Influencia del desequilibrio sagital en el resultado clínico de la vertebroplastia en pacientes con fractura por compresión vertebral osteoporótica / Sagittal imbalance influences outcome of vertebroplasty in patients with osteoporotic vertebral compression fracture

Antecedentes y objetivos: La correlación entre el equilibrio sagital de la columna y el resultado clínico tras una vertebroplastia (VP) en pacientes con fractura vertebral osteoporótica por compresión (FVOC) ha sido poco estudiada. Analizamos el resultado clínico de la VP en pacientes con FVOC teniendo en cuenta el equilibrio sagital. Material y método: El objetivo primario es valorar el cambio en el dolor axial, la discapacidad y la calidad de vida relacionada con la salud mediante la escala analógica visual (VAS), índice de discapacidad de Oswestry (ODI) y el test SF-36, respectivamente. Todo ello, correlacionado con el eje sagital vertical (SVA) que define el equilibrio sagital. El estudio radiográfico consistió en una radiografía de perfil de columna completa en bipedestación. Los controles clínico-radiológicos se realizaron pre- y post-VP (1, 3 y 12 meses). Resultados: Se incluyeron 51 pacientes con un total de 113 FVOC; 30 pacientes (60,7%) presentaron múltiples FVOC. Comparando la evolución del resultado del VAS y del ODI durante el seguimiento no se observaron diferencias significativas entre los grupos de pacientes con SVA<50mm y >50mm (p>0,05). Por el contrario, en pre-VP los resultados del SF-36 presentaban peores puntuaciones en el grupo de SVA>50mm en la sección de función física (FP) (p<0,05) y de componente de salud física (CSF) (p<0,05). Estas diferencias se mantenían hasta los 3 meses de seguimientos en CSF y hasta el final del seguimiento en la sección de FP (p<0,05). Conclusiones: Los pacientes con SVA>50mm presentan una recuperación más lenta de su calidad de vida relacionada con la salud tras VP por FVOC, pero sin diferencias significativas con respecto al dolor o discapacidad cuando se comparan con pacientes con SVA<50mm.(AU)

Background and objectives: The correlation between sagittal balance of the spine and clinical outcome after vertebroplasty (VP) in patients with osteoporotic vertebral compression fractures (OVCF) is poorly investigated. We analysed the clinical outcome of patients with OVCF undergoing VP taking into account sagittal balance. Material and method: The primary endpoint was the change in axial back pain, disability and health-related quality-of-life using Visual Analogue Scale (VAS), Oswestry Disability Index (ODI) and SF-36, respectively, in correlation to the parameters that define sagittal balance (SVA). Radiographic assessment included full spine standing lateral films. Imaging and clinical data were collected pre- and post-procedure at 1, 3 and 12 months. Results: 51 patients were included presenting a total of 113 OVCF. 30 patients (60.7%) had multiple OVCF. Comparing the evolution of VAS and ODI throughout the follow-up it does not seem that there are significant differences in their behaviour between the SVA>50mm and the SVA<50mm groups (p>0.05). On the contrary, pre-VP SF-36 scores showed worst results in the SVA>50mm group in the physical functioning (PF) section (p<0.05) and in the physical component score (PCS) (p<0.05). These differences were maintained until 3 months of follow-up in the case of the PCS and until the end of follow-up in the case of the PF (p<0.05). Conclusions: Patients with a SVA>50mm showed a slower recovery of their quality-of-life after VP for OVCF, but without significant differences with respect to pain or disability, when compared patients with SVA<50mm.(AU)

[Translated article] Sagittal imbalance influences outcome of vertebroplasty in patients with osteoporotic vertebral compression fracture / Influencia del desequilibrio sagital en el resultado clínico de la vertebroplastia en pacientes con fractura por compresión vertebral osteoporótica

Antecedentes y objetivos: La correlación entre el equilibrio sagital de la columna y el resultado clínico tras una vertebroplastia (VP) en pacientes con fractura vertebral osteoporótica por compresión (FVOC) ha sido poco estudiada. Analizamos el resultado clínico de la VP en pacientes con FVOC teniendo en cuenta el equilibrio sagital. Material y método: El objetivo primario es valorar el cambio en el dolor axial, la discapacidad y la calidad de vida relacionada con la salud mediante la escala analógica visual (VAS), índice de discapacidad de Oswestry (ODI) y el test SF-36, respectivamente. Todo ello, correlacionado con el eje sagital vertical (SVA) que define el equilibrio sagital. El estudio radiográfico consistió en una radiografía de perfil de columna completa en bipedestación. Los controles clínico-radiológicos se realizaron pre- y post-VP (1, 3 y 12 meses). Resultados: Se incluyeron 51 pacientes con un total de 113 FVOC; 30 pacientes (60,7%) presentaron múltiples FVOC. Comparando la evolución del resultado del VAS y del ODI durante el seguimiento no se observaron diferencias significativas entre los grupos de pacientes con SVA<50mm y >50mm (p>0,05). Por el contrario, en pre-VP los resultados del SF-36 presentaban peores puntuaciones en el grupo de SVA>50mm en la sección de función física (FP) (p<0,05) y de componente de salud física (CSF) (p<0,05). Estas diferencias se mantenían hasta los 3 meses de seguimientos en CSF y hasta el final del seguimiento en la sección de FP (p<0,05). Conclusiones: Los pacientes con SVA>50mm presentan una recuperación más lenta de su calidad de vida relacionada con la salud tras VP por FVOC, pero sin diferencias significativas con respecto al dolor o discapacidad cuando se comparan con pacientes con SVA<50mm.(AU)

