Comparative cryopreservation of bovine and porcine primary hepatocytes.

The isolation of primary hepatocytes from liver tissue of farm animals yields a very high number of cells, and a part of them can be stored by cryopreservation for future experiments. As no experience exists with the cryopreservation of hepatocytes from cattle, our study aimed at the cryopreservation of bovine hepatocytes by use of different protocols compared with the cryopreservation of hepatocytes from pig. We tested different freezing media (William's Medium E vs. University of Wisconsin solution), cryoprotectants (dimethyl sulfoxide with vs. without trehalose as additional additive), freezing systems (standard freezing container vs. controlled-rate freezer) and freezing times (4 vs. 28 d). These tests identified a general influence of species and freezing systems, whereas the influence of freezing media, trehalose additive and freezing time was less or not obvious. In this regard, we determined a mean recovery of 30% of bovine hepatocytes and 55% of porcine hepatocytes cryopreserved in a controlled-rate freezer, whereas the rates were about 10% less when hepatocytes were frozen in a standard freezing container. In accordance with this observation, the cultivation of cryopreserved hepatocytes from cattle was less effective than that of porcine hepatocytes. Hepatocytes from cattle can be successfully cryopreserved and partially cultured after cryopreservation but with lower percentage than porcine hepatocytes.

Increased flexibility in methadone take-home scheduling during the COVID-19 pandemic: Should this practice be incorporated into routine clinical care?

In the context of the COVID-19 pandemic and the state of emergency that the government of Spain declared, the rapid adaptation of health services is of paramount importance to preserve access to and continuity of service delivery. This research note underscores the importance of ensuring a sufficient quantity of methadone take-home doses for patients on methadone maintenance treatment (MMT) to maximize their adherence to government-imposed lockdown restrictions and social distancing measures designed to curtail the spread of SARS-CoV-2. We evaluate the impact of COVID-19 on take-home medication (number of days provided) in a methadone clinic in Barcelona (Catalonia, Spain). This work conveys that we should consider maintaining the take-home practices that we adopted in response to the pandemic, even after the pandemic has abated.

Effectiveness of Integrative Laughter Therapy to Reduce Anxiety, Improve Self-Esteem and Increase Happiness: A Naturalistic Study at a Day Hospital for Addictive Disorders.

Little is known about the effectiveness of laughter therapy as an adjunctive treatment for patients with addictive disorders. This study aims to evaluate the benefits of integrative laughter therapy (ILT) on levels of self-esteem, anxiety, and happiness in patients treated for addiction at a day hospital (DH). A prospective, naturalistic study with a pre-post design was conducted. All 185 participants received the standard, multicomponent treatment at the DH (treatment as usual; TAU). The participants were also invited to attend weekly ILT sessions. Upon completion of the 2-month DH treatment program, patients were classified according to their attendance at the ILT sessions: patients who attended ≥80% constituted the experimental group (TAU + ILT group) while those who attended <80% were considered controls. Although both groups achieved statistically significant increases in self-esteem and happiness with a decrease in trait anxiety, the improvement on these variables was significantly greater in the TAU + ILT group. Subject to the limitations inherent to quasi-experimental research, the findings of the present study suggest that the addition of an ILT module to the standard treatment in a DH for addictive disorders may yield greater improvement in self-esteem, anxiety, and happiness compared to TAU.

Advanced survey of tuberculosis transmission in a complex socioepidemiologic scenario with a high proportion of cases in immigrants.

