Inter-observer variability with five computed tomography severity scales for COVID-19 pneumonia assessment.

INTRODUCTION: By the end 2019 there was an outbreak of pneumonia caused by a new coronavirus, a disease that was called coronavirus disease 2019 (COVID-19). Computed tomography (CT) has played an important role in the diagnosis of COVID-19 patients. OBJECTIVE: To demonstrate inter-observer variability with five scales proposed for measuring the extent of COVID-19 pneumonia on tomography. METHODS: Thirty five initial chest CT scans of patients who attended respiratory triage for suspected COVID-19 pneumonia were analyzed. Three radiologists classified the tomographic images according to the severity scales proposed by Yang (1), Yuan (2), Chun (3), Wang (4) and Instituto Nacional de Enfermedades Respiratorias-Chung-Pan (5). The percentage of agreement between the evaluators for each scale was calculated using the intra-class correlation index. RESULTS: In most patients were five pulmonary lobes compromised (77.1% of the patients). Scales 1, 2, 4 and 5 showed an intra-class correlation > 0.91 (p < 0.0001), with agreement thus being almost perfect. CONCLUSIONS: Scale 4 (proposed by Wang) showed the best inter-observer agreement, with a coefficient of 0.964 (p = 0.001).

INTRODUCCIÓN: A finales de 2019 se presentó un brote de neumonía causada por un nuevo coronavirus, enfermedad a la que se denominó COVID-19. La tomografía computarizada ha desempeñado un papel importante en el diagnóstico de los pacientes con COVID-19. OBJETIVO: Demostrar la variabilidad interobservador con cinco escalas propuestas para la medición de la extensión de la neumonía ocasionada por COVID-19 mediante tomografía. MÉTODOS: Se analizaron 35 tomografías de tórax iniciales de pacientes que asistieron al triaje respiratorio por sospecha de neumonía por COVID-19. Tres radiólogos realizaron la clasificación de las imágenes tomográficas de acuerdo con las escalas de severidad propuestas por Yang (1), Yuan (2), Chun (3), Wang (4) e INER-Chung-Pan (5). Se calculó el porcentaje de concordancia entre los evaluadores para cada escala con el índice de correlación intraclase. RESULTADOS: La mayoría de los pacientes presentó afección de cinco lóbulos pulmonares (77.1 % de los pacientes). Las escalas 1, 2, 4 y 5 mostraron una correlación intraclase > 0.91, con p < 0.0001, por lo que la concordancia fue casi perfecta. CONCLUSIONES: La escala 4 (de Wang) mostró la mejor concordancia interobservador, con un coeficiente de 0.964 (p = 0.001).

[Morphine induced biliary pain. Case report]. / Cólico biliar inducido por morfina.

Morphine produces contraction of Oddi’s sphincter, which can be severe and of longer duration in some pathological conditions. This exaggerated response can manifest as a colicky biliary pain, frequently accompanied by a dramatic increase in hepatic enzymes. We report a 32 years old female who consulted in the emergency room for severe low abdominal pain of gynecologic origin, which was completely controlled by morphine. However, she presented a sudden epigastric colicky pain irradiating in the back, which persisted for several hours in spite of the repeated administration of analgesics. Transaminases elevated from previously normal value to over 1,000 U/L, and returned to the normal level without further treatment after several days. Magnetic resonance cholangiography showed normal fine bile duct, without stones. This transient increase in hepatic enzymes was considered as a consequence of high biliary pressure secondary to morphine-induced spastic contraction of Oddi’s sphincter and a consecutive hepatocellular necrosis.

The 3,4-dihydroxyphenylacetic acid catabolon, a catabolic unit for degradation of biogenic amines tyramine and dopamine in Pseudomonas putida U.