Background and objectives: The correlation between sagittal balance of the spine and clinical outcome after vertebroplasty (VP) in patients with osteoporotic vertebral compression fractures (OVCF) is poorly investigated. We analysed the clinical outcome of patients with OVCF undergoing VP taking into account sagittal balance. Material and method: The primary endpoint was the change in axial back pain, disability and health-related quality-of-life using Visual Analogue Scale (VAS), Oswestry Disability Index (ODI) and SF-36, respectively, in correlation to the parameters that define sagittal balance (SVA). Radiographic assessment included full spine standing lateral films. Imaging and clinical data were collected pre- and post-procedure at 1, 3 and 12 months. Results: 51 patients were included presenting a total of 113 OVCF. 30 patients (60.7%) had multiple OVCF. Comparing the evolution of VAS and ODI throughout the follow-up it does not seem that there are significant differences in their behaviour between the SVA>50mm and the SVA<50mm groups (p>0.05). On the contrary, pre-VP SF-36 scores showed worst results in the SVA>50mm group in the physical functioning (PF) section (p<0.05) and in the physical component score (PCS) (p<0.05). These differences were maintained until 3 months of follow-up in the case of the PCS and until the end of follow-up in the case of the PF (p<0.05). Conclusions: Patients with a SVA>50mm showed a slower recovery of their quality-of-life after VP for OVCF, but without significant differences with respect to pain or disability, when compared patients with SVA<50mm.(AU)

Genetics: Is LADA just late onset type 1 diabetes?

Background: There is a controversy regarding Latent Autoimmune Diabetes in Adults (LADA) classification and whether it should be considered a slowly progressing form of type 1 (T1) diabetes (DM) or a distinct type of DM altogether. Methods: This cross-sectional study assessed major genes associated with T1DM (class II HLA, PTPN22 [rs2476601] and INS [rs689]) in patients with LADA, as compared with participants with T1DM (stratified according to age of diagnosis before or after 30) and T2DM. HLA genotyping of the DRB1, DQA1 and DQB1 loci was performed by reverse PCR sequence-specific oligonucleotides. HLA haplotypes were assigned according to those most frequently described in the European population. INS and PTPN22 SNPs were genotyped by real-time PCR. Results: A total of 578 participants were included: 248 with T1DM (70 diagnosed after the age of 30), 256 with T2DM and 74 with LADA. High risk HLA alleles were significantly more frequent in LADA than in T2DM, whereas the opposite was true for protective alleles. We found a lower frequency of the high-risk DRB1*04-DQB1*03:02-DQA1*03:01 haplotype in LADA (21.1%) than in the overall T1DM (34.7%) (p<0.05), whereas no differences were found between these groups for DRB1*03-DQB1*02:01-DQA1*05:01 or for protective alleles. Only 12% the overall T1DM group had no risk alleles vs 30% of LADA (p<0.0005). However, HLA allele distribution was similar in LADA and T1DM diagnosed after the age of 30. A total of 506 individuals (195 with T1DM [21 diagnosed after age 30] 253 with T2DM and 58 with LADA) were genotyped for the PTPN22 and INS SNPs. The G/A genotype of the PTPN22 rs2476601 was more frequent and the T/T genotype of the INS SNP rs689 was less frequent in T1DM compared to LADA. We did not find any significant differences in the frequency of the mentioned SNPs between LADA and T2DM, or between LADA and T1DM diagnosed after the age of 30. Conclusion: In this relatively small cross-sectional study, the genetic profile of subjects with LADA showed a similar T1DM-related risk allele distribution as in participants with T1DM diagnosed after the age of 30, but fewer risk alleles than those diagnosed before 30. Differences were present for HLA, as well as PTPN22 and INS genes.

[Translated article] Sagittal imbalance influences outcome of vertebroplasty in patients with osteoporotic vertebral compression fracture.

