In-depth characterization of a new patient-derived xenograft model for metaplastic breast carcinoma to identify viable biologic targets and patterns of matrix evolution within rare tumor types.

Metaplastic breast carcinoma (MBC) is a rare breast cancer subtype with rapid growth, high rates of metastasis, recurrence and drug resistance, and diverse molecular and histological heterogeneity. Patient-derived xenografts (PDXs) provide a translational tool and physiologically relevant system to evaluate tumor biology of rare subtypes. Here, we provide an in-depth comprehensive characterization of a new PDX model for MBC, TU-BcX-4IC. TU-BcX-4IC is a clinically aggressive tumor exhibiting rapid growth in vivo, spontaneous metastases, and elevated levels of cell-free DNA and circulating tumor cell DNA. Relative chemosensitivity of primary cells derived from TU-BcX-4IC was performed using the National Cancer Institute (NCI) oncology drug set, crystal violet staining, and cytotoxic live/dead immunofluorescence stains in adherent and organoid culture conditions. We employed novel spheroid/organoid incubation methods (Pu·MA system) to demonstrate that TU-BcX-4IC is resistant to paclitaxel. An innovative physiologically relevant system using human adipose tissue was used to evaluate presence of cancer stem cell-like populations ex vivo. Tissue decellularization, cryogenic-scanning electron microscopy imaging and rheometry revealed consistent matrix architecture and stiffness were consistent despite serial transplantation. Matrix-associated gene pathways were essentially unchanged with serial passages, as determined by qPCR and RNA sequencing, suggesting utility of decellularized PDXs for in vitro screens. We determined type V collagen to be present throughout all serial passage of TU-BcX-4IC tumor, suggesting it is required for tumor maintenance and is a potential viable target for MBC. In this study we introduce an innovative and translational model system to study cell-matrix interactions in rare cancer types using higher passage PDX tissue.

Racial differences in intracerebral haemorrhage outcomes in patients with obesity.

OBJECTIVE: This study was conducted to determine the role of obesity and race in intracerebral haemorrhage (ICH) outcomes. METHODS: The Get with the guideline-Stroke database was queried for all admitted patients with spontaneous ICH. Secondary causes of ICH were excluded. Body mass index (BMI) was classified using the Center for Disease Control guidelines. Race was classified as White or non-White. Demographics, clinical, imaging data were retrieved. Outcome measures were hematoma expansion at 24 h and discharge disposition. RESULTS: A total of 428 patients were included in our analysis. Female gender, past history of congestive heart failure, diabetes mellitus, HbA1c, blood pressure, ICH volume, ICH location, intraventricular haemorrhage and hospital length of stay deferred across BMI categories. On multivariate analysis, along with obese categories, age, ICH location and ICH volume were independent predictors of poor outcomes (hematoma expansion and poor discharge disposition). After adjusting for these variables, obesity remained a predictor of poor disposition outcome compared with normal and overweight subjects; Normal vs. Obese OR 0.26 CI 0.115-0.593 p = 0.0014; Obese vs. Overweight OR 3.79 CI 1.68-8.52 p = 0.0013. Nonetheless, obesity did not influence hematoma expansion. Overall, BMI-race classification did not influence outcomes. However, among non-Whites, the obese category had higher odds of a poor disposition outcome than normal (OR 6.84 CI 2.12-22.22 p = 0.0013) or overweight (OR 8.45 CI 2.6-27.49 p = 0.0004) categories. CONCLUSION: An obesity paradox in ICH was not observed in our cohort. In the non-White population, patients with obesity were likely to be associated with poor disposition outcome. Similar findings were not observed in White population.