BACKGROUND: An increase in the incidence of tuberculosis (TB) in immigrants has changed the socioepidemiologic scenario in Spain. It is generally assumed that TB in immigrants is the result of importation of infection, but the role of recent transmission is rarely considered. Standard contact tracing is not suitable for the survey of transmission in this complex scenario. METHODS: During the study period (2003-2006), we genotyped 356 (90.4%) of 394 isolates from patients with microbiologically confirmed TB in Almería, the province with the highest percentage of TB cases among immigrants in Spain. The epidemiologic survey of TB transmission was performed by active data collection using standardized interviews of the patients with TB and subsequent interviews of the clustered patients (who were clustered on the basis of the restriction fragment-length polymorphism types of their isolates) to identify transmission locations (supported by nominal and/or photographic recognition by the clustered patients). RESULTS: Of all 356 genotyped isolates, 131 (36.8%) were clustered, suggesting recent transmission. The difference between the clustering rate for immigrants (32.8%) and that for native patients (41.6%) was not statistically significant (P = .087); of the 45 clusters, 15 (33.3%) involved only immigrants, 17 (37.8%) involved only autochthonous patients, and 13 (28.9%) involved both immigrants and autochthonous patients. The advanced system to investigate the clustered patients succeeded in detecting links in 10 of the 12 clusters that involved >4 patients, whereas the conventional approach, based on contact tracing, could detect links in only 2 clusters. CONCLUSIONS: Recent transmission among immigrants and transmission permeability between the immigrant and autochthonous populations were found. Epidemiologic strategies that combine universal genotyping and refined surveys of the clustered patients are needed to investigate transmission patterns in complex scenarios.

Swallowing disorders in Parkinson's disease. / Trastornos de la deglución en la enfermedad de Parkinson.

INTRODUCTION: Parkinson's disease is a type of chronic neurodegenerative pathology with a typical movement pattern, as well as different, less studied symptoms such as dysphagia. Disease-related disorders in efficacy or safety in the process of swallowing usually lead to malnutrition, dehydration or pneumonias. The aim of this study was identifying and analyzing swallowing disorders in Parkinson's disease. SUBJECTS AND METHODS: The initial sample consisted of 52 subjects with Parkinson's disease to whom the specific test for dysphagia SDQ was applied. Nineteen participants (36.5%) with some degree of dysphagia in the SDQ test were selected to be evaluated by volume-viscosity clinical exploration method and fiberoptic endoscopic evaluation of swallowing. RESULTS: Disorders in swallowing efficiency and safety were detected in 94.7% of the selected sample. With regards to efficiency, disorders were found in food transport (89.5%), insufficient labial closing (68.4%) and oral residues (47.4%), relating to duration of ingestion. Alterations in security were also observed: pharynx residues (52.7%), coughing (47.4%), penetration (31.64%), aspiration and decrease of SaO2 (5.3%), relating to the diagnosis of respiratory pathology in the previous year. CONCLUSION: The SDQ test detected swallowing disorders in 36.5% of the subjects with Parkinson's disease. Disorders in swallowing efficiency and safety were demonstrated in 94.7% of this subset. Disorders of efficiency were more frequent than those of safety, establishing a relationship with greater time in ingestion and the appearance of respiratory pathology and pneumonias.

Complex clonal features in an mycobacterium tuberculosis infection in a two-year-old child.

The focus of this report is to describe the highly complex clonal features of the Mycobacterium tuberculosis (MTB) isolates cultured from a 2-year-old child with no risk factors for tuberculosis. Molecular analysis showed that she was coinfected by 2 MTB isolates. The coinfecting isolates showed only subtle genetic differences and shared equivalent infectivity in vitro; however, they were infecting independent anatomic sites (compartmentalized infection). One of the MTB isolates was cultured only from respiratory specimens, whereas the other was isolated from extrarespiratory site (central nervous system).

Trastornos de la deglución en la enfermedad de Parkinson / Swallowing disorders in Parkinson's disease

Introducción. La enfermedad de Parkinson es una patología neurodegenerativa crónica, con síntomas motores característicos y otros menos estudiados como la disfagia. Suele relacionarse con desnutrición, deshidratación o neumonías por aspiración por trastornos en la eficacia y seguridad en la deglución. El objetivo de este trabajo es identificar y analizar los trastornos deglutorios en la enfermedad de Parkinson. Sujetos y métodos. Muestra inicial de 52 sujetos con enfermedad de Parkinson a los que se aplicó el test SDQ, específico para la disfagia. Se seleccionaron 19 (36,5%) participantes con algún grado de disfagia en dicho test para ser evaluados con el método de exploración clínica volumen-viscosidad y la videoendoscopia de la deglución. Resultados. Se detectaron trastornos de eficacia y seguridad de la deglución en el 94,7%, siendo las alteraciones de la eficacia: dificultad en el transporte de alimento (89,5%), deglución fraccionada (78,9%), mal sello labial (68,4%) y residuos orales (47,4%), relacionándose con el tiempo que dura la ingesta. Se observaron también alteraciones en la seguridad como residuos faríngeos (52,7%), tos (47,4%), penetración (31,64%), aspiración y descenso de SaO2 (5,3%), relacionándose con el diagnóstico de patología respiratoria en el último año. Conclusiones. El test SDQ detectó trastornos deglutorios en el 36,5% de los pacientes con enfermedad de Parkinson. Las alteraciones en la eficacia y seguridad de la deglución se demostraron en el 94,7% de ellos, siendo más frecuentes los trastornos de eficacia que los de seguridad, estableciendo relación con el mayor tiempo en la ingesta y la aparición de patología respiratoria y neumonías (AU)

