Surgical Outcomes in a Lung Cancer-Screening Program Using Low Dose Computed Tomography.

OBJECTIVE: Lung cancer (LC) is the leading cause of death from cancer worldwide. More than 27,000 LCs are diagnosed annually in Spain, and most are unresectable. Early detection and treatment reduce LC mortality. This study describes surgical outcomes in a longstanding LC screening cohort in Spain. METHODS: We conducted a retrospective study of surgical outcomes in a LC screening (LCS) program using low dose computed tomography (LDCT) since the year 2000. A descriptive analysis of clinical and radiological parameters, presence or absence of a preoperative diagnosis, pathological staging, morbidity, mortality, and survival was performed. RESULTS: Ninety-seven (2.5%) LC were diagnosed in 3825 screened. Twenty individuals with LC had no surgery due to advanced stage or small cell histology. Eighty-seven surgical procedures were carried out for suspected or biopsy proven LC, detected by LDCT. Most operated patients were male (57[85%]) aged 64±9.1 years. Nine patients underwent a second operation for a metachronous primary lung cancer. Mean tumor size was 15.2±7.6mm. Eight nodules were benign (9.2%). Lobectomy was performed in 56 cases (83.6%). Adenocarcinoma (n=39; 58.2%) was the most frequent histological type followed by squamous cell carcinoma (n=17; 25.4%). Fifty-nine (88%) tumors were in Stage I. Thirteen patients (15.4%) had 16 complications. The estimated survival rates at 5 and 10 years for stage I were 93% (95% CI: 79%-98%) and 83% (95% CI: 65%-92%), respectively. CONCLUSION: Lung cancer screening was associated with excellent surgical outcomes with 5 and 10-year survival rates exceeding 90 and 80%, respectively.

Genome-wide profiling of non-smoking-related lung cancer cells reveals common RB1 rearrangements associated with histopathologic transformation in EGFR-mutant tumors.

BACKGROUND: The etiology and the molecular basis of lung adenocarcinomas (LuADs) in nonsmokers are currently unknown. Furthermore, the scarcity of available primary cultures continues to hamper our biological understanding of non-smoking-related lung adenocarcinomas (NSK-LuADs). PATIENTS AND METHODS: We established patient-derived cancer cell (PDC) cultures from metastatic NSK-LuADs, including two pairs of matched EGFR-mutant PDCs before and after resistance to tyrosine kinase inhibitors (TKIs), and then performed whole-exome and RNA sequencing to delineate their genomic architecture. For validation, we analyzed independent cohorts of primary LuADs. RESULTS: In addition to known non-smoker-associated alterations (e.g. RET, ALK, EGFR, and ERBB2), we discovered novel fusions and recurrently mutated genes, including ATF7IP, a regulator of gene expression, that was inactivated in 5% of primary LuAD cases. We also found germline mutations at dominant familiar-cancer genes, highlighting the importance of genetic predisposition in the origin of a subset of NSK-LuADs. Furthermore, there was an over-representation of inactivating alterations at RB1, mostly through complex intragenic rearrangements, in treatment-naive EGFR-mutant LuADs. Three EGFR-mutant and one EGFR-wild-type tumors acquired resistance to EGFR-TKIs and chemotherapy, respectively, and histology on re-biopsies revealed the development of small-cell lung cancer/squamous cell carcinoma (SCLC/LuSCC) transformation. These features were consistent with RB1 inactivation and acquired EGFR-T790M mutation or FGFR3-TACC3 fusion in EGFR-mutant tumors. CONCLUSIONS: We found recurrent alterations in LuADs that deserve further exploration. Our work also demonstrates that a subset of NSK-LuADs arises within cancer-predisposition syndromes. The preferential occurrence of RB1 inactivation, via complex rearrangements, found in EGFR-mutant tumors appears to favor SCLC/LuSCC transformation under growth-inhibition pressures. Thus RB1 inactivation may predict the risk of LuAD transformation to a more aggressive type of lung cancer, and may need to be considered as a part of the clinical management of NSK-LuADs patients.