Guía de práctica clínica de la SECT sobre el manejo de pacientes con neumotórax espontáneo / SECT Clinical practice guideline on the management of patients with spontaneous pneumothorax

Esta guía de práctica clínica (GPC) surge como iniciativa del comité científico de la Sociedad Española de Cirugía Torácica. Para elaborar dicha GPC se han formulado las preguntas PICO (paciente, intervención, comparación y outcome o variable resultado) sobre distintos aspectos del neumotórax espontáneo. Para la evaluación de la calidad de la evidencia y elaboración de las recomendaciones se han seguido las directrices del grupo de trabajo Grading of Recommendations, Assessent, Development and Evaluation (GRADE).(AU)

This clinical practice guideline (CPG) emerges as an initiative of the scientific committee of the Spanish Society of Thoracic Surgery. We formulated PICO (patient, intervention, comparison, and outcome) questions on various aspects of spontaneous pneumothorax. For the evaluation of the quality of evidence and preparation of recommendations we followed the guidelines of the Grading of recommendations, Assessment, Development and Evaluation (GRADE) working group.(AU)

SECT Clinical practice guideline on the management of patients with spontaneous pneumothorax

This clinical practice guideline (CPG) emerges as an initiative of the scientific committee of the Spanish Society of Thoracic Surgery. We formulated PICO (patient, intervention, comparison, and outcome) questions on various aspects of spontaneous pneumothorax. For the evaluation of the quality of evidence and preparation of recommendations we followed the guidelines of the Grading of recommendations, Assessment, Development and Evaluation (GRADE) working group.

Electrical transport measurements of thin film samples under high hydrostatic pressure.

We present a method to perform electrical measurements of epitaxial films and heterostructures a few nanometers thick under high hydrostatic pressures in a diamond anvil cell (DAC). Hydrostatic pressure offers the possibility to tune the rich landscape of properties shown by epitaxial heterostructures, systems in which the combination of different materials, performed with atomic precision, can give rise to properties not present in their individual constituents. Measuring electrical conductivity under hydrostatic pressure in these systems requires a robust method that can address all the challenges: the preparation of the sample with side length and thickness that fits in the DAC setup, a contacting method compatible with liquid media, a gasket insulation that resists high forces, as well as an accurate procedure to place the sample in the pressure chamber. We prove the robustness of the method described by measuring the resistance of a two dimensional electron system buried at the interface between two insulating oxides under hydrostatic conditions up to ∼5 GPa. The setup remains intact until ∼10 GPa, where large pressure gradients affect the two dimensional conductivity.

Comparison of two concentrations of bupivacaine by continuous paravertebral infusion after thoracotomy with pulmonary resection: a double-blind, randomized clinical trial

Background and objectives: Post-thoracotomy pain management should be based on a multimodal approach that includes continuous regional analgesia. The objective of this study was to compare the analgesic efficacy of two concentrations of bupivacaine (0.2 % and 0.3 %) through a paravertebral catheter, both group plus fentanyl 2 mcg/ml. Methods: We conducted a randomized double-blind clinical trial to compare these two concentrations in patients undergoing pulmonary resection by thoracotomy in Donostia University Hospital between November 2010 and May 2011 (n = 59). The paravertebral catheter was placed prior to the surgical intervention, with the patient awake and sitting upright. Data were analyzed on an intention-to-treat basis. The Chi-squared test was used for qualitative variables and Student’s t-tests or Mann-Whitney-Wilcoxon tests for quantitative variables, depending on the distribution of the variables. Statistical analysis was performed using IBM SPSS software (Version 17). Results: We did not find statistically significant differences in postoperative pulmonary function (p = 0.49), self-perceived pain (VAS; p = 0.28) or cumulative morphine consumption (p = 0.101) in the two groups. We observed adverse effects in 8 patients in group 1 (29.6 %) and in 12 patients (37.5 %) in group 2, the difference not being statistically significant (p = 0.52). Conclusions: Continuous thoracic paravertebral block for 48 hours is a good technique for the management of postoperative pain after pulmonary resection by thoracotomy. With moderate doses of local anesthetics (bupivacaine 0.20 %) we achieved good pain control and observed few systemic complications than major doses (bupivacaine 0.30 %) (AU)