Introduction. Parkinson's disease is a type of chronic neurodegenerative pathology with a typical movement pattern, as well as different, less studied symptoms such as dysphagia. Disease-related disorders in efficacy or safety in the process of swallowing usually lead to malnutrition, dehydration or pneumonias. The aim of this study was identifying and analyzing swallowing disorders in Parkinson's disease. Subjects and methods. The initial sample consisted of 52 subjects with Parkinson's disease to whom the specific test for dysphagia SDQ was applied. Nineteen participants (36.5%) with some degree of dysphagia in the SDQ test were selected to be evaluated by volume-viscosity clinical exploration method and fiberoptic endoscopic evaluation of swallowing. Results. Disorders in swallowing efficiency and safety were detected in 94.7% of the selected sample. With regards to efficiency, disorders were found in food transport (89.5%), insufficient labial closing (68.4%) and oral residues (47.4%), relating to duration of ingestion. Alterations in security were also observed: pharynx residues (52.7%), coughing (47.4%), penetration (31.64%), aspiration and decrease of SaO2 (5.3%), relating to the diagnosis of respiratory pathology in the previous year. Conclusion. The SDQ test detected swallowing disorders in 36.5% of the subjects with Parkinson's disease. Disorders in swallowing efficiency and safety were demonstrated in 94.7% of this subset. Disorders of efficiency were more frequent than those of safety, establishing a relationship with greater time in ingestion and the appearance of respiratory pathology and pneumonias (AU)

Analysis of changes in recent tuberculosis transmission patterns after a sharp increase in immigration.

We conducted a population-based molecular epidemiological study of tuberculosis (TB) in Madrid, Spain (2002 to 2004), to define transmission patterns and factors associated with clustering. We particularly focused on examining how the increase in TB cases among immigrants in recent years (2.8% in 1997 to 1999 to 36.2% during the current study) was modifying transmission patterns. Mycobacterium tuberculosis isolates obtained from patients living in nine districts of Madrid (1,459,232 inhabitants) were genotyped. The TB case rate among foreign-born people was three to four times that of Spanish-born people, and the median time from arrival to the onset of treatment was 22.4 months. During the study period, 227 (36.3%) patients were grouped in 64 clusters, and 115 (50.7%) of them were in 21 clusters with mixed Spanish-born and foreign-born patients. Three of the 21 mixed clusters accounted for 21.1% of clustered patients. Twenty-two of 38 (57.9%) immigrants in mixed clusters were infected with TB strains that had already been identified in the native population in 1997 to 1999, including the three most prevalent strains. Factors identified as independent predictors of clustering were homelessness (odds ratio [OR], 2.3; 95% confidence interval [95% CI], 1.2 to 4.5; P = 0.011) and to be born in Spain (OR, 1.8; 95% CI, 1.2 to 2.6; P = 0.002). The results indicated that (i) TB transmission was higher in Spanish-born people, associated mainly with homelessness, (ii) that foreign-born people were much less likely to be clustered, suggesting a higher percentage of infection before arriving in Spain, and (iii) that an extensive transmission between Spanish- and foreign-born populations, caused mainly by autochthonous strains, was taking place in Madrid.