BACKGROUND AND OBJECTIVES: The correlation between sagittal balance of the spine and clinical outcome after vertebroplasty (VP) in patients with osteoporotic vertebral compression fractures (OVCF) is poorly investigated. We analysed the clinical outcome of patients with OVCF undergoing VP taking into account sagittal balance. METHODS: The primary endpoint was the change in axial back pain; disability and health-related quality of life using VAS, ODI and SF-36 respectively in correlation to the parameters that define sagittal balance (SVA). Radiographic assessment included full spine standing lateral films. Imaging and clinical data were collected pre and post procedure at 1, 3 and 12 months. RESULTS: Fifty-one patients were included presenting a total of 113 OVCF. Thirty patients (60.7%) had multiple OVCF. Comparing the evolution of VAS and ODI throughout the follow-up it does not seem that there are significant differences in their behaviour between the SVA>50mm and the SVA<50mm groups (p>0.05). On the contrary, preVP SF-36 scores showed worst results in the SVA>50mm group in the physical functioning section (PF) (p<0.05) and in the physical component score (PCS) (p<0.05). These differences were maintained until 3 months of follow-up in the case of the PCS and until the end of follow-up in the case of the PF (p<0.05). CONCLUSIONS: Patients with a SVA>50mm showed a slower recovery of their quality of life after VP for OVCF, but without significant differences with respect to pain or disability, when compared with patients with SVA<50mm.

Sagittal imbalance influences outcome of vertebroplasty in patients with osteoporotic vertebral compression fracture. / Influencia del desequilibrio sagital en el resultado clínico de la vertebroplastia en pacientes con fractura por compresión vertebral osteoporótica.

BACKGROUND AND OBJECTIVES: The correlation between sagittal balance of the spine and clinical outcome after vertebroplasty (VP) in patients with osteoporotic vertebral compression fractures (OVCF) is poorly investigated. We analysed the clinical outcome of patients with OVCF undergoing VP taking into account sagittal balance. MATERIAL AND METHOD: The primary endpoint was the change in axial back pain, disability and health-related quality-of-life using Visual Analogue Scale (VAS), Oswestry Disability Index (ODI) and SF-36, respectively, in correlation to the parameters that define sagittal balance (SVA). Radiographic assessment included full spine standing lateral films. Imaging and clinical data were collected pre- and post-procedure at 1, 3 and 12 months. RESULTS: 51 patients were included presenting a total of 113 OVCF. 30 patients (60.7%) had multiple OVCF. Comparing the evolution of VAS and ODI throughout the follow-up it does not seem that there are significant differences in their behaviour between the SVA>50mm and the SVA<50mm groups (p>0.05). On the contrary, pre-VP SF-36 scores showed worst results in the SVA>50mm group in the physical functioning (PF) section (p<0.05) and in the physical component score (PCS) (p<0.05). These differences were maintained until 3 months of follow-up in the case of the PCS and until the end of follow-up in the case of the PF (p<0.05). CONCLUSIONS: Patients with a SVA>50mm showed a slower recovery of their quality-of-life after VP for OVCF, but without significant differences with respect to pain or disability, when compared patients with SVA<50mm.

Essential oils microemulsions prepared with high-frequency ultrasound: physical properties and antimicrobial activity.

Essential oils (EOs) have demonstrated antimicrobial activity against bacteria due to the effects of their major components. The direct application of EOs may present a rapid volatilization of its components and can decrease their effectiveness. Encapsulation by means of emulsification can provide protection to lipid compounds on a microscale. The aim of this study was to characterize microemulsions of cinnamon essential oil (CEO), oregano essential oil (OEO), and rosemary essential oil (REO) prepared by high-frequency ultrasound and evaluate their antimicrobial activities against Escherichia coli and Listeria monocytogenes. The microemulsions (oil-in-water, O/W) of EOs were prepared using high-frequency ultrasound, applying a wave amplitude of 84 µm for 15 min (REO and CEO) or 30 min (OEO). The antimicrobial activity was determined by inoculating 108 CFU/mL of bacteria. Nonsurvival of the bacteria was confirmed by plate count in tryptic soy agar, determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The microemulsions exhibited droplet size diameters of 1.98 to 5.46 µm, showing high encapsulation efficiencies (79.91-81.97%) and low separation rates (2.50-6.67%). The MIC and MBC for the microemulsions for both bacteria were 20-75% less than values obtained for the non-encapsulated EOs. This study demonstrates that high-frequency ultrasound is a suitable technique for obtaining stable microemulsions to deliver natural antimicrobials that can be applied to control bacteria of high relevance in food safety.