Antecedentes y objetivos: El manejo del dolor post-toracotomía debe basarse en un enfoque multimodal que incluye la analgesia regional continua. El objetivo de este estudio fue comparar la eficacia analgésica de dos concentraciones de bupivacaína (0,2 y 0,3 %) a través de un catéter paravertebral, ambos grupos más fentanilo 2 mcg/ml. Material y métodos: Se realizó un ensayo clínico aleatorizado, doble ciego, para comparar estas dos concentraciones en pacientes sometidos a resección pulmonar por toracotomía en el Hospital Universitario Donostia entre noviembre de 2010 y mayo de 2011 (n = 59). El catéter paravertebral se colocó antes de la intervención quirúrgica, con el paciente despierto en posición sentada. Los datos se analizaron sobre la base de intención de tratar. Se utilizó la prueba de Chi cuadrado para variables cualitativas y la t de Student o pruebas de Mann-Whitney-Wilcoxon para las variables cuantitativas, en función de la distribución de las variables. El análisis estadístico se realizó utilizando el software de IBM SPSS (versión 17). Resultados: No se encontraron diferencias estadísticamente significativas en la función pulmonar postoperatoria (p = 0,49), la percepción subjetiva de dolor (VAS; p = 0,28) o el consumo Antecedentes y objetivos: El manejo del dolor post-toracotomía debe basarse en un enfoque multimodal que incluye la analgesia regional continua. El objetivo de este estudio fue comparar la eficacia analgésica de dos concentraciones de bupivacaína (0,2 y 0,3 %) a través de un catéter paravertebral, ambos grupos más fentanilo 2 mcg/ml. Material y métodos: Se realizó un ensayo clínico aleatorizado, doble ciego, para comparar estas dos concentraciones en pacientes sometidos a resección pulmonar por toracotomía en el Hospital Universitario Donostia entre noviembre de 2010 y mayo de 2011 (n = 59). El catéter paravertebral se colocó antes de la intervención quirúrgica, con el paciente despierto en posición sentada. Los datos se analizaron sobre la base de intención de tratar. Se utilizó la prueba de Chi cuadrado para variables cualitativas y la t de Student o pruebas de Mann-Whitney-Wilcoxon para las variables cuantitativas, en función de la distribución de las variables. El análisis estadístico se realizó utilizando el software de IBM SPSS (versión 17). Resultados: No se encontraron diferencias estadísticamente significativas en la función pulmonar postoperatoria (p = 0,49), la percepción subjetiva de dolor (VAS; p = 0,28) o el consumo de morfina acumulada (p = 0,101) en los dos grupos. Hemos observado efectos adversos en 8 pacientes del grupo 1 (29,6 %) y en 12 pacientes (37,5 %) del grupo 2; la diferencia no fue estadísticamente significativa (p = 0,52). Conclusiones: El bloqueo paravertebral torácico continuo durante 48 horas es una buena técnica para el manejo del dolor postoperatorio después de la resección pulmonar por toracotomía. Con dosis moderadas de anestésicos locales (bupivacaína 0,20 %) se logró un buen control del dolor y observamos un número menor de complicaciones sistémicas que el grupo de dosis mayores (bupivacaína 0,30 %) (AU)

Comparative transcriptomic analysis reveals the oncogenic fusion protein PAX3-FOXO1 globally alters mRNA and miRNA to enhance myoblast invasion.

Rhabdomyosarcoma, one of the most common childhood sarcomas, is comprised of two main subtypes, embryonal and alveolar (ARMS). ARMS, the more aggressive subtype, is primarily characterized by the t(2;13)(p35;p14) chromosomal translocation, which fuses two transcription factors, PAX3 and FOXO1 to generate the oncogenic fusion protein PAX3-FOXO1. Patients with PAX3-FOXO1-postitive tumors have a poor prognosis, in part due to the enhanced local invasive capacity of these cells, which leads to the increased metastatic potential for this tumor. Despite this knowledge, little is known about the role that the oncogenic fusion protein has in this increased invasive potential. In this report we use large-scale comparative transcriptomic analyses in physiologically relevant primary myoblasts to demonstrate that the presence of PAX3-FOXO1 is sufficient to alter the expression of 70 mRNA and 27 miRNA in a manner predicted to promote cellular invasion. In contrast the expression of PAX3 alters 60 mRNA and 23 miRNA in a manner predicted to inhibit invasion. We demonstrate that these alterations in mRNA and miRNA translate into changes in the invasive potential of primary myoblasts with PAX3-FOXO1 increasing invasion nearly 2-fold while PAX3 decreases invasion nearly 4-fold. Taken together, these results allow us to build off of previous reports and develop a more expansive molecular model by which the presence of PAX3-FOXO1 alters global gene regulatory networks to enhance the local invasiveness of cells. Further, the global nature of our observed changes highlights the fact that instead of focusing on a single-gene target, we must develop multi-faceted treatment regimens targeting multiple genes of a single oncogenic phenotype or multiple genes that target different oncogenic phenotypes for tumor progression.

