Generation and characterization of avian IgY antibodies for detecting beta-casein A1 in bovine milk.

Beta-casein is a primary milk protein that constitutes approximately 30% of the casein in bovine milk, with the two most common types in cattle being A1 and A2. The A2 protein differs from the A1 version due to a mutation in the codon at position 67, resulting in a histidine to proline substitution. However, the bioactive peptide, beta-casomorphine-7 (BCM7), which originates from partial proteolysis of the A1 variant, has been linked to several gastrointestinal disorders in humans. Production of A1 beta casein-free products is increasing demand in the milk market, worldwide. This study generated and characterized a polyclonal IgY antibody that specifically recognizes the A1 beta-casein protein present in cow's milk. A commercially available IgY anti-A1 antibody was used as a positive control, and the sensitivity and specificity of both the commercial and produced anti-A1 antibodies were evaluated. The results showed 100% sensitivity and specificity of 100% of the commercial IgY anti-A1. The in-house produced anti-A1 antibody demonstrated a sensitivity of 95.2% and a specificity of 100%, indicating its potential as a reliable and cost effective tool for detecting A1 beta-casein protein in milk samples.

Applying the Haddon Matrix to Frontline Care Preparedness and Response in Asymmetric Warfare.

INTRODUCTION: Asymmetric warfare and the reaction to its threats have implications in the way far-forward medical assistance is provided in such settings. Investments in far-forward emergency resuscitation and stabilization can contribute to saving lives and increase the resilience of health systems. Thus, it is proposed to extend the use of the Haddon Matrix to determine a set of strategies to better understand and prioritize activities to prepare for and set-up frontline care in the form of Trauma Stabilization Points (TSPs). METHODS: An expert consensus methodology was used to achieve the research aim. A small subject matter experts' group was convened to create and validate the content of the Haddon Matrix. RESULTS: The result of the expert group consultations presented an overview of TSP Preparedness and Operational Readiness activities within a Haddon Matrix framework. Main strategies to be adopted within the cycle from pre- to post-event had been identified and presented considering the identified opportunities in the context of the possibility of implementation. Of particular importance was the revision of a curriculum that fits the civilian medical system and facilitates its adaptation to the context and available resources. CONCLUSION: The new framework to enhance frontline care preparedness and response using the Haddon Matrix facilitated the identification of a set of strategies to support frontline health care workers in a more efficient manner. Since the existing approach and tools are insufficient for modern warfare, additional research is needed.

Targeting Trypanosoma evansi with disulphide-rich peptides derived from a phage display library.

A phage-display library was generated using a Bus thalamus scorpion toxin (BTK-2) as a peptide scaffold. BTK-2 belongs to the disulfide-rich family of proteins with pronounced structural stability due to the presence of three disulfide bridges that connects antiparallel beta-sheets and one alpha helix. Using BTK-2 as a phage display scaffold, we introduced mutations in five residues located in the alpha-helix and two residues located in the smaller loop, keeping intact the disulfide bridges to create a peptide phage-displayed library with disulfide-rich family properties. The library was subjected to in vivo and in vitro phage display selections against Trypanosoma evansi, the etiological agent of "Surra", a disease that affects a wide range of mammals. The development of T. evansi specific biomarkers is essential to improve diagnostic methods and epidemiological studies leading to a more accurate clinical decision for the treatment of this disease of economic impact for commercial livestock production. In this study, we identified two disulfide-rich peptides targeting T. evansi parasites. Further specificity studies are necessary to investigate the potential of selected peptides as new biomarkers to aid diagnostic and treatment procedures of T. evansi infections.

Xpert MTB/RIF test performance assay in respiratory samples at real work settings in a developing country.