Toxicidad del formaldehido en trabajadores profesionalmente expuestos. Revisión bibliográfica / Scoping review about formaldehyde toxicity on occupationally exposed workers

La presencia del formaldehido en el ámbito laboral sigue siendo importante. La exposición produce irritación local de mucosas oculares, nasales y del tracto respiratorio superior, y crónicamente se ha asociado con mayor riesgo de desarrollar cáncer a nivel de senos paranasales, naso-orofaringe y pulmón. Esta revisión tiene por objetivo actualizar la bibliografía y categorizar la evidencia científica más actualizada de los efectos que el formaldehido produce sobre el organismo humano. Búsqueda bibliográfica en la base de datos electrónica Medline/PubMed, limitada a los últimos 10 años mediante combinación de lenguaje libre y controlado. Se revisaron 185 artículos con inclusión final de 54 tras descartar por duplicidad, idioma y criterios de inclusión. Se observa un elevado grado de evidencia respecto a la genotoxicidad, evidencia contradictoria, inconsistente o limitada respecto a patologías neoplásicas de origen hematopoyético, laringe, naso-sinusales o de pulmón y falta de evidencia sobre la relación con el asma bronquial. Es preciso efectuar nuevos estudios, especialmente con carácter longitudinal y mayor potencia epidemiológica, para generar nuevo conocimiento sobre el comportamiento de este tóxico

The presence of formaldehyde at workplace remains significant. Exposure to it results in local irritation of the eye, nose and upper respiratory tract mucous membranes, and it has been chronically related to a higher risk of cancer development at the paranasal sinuses, naso-oropharynx and lungs. The aim of our work has been the updating of the bibliography and the categorization of the most up to date scientific evidence of formaldehyde effects on human body. Bibliographic search on the electronic database Medline / PubMed, restricted to the last 10 years through a combination of free and controlled language. Review of 185 scientific articles, finally including 54 due to duplicity, language, and inclusion criteria. We find among the main results a major evidence regarding genotoxicity; limited, inconsistent, and contradictory evidence regarding various neoplastic pathologies; and lack of evidence regarding bronchial asthma. Further studies have to be carried out, especially longitudinal studies and greater epidemiological power, to generate new knowledge about the behavior of this toxic

La presència del formaldehid a l'àmbit laboral segueix sent important. L'exposició produeix irritació local de mucoses, com ara les oculars, nasals i del tracte respiratori superior i de manera crònica s'ha associat amb un major risc de desenvolupar càncer a nivell de mucosa i sinus paranasals, naso-orofaringe i pulmó. L'objectiu del nostre treball ha estat actualitzar la bibliografia i categoritzar l'evidència científica més actualitzada dels efectes que el formaldehid produeix sobre l'organisme humà. Mètode: Revisió de la literatura utilitzant la metodologia "scoping review". Cerca bibliogràfica a la base de dades electrònica Medline/PubMed, limitada als darrers 10 anys mitjançant combinació de llenguatge lliure i controlat. Anàlisi efectuada per un col.lectiu de Metges especialistes en medicina del treball, pertanyents a l'àmbit sanitari i que també han comptat amb la participació d'higienistes i altres professionals de la prevenció de riscos. Resultats: Revisió de 185 articles, amb inclusió final de 54 un cop descartats per duplicitat, idioma i criteris d'inclusió. Entre els principals resultats es troba un elevat grau d'evidència referida a genotoxicitat, evidència contradictòria, inconsistent o limitada pel que fa a diverses patologies neoplàsiques d'origen hematopoiètic, laringe, naso-sinusals o de pulmó i manca d'evidència sobre la relació ambl'asma bronquial. Conclusions: La revisió mostra un canvi de paradigma dels efectes del formaldehid sobre l'organisme humà, amb dades que qüestionen els efectes tòxics clàssicament admesos. Cal efectuar nous estudis, especialment amb caràcter longitudinal i major potència epidemiològica, per generar nou coneixement sobre el comportament d'aquest tòxic

[Scoping review on formaldehyde toxicity in occupationally exposed workers]. / Toxicidad del formaldehido en trabajadores profesionalmente expuestos. Revisión bibliográfica.

The presence of formaldehyde at workplace remains significant. Exposure to it results in local irritation of the eye, nose and upper respiratory tract mucous membranes, and it has been chronically related to a higher risk of cancer development at the paranasal sinuses, naso-oropharynx and lungs. The aim of our work has been the updating of the bibliography and the categorization of the most up to date scientific evidence of formaldehyde effects on human body. Bibliographic search on the electronic database Medline / PubMed, restricted to the last 10 years through a combination of free and controlled language.Review of 185 scientific articles, finally including 54 due to duplicity, language, and inclusion criteria. We find among the main results a major evidence regarding genotoxicity; limited, inconsistent, and contradictory evidence regarding various neoplastic pathologies; and lack of evidence regarding bronchial asthma. Further studies have to be carried out, especially longitudinal studies and greater epidemiological power, to generate new knowledge about the behavior of this toxic.