Injerto costal para reconstrucción nasal: experiencia de 6 años en el Hospital San Juan de Dios / Costal graft for nasal reconstruction: 6 years experience in San Juan de Dios Hospital

Introducción: El injerto de preferencia para cirugía nasal es el cartílago septal. El Injerto de cartílago costal otorga gran material para trabajo ya que posee características similares al cartílago septal. Este injerto se utiliza generalmente para cirugías de dorso nasal, nariz en silla de montar, laterorrinias, disfunciones de la válvula nasal interna y/o externa, poca proyección nasal y rinoplastías de revisión. Objetivo: Conocer las características, resultados y complicaciones de las rinoplastías con injerto de cartílago costal realizadas en el Hospital San Juan de Dios (HSJD) entre los años 2010 y 2016. Material y método: Estudio retrospectivo con revisión de fichas clínicas de pacientes sometidos a cirugías de reconstrucción nasal con injerto de cartílago costal en el HSJD entre enero 2010 y agosto 2016. Resultado: Entre los años 2010 y 2016 se realizaron 20 rinoplastías con uso de cartílago costal: 14 eran hombres (70%) y 6 eran mujeres (30%). En 14 pacientes se usó el cartílago costal en la primera cirugía. No se presentaron complicaciones a corto, mediano y largo plazo. Conclusión: El injerto de cartílago costal es una herramienta útil a la hora de enfrentarse a cirugías complicadas. La técnica quirúrgica no es dificultosa y tiene una baja tasa de complicaciones.

Introduction: The ideal graft for nasal surgery is septal cartilage. The costal cartilage has similar characferistics to the septal cartilage. This particular graft is generally used for nasal dorsum, saddle nose, nasal deviations, dysfunctions of the internal or external nasal valve, poor nasal projection and revision rhinoplasty. Aim: Determine the characteristics, outcomes and complications of rhinoplasty with costal cartilage graft made in the San Juan de Dios Hospital (HSJD) between 2010 and 2016. Material and method: Retrospective study with reviewing medical records of all patients underwent nasal surgery reconstruction with costal cartilage graft in the HSJD between January 2010 and August 2016. Results: Between 2010 and 2016 a total of 20 rhinoplasties were performed with use of costal cartilage. Of these 14 were men (70%) and 6 were women (30%). In 14 patients we used the costal cartilage in the first surgery. No complications in short, medium and long term were presented. Conclusion: The rib cartilage graft is a useful tool when dealing with complicated surgeries. The surgical technique is not difficult and has a low complication rate.

Total and mutated EGFR quantification in cell-free DNA from non-small cell lung cancer patients detects tumor heterogeneity and presents prognostic value.

Mutation analysis of epidermal growth factor receptor (EGFR) gene is essential for treatment selection in non-small cell lung cancer (NSCLC). Analysis is usually performed in tumor samples. We evaluated the clinical utility of EGFR analysis in plasma cell-free DNA (cfDNA) from patients under treatment with EGFR inhibitors. We selected 36 patients with NSCLC and EGFR-activating mutations. Blood samples were collected at baseline and during treatment with EGFR inhibitors. Wild-type EGFR, L858R, delE746-A750, and T790M mutations were quantified in cfDNA by droplet digital PCR. Stage IV patients had higher total circulating EGFR copy levels than stage I (3523 vs. 1003 copies/mL; p < 0.01). There was high agreement for activating mutations between baseline cfDNA and tumor samples, especially for L858R mutation (kappa index = 0.679; p = 0.001). In 34 % of advanced NSCLC patients, we detected mutations in cfDNA not previously detected in tumor samples and double mutations in 17 %. Patients with baseline total EGFR copy levels above the median presented decreased overall survival (OS) (341 vs. 870 days, p < 0.05) and progression-free survival (PFS) (238 vs. 783 days; p < 0.05) compared with those with total EGFR copy levels below the median. Patients with baseline concentrations of activating mutations above the median (94 copies/mL) had lower OS (317 vs. 805 days; p < 0.05) and PFS (195 vs. 724 days; p < 0.05). During follow-up, T790M resistance mutation was detected in 53 % of patients. Total and mutated EGFR analysis in cfDNA seems a relevant tool to characterize the molecular profile and prognosis of NSCLC patients harboring EGFR mutations.