Norovirus as the cause of medically attended gastroenteritis: a hospital-based experience.

Gastroenteritis remains an important cause of morbidity and mortality worldwide. With the introduction of vaccines against rotavirus, interest has shifted to understanding the epidemiology of norovirus (NoV). While the importance of NoV in gastroenteritis outbreaks is well established, its role in sporadic gastroenteritis is less known. To better define the role of NoV as a cause of sporadic gastroenteritis we investigated its prevalence in the patients seen in our paediatric hospital with special emphasis on its seasonal and age distribution. Over a 12-month period discarded stool specimens submitted to our paediatric hospital for testing of an infectious aetiology were retrieved and additionally tested for NoV by real-time reverse transcriptase-polymerase chain reaction; demographical and clinical information were also obtained. Overall, NoV was the single most commonly identified pathogen and found in 68/892 (7·6%) total specimens or 68/258 (26%) of pathogen-positive specimens. The highest rates of NoV were detected in children aged 6 months to 4 years (50/332, 15·1%) and presenting between October and January (46/314, 14·7%). NoV has become the main cause of gastroenteritis in our paediatric population.

Chemical solution route to self-assembled epitaxial oxide nanostructures.

Self-assembly of oxides as a bottom-up approach to functional nanostructures goes beyond the conventional nanostructure formation based on lithographic techniques. Particularly, chemical solution deposition (CSD) is an ex situ growth approach very promising for high throughput nanofabrication at low cost. Whereas strain engineering as a strategy to define nanostructures with tight control of size, shape and orientation has been widely used in metals and semiconductors, it has been rarely explored in the emergent field of functional complex oxides. Here we will show that thermodynamic modeling can be very useful to understand the principles controlling the growth of oxide nanostructures by CSD, and some attractive kinetic features will also be presented. The methodology of strain engineering is applied in a high degree of detail to form different sorts of nanostructures (nanodots, nanowires) of the oxide CeO2 with fluorite structure which then is used as a model system to identify the principles controlling self-assembly and self-organization in CSD grown oxides. We also present, more briefly, the application of these ideas to other oxides such as manganites or BaZrO3. We will show that the nucleation and growth steps are essentially understood and manipulated while the kinetic phenomena underlying the evolution of the self-organized networks are still less widely explored, even if very appealing effects have been already observed. Overall, our investigation based on a CSD approach has opened a new strategy towards a general use of self-assembly and self-organization which can now be widely spread to many functional oxide materials.

Inverse correlation of serum inflammatory markers with metabolic parameters in healthy, Black and White prepubertal youth.

OBJECTIVE: To examine for the first time the associations between pro-inflammatory cytokines and obesity-related metabolic biomarkers in, exclusively prepubertal, otherwise healthy obese and non-obese Black and White children, 7-9 years of age. DESIGN AND METHODS: Body mass index (BMI), homeostasis model assessment-estimated insulin resistance, visceral adipose tissue and subcutaneous adipose tissue (SAT (magnetic resonance imaging)); total body fat (dual-energy X-ray absorptiometry), ectopic, intrahepatic lipid (IHL) and intramyocellular lipid (IMCL) fat (proton magnetic resonance spectroscopy) and serum levels of interleukin (IL)-1, IL-6, IL-8, tumor necrosis factor alpha (TNF-α) and monocyte chemoattractant protein-1 were measured in 40 obese and non-obese children. Relationships between inflammatory cytokines and obesity were assessed by analysis of variance and Spearman's rank correlation. RESULTS: Significant inverse correlations were found between BMI z-score, SAT, total BF, and IHL and levels of TNF-α (Spearman's ρ=-0.36, -0.39, -0.43 and -0.39, respectively; P<0.05). Levels of IL-8 were significantly and inversely correlated with IMCL (-0.39; P=0.03) and remained significant after adjusting for race. IMCL was inversely associated with TNF-α only after adjusting for race (-0.37; P=0.04). CONCLUSIONS: Relationships between pro-inflammatory and metabolic markers commonly observed in adults are reversed in healthy, Black and White children before puberty. Prospective studies are warranted to determine how these inverse relationships modify chronic disease risk later in life.

