A Real-Time PCR Assay for Detection of Low Pneumocystis jirovecii Levels.

Here we report a new real-time PCR assay using SYBR Green which provides higher sensitivity for the specific detection of low levels of Pneumocystis jirovecii. To do so, two primer sets were designed, targeting the family of genes that code for the most abundant surface protein of Pneumocystis spp., namely the major surface glycoproteins (Msg), and the mitochondrial large subunit rRNA (mtLSUrRNA) multicopy gene, simultaneously detecting two regions. PCR methods are instrumental in detecting these low levels; however, current nested-PCR methods are time-consuming and complex. To validate our new real-time Msg-A/mtLSUrRNA PCR protocol, we compared it with nested-PCR based on the detection of Pneumocystis mitochondrial large subunit rRNA (mtLSUrRNA), one of the main targets used to detect this pathogen. All samples identified as positive by the nested-PCR method were found positive using our new real-time PCR protocol, which also detected P. jirovecii in three nasal aspirate samples that were negative for both rounds of nested-PCR. Furthermore, we read both rounds of the nested-PCR results for comparison and found that some samples with no PCR amplification, or with a feeble band in the first round, correlated with higher Ct values in our real-time Msg-A/mtLSUrRNA PCR. This finding demonstrates the ability of this new single-round protocol to detect low Pneumocystis levels. This new assay provides a valuable alternative for P. jirovecii detection, as it is both rapid and sensitive.

Increase in secreted airway mucins and partial Muc5b STAT6/FoxA2 regulation during Pneumocystis primary infection.

Airway mucus responses to subclinical infections may explain variations in progression of chronic lung diseases and differences in clinical expression of respiratory infections across individuals. Pneumocystis associates to more severe Chronic Obstructive Pulmonary Disease (COPD), asthma, respiratory distress of premature newborns, and is a consistent subclinical infection between 2 and 5 months of age when hospitalizations for respiratory cause and infant mortality are higher. This atypical fungus associates to increased mucin 5AC (MUC5AC), a central effector of Th2-type allergic inflammation, in infant lungs. However, mucus progression, expression of MUC5B essential for airway defense, and potential for pharmacologic modulation of mucus during Pneumocystis infection remain unknown. We measured MUC5B and Pneumocystis in infant lungs, and progression of mucin levels and effect of inhibition of the STAT6/FoxA2 mucus pathway using Kaempferol, a JAK/STAT6 inhibitor, in immunocompetent rats during Pneumocystis primary infection. Pneumocystis associated to increased MUC5B in infant lungs. Muc5b increased earlier and more abundantly than Muc5ac during experimental primary infection suggesting an acute defensive response against Pneumocystis as described against bacteria, while increased Muc5ac levels supports an ongoing allergic, Th2 lymphocyte-type response during primary Pneumocystis infection. Kaempferol partly reversed Muc5b stimulation suggesting limited potential for pharmacological modulation via the STAT6-FoxA2 pathway.

Muerte repentina posterior a delirio agitado y contención policial en una persona con pancreatitis aguda hemorrágica / Sudden death due to agitated delirium and police restrain in a person with acute hemorhagic pancreatitis

ResumenSe expone un complejo caso de muerte repentina, por delirio agitado secundario a intoxicación aguda por alcohol y cocaína, posterior a contención policial. En la autopsia se encontraron inusuales hemorragias en el sistema digestivo y una pancreatitis aguda hemorrágica, hallazgos que plantean interrogantes sobre la causa de la muerte y el efecto combinado de los tóxicos de abuso.

AbstractSudden death of an individual, in agitated delirium secondary to alcohol and cocaine acute intoxication, due to police restriction is presented. The autopsy findings included gastrointestinal and acute hemorrhagic pancreatitis, findings that raise questions about the cause of death and the combined effect of the toxic substances.

Suicidio por interrupción de hemodiálisis en un paciente con nefropatía crónica: análisis de un caso / Suicide by interruption of hemodialysis, in a patient with chronic renal failure: Analysis of one case

Background: In Chile, interruption of hemodialysis is cause of death in 0.37% of patients under this treatment. We report a 83-year-old woman suffering from chronic renal failure who withdrew from prolonged hemodialysis treatment. Biographical aspects, circumstances and the cause of death are analyzed from forensic and psychiatric points of view, highlighting attributes that allow classification of the death of this patient as suicide. Risk factors involved in this type of patients, as well as possible preventive measures are discussed.

[Suicide by interruption of hemodialysis, in a patient with chronic renal failure. Analysis of one case]. / Suicidio por interrupción de hemodiálisis en un paciente con nefropatía crónica: análisis de un caso.

In Chile, interruption of hemodialysis is cause of death in 0.37% of patients under this treatment. We report a 83-year-old woman suffering from chronic renal failure who withdrew from prolonged hemodialysis treatment. Biographical aspects, circumstances and the cause of death are analyzed from forensic and psychiatric points of view, highlighting attributes that allow classification of the death of this patient as suicide. Risk factors involved in this type of patients, as well as possible preventive measures are discussed.

Síndrome de Münchausen by proxy: enfoque forense / Munchausen syndrome by proxy: forensic approach

El síndrome de Münchausen by proxy es una forma de maltrato infantil cuyas consecuencias pueden ser tan graves como la muerte del niño, víctima de enfermedades ficticias provocadas por un cuidador del que es muy dificil sospechar.

Bacteriocin like substance production by Carnobacterium piscicola in a continuous system with three culture broths. Study of antagonism against Listeria monocytogenes on vacuum packaged salmon

Três meios de cultura foram avaliados quanto a produção de substâncias semelhantes a bacteriocinas (SSB) por Carnobacterium piscicola L 103, utilizando um sistema contínuo de cultura. A eficácia da substância antagonista contra Listeria monocytogenes foi testada em salmão embalado a vácuo. As SSB foram produzidas em bioreator de 1.0 L, em cultura contínua e em cultura estacionária, utilizando D-MRS, mod. D-MRS e APT como meios de cultivo. Filés de salmão foram inoculados com SSB (200 AU mL-1 e 800 AU mL-1) e L. monocytogenes (8.0 x 101 ufc cm-2) e mantidos a 4ºC. O crescimento de L. monocytogenes foi verificado a cada 5 dias, durante 15 dias. C. piscicola L 103 antigiu a fase estacionária depois de 12 h de incubação em cultivo em batelada, sendo a atividade SSB de 800 AU mL-1 nos meios D-MRS e mod. D-MRS e de 400 AU mL-1 no caldo APT. Durante a cultura continua, a atividade SSB aumentou até 6400 AU mL-1 nos dois tipos de caldo MRS, ao passo que no caldo APT esta atividade diminuiu a 50 AU mL-1, indicando uma clara vantagem de uso dos dois primeiros meios de cultura e do sistema de crescimento contínuo. SSB mostrou efeito bacteriostático sobre L. monocytogenes quando inoculada em salmão, com contagens de 6,0 x 103 ufc cm-2 após 15 dias. Não foram encontradas diferenças significantes entre as duas atividades utilizadas. No ensaio controle, sem SSB, a contagem de L. monocytogenes aumentou até 1,0 x 106 ufc cm-2 após 15 dias de estocagem.