Presencia de síndrome de burnout en una muestra de residentes y otorrinolaringólogos de Chile / Burnout syndrome in a sample of residents and otolaryngologists from Chile

Introducción: El síndrome de Burnout o desgaste profesional relacionado con el trabajo se genera frente a un estrés laboral crónico en individuos que trabajan con personas. Se caracteriza por un agotamiento emocional, despersonalización y baja realización personal. Objetivo: Evaluar el síndrome de Burnout en los médicos residentes y otorrinolaringólogos (ORL) del país y establecer una relación entre las características sociodemográficas. Material y método: Se realizó un estudio de corte transversal. Se envió el inventario Burnout de Maslach (MBI) mediante correo electrónico a todos los residentes y ORL miembros de la Sociedad Chilena de Otorrinolaringología, Medicina y Cirugía de Cabeza y Cuello (SOCHIORL). Se utilizó test exacto de Fisher para evaluar asociación entre variables cualitativas. Resultados: De un total de 432 miembros de la SOCHIORL se recibieron 133 encuestas, de las cuales se analizaron 117 contestadas correctamente. El 100% del total de los encuestados presentó Burnout ya sea de alto grado o intermedio. Los que presentaron mayor porcentaje de alto grado de Burnout son aquellos entre 25 y 35 años, con 53% (p =0,03). El mayor grado de Burnout se observó en los residentes (57%) (p =0,03). Conclusión: El síndrome de Burnout tiene una alta prevalencia entre los ORL del país, especialmente en residentes y jóvenes ORL y quienes llevan pocos años en el desarrollo de la especialidad. Es necesario buscar herramientas para enfrentar y combatir esta realidad.

Introduction: Burnout syndrome is generated against a chronic work stress on individuals working with people. It is characterized by emotional exhaustion, depersonalization and reduced personal accomplishment. Aim: To assess Burnout in residents and otolaryngologists (ORL) in the country and establish a relationship between sociodemographic characteristics. Material and method: A cross sectional study was conducted. Maslach Burnout Inventory (MBI) was sent via email to all residents and ORL members of the Chilean Society of Otolaryngology, Medicine and Head and Neck Surgery (SOCHIORL). Fisher exact test was used to assess the association between qualitative variables. Results: Of a total of 432 members of SOCHIORL, 133 surveys were received, of which 117 were analysed. 100% of respondents had Burnout, either high or intermediate degree. Those with higher percentage of high Burnout are those between 25 and 35 years (53%) (p =0.03). The greatest degree of Burnout was observed in residents (57%) (p =0.03). Conclusion: Burnout syndrome is highly prevalent among ORL in Chile, especially in residents and young ORL with few years in the medical practice of the specialty. It is necessary to search for tools to confront and combat this reality.

Complement activation product C4d in oral and oropharyngeal squamous cell carcinoma.

OBJECTIVE: Complement C4d-containing fragments have been proposed as diagnostic markers for lung cancer. The purpose of this study was to evaluate the presence of C4d in oropharyngeal (OPSCC) and oral (OSCC) squamous cell carcinomas. SUBJECTS AND METHODS: C4d staining was analyzed by immunohistochemistry in 244 OPSCC surgical specimens. C4d levels were quantified by ELISA in resting saliva samples from 48 patients with oral leukoplakia and 62 with OSCC. Plasma samples from 21 patients with leukoplakia and 30 with oral carcinoma were also studied. RESULTS: C4d staining in OPSCC specimens was associated with nodal invasion (P = 0.001), histopathologic grade (P = 0.014), disease stage (P = 0.040), and focal-adhesion kinase expression (P < 0.001). No association was found between C4d and prognosis. Saliva C4d levels were higher in patients with oral cancer than in subjects with leukoplakia (0.07 ± 0.07 vs 0.04 ± 0.03 µg ml(-1) , P = 0.003). The area under the ROC curve was 0.63 (95%CI: 0.55-0.71). Salivary C4d levels in stage IV patients were higher than in patients with earlier stages (P = 0.028) and correlated with tumor size (P = 0.045). Plasma C4d levels also correlated with salivary C4d levels (P = 0.041), but differences between patients with oral cancer and subjects with leukoplakia were not significant (1.26 ± 0.59 vs 1.09 ± 0.39 µg ml(-1) , P = 0.232). CONCLUSION: C4d-containing fragments are detected in oral primary tumors and are increased in saliva from patients with OSCC.