Estudio epidemiológico multivariable del paciente con fractura osteoporótica de fémur proximal / Multivariate epidemiological study of the patient with osteoporotic proximal femoral fracture

Objetivo: Revisar las fracturas de cadera en personas mayores de 50 años para valorar cómo afectan las comorbilidades asociadas al pronóstico. Material y método: Estudiamos 200 pacientes mayores de 50 años tratados de fractura de fémur proximal mediante un análisis retrospectivo de las variables, con el fin de valorar el pronóstico y la calidad de vida residual. Resultados: Observamos una tendencia, sin significación estadística, de a mayor edad del paciente, mayor probabilidad de que su fractura sea extracapsular. El índice de Singh no se correlacionó con el tipo de fractura. La carencia de vitamina D fue una constante. Hallamos una correlación entre la comorbilidad y el grado de insuficiencia renal (creatinina). No hubo incremento de estancia media en pacientes con mayor número de patologías. La demora quirúrgica no prolongó el tiempo de hospitalización postquirúrgica. Conclusión: El paciente osteoporótico con fractura de cadera ha de ser tratado de una manera multidisciplinar. Son muchos los factores que condicionarán su pronóstico y situación funcional final (AU)

Objective: To review hip fractures in patients over 50 years of age, with a view to assessing how they affect the comorbidities associated to the prognosis. Material and method: The study comprised a total of 200 patients over 50 years of age treated for proximal femoral fractures, based on a retrospective analysis of the variables, in order to define the prognosis and residual quality of life. Results: Increased patient age was found to be associated to an increased probability of extracapsular fracture, though statistical significance was not reached. The Singh index was not correlated to the type of fracture. Vitamin D deficiency was a constant finding. A correlation was observed between comorbidity and the degree of renal failure (creatinine). There was no increase in hospital stay in patients with a larger number of disorders. Surgical delay did not prolong postoperative hospital stay. Conclusion: Osteoporotic hip fracture patients require multidisciplinary management. Many factors condition the prognosis and ultimate functional outcome (AU)

Enfermedad de Castleman y embarazo. ¿Casualidad o causalidad? / Castleman’s disease and pregnancy. Cause or coincidence?

La enfermedad de Castleman (EC) es un trastorno linfoproliferativo poco frecuente y clínicamente se diferencian la forma clínica localizada y la forma clínica multicéntrica. A pesar de su baja incidencia se han publicado otros casos de enfermedad de Castleman en mujeres embarazadas, sugiriendo una relación entre el embarazo y la enfermedad de Castleman. En este trabajo presentamos un caso de enfermedad de Castleman en una mujer embarazada tratada en nuestro hospital y revisamos la bibliografía relacionada con esta infrecuente entidad (AU)

Castleman’s disease is an infrequent lymphoproliferative disorder. Clinically, the localized form is distinguished from the multicentric form. Despite the low incidence of this entity, other cases of Castleman’s disease in pregnant woman have been published, suggesting that there may be a relationship between pregnancy and this disorder. We present a case of Castleman’s disease in a pregnant woman treated in our hospital and review the literature on this uncommon entity (AU)

Colorectal cancer pulmonary oligometastases: pooled analysis and construction of a clinical lung metastasectomy prognostic model.