INTRODUCTION: The Xpert MTB/RIF test detects DNA from Mycobacterium tuberculosis complex and susceptibility to rifampin. It has been evaluated repeatedly under "ideal" conditions including centrifugation of sputum and bronchoalveolar lavage, Ziehl Neelsen (ZN) and auramine/rhodamine staining, as well as with solid and liquid automated culture methods. Results from such evaluations cannot be extrapolated to low-income countries that do not routinely use all these processes. OBJECTIVE: To assess the performance of the Xpert MTB/RIF test in respiratory samples under "real" conditions of work in a low-income country and its correlation with phenotypic susceptibility testing. MATERIALS AND METHODS: We conducted a cross-sectional study to assess the performance of the Xpert MTB/RIF test in =12 year-old patients with suspected pulmonary tuberculosis. In routine sample processing at the Hospital we do not use sputum centrifugation, staining with auramine/rhodamine or automated liquid culture. RESULTS: We screened 152 patients of whom 108 were eligible for the study and 103 were included in the analysis; 34% of the samples were positive. The overall test sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 91%, 92%, 83% and 96%, respectively. In ZN-negative samples the sensitivity, specificity, PPV and NPV were 87%, 91%, 68% and 97%, respectively. The results of sensitivity and resistance to rifampin were concordant with susceptibility testing using the multiple proportions method (kappa=1, p<0.0001). CONCLUSIONS: The Xpert MTB/RIF test overall performance was similar to the one achieved under ideal conditions. Its performance in ZN-negative samples was better under "real" conditions of work in a low-income country.

Xpert MTB/RIF test performance assay in respiratory samples at real work settings in a developing country / Rendimiento de la prueba Xpert MTB/RIF en muestras respiratorias en el escenario real de trabajo en un país en desarrollo

Introduction: The Xpert MTB/RIF test detects DNA from Mycobacterium tuberculosis complex and susceptibility to rifampin. It has been evaluated repeatedly under "ideal" conditions including centrifugation of sputum and bronchoalveolar lavage, Ziehl Neelsen (ZN) and auramine/rhodamine staining, as well as with solid and liquid automated culture methods. Results from such evaluations cannot be extrapolated to low-income countries that do not routinely use all these processes. Objective: To assess the performance of the Xpert MTB/RIF test in respiratory samples under "real" conditions of work in a low-income country and its correlation with phenotypic susceptibility testing. Materials and methods: We conducted a cross-sectional study to assess the performance of the Xpert MTB/RIF test in =12 year-old patients with suspected pulmonary tuberculosis. In routine sample processing at the Hospital we do not use sputum centrifugation, staining with auramine/rhodamine or automated liquid culture. Results: We screened 152 patients of whom 108 were eligible for the study and 103 were included in the analysis; 34% of the samples were positive. The overall test sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 91%, 92%, 83% and 96%, respectively. In ZN-negative samples the sensitivity, specificity, PPV and NPV were 87%, 91%, 68% and 97%, respectively. The results of sensitivity and resistance to rifampin were concordant with susceptibility testing using the multiple proportions method (kappa=1, p<0.0001). Conclusions: The Xpert MTB/RIF test overall performance was similar to the one achieved under ideal conditions. Its performance in ZN-negative samples was better under "real" conditions of work in a low-income country.

Introducción. La prueba Xpert MTB/RIF detecta el ADN del complejo Mycobacterium tuberculosis y la sensibilidad a rifampicina. La prueba ha sido evaluada en condiciones "ideales" que incluyen la centrifugación de esputo y el lavado broncoalveolar, la tinción de Ziehl Neelsen (ZN) y de auramina-rodamina y los métodos de cultivo sólido y de cultivo líquido automatizado. Los resultados de tales evaluaciones no pueden extrapolarse a países de bajos ingresos que no utilizan habitualmente todos estos procesos. Objetivo. Evaluar el rendimiento de la prueba Xpert MTB/RIF en muestras respiratorias bajo condiciones "reales" de trabajo y su correlación con las pruebas fenotípicas de sensibilidad. Materiales y métodos. Se llevó a cabo un estudio transversal para evaluar el rendimiento de la prueba Xpert MTB/RIF en pacientes =12 años con sospecha de tuberculosis pulmonar. En el procesamiento rutinario de muestras en el Hospital del estudio no se usa la centrifugación del esputo, la tinción con auramina-rodamina ni el cultivo líquido automatizado. Resultados. Se incluyeron 152 pacientes, de los cuales 108 eran elegibles y 103 se incluyeron en el análisis. El 34 % de las muestras fueron positivas; la sensibilidad de la prueba fue de 91 %, la especificidad de 92 %, el valor diagnóstico positivo de 83 % y el valor diagnóstico negativo global de 96 %. En las muestras negativas con Ziehl Neelsen, la sensibilidad fue de 87 %, la especificidad de 91 % y los valores diagnósticos positivo y negativo alcanzaron 68 y 97 %, respectivamente. Los resultados de sensibilidad o resistencia a la rifampicina concordaron con los de la prueba fenotípica de sensibilidad (valor de kappa=1, p<0,0001). Conclusiones. El rendimiento global de la prueba fue similar al obtenido bajo condiciones "ideales". En las muestras negativas con Ziehl Neelsen se obtuvo un mejor rendimiento en las condiciones "reales" de trabajo de un país de bajos ingresos.