Early Growth Response 3 regulates genes of inflammation and directly activates IL6 and IL8 expression in prostate cancer.

BACKGROUND: Transcription factor EGR3 (Early Growth Response 3) is a little-studied member of the EGR family that is highly expressed in human prostate tumours compared with normal tissue. Its function in prostate cancer, however, is unknown. METHODS: Stable shRNA silencing was achieved in naturally overexpressing prostate cancer cells, followed by Affymetrix expression analysis. Fold changes of ⩾2 and ⩽-2 were considered valid and t-tests P-values of ⩽0.01 were considered statistically significant. Potential EGR3 target genes were validated by real-time qPCR, chromatin immunoprecipitation, and gain-of-function experiments. Promoter analysis confirmed the presence of consensus binding sites in the promoters of target genes. RESULTS: Early Growth Response 3 regulates the expression of ∼330 genes, 35% of which are involved in immune responses and inflammatory processes, and 15% crosstalk with the NF-κB signalling pathway. In particular, EGR3 induces the expression of over 50 secreted cytokines, growth factors, and matrix remodelling factors. Two interleukins of great relevance to prostate cancer, IL6 and IL8, were further validated as EGR3 target genes: both promoters contain EGR consensus binding sites and are pulled down in intact cells by EGR3 chromatin immunoprecipitation. Silencing of EGR3 decreased IL6 and IL8 expression, whereas overexpression of EGR3 in nontransformed cells induced IL6 and IL8 expression. CONCLUSIONS: Chronic inflammation plays a critical role in prostate cancer and elevated production of pro-inflammatory cytokines IL8 and IL6, in particular, contributes to disease progression and to the onset of castration resistance. It is shown for the first time that EGR3 is involved in the upregulation of both IL6 and IL8. Together with our previous observation that EGR3 is highly expressed in prostate tumours compared with normal tissue and strongly correlates with IL6 and IL8 expression in clinical samples, the present study suggests that EGR3 promotes excessive production of IL6 and IL8 observed during the progression of prostate cancer.

Análisis de los pacientes con diabetes controlados a nivel primaria en el Servicio de Salud Concepción / Analysis of diabetic patients controlled at primary level in Concepcion Health Service

To evaluate diabetic patients in control in Concepcion Health Service, evaluating coverage, metabolic control and complications. Material: A retrospective analysis of the statistical record of 2011 (REM) using the CIE-10 classification was done. Estimation is performed with the National Health Survey of 2010. Results: 26,638 patients are controlled, achieving a 67.9 percent coverage (point estimate based on the National Health Survey of 2010), being lower in the group between 15 and 44 years old (34 percent). 38.4 percent had good metabolic control (HbA1C less than 7) and 20.8 percent poor control (HbA1C greater than 9). According to age, the group over 65 had better control and the group between 15 and 44 years worse control. The complication described was diabetic foot (51.9 percent), retinopathy (5 percent) and nephropathy (4.6 percent). Discussion: The prevalence of diabetes is increasing every year in Chile. There is acceptable coverage and early diagnosis, but still are poorly controlled cases and require a multifactorial management that begins with a self-care of their diabetes...

TGFBI expression is an independent predictor of survival in adjuvant-treated lung squamous cell carcinoma patients.