La presencia del formaldehido en el ámbito laboral sigue siendo importante. La exposición produce irritación local de mucosas oculares, nasales y del tracto respiratorio superior, y crónicamente se ha asociado con mayor riesgo de desarrollar cáncer a nivel de senos paranasales, naso-orofaringe y pulmón. Esta revisión tiene por objetivo actualizar la bibliografía y categorizar la evidencia científica más actualizada de los efectos que el formaldehido produce sobre el organismo humano. Búsqueda bibliográfica en la base de datos electrónica Medline/PubMed, limitada a los últimos 10 años mediante combinación de lenguaje libre y controlado. Se revisaron 185 artículos con inclusión final de 54 tras descartar por duplicidad, idioma y criterios de inclusión. Se observa un elevado grado de evidencia respecto a la genotoxicidad, evidencia contradictoria, inconsistente o limitada respecto a patologías neoplásicas de origen hematopoyético, laringe, naso-sinusales o de pulmón y falta de evidencia sobre la relación con el asma bronquial. Es preciso efectuar nuevos estudios, especialmente con carácter longitudinal y mayor potencia epidemiológica, para generar nuevo conocimiento sobre el comportamiento de este tóxico.

Report From the First and Second Spanish Killer Immunoglobulin-Like Receptor Genotyping Workshops: External Quality Control for Natural Killer Alloreactive Donor Selection in Haploidentical Stem Cell Transplantation.

An important factor affecting the success in the setting of related haploidentical hematopoietic stem cell transplantation (HSCT) is the graft-versus-leukemia effect mediated by natural killer (NK) cells when the donor displays NK alloreactivity versus the recipient. NK cell function is regulated by killer immunoglobulin-like receptors (KIR) and it has been described that donor KIR genotype influences transplantation outcome. This has led to a requirement of laboratories to have a quality assurance program for validation and control of their KIR genotyping methods. The goal of the 1st and 2nd Spanish KIR Genotyping Workshops was to provide an external proficiency testing program in KIR genotyping for Spanish immunology and transplant laboratories. These workshops were conducted during the years 2014-2016 and consisted of 17 participating laboratories typing a set of 20 samples. The presence/absence of 16 mandatory KIR loci (2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 2DP1, 3DL1, 3DL2, 3DL3, 3DS1, and 3DP1) was evaluated per sample. Methods for KIR genotyping included polymerase chain reaction with the use of sequence-specific primers and sequence-specific oligoprobes. Consensus typing was reached in all samples, and the performance of laboratories in external proficiency testing was satisfactory in all cases. The polymorphism detected in the small sample studied in both workshops is indicative of an ample variety of KIR gene profiles in the Spanish population.

Corps étrangers laryngo-trachéo-bronchiques : à propos de 130 cas

Objectifs : Les objectifs de notre étude étaient de décrire les aspects épidémiologiques, diagnostiques et thérapeutiques des corps étrangers des voies respiratoires inférieures à la clinique d'ORL et de chirurgie cervico-faciale du centre Hospitalier National et Universitaire de Fann. Patients et méthodes : Il s'agit d'une étude rétrospective, couvrant la période du 1er janvier 2011 au 31 décembre 2013. Nous avons colligé, durant cette période, 130 dossiers de patients âgés de moins de 18 ans qui étaient reçus pour inhalation de corps étrangers. Les endoscopies blanches étaient exclues. Les paramètres étudiés étaient : l'âge, le sexe, la présence ou non d'un syndrome de pénétration, les signes fonctionnels, le délai de consultation, le traitement effectué, la localisation endoscopique, la nature du corps étranger, les données de l'imagerie, l'indication de la trachéotomie et les complications. L'analyse des données était effectuée avec le logiciel Sphinx Lexica V 4.5 2003. Résultats : L'âge moyen de nos patients était de 3,84 ans avec des extrêmes allant de 4 mois à 17 ans. Une prédominance féminine était notée avec un sex-ratio de 0,91. Les patients évacués d'autres pays représen- taient 6,9% des cas. Le délai moyen de consultation était de 8,05 jours. Le syndrome de pénétration était retrouvé dans 78,5% des cas. La radiographie standard mettait en évidence un corps étranger radio-opaque dans 33,1% des cas. La localisation laryngée était la plus fréquente, elle représentait 26,7% des cas. La graine d'arachide était le corps étranger le plus fréquemment retrouvé soit 39,2%. Onze patients étaient trachéotomisés. Quatre cas de décès ont été rapportés soit 3% des cas.Conclusion : L'inhalation d'un corps étranger est un problème de santé publique. L'augmentation de l'incidence de cet accident et la transition épidémiologique impose une meilleure prévention

Desensitization Before Living Donor Kidney Transplantation in Highly HLA-Sensitized Patients: A Single-Center Study.