Brote hospitalario de Achromobacter denitrificans relacionado con el uso de clorhexidina contaminada / Achromobacter denitrificans hospital outbreak associated with the use of contaminated chlorhexidine

El objetivo de este artículo es describir una serie de casos de pacientes con pseudobacteriemia causada por Achromobacter denitrificans, germen aislado a partir de los hemocultivos de 14 pacientes hospitalizados en una institución de alta complejidad de Medellín. Los casos estuvieron asociados con el uso de clorhexidina al 4 % en jabón contaminado. Todos los pacientes recibieron antibióticos con resultados favorables, posiblemente debido a la baja virulencia de este microorganismo y a las medidas oportunas adoptadas por el Grupo de Control de Infecciones del hospital.

The aim of this paper is to describe a case series of pseudobacteremia caused by Achromobacter denitrificans, isolated from blood cultures from 14 hospitalized patients in a tertiary care institution in Medellín. The cases were associated with the use of contaminated 4% chlorhexidine soap. All patients received antibiotic treatment and had favorable results, possibly because of the low virulence of this organism and the timely measures taken by the hospital infections control group.

Leptospirosis con inmunoglobulina M positiva en pacientes hospitalizados en una institución de tercer nivel de Medellín, Colombia, en 2009 / Leptospirosis and positive IgM in hospitalized patients in a tertiary care institution in Medellín, Colombia, 2009. Case report and literature review

La leptospirosis es una enfermedad zoonótica que debe ser incluida entre los diagnósticos infecciosos por descartar en nuestros pacientes, ya que es subdiagnosticada y puede causar complicaciones graves. Ocurre en poblaciones desarrolladas y en regiones tropicales, tanto a nivel urbano como rural, pudiendo causar epidemias o endemias. La mayoría de las veces los pacientes cursan con cuadros clínicos benignos o, incluso, asintomáticos, pudiendo asemejarse a otras enfermedades infecciosas, lo cual dificulta la sospecha diagnóstica, pero se debe tener en cuenta que en algunos casos ocasiona cursos clínicos graves y complicaciones fatales. El objetivo de esta comunicación es recordar la relevancia de tener una sospecha clínica para hacer un diagnóstico oportuno de esta entidad, reportando una serie de 14 casos con clínica indicativa de leptospirosis, que presentaron resultado positivo de serología para Leptospira spp., lo cual apoyaba el diagnóstico. De ellos, seis pacientes tuvieron complicaciones graves y dos tuvieron desenlaces fatales. Los datos se obtuvieron de pacientes procedentes de diferentes municipios de Antioquia y Chocó, que consultaron y fueron hospitalizados en una institución de tercer nivel de Medellín durante el año 2009.

Due to the fact that leptospirosis is a zoonotic disease that goes underdiagnosed and may cause serious complications, it must be included among the infectious diagnoses to rule out for infected patients. It occurs in developed populations and in tropical regions, both in urban and rural areas, causing epidemics or becoming endemic. Patients usually show benign symptoms or are even asymptomatic, and it may resemble other infectious diseases, which complicates its diagnosis; however, in some cases, it causes severe and fatal complications. The purpose of this paper is to remark the importance of having a high clinical suspicion to make a timely diagnosis of this disease, reporting a series of 14 patients with symptoms suggestive of leptospirosis, who had a positive serology for Leptospira spp., supporting the clinical diagnosis. Out of those patients, six had serious complications and two had fatal outcomes. Data were obtained from patients from different municipalities of Antioquia and Chocó, who consulted and were hospitalized in a tertiary care institution in Medellín, Colombia, during 2009.