BACKGROUND: Transforming growth factor ß-induced protein (TGFBI) is a secreted protein that mediates cell anchoring to the extracellular matrix. This protein is downregulated in lung cancer, and when overexpressed, contributes to apoptotic cell death. Using a small series of stage IV non-small cell lung cancer (NSCLC) patients, we previously suggested the usefulness of TGFBI as a prognostic and predictive factor in chemotherapy-treated late-stage NSCLC. In order to validate and extend these results, we broaden the analysis and studied TGFBI expression in a large series of samples obtained from stage I-IV NSCLC patients. METHODS: TGFBI expression was assessed by immunohistochemistry in 364 completely resected primary NSCLC samples: 242 adenocarcinomas (ADCs) and 122 squamous cell carcinomas (SCCs). Kaplan-Meier curves, log-rank tests and the Cox proportional hazards model were used to analyse the association between TGFBI expression and survival. RESULTS: High TGFBI levels were associated with longer overall survival (OS, P<0.001) and progression-free survival (PFS, P<0.001) in SCC patients who received adjuvant platinium-based chemotherapy. Moreover, multivariate analysis demonstrated that high TGFBI expression is an independent predictor of better survival in patients (OS: P=0.030 and PFS: P=0.026). CONCLUSIONS: TGFBI may be useful for the identification of a subset of NSCLC who may benefit from adjuvant therapy.

[Mucopenetrating nanoparticles: vehicles for the oral administration of paclitaxel]. / Nanoparticules mucopénétrantes: véhicules pour l'administration orale du paclitaxel.

Paclitaxel is an anticancer drug used as solution for perfusion for the treatment of certain types of cancers. In the last years, a number of strategies have been proposed for the development of an oral formulation of this drug. However, this task is quite complicated due to the poor aqueous solubility of paclitaxel as well as the fact that this compound is substrate of the intestinal P-glycoprotein and the cytochrome P450 enzymatic complex. In this work, we have developed pegylated nanoparticles with mucopenetrating properties in order to conduct paclitaxel onto the surface of the enterocyte. These nanoparticles displayed a size of about 180 nm and a drug loading close to 15% by weight. The pharmacokinetic study in mice has shown that these nanoparticles were capable to offer therapeutic plasma levels of paclitaxel up to 72 hours. In addition, the oral relative bioavailability of paclitaxel when loaded in nanoparticles pegylated with poly(ethylene glycol) 2000 (PEG) was found to be 85%. In a subcutaneous model of tumour in mice, these pegylated nanoparticles administered orally every 3 days have demonstrated a similar efficacy than Taxol® administered intravenously every day during 9 days. All of these results suggested that these pegylated nanoparticles were capable to cross the mucus layer of the gut and, then, reach the surface of the enterocytes. The PEG molecules would facilitate the adhesion of nanoparticles to this epithelial surface, minimise the pre-systemic metabolism of paclitaxel and, thus, promote its absorption.

Miocardiopatía dilatada en paciente con lupus eritematoso sistémico: reporte de un caso / Dilated cardiomyopathy in patient with systemic lupus erythematosus: case report