BACKGROUND: Renal transplantation in highly sensitized patients represents a major clinical challenge leading to long periods on the waiting list. When a living donor is available, the use of different strategies to desensitize recipients with preformed human leukocyte antigen antibodies can allow a successful transplantation. METHODS: We performed a retrospective observational study including all living donor kidney transplantation (LDKT) with desensitization (DS) from 2008 to 2014 in our transplant unit. The rates of rejection and graft survival were evaluated. DS consisted of plasma exchange (PE), rituximab (RTX), and intravenous immunoglobulin (IVIG) induction with thymoglobulin and maintenance immunosuppression with tacrolimus, corticosteroids, and mycophenolate mofetil. RESULTS: From 2008 to 2014, we performed 368 LDKT, with 31 receiving desensitization. Seven cases from a clinical trial were excluded. Demographic data and outcomes were recorded. All of the patients received RTX + PE + IVIG. DS was performed for positive complement-dependent cytotoxicity cross-match (4.2%), T-cell- and/or B-cell-positive flow cytometry cross-match (87.5%) and presence of donor-specific antibodies alone (8.3%). We identified 23 episodes of rejection in 12 patients (50%); 79% were antibody-mediated rejections (AMR). Graft failure was 12.5%, with a mean time to graft loss of 229 ± 203 days. Mean follow-up was 37 ± 27 months, and graft survival was 91% and 86% at 1 and 5 years, respectively. CONCLUSIONS: Desensitization in LDKT appears to offer an acceptable option for highly sensitized patients. In our series, 41% presented an AMR and 12.5% showed transplant glomerulopathy in protocol and/or indication biopsies. However, short-term outcomes and graft survival were satisfactory.

HLA-DQ2/DQ8 and HLA-DQB1*02 homozygosity typing by real-time polymerase chain reaction for the assessment of celiac disease genetic risk: evaluation of a Spanish celiac population.

Celiac disease (CD) is a complex autoimmune disorder caused by ingestion of gluten in genetically susceptible individuals. Different genetic risk factors have been identified, but virtually all patients are human leukocyte antigen (HLA)-DQ2 and/or HLA-DQ8 positive. We describe a new, fast, accurate and simple real-time polymerase chain reaction (PCR)-based assay for the genotyping and homozygosity analysis of the CD-related HLA alleles. The assay overcomes the major limitations of protocols currently in use, allowing HLA-DQ2/DQ8 genotyping by using only three real-time PCR reactions. For the appraisal of DQ2 homozygosity, only one more reaction is needed. These reactions are easily automated and suitable for large screening studies in diagnostic procedures, as it is demonstrated by their successful application in our HLA diagnostic laboratory. Finally, we assessed the clinical relevance of this real-time PCR-based assay by studying a cohort of fully characterized patients. As expected, all CD patients had at least one of the CD-associated alleles, and the highest CD risk was indicated by the presence of the HLA-DQ2.5 heterodimer (HLA-DQA1*05-DQB1*02) with HLA-DQB1*02 in homozygosity.

Probiotic viability and storage stability of yogurts and fermented milks prepared with several mixtures of lactic acid bacteria.

Currently, the food industry wants to expand the range of probiotic yogurts but each probiotic bacteria offers different and specific health benefits. Little information exists on the influence of probiotic strains on physicochemical properties and sensory characteristics of yogurts and fermented milks. Six probiotic yogurts or fermented milks and 1 control yogurt were prepared, and we evaluated several physicochemical properties (pH, titratable acidity, texture, color, and syneresis), microbial viability of starter cultures (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and probiotics (Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus reuteri) during fermentation and storage (35 d at 5°C), as well as sensory preference among them. Decreases in pH (0.17 to 0.50 units) and increases in titratable acidity (0.09 to 0.29%) were observed during storage. Only the yogurt with S. thermophilus, L. delbrueckii ssp. bulgaricus, and L. reuteri differed in firmness. No differences in adhesiveness were determined among the tested yogurts, fermented milks, and the control. Syneresis was in the range of 45 to 58%. No changes in color during storage were observed and no color differences were detected among the evaluated fermented milk products. Counts of S. thermophilus decreased from 1.8 to 3.5 log during storage. Counts of L. delbrueckii ssp. bulgaricus also decreased in probiotic yogurts and varied from 30 to 50% of initial population. Probiotic bacteria also lost viability throughout storage, although the 3 probiotic fermented milks maintained counts ≥ 10(7)cfu/mL for 3 wk. Probiotic bacteria had variable viability in yogurts, maintaining counts of L. acidophilus ≥ 10(7) cfu/mL for 35 d, of L. casei for 7d, and of L. reuteri for 14 d. We found no significant sensory preference among the 6 probiotic yogurts and fermented milks or the control. However, the yogurt and fermented milk made with L. casei were better accepted. This study presents relevant information on physicochemical, sensory, and microbial properties of probiotic yogurts and fermented milks, which could guide the dairy industry in developing new probiotic products.

Aspergillus niger time to growth in dried tomatoes.

