazole, and surgery.

Determination of glutathione peroxidase activity in human milk.

An analytical method for determining glutathione peroxidase (GPx) (EC 1.11.1.9) activity in whole blood has been adapted to human milk samples. The values obtained for precision (relative standard deviation: 8.4%), linearity and accuracy (recovery: 90.4%) indicate the adequacy of the method for the mentioned purpose. The method was applied to 11 human milk samples obtained in the range from 6 to 135 days post partum. The resulting GPx activities (83.3 +/- 23.6 U/L) did not correlate to the selenium content of the samples, though a reciprocal correlation was found between the duration of lactation and milk GPx activity.