INTRODUCCIÓN: El Lupus Eritematoso Sistémico (LES) es una enfermedad autoinmune de etiología desconocida, cuyo cuadro clínico incluye diversas manifestaciones cardiovasculares, lasque se pueden presentar entre el 50 y 60 por ciento de los pacientes. La miocarditis es infrecuente (10 por ciento), pudiendo evolucionar tanto a miocardiopatía dilatada como a la mejoría. PRESENTACIÓN DEL CASO: Mujer de 40 años con antecedentes de LES diagnosticado el año 2008 y hospitalización anterior por cuadro de síndrome nefrótico, consulta en la urgencia del Hospital Guillermo Grant Benavente el por cuadro clínico de dos semanas de evolución caracterizado por disnea de mínimo esfuerzo, oliguria y edema continuo de extremidades inferiores. Destacan dentro de sus exámenes de ingreso: creatinina de 1,8 mg/dl, PCR <5 mg/l e índice proteinuria/creatininuria aislada >5; radiografía de tórax muestra cardiomegalia. Se hospitaliza en servicio de medicina interna para manejo. Evoluciona con mayor disnea ydolor torácico, por lo que se realiza ecocardiograma que revela miocardiopatía dilatada y disfunción sistólica severa. Se añade al tratamiento esteroidal previo, furosemida, carvedilol, enalapril y bolos de metilprednisolona y ciclofosfamida. Paciente evoluciona satisfactoriamente, con disminución de la disnea y leve edema de las extremidades inferiores, aunque permanece con valores de creatinina alterados. Luego de un mes es dada de alta. DISCUSIÓN: Señalamos que la miocardiopatía dilatada es una complicación infrecuente del LES, y si bien es cierto es la principal explicación para el cuadro clínico de insuficiencia cardiaca en esta paciente, no explica por si sola la sintomatología, considerando el antecedente de compromiso renal.

INTRODUCTION: Systemic Lupus Erythematosus is an autoimmune disease of unknown etiology, whose clinical picture includes various cardiovascular manifestations, which occur in between the 50 and 60 percent of the patients. Myocarditis is a rare complication (10 percent) and may evolve to dilated cardiomyopathy, or to the improvement. CASE REPORT: Forty year old woman with a history of Systemic Lupus Erythematosus diagnosed in 2008 and previous hospitalization for nephrotic syndrome, consultates in the urgency of Guillermo Grant Benavente Hospital because of a two weeks clinical picture characterized by small effort dyspnea, oliguria and continuous edema of lower extremities.Stands in their entrance examinations: creatinine 1.8 mg / dl, CRP <5 mg/l, proteinuria /creatinine index >5; X-ray shows cardiomegaly. It is decided to hospitalized the patient. Evolves with increasing respiratory distress and chest pain, so it is decided to do echocardiogram, wich revealed dilated cardiomyopathy and severe systolic dysfunction. It´s added to her prevoiuos steroid treatment, furosemide, carvedilol, enalapril, methylprednisolone and cyclophosphamide. The patient has a satisfactory evolution, with decreased dyspnea and mild edema of the lower extremities, although it remains with altered values of creatinine. The patient is sent home and referred to early control. DISCUSSION: We propose that dilated cardiomyopathy is a rare complication of Systemic Lupus Erythematosus, and though we can say that it is the main explanation for the clinical picture of this patient, it does not explain by itself the symptomatology, considering the history of renal involvement.

Overexpression of TMPRSS4 in non-small cell lung cancer is associated with poor prognosis in patients with squamous histology.

BACKGROUND: Mortality rates in lung cancer patients have not decreased significantly in recent years, even with the implementation of new therapeutic regimens. One of the main problems is that a large proportion of patients present local or distant metastasis at the time of diagnosis. The need for identification of novel biomarkers and therapeutic targets for a more effective management of lung cancer led us to investigate TMPRSS4, a protease reported to promote tumour growth and metastasis. MATERIAL AND METHODS: In all, 34 lung cancer cell lines were used to evaluate the TMPRSS4 expression. Cell migration and clonogenic assays, and an in-vivo lung metastasis model were used for functional analysis of the TMPRSS4 downregulation in H358, H441 and H2170 cell lines. The TMPRSS4 expression analysis in normal and malignant lung tissue samples was performed by qPCR. Five different microarray-based publicly available expression databases were used to validate our results and to study prognosis. RESULTS: The TMPRSS4 knock down in H358, H441 and H2170 cells resulted in a significant reduction in proliferation, clonogenic capacity and invasion. A significant (P<0.05) decrease in the lung colonisation and growth was found when mice were injected with TMPRSS4-depleated H358-derived clones, as compared with controls. Expression of TMPRSS4 showed a >30-fold increase (P<0.001) in tumours in comparison with non-malignant samples. Levels in tumours with squamous cell carcinoma (SCC) histology were found to be significantly higher (P<0.001) than those with adenocarcinoma (AC) histology, which was confirmed in data retrieved from the microarrays. Kaplan-Meier curves demonstrated that high levels of TMPRSS4 were significantly associated (P=0.017) with reduced overall survival in the patients with SCC histology, whereas no correlation was found for the AC histology. CONCLUSION: Our results demonstrate that TMPRSS4 has a role in the lung cancer development. The potential use of TMPRSS4 as a biomarker for lung cancer detection or as a predictor of patient's outcome warrants further investigation.