Individual and combined effects of aw and incorporation of selected concentrations of Mexican oregano essential oil on the time to growth (TTG) of Aspergillus niger intentionally inoculated into dried tomatoes were studied during storage at 25°C for 100 days. For aw 0.96, 1,000 ppm of Mexican oregano essential oil inhibited A. niger growth during 100 days, whereas 500 ppm were sufficient at aw 0.91 and 250 ppm for tomatoes with aw 0.78. A. niger growth was evident at different incubation times depending on tested tomato aw and concentration of essential oil; these data were utilized to model TTG. Regression analysis revealed good agreement between experimental and predicted data with a correlation coefficient higher than 0.98. Analysis of mold growth data through TTG models makes possible to include observations detected as no growth and can be utilized to predict mold time to growth for specific preservation factor combinations or to select preservation factor levels for an expected shelf-life based on A. niger growth.

Zygosaccharomyces bailii inactivation by means of UV light and low-frequency ultrasound treatments.

Zygosaccharomyces bailii inactivation suspended in apple juice was evaluated under the effects of selected treatments: short-wave UV light (UVC, using one or two lamps), or low-frequency ultrasound (US), or their simultaneous combination. US treatments (20 kHz, 120-µm wave amplitude) were performed at 35°C in a double-wall vessel by using a 13-mm probe. The UVC device consists of two 90-cm-long stainless steel tubes with 40-W UVC lamps covered with quartz tubes, each one inside a stainless steel tube (annular inside diameter of 2.6 cm) connected to a peristaltic pump. Inoculated systems were recirculated through individual or simultaneous US and UVC treatments, samples were taken periodically, and yeast survivors were determined by the plate-count technique. Yeast survival curves demonstrated that UVC alone or in combination with US produced higher inactivation than US alone. Survival curves were appropriately described by the Weibull distribution of resistances model, obtaining model parameter values that adequately reflected the effect of the studied treatments. For every tested case, the distribution of resistances model revealed an absence of mode, while mean values and variances decreased when simultaneous UVC irradiation with two lamps and US were applied, reaching a 7-log cycle reduction after 40 min of treatment. Combined treatment was more effective than individual US or UVC treatments.

Prevalence of drug resistance and importance of viral load measurements in Honduran HIV-infected patients failing antiretroviral treatment.

OBJECTIVE: The Honduran HIV/AIDS Program began to scale up access to HIV therapy in 2002. Up to May 2008, more than 6000 patients received combination antiretroviral therapy (cART). As HIV drug resistance is the major obstacle for effective treatment, the purpose of this study was to assess the prevalence of antiretroviral drug resistance in Honduran HIV-1-infected individuals. METHODS: We collected samples from 138 individuals (97 adults and 41 children) on cART with virological, immunological or clinical signs of treatment failure. HIV-1 pol sequences were obtained using an in-house method. Resistance mutations were identified according to the 2007 International AIDS Society (IAS)-USA list and predicted susceptibility to cART was scored using the ANRS algorithm. RESULTS: Resistance mutations were detected in 112 patients (81%), 74% in adults and 98% in children. Triple-, dual- and single-class drug resistance was documented in 27%, 43% and 11% of the study subjects, respectively. Multiple logistic regression showed that resistance was independently associated with type of treatment failure [virological failure (odds ratio (OR) = 1) vs. immunological failure (OR = 0.11; 95% confidence interval (CI) 0.030-0.43) vs. clinical failure (OR = 0.037; 95% CI 0.0063-0.22)], route of transmission (OR = 42.8; 95% CI 3.73-491), and years on therapy (OR = 1.81; 95% CI 1.11-2.93). CONCLUSION: The prevalence of antiretroviral resistance was high in Honduran HIV-infected patients with signs of treatment failure. A majority of study subjects showed dual- or triple-class resistance to nucleoside reverse transcriptase inhibitors, nonnucleoside reverse transcriptase inhibitors and protease inhibitors. Virologically defined treatment failure was a strong predictor of resistance, indicating that viral load testing is needed to correctly identify patients with treatment failure attributable to resistance.

High resolution definition of HLA-DRB haplotypes by a simplified microsatellite typing technique.

In humans, the region configurations DR1, DR8, DR51, DR52 and DR53 are known to display copy number as well as allelic variation, rendering high resolution typing of HLA-DRB haplotypes cumbersome. Advantage was taken of microsatellite D6S2878, present in all DRB genes/pseudogenes with an intact exon 2-intron 2 segment. This DRB-STR is highly polymorphic in composition and length. Recently, it was proven that all exon 2 sequences could be linked to a certain DRB-STR that segregates with the respective DRB allele. Because haplotypes show differential copy numbers and compositions of exon 2-positive DRB genes/pseudogenes, unique DRB-STR patterns could be described that appear to be specific for a particular DRB haplotype. The aim of this workshop project was to approve and to qualify this simple typing protocol in a larger panel covering different European populations. All participants succeeded in correctly defining the DRB-STR amplicons varying from 135 to 222 base pair (bp) lengths. The panel of 101 samples covered 50 DRB alleles distributed over 37 different haplotypes as defined by exon 2 sequence-based typing. These haplotypes could be refined into 105 haplotypes by DRB-STR typing. Thus, discrimination of exon 2-identical DRB alleles was feasible, as well as the exact description of three different crossing-over events that resulted in the generation of hybrid DR region configurations. This typing procedure appears to be a quick and highly robust technique that can easily be performed by different laboratories, even without experience in microsatellite typing; thus, it is suitable for a variety of researchers in diverse research areas.