Degradation of tyramine and dopamine by Pseudomonas putida U involves the participation of twenty one proteins organized in two coupled catabolic pathways, Tyn (tynABFEC tynG tynR tynD, 12 338 bp) and Hpa (hpaR hpaBC hpaHI hpaX hpaG1G2EDF hpaA hpaY, 12 722 bp). The Tyn pathway catalyses the conversion of tyramine and dopamine into 4-hydroxyphenylacetic acid (4HPA) and 3,4-dihydroxyphenylacetic acid (3,4HPA) respectively. Together, the Tyn and Hpa pathways constitute a complex catabolic unit (the 3,4HPA catabolon) in which 3,4HPA is the central intermediate. The genes encoding Tyn proteins are organized in four consecutive transcriptional units (tynABFEC, tynG, tynR and tynD), whereas those encoding Hpa proteins constitute consecutive operons (hpaBC, hpaG1G2EDF, hpaX, hpaHI) and three independent units (hpaA, hpaR and hpaY). Genetic engineering approaches were used to clone tyn and hpa genes and then express them, either individually or in tandem, in plasmids and/or bacterial chromosomes, resulting in recombinant bacterial strains able to eliminate tyramine and dopamine from different media. These results enlarge our biochemical and genetic knowledge of the microbial catabolic routes involved in the degradation of aromatic bioamines. Furthermore, they provide potent biotechnological tools to be used in food processing and fermentation as well as new strategies that could be used for pharmacological and gene therapeutic applications in the near future.

Temperature has reciprocal effects on colanic acid and polysialic acid biosynthesis in E. coli K92.

Escherichia coli K92 is an opportunistic pathogen bacterium able to produce polysialic acid (PA) capsules when grows at 37 degrees C. PA polysaccharides are cell-associated homopolymers tailored from acid sialic monomers that function as virulence factors in different neuroinvasive diseases caused by certain Enterobacteriaceae. Conversely, when grows at 19 degrees C (restrictive conditions), PA synthesis was negligible, whereas in such condition, a slimy substance started to be accumulated in the culture broths. Analysis by uronic acids colorimetric determinations, gas chromatography-mass spectrometry, and Fourier transform infrared spectroscopy allowed the isolation and identification of mucoid substance as colanic acid (CA). CA is a heteropolymer containing glucose, galactose, fucose, and glucuronic acid as monomers which seems to be involved in the protection of this bacterium against environment assaults. The study of physicochemical conditions required for CA synthesis revealed that in E. coli K92, nutrient (carbon and nitrogen sources) modulates CA production, reaching the maximal values when glucose and proline were as carbon and nitrogen sources, respectively. Furthermore, we have found that E. coli K92 is able to produce CA at all temperatures tested (from 42 degrees C to 15 degrees C), whereas PA synthesis only occurred when bacteria were cultured at temperatures higher than 25 degrees C. Additionally, genetic engineering approaches revealed that the CA cluster including several genes required for synthesis was placed into a DNA fragment of 100 kb using polymerase chain reaction methodology.

Genetic analyses and molecular characterization of the pathways involved in the conversion of 2-phenylethylamine and 2-phenylethanol into phenylacetic acid in Pseudomonas putida U.

In Pseudomonas putida U two different pathways (Pea, Ped) are required for the conversion of 2-phenylethylamine and 2-phenylethanol into phenylacetic acid. The 2-phenylethylamine pathway (PeaABCDEFGHR) catalyses the transport of this amine, its deamination to phenylacetaldehyde by a quinohaemoprotein amine dehydrogenase and the oxidation of this compound through a reaction catalysed by a phenylacetaldehyde dehydrogenase. Another catabolic route (PedS(1)R(1)ABCS(2)R(2)DEFGHI) is needed for the uptake of 2-phenylethanol and for its oxidation to phenylacetic acid via phenylacetaldehyde. This implies the participation of two different two-component signal-transducing systems, two quinoprotein alcohol dehydrogenases, a cytochrome c, a periplasmic binding protein, an aldehyde dehydrogenase, a pentapeptide repeat protein and an ABC efflux system. Additionally, two accessory sets of elements (PqqABCDEF and CcmABCDEFGHI) are necessary for the operation of the main pathways (Pea and Ped). PqqABCDEF is required for the biosynthesis of pyrroloquinoline quinone (PQQ), a prosthetic group of certain alcohol dehydrogenases that transfers electrons to an independent cytochrome c; whereas CcmABCDEFGHI is required for cytochrome c maturation. Our data show that the degradation of phenylethylamine and phenylethanol in P. putida U is quite different from that reported in Escherichia coli, and they demonstrate that PeaABCDEFGHR and PedS(1)R(1)ABCS(2)R(2)DEFGHI are two upper routes belonging to the phenylacetyl-CoA catabolon.

[Delusional parasitosis intestinal and dermatological: clinical cases]. / Parasitosis ilusoria intestinal y dermatológica: casos clínicos.