La voz del laringectomizado: incapacidad, percepción y análisis acústico / Laryngectomized voice rehabilitation: handicap, perception and acoustic analysis

Los pacientes laringectomizados tienen una limitación muy importante al perder la función vocal. La rehabilitación de la voz se realiza por erigmofonía o prótesis fonatoria. Ambos procedimientos son útiles pero tienen diferencias evidentes en la calidad vocal obtenida. Objetivo. Determinar cómo es percibida la voz rehabilitada con ambos procedimientos por los pacientes y observadores, completando el estudio con su análisis acústico. Material y método. Pacientes laringectomizados (n = 18) con voz rehabilitada con erigmofonía (n = 10) y prótesis fonatoria (n = 8). Estudio por medio de VHI-30, análisis acústico (F0, TMF, HNR, «speech-rate», intensidad y espectrograma) y evaluación perceptual (GRBAS y CAPE-V). Resultados. El VHI-30 presenta valores similares en ambos tipos de voz rehabilitada. El análisis acústico muestra diferencias importantes entre ambos tipos de voces, siendo los valores obtenidos con prótesis fonatoria más parecidos a los de la voz laríngea. La evaluación perceptual indica mayor grado de afectación en la voz erigmofónica que en la prótesis fonatoria. No se observa relación significativa entre lo percibido por los pacientes con ambos tipos de voces rehabilitadas y lo valorado por el observador. Los estudios acústicos y perceptuales muestran valores más próximos a la normalidad en la voz de prótesis fonatoria (AU)

Laryngectomized patients have a very important limitation when they lose the vocal function. Voice rehabilitation is fulfilled by means of the esophageal voice or by means of a phonatory prosthesis. Both methods are useful but they have evident differences concerning the vocal quality obtained. Objective. Determine how rehabilitated voice is perceived with both methods by patients and observers, completing the study with its acoustic analysis. Material and method. Laryngectomized patients (n = 18) with rehabilitated voice by means of esophageal voice (n = 10) and phonatory prosthesis (n = 8). A study was made through the VHI-30, acoustic analysis (F0, TMF,HNR, «speech rate», intensity and spectrogram) and perceptual evaluation (GRBAS and CAPE-V). Results. VHI-30 displays similar values in both types of rehabilitated voice. The acoustic analysis shows differences between both types of rehabilitated voices and the usual laryngeal voice. The perceptual study presents a higher degree of affectation on the esophageal voice than on the PF one. There is no relation between what the patient perceives and what the observer values. The acoustic and perceptual studies show closer values to normality in the phonatory prosthesis voice (AU)

Impact of laboratory cross-contamination on molecular epidemiology studies of tuberculosis.

Laboratory cross-contamination by Mycobacterium tuberculosis is known to be responsible for the misdiagnosis of tuberculosis, but its impact on other contexts has not been analyzed. We present the findings of a molecular epidemiology analysis in which the recent transmission events identified by a genotyping reference center were overestimated as a result of unnoticed laboratory cross-contamination in the original diagnostic laboratories.

Characterization of clonal complexity in tuberculosis by mycobacterial interspersed repetitive unit-variable-number tandem repeat typing.

In recent years, the application of molecular tools has shown us that clonal complexity in infection by Mycobacterium tuberculosis is not anecdotal. Exogenous reinfections, mixed infections, compartmentalization, and microevolution are different aspects of this issue. The detection and characterization of clonal variants of M. tuberculosis by standard genotyping methods is laborious and frequently requires expertise. Our aim was to evaluate a new genotyping PCR-based method for M. tuberculosis, mycobacterial interspersed repetitive unit-variable-number tandem repeat typing (MIRU-VNTR), as a potential tool to simplify and optimize the clonal analysis of tuberculosis. MIRU-VNTR was able to detect mixed clonal variants in vitro, even for clones at low ratios (1:99). This technique was prospectively applied to search for cases infected by more than one clone. Clonal variants within the same host were detected in 3 out of 115 cases (2.6%), including cases with clones which were indistinguishable by restriction fragment length polymorphism or spoligotyping. In one case, coinfecting clonal variants differed in antibiotic susceptibilities. MIRU-VNTR was applied to cases with proven polyclonal infection, and it succeeded in detecting the coinfecting strains and proved useful in confirming cases of compartmentalized infection. MIRU-VNTR is a simple, rapid, and sensitive method which could facilitate and optimize the identification and characterization of clonal complexity in M. tuberculosis infection.