Copy number variation in the CCL4L gene is associated with susceptibility to acute rejection in lung transplantation.

Lung transplantation (LT) has become an accepted therapy for selected patients with advanced lung disease. One of the main limitations to successful LT is rejection of the transplanted organ where chemokines are pivotal mediators. Here, we test the relationship between copy number variation (CNV) in the CCL4L chemokine gene and rejection risk in LT patients (n=161). Patients with no acute rejection showed a significantly lower mean number of CCL4L copies than patients that showed acute rejection (1.66 vs 1.96, P=0.014), with an even greater number of gene copies seen in patients with more than one episode of acute rejection (1.66 vs 2.30, P=0.001). Additionally, patients with > or =2 CCL4L copies had a significantly higher risk of acute rejection compared with patients that had 0-1 CCL4L copies (odds ratio 2.65; 95% confidence interval, 1.33-5.28; P=0.0046). A combined analysis of CCL4L CNV and the rs4796195 CCL4L single nucleotide polymorphism demonstrated that the effect of CCL4L copy number in acute rejection is mainly because of the number of copies of the CCL4L1 allelic variant. This finding constitutes the first report of CNV as a correlate factor in allograft rejection.

Susceptibility of food-borne bacteria to binary combinations of antimicrobials at selected a(w) and pH.

AIM: To evaluate the antibacterial susceptibilities of food-borne bacteria to individual and binary mixtures of a synthetic antimicrobial agent with a natural phenolic compound. METHODS AND RESULTS: Antibacterial susceptibilities of Escherichia coli, Listeria innocua, Salmonella Typhimurium and Staphylococcus aureus to individual and binary mixtures of potassium sorbate with a phenolic compound (thymol, carvacrol, or eugenol) were evaluated, at selected water activity (a(w); 0.99 or 0.97) and pH (5.5 or 4.5). The bacteria studied were susceptible to the action of the antimicrobials individually with minimal inhibitory concentrations that varied from 800-ppm potassium sorbate for Staph. aureus at a(w) 0.99, and pH 5.5 to 100-ppm thymol or carvacrol for the four studied bacteria at a(w) 0.97 and pH 4.5. Several binary mixtures of potassium sorbate with thymol, carvacrol or eugenol inhibited bacterial growth. Antimicrobial agent inhibitory concentrations in the mixture varied among bacteria, additionally depending on the a(w) and the pH tested. CONCLUSIONS: Synergistic binary mixtures with fractional inhibitory concentration index <0.6 include 100- or 200-ppm potassium sorbate with 50- or 100-ppm thymol, carvacrol or eugenol. SIGNIFICANCE AND IMPACT OF THE STUDY: The synergistic combinations could be useful in reducing the amounts of antimicrobials needed to inhibit growth, thus diminishing consumer concerns regarding chemical preservatives.

Analysis of INF-gamma, TNF-alpha and dendritic cells to predict hepatitis C virus recurrence in liver transplant patients.

INTRODUCTION: Hepatitis C virus (HCV) infection is one of the leading causes of chronic liver disease and the reason for more than 50% of liver transplantations (OLT). Recurrent HCV infection occurs in almost all transplant recipients and has an unfavorable course. Although immunosuppressive agents are necessary to avoid allograft rejection, these drugs may favor viral replication facilitating viral-mediated graft injury. METHODS: To predict the evolution of two HCV(+) patients who underwent OLT, we studied INF-gamma and TNF-alpha production and the maturation capacity of dendritic cells (DCs) at three time points: before transplantation (Pre-Tx) and at 2 (2M) and 6 (6M) months after transplantation. Cytometric bead assays were used to quantify INF-gamma and TNF-alpha production in the supernates of mixed leukocyte reactions (MLR) between spleen cells from the liver donor and CD4(+) cells from the recipients. Immature and mature DCs were generated in vitro from patient monocytes. RESULTS: The one patient who experienced recurrent HCV showed loss of CD4(+) responses to donor antigens and INF-gamma and TNF-alpha production after OLT. In contrast, the other patient maintained detectable levels of these cytokines after OLT. It was possible to generate mature DCs from monocytes with the aid of CD40L in both cases, but decreased expression of HLA-DR, CD80, and CD86 markers was observed upon posttransplantation analyses in the patient with recurrent HCV. CONCLUSION: Loss of the proliferative response as well as INF-gamma and TNF-alpha production, together with a decreased HLA-DR, CD80, and CD86 (markers of mature DCs), indicated an inadequate immune response to viral progression in the liver transplant recipient with relapsing HCV infection.