Illusory parasitosis, better known as delusional parasitosis, is a neuropsychiatric syndrome in which patients have the belief of suffering a parasitic disease, that can not be demonstrated after an exhaustive medical study. These patients are characterized by being polyconsultants in different medical specialties and, many of them, have antecedents of psychiatric disorders, some of them undiagnosed. Knowing the existence of the clinical picture, diagnosing early and empathizing with the patient, could give to clinician some clues for a timely and assertive psychiatric referral, and improve patient adherence to the proposed treatment.

A genetically engineered strain of Pseudomonas putida as a useful tool for identifying new therapeutic herbicides.

A genetically engineered strain of Pseudomonas putida U designed for the identification of new therapeutic herbicides has been obtained. In this bacterium, deletion of the homogentisate gene cluster (hmgRABC) confers upon this mutant huge biotechnological possibilities since it can be used: (i) as a target for testing new specific herbicides (p-hydroxy-phenylpyruvate dioxygenase inhibitors); (ii) to identify new therapeutic drugs-effective in the treatment of alkaptonuria and other related tyrosinemia - and (iii) as a source of homogentisic acid in a plant-bacterium association.

Sero- and genotyping of Salmonella in slaughter pigs, from farm to cutting plant, with a focus on the slaughter process.

The objective of this study was to investigate the role of the slaughtering process in Salmonella carcass contamination by typing isolates recovered previously in a double study of the following: (1) a tracking survey from the farm to the slaughterhouse and (2) a survey of the slaughterhouse environment (i.e., lairage area, slaughter line, cutting plant and carcasses). The Salmonella serotypes identified on the carcasses of the 16 tracked batches were frequently linked to lairage, whereas the serotypes detected at the farm, transport or pig-related samples (i.e., caecum content and lymph nodes) were only occasionally detected at the carcass level. Multi-locus variable-number tandem repeats (MLVA) of 77 Salmonella enterica ser. Typhimurium isolates from seven of these batches confirmed the link between the isolates recovered from carcasses and holding pens. Only four of the 16 positive carcasses had profiles previously isolated from lymph nodes or caecal content. In the second part of the study, a total of 131 S. enterica ser. Typhimurium and 74 S. enterica ser. Derby isolates were further characterised by MLVA and Pulsed Field Gel Electrophoresis (PFGE), respectively. The MLVA profiles identified in carcasses varied throughout the working day and were frequently linked to those identified in samples from the slaughter line points collected close in time. PFGE and MLVA profiles identified at lairage were also detected in later processing facilities (i.e., slaughter line and cutting plant) as well as in carcasses. Finally, most of the profiles found at the cutting plants were previously identified in the slaughter line or carcass samples. The results from this study show that Salmonella contamination in pigs entering the slaughterhouse can be attributed to several sources. Typing of isolates by MLVA and PFGE clarified the sources of carcass contamination and improved the accuracy of cross-contamination attributable values. Without obviating the relevant role of infected pigs entering the slaughterhouse, the present study highlights the lairage and slaughtering as important sources of carcass contamination.

Cólico biliar inducido por morfina / Morphine induced biliary pain: case report

Morphine produces contraction of Oddi’s sphincter, which can be severe and of longer duration in some pathological conditions. This exaggerated response can manifest as a colicky biliary pain, frequently accompanied by a dramatic increase in hepatic enzymes. We report a 32 years old female who consulted in the emergency room for severe low abdominal pain of gynecologic origin, which was completely controlled by morphine. However, she presented a sudden epigastric colicky pain irradiating in the back, which persisted for several hours in spite of the repeated administration of analgesics. Transaminases elevated from previously normal value to over 1,000 U/L, and returned to the normal level without further treatment after several days. Magnetic resonance cholangiography showed normal fine bile duct, without stones. This transient increase in hepatic enzymes was considered as a consequence of high biliary pressure secondary to morphine-induced spastic contraction of Oddi’s sphincter and a consecutive hepatocellular necrosis.