BACKGROUND: Although resecting colorectal cancer (CRC) pulmonary metastasis is associated with long-term survival, identification of prognostic groups is needed for future randomized trials, and construction of a lung metastasectomy prognostic model (LMPM) is warranted. PATIENTS AND METHODS: We searched the PubMed database for retrospective studies evaluating prognostic factors following resecting CRC lung metastasis. Individual patient data were analyzed. Independent prognostic factors were used to construct an LMPM. RESULTS: Between 1983 and 2008, 1112 metastasectomies were carried out on 927 patients included in eight studies. Five-year survival rate was 54.3% following the first lung resection. Multivariate analysis identified three independently poor prognostic factors: pre-thoracotomy carcinoembryonic antigen ≥5 ng/ml, disease-free interval <36 months, and more than one metastatic lesion. Patients with good-, intermediate-, and high-risk groups according to the LMPM had a 5-year survival of 68.2%, 46.4%, and 26.1%, respectively (P < 0.001). Perioperative chemotherapy and previously resected liver metastasis had no influence on survival. CONCLUSIONS: The low- and intermediate-risk groups have a good chance of long-term survival following metastasectomy. However, more studies are needed to investigate whether surgery offers any advantage over systemic therapy for the poor-risk group.

Survival after lung metastasectomy for colorectal cancer: importance of previous liver metastasis as a prognostic factor.

AIMS: To analyse patient survival after the resection of lung metastases from colorectal carcinoma and specifically to verify whether presence of liver metastasis prior to lung metastasectomy affects survival. METHODS: All patients who, between 1998 and 2008, underwent lung metastasectomy due to colorectal cancer were included in the study. Kaplan-Meier survival analysis was performed with the log-rank test and Cox regression multivariate analysis. RESULTS: During this period, 101 metastasectomies were performed on 84 patients. The median age of patients was 65.4 years, and 60% of patients were male. The 30-day mortality rate was 2%, and incidence of complications was 7%. The overall survival was 72 months, with 3-and 5-year survival rates of 70% and 54%, respectively. A total of 17 patients (20%) had previously undergone resection of liver metastasis. No significant differences were found in the distribution of what were supposed to be the main variables between patients with and without previous hepatic metastases. Multivariate analysis identified the following statistically significant factors affecting survival: previous liver metastasectomy (p = 0.03), tumour-infiltrated pulmonary lymph nodes (p = 0.04), disease-free interval ≥ 48 months (p = 0.03), and presence of more than one lung metastasis (p < 0.01). In patients with previous liver metastasis, the shorter the time between primary colorectal surgery and the hepatectomy, the lower the survival rate after pulmonary metastasectomy (p = 0.048). CONCLUSIONS: A previous history of liver metastasis shortens survival after lung metastasectomy. The time between hepatic resection and lung metastasectomy does not affect survival; however, patients with synchronous liver metastasis and colorectal neoplasia have poorer survival rates than those with metachronous disease.

[Ovarian hyperstimulation syndrome]. / Síndrome de hiperestimulación ovárica.

Experience with ovulation induction treatment and understanding of the physiopathology of ovarian hyperstimulation syndrome (OHS), the risk factors and the clinical characteristics are of key importance for preventing and managing OHS. Light manifestations ofOHS are fairly common, occurring in up to a third of the cycles with superovulation induced by exogenous gonadotropins. A worsening of the symptoms of OHS can still normally be managed in out-patient form, but frequent monitoring and evaluation are essential. Serious disease resulting from OHS is much less common, but can involve risk to life. Hospitalisation might be necessary in some cases.

Síndrome de hiperestimulación ovárica / Ovarian hyperstimulation syndrome

La experiencia con el tratamiento inductor de laovulación y el conocimiento de la fisiopatología del síndromede hiperestimulación ovárica (SHO), los factoresde riesgo y las características clínicas son claves parala prevención y el manejo del SHO. Las manifestacionesleves del SHO son bastante comunes, ocurriendo enhasta un tercio de los ciclos con superovulación inducidospor gonadotropinas exógenas.El empeoramiento de los síntomas del SHO normalmentetodavía se puede manejar de forma ambulatoria,pero es esencial la monitorización y evaluación frecuente.La enfermedad grave resultante del SHO es muchomenos común, pero puede conllevar riesgo vital. Lahospitalización puede ser necesaria, en algunos casos(AU)