Bacteriemia por Elizabethkingia meningoseptica en paciente con leucemia linfoblástica aguda / Bacteriemia by Elizabethkingia meningoseptica in acute linfoblastic leukemia patients

Se reporta un caso de bacteriemia por Elizabethkingia meningoseptica en un paciente inmunocomprometido, con antecedentes de leucemia linfoblástica aguda, que ingresa por infiltración al sistema nervioso central y sospecha de proceso infeccioso. Elizabethkingia meningoseptica es una bacteria oportunista en infecciones intrahospitalarias, con notable resistencia antimicrobiana, la cual es un inusual patógeno en humanos.

A case report of bacteraemia by Elizabethkingia meningoseptica in an immunocompromised patient, who was admitted to the hospital with an acute Linfoblastic leukemia and infection diagnosis, is presented. E. meningoseptica is an emerging bacterium in nosocomial infections with remarkable antimicrobial resistance and an unusual pathogen in humans.

Alteraciones metabólicas con terapia antirretroviral altamente efectiva en niños positivos para VIH, Cali, Colombia / Metabolic alterations with anti-retroviral highly effective therapy in positive children for VIH, Cali, Colombia

Objetivos: describir las alteraciones metabólicas en niños con diagnóstico de VIH y en tratamiento con terapia antirretroviral altamente efectiva (Highly Active Antiretroviral Therapy, HAART). Métodos: se realizó una primera fase descriptiva de los valores de lípidos y glucemia en una cohorte de niños positivos para VIH. De una clínica pediátrica se reclutaron, entre junio de 2003 y junio de 2005, niños mayores de un mes y menores de 16 años en terapia HAART. Estos resultados se compararon con valores de la población. En una segunda fase, se estudió la densidad ósea en estos niños, utilizando DEXA (dual energy X-ray absorptiometry) y antropometría, y se comparó la de controles sanos. Resultados: se incluyeron 38 niños positivos para VIH. En 59,5% de los niños se clasificaron con displidemia. Al compararlo con la población de referencia, el grupo positivo para VIH presentó una prevalencia mayor de hipertrigliceridemia y HDLc (highdensity lipoprotein) anormalmente bajo. Tomando en cuenta la variación por edad, los valores de colesterol total y LDLc (lowdensity lipoprotein), mostraron un aumento en el grupo que recibía inhibidores de proteasa (IP) contra el que no. La diferencia del puntaje Z de BMD (bone mineral density) entre los grupos fue de 0,56 (IC95%: 0,1- 1,0), teniendo un menor puntaje Z el grupo positivo para VIH. El puntaje Z de la densidad de masa ósea mostró un declive con el tiempo de exposición, que no fue evidente en el grupo control. Conclusiones: encontramos alteraciones en los lípidos similares a las descritas en el adulto seropositivo. En el grupo con IP se encontraron alteraciones del colesterol que cambiaban según la edad. Se encontró una pérdida de la densidad ósea, progresiva con el tiempo de exposición e independiente de la edad. Consideramos que esta relación podría ser de origen multifactorial, incluyendo los efectos de la infección y del tratamiento.

Objectives: our goal is to describe metabolic alterations in children with HIV and under highly effective anti-retroviral treatment (HAART). Methodology: a first descriptive phase of lipid levels and glucemia was carried out in a cohort of HIV positive children. In a paediatric hospital, children >1 month and <16 years old under HAART were recruited from June, 2003 to June, 2005. The results were compared to population values. During the second phase, bone density was studied in these children using DEXA and anthropometric values and compared to healthy control subjects. Results: thirty eight positive children were included. 59.5% of the children were classified as having dyslipidemia. Upon comparison to the reference population, the HIV(+) group showed larger hypertrigliceridemia prevalence and abnormally low cHDL. Taking into account age variations, total colesterol values and cLDL showed increase in the group that received PI against those that did not. The difference of the BMD Z-Score among the groups was 0,56 (CI95%: 0.1, 1.0), the HIV(+) group having a smaller Z-Score. The bone mass density Z-Score showed a decline according exposition time, which was not evident in the control group. Conclusions: alterations in lipids similar to those described in the seropositive adult were found. The group with PI showed cholesterol alterations that changed according to age. Progressive bone density loss according to exposition time was found regardless of age. It is considered that this relationship could have multifactorial origin, including infection and treatment effects.