Parasitosis ilusoria intestinal y dermatológica: casos clínicos / Delusional parasitosis intestinal and dermatological: clinical cases

Illusory parasitosis, better known as delusional parasitosis, is a neuropsychiatric syndrome in which patients have the belief of suffering a parasitic disease, that can not be demonstrated after an exhaustive medical study. These patients are characterized by being polyconsultants in different medical specialties and, many of them, have antecedents of psychiatric disorders, some of them undiagnosed. Knowing the existence of the clinical picture, diagnosing early and empathizing with the patient, could give to clinician some clues for a timely and assertive psychiatric referral, and improve patient adherence to the proposed treatment.

La parasitosis ilusoria, más conocida como delusión parasitaria, es un síndrome neuropsiquiátrico donde los pacientes tienen el convencimiento de padecer una infestación parasitaria, que no puede ser demostrada tras un exhaustivo estudio médico. Estos pacientes se caracterizan por ser policonsultantes en distintas especialidades médicas y, muchos de ellos, poseen antecedentes de trastornos psiquiátricos, algunos de ellos no diagnosticados. Conocer la existencia del cuadro, diagnosticar precozmente y empatizar con el paciente, pueden dar al médico clínico algunas claves para una derivación psiquiátrica oportuna y asertiva, y mejorar la adherencia del paciente al tratamiento propuesto. Se presentan cuatro casos clínicos que consultaron por esta extraña condición.

Poly-3-hydroxyalkanoate synthases from Pseudomonas putida U: substrate specificity and ultrastructural studies.

The substrate specificity of the two polymerases (PhaC1 and PhaC2) involved in the biosynthesis of medium-chain-length poly-hydroxyalkanoates (mcl PHAs) in Pseudomonas putida U has been studied in vivo. For these kind of experiments, two recombinant strains derived from a genetically engineered mutant in which the whole pha locus had been deleted (P. putida U Δpha) were employed. These bacteria, which expresses only phaC1 (P. putida U Δpha pMC-phaC1) or only phaC2 (P. putida U Δpha pMC-phaC2), accumulated different PHAs in function of the precursor supplemented to the culture broth. Thus, the P. putida U Δpha pMC-phaC1 strain was able to synthesize several aliphatic and aromatic PHAs when hexanoic, heptanoic, octanoic decanoic, 5-phenylvaleric, 6-phenylhexanoic, 7-phenylheptanoic, 8-phenyloctanoic or 9-phenylnonanoic acid were used as precursors; the highest accumulation of polymers was observed when the precursor used were decanoic acid (aliphatic PHAs) or 6-phenylhexanoic acid (aromatic PHAs). However, although it synthesizes similar aliphatic PHAs (the highest accumulation was observed when hexanoic acid was the precursor) the other recombinant strain (P. putida U Δpha pMC-phaC2) only accumulated aromatic PHAs when the monomer to be polymerized was 3-hydroxy-5-phenylvaleryl-CoA. The possible influence of the putative three-dimensional structures on the different catalytic behaviour of PhaC1 and PhaC2 is discussed.

Genetic and ultrastructural analysis of different mutants of Pseudomonas putida affected in the poly-3-hydroxy-n-alkanoate gene cluster.

Functional analyses of the different proteins involved in the synthesis and accumulation of polyhydroxyalkanoates (PHAs) in P. putida U were performed using a mutant in which the pha locus had been deleted (PpUDeltapha). These studies showed that: (i) Pha enzymes cannot be replaced by other proteins in this bacterium, (ii) the transformation of PpDeltapha with a plasmid containing the locus pha fully restores the synthesis of bioplastics, (iii) the transformation of PpDeltapha with a plasmid harbouring the gene encoding the polymerase PhaC1 (pMCphaC1) permits the synthesis of polyesters (even in absence of phaC2ZDFI); however, in this strain (PpUDeltapha-pMCphaC1) the number of PHAs granules was higher than in the wild type, (iv) the expression of phaF in PpUDeltapha-pMCphaC1 restores the original phenotype, showing that PhaF is involved in the coalescence of the PHAs granules. Furthermore, the deletion of the phaDFI genes in P. putida U considerably decreases (> 70%) the biosynthesis of PHAs consisting of hydroxyalkanoates with aliphatic constituents, and completely prevents the synthesis of those ones containing aromatic monomers. Additional experiments revealed that the deletion of phaD in P. putida U strongly reduces the synthesis of PHA, this effect being restored by PhaF. Moreover, the overexpression of phaF in P. putida U, or in its DeltafadBA mutant, led to the collection of PHA over-producer strains.