Experience with ovulation induction treatmentand understanding of the physiopathology of ovarianhyperstimulation syndrome (OHS), the risk factors andthe clinical characteristics are of key importance forpreventing and managing OHS. Light manifestations ofOHS are fairly common, occurring in up to a third ofthe cycles with superovulation induced by exogenousgonadotropins.A worsening of the symptoms of OHS can still normallybe managed in out-patient form, but frequentmonitoring and evaluation are essential. Serious diseaseresulting from OHS is much less common, but caninvolve risk to life. Hospitalisation might be necessaryin some cases(AU)

Etanercept y embarazo / Etanercept and pregnancy

Presentamos el caso de una paciente afecta de artritis juvenil en tratamiento con etanercept que tras quedar embarazada mantiene el tratamiento a dosis estándar con dicho fármaco 25 mg dos veces por semana, sin presentar actividad ni complicaciones y con embarazo normal a término

We present the case of a pregnant woman with arthritis in treatment with etanercept. After becoming pregnant she continued treatment with standard doses of this drug, 25 mg twice a week, without complications

[Etanercept and pregnancy]. / Etanercept y embarazo.

We present the case of a pregnant woman with arthritis in treatment with etanercept. After becoming pregnant she continued treatment with standard doses of this drug, 25 mg twice a week, without complications.

Intragraft cytokine expression in heart transplants with mild or no histological rejection.

UNLABELLED: The study of pro-inflammatory cytokines produced in situ in heart allografts may help to understand the mechanisms of rejection and open new possibilities to control graft rejection. METHODS: A total of 23 endomyocardial biopsies obtained from 16 transplanted patients treated with triple-drug therapy (azathioprine, prednisone, and cyclosporine) were studied. mRNA expression for tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, IL-6, IL-10, IL-12, IL-15, transforming growth factor (TGF)-beta, and beta-actin was determined by reverse transcription polymerase chain reaction (RT-PCR) and Southern blotting. Semiquantitative analysis was done by establishing the ratio between densitometric integrated value of each cytokine with the beta-actin and correlated with the histopathologic findings. RESULTS: Three groups of biopsies were determined according to the International Society for Heart and Lung Transplantation criteria: grade 0 (control group, n=12), grade 1A (sub-clinical rejection, n=6) and 'quilty effect' (n=5). An increased expression of mRNA for TNF-alpha and IL-6 (p=0.0091 and 0.0075, respectively) was found associated with rejection grade 1A episodes, mRNA for IL-1 beta was nonspecifically expressed in all the study groups, while IL-10 mRNA was not detected in any of the biopsies studied. mRNA for IL-12 and IL-15 was not associated with rejection. Interestingly, TGF-beta was not detected in any of the biopsies with the 'quilty pattern'. CONCLUSION: The association of TNF-alpha and IL-6 mRNA in situ expression with mild histologically probed rejection episodes may be used in the monitoring of heart transplants.

Diminished adherence and/or ingestion of virulent Mycobacterium tuberculosis by monocyte-derived macrophages from patients with tuberculosis.

The interaction between the macrophage and Mycobacterium tuberculosis is mediated by a variety of macrophage membrane-associated proteins. Complement receptors have been implicated in the adherence of M. tuberculosis to macrophages. In the present work, the adherence and/or ingestion of M. tuberculosis H37Rv to human monocyte-derived macrophages (MDM) from patients with tuberculosis (TB) and healthy controls was measured by microscopical examination, [3H]uracil incorporation, and CFU. The adherence and/or ingestion was enhanced by fresh serum and inhibited by heat inactivation, EDTA treatment, and anti-CR1 and anti-CR3 antibodies. Comparison of MDM from TB patients and healthy controls showed that the former exhibited a significantly decreased capacity to adhere and/or ingest M. tuberculosis, as determined by the number of CFU and 3H incorporation. The expression of CR1 (CD35) and CR3 (CD11b/CD18) on MDM from TB patients and healthy controls, as determined by flow cytometry, did not show significant differences. These results suggest that the lower ingestion of M. tuberculosis by MDM from TB patients is not due to defects in complement receptors, and therefore, there might be other molecules involved in the adherence and/or ingestion process that render MDM from TB patients ingest less mycobacteria than those from healthy controls.