Sensitization to horse allergens in Italy: a multicentre study in urban atopic subjects without occupational exposure.

BACKGROUND: Horses play a significant role in people's leisure time in Italy and other countries, but few data are available on IgE-mediated sensitization to horse allergens in patients without occupational exposure. We assessed, in a multicentric survey, the prevalence of horse sensitization in atopic subjects and its clinical characteristics. METHODS: Allergists from the whole Italian territory were required to collect the results of skin prick tests from at least 100 consecutive subjects. Those patients with a positive skin test to horse dander underwent a detailed interview concerning clinical history, pet ownership and possible exposure. RESULTS: Data from 3,235 outpatients were collected and 2,097 had at least 1 skin positivity. Among them, 113 (5.38%) were sensitized to horse dander (9 monosensitized). Thirty patients reported direct horse contact (4 owners and 26 for riding or occasional contact), 23 patients were sometimes in contact with horse owners and 60 subjects denied any direct or indirect exposure. Among 9 horse monosensitized patients, 6 had intermittent and mild rhinitis and 3 persistent moderate/severe rhinitis plus asthma. Three of them were horse owners or riders and the remaining had no contact with the animal. CONCLUSIONS: Our data evidence that the rate of sensitization to horse dander is not negligible and probably underestimated. In susceptible, not occupationally exposed individuals, horse contact, but also indirect or no apparent exposure, may induce sensitization. We recommend inclusion of horse allergen in the routine panel for the diagnosis of respiratory allergy.

Molecular characterization of small peripheral lung tumors based on the analysis of fine needle aspirates.

The computed tomography (CT)-based early lung cancer diagnostic technologies allow the detection of very small stage I lung tumors. As part of these screening protocols any suspicious nodule has to be diagnosed morphologically, which requires CT-guided Fine Needle Aspiration, open biopsy or surgery. Fine Needle Aspiration (FNA) cytology is a well-recognised method for a rapid and accurate diagnosis of small lung tumors. Molecular analysis of the FNA specimens could complement cytology diagnosis by the characterization of the biological traits at the preoperative stage. In this study, we aimed to characterize the biological profile of 33 paraffin-embedded transthoracic FNA samples obtained from three groups of lung cancer patients: two groups of small early-detected lung adenocarcinomas (radiologically subsolid and solid nodules) and a third group of small metastatic adenocarcinomas. Genetic analysis was performed by fluorescence in situ hybridization using the four-color LAVysion probe. p53 and Ki-67 protein expression was also evaluated by immunocytochemistry. The samples showed gains for all targets analyzed; two cases had EGFR gene amplification and two cases had MYC amplification. There were no significant differences in the percentage of genetically malignant cells and the expression of Ki-67 among the three groups. However, p53 accumulation was significantly higher in the metastatic group compared to the subsolid early-detected group (P = 0.001). In conclusion, molecular analysis of FNA specimens may provide useful information at preoperative stages. In our series, a good prognostic profile in subsolid early detected adenocarcinomas is suggested.

[FICTION as a new tool to early lung cancer diagnosis]. / Desarrollo de la técnica de FICTION como nueva herramienta para el diagnóstico precoz de cáncer de pulmón.

Lung cancer is one of the most frequent causes of cancer death in Western countries. Overall 5-year survival rate is lower than 15% mainly due to the late diagnosis of the disease. Primary prevention (reduction of tobacco consumption) and more effective methods for early detection are needed. Some studies have recently shown that low-dose spiral computed tomography (CT) is a useful technique to the detection of pulmonary malignant nodules in early stages. Studies are developing to evaluate its efficacy in series of high-risk patients. A new cytogenetic technique has been developed: the FICTION technique (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms). This technique allows the simultaneous study of immunophenotypic markers and genetic abnormalities present in tumour cells. The goal of our project is optimise this technique in sputum and bronchoalveolar lavage specimens from lung cancer patients. The overall goal of this project is evaluate the usefulness of this technique, together with the new radiological techniques, in early detection programs of lung cancer in high-risk patients. In the present study we review the cytogenetic studies on lung cancer carried out in the recent years. We also introduce the basic methodological aspects that will be developed in our project.

Expression of the adrenomedullin binding protein, complement factor H, in the pancreas and its physiological impact on insulin secretion.

Adrenomedullin (AM) is a ubiquitous peptide hormone which, among other functional roles, reduces insulin secretion in the pancreas. Recently we have described the interaction between AM and the complement regulator protein factor H, which results in mutual modulation of their respective functions. Here we identify the expression of factor H in the beta cells of the rat pancreatic islets by immunohistochemistry and multiple immunofluorescence followed by confocal microscopy. In addition, double immunogold staining under the electron microscope showed coexistence of insulin and factor H immunoreactivities within the same secretory granules; interestingly, factor H staining was found in the electron-lucent haloes whereas the insulin antibody labeled preferentially the dense cores. The existence of factor H mRNA in the pancreas was confirmed by RT-PCR and in situ hybridization. The function of factor H in the pancreas was investigated with an insulin secretion assay. Addition of factor H to freshly isolated islets in the presence of AM resulted in a further reduction in insulin secretion with a concomitant elevation of cAMP, suggesting that factor H increases AM function in glucose homeostasis. The expression of factor H in the pancreas may play other important roles such as protection against complement-mediated cell lysis.

Complement factor H is a serum-binding protein for adrenomedullin, and the resulting complex modulates the bioactivities of both partners.

Adrenomedullin (AM) is an important regulatory peptide involved in both physiological and pathological states. We have previously demonstrated the existence of a specific AM-binding protein (AMBP-1) in human plasma. In the present study, we developed a nonradioactive ligand blotting assay, which, together with high pressure liquid chromatography/SDS-polyacrylamide gel electrophoresis purification techniques, allowed us to isolate AMBP-1 to homogeneity. The purified protein was identified as human complement factor H. We show that AM/factor H interaction interferes with the established methodology for quantification of circulating AM. Our data suggest that this routine procedure does not take into account the AM bound to its binding protein. In addition, we show that factor H affects AM in vitro functions. It enhances AM-mediated induction of cAMP in fibroblasts, augments the AM-mediated growth of a cancer cell line, and suppresses the bactericidal capability of AM on Escherichia coli. Reciprocally, AM influences the complement regulatory function of factor H by enhancing the cleavage of C3b via factor I. In summary, we report on a potentially new regulatory mechanism of AM biology, the influence of factor H on radioimmunoassay quantification of AM, and the possible involvement of AM as a regulator of the complement cascade.

Cancer and diabetes: two pathological conditions in which adrenomedullin may be involved.

Adrenomedullin (AM) is a regulatory peptide involved in several physiological processes. Among them, AM has been implicated in the regulation of growth, both with mitogenic and antiproliferative activities on normal cells. AM is widely expressed during embryogenesis and may have a significant role in the proliferation and differentiation processes associated with development. AM is also expressed by cancer cell lines and tumors and has been implicated in the growth of malignant cells. Some additional activities associated with AM (antiapoptotic capabilities, angiogenic potential, and upregulation in hypoxic conditions), together with its wide distribution in cancer, suggest that AM may be an important factor in carcinogenesis. Besides its implication in growth, embryogenesis and tumor biology, AM is also involved in pancreatic regulation and diabetes. AM regulates insulin secretion and is overexpressed in the plasma of diabetic patients. Several findings indicate that AM may participate in the pathogenesis and/or clinical complications of this disease.