Overall survival prediction in metastatic castration-resistant prostate cancer treated with radium-223.

OBJECTIVE: Radium-223(223Ra) is indicated for patients (p) with metastatic castration resistant prostate cancer (mCRCP). OBJECTIVES: The aim of this study was to evaluate the role of baseline clinical variables associated with overall survival (OS) and toxicity of 223Ra. Its purpose was to identify the factors that can predict a better response to treatment and provide information regarding the most appropriate time for the application of 223Ra. MATERIALS AND METHODS: Prospective study in 40p with mCRPC treated with 223Ra. End points were OS, progression-free survival and time to progression. The follow-up parameters were: doses received, hemoglobin (Hb), absolute neutrophil count (ANC), platelet count (PC), prostate specific antigen (PSA), alkaline phosphatase (ALP), Visual Analogue Scale for pain, Eastern Cooperative Oncology Group (ECOG) and WHO's Cancer Pain Ladder. The use of other treatments was also evaluated. RESULTS: Median OS was 17.1 months(mo) (CI95%6.5-27.7); 26/40p received complete treatment of 223Ra, without reaching a median OS and 14p received incomplete treatment with a median OS 13.6mo(CI95%1.6-25.6). Median follow-up was 11.2mo (range:1.3-45.2). The univariate analysis showed that factors as VAS, ECOG, Hb and ALP values were independently associated with OS. First line treatment with 223Ra was started in 11/40p, while 19p had been heavily pre-treated and 13p received concomitant treatment. CONCLUSIONS: 223Ra therapy require an adequate selection of patients to obtain the greatest clinical benefit. Low basal Hb, hight basal ALP, bone marrow involvement and an altered ECOG were the main factors that decreased OS in our patients. 223Ra should be considered relatively early in the course of treatment. Available at. https://www.intbrazjurol.com.br/pdf/aop/2019-0343OA.pdf.

Does the Capsule Interfere with Performance of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of Cryptococcus neoformans and Cryptococcus gattii?

We described the impact of the capsule size for Cryptococcus neoformans and Cryptococcus gattii identification at the species level by Bruker matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). After experimental capsule size modulation, we observed that reducing the capsule size resulted in improved identification by Bruker MALDI-TOF MS across all of the reference strains analyzed.

Correlation between theoretical anatomical patterns of lymphatic drainage and lymphoscintigraphy findings during sentinel node detection in head and neck melanomas.

PURPOSE: In the diagnosis of head and neck melanoma, lymphatic drainage is complex and highly variable. As regional lymph node metastasis is one of the most important prognostic factors, lymphoscintigraphy can help map individual drainage patterns. The aim of this study was to compare the results of lymphoscintigraphy and sentinel lymph node (SLN) detection with theoretical anatomical patterns of lymphatic drainage based on the location of the primary tumour lesion in patients with head and neck melanoma. We also determined the percentage of discrepancies between our lymphoscintigraphy and the theoretical location of nodal drainage predicted by a large lymphoscintigraphic database, in order to explain recurrence and false-negative SLN biopsies. METHODS: In this retrospective study of 152 patients with head and neck melanoma, the locations of the SLNs on lymphoscintigraphy and detected intraoperatively were compared with the lymphatic drainage predicted by on-line software based on a large melanoma database. RESULTS: All patients showed lymphatic drainage and in all patients at least one SLN was identified by lymphoscintigraphy. Of the 152 patients, 4 had a primary lesion in areas that were not described in the Sydney Melanoma Unit database, so agreement could only be evaluated in 148 patients. Agreement between lymphoscintigraphic findings and the theoretical lymphatic drainage predicted by the software was completely concordant in 119 of the 148 patients (80.4 %, 95 % CI 73.3 - 86 %). However, this concordance was partial (some concordant nodes and others not) in 18 patients (12.2 %, 95 % CI 7.8 - 18.4 %). Discordance was complete in 11 patients (7.4 %, 95 % CI 4.2 - 12.8 %). CONCLUSION: In melanoma of the head and neck there is a high correlation between lymphatic drainage found by lymphoscintigraphy and the predicted drainage pattern and basins provided by a large reference database. Due to unpredictable drainage, preoperative lymphoscintigraphy is essential to accurately detect the SLNs in head and neck melanoma.

Matrix-assisted laser desorption ionization-time of flight mass spectrometry for differentiation of the dimorphic fungal species Paracoccidioides brasiliensis and Paracoccidioides lutzii.

Isolates of Paracoccidioides brasiliensis and Paracoccidioides lutzii, previously characterized by molecular techniques, were identified for the first time by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). All isolates were correctly identified, with log score values of >2.0. Thus, MALDI-TOF MS is a new tool for differentiating species of the genus Paracoccidioides.

Genetic and forensic implications in epilepsy and cardiac arrhythmias: a case series.

Epilepsy affects approximately 3% of the world's population, and sudden death is a significant cause of death in this population. Sudden unexpected death in epilepsy (SUDEP) accounts for up to 17% of all these cases, which increases the rate of sudden death by 24-fold as compared to the general population. The underlying mechanisms are still not elucidated, but recent studies suggest the possibility that a common genetic channelopathy might contribute to both epilepsy and cardiac disease to increase the incidence of death via a lethal cardiac arrhythmia. We performed genetic testing in a large cohort of individuals with epilepsy and cardiac conduction disorders in order to identify genetic mutations that could play a role in the mechanism of sudden death. Putative pathogenic disease-causing mutations in genes encoding cardiac ion channel were detected in 24% of unrelated individuals with epilepsy. Segregation analysis through genetic screening of the available family members and functional studies are crucial tasks to understand and to prove the possible pathogenicity of the variant, but in our cohort, only two families were available. Despite further research should be performed to clarify the mechanism of coexistence of both clinical conditions, genetic analysis, applied also in post-mortem setting, could be very useful to identify genetic factors that predispose epileptic patients to sudden death, helping to prevent sudden death in patients with epilepsy.

Validation of N Protein Antibodies to Diagnose Previous SARS-CoV-2 Infection in a Large Cohort of Healthcare Workers: Use of Roche Elecsys® Immunoassay in the S Protein Vaccination Era.

The aim of this study was to validate the detection of anti-nucleocapsid protein (N protein) antibodies for the diagnosis of SARS-CoV-2 infection in light of the fact that most COVID-19 vaccines use the spike (S) protein as the antigen. Here, 3550 healthcare workers (HCWs) were enrolled from May 2020 (when no S protein vaccines were available). We defined SARS-CoV-2 infection if HCWs were found to be positive by RT-PCR or found to be positive in at least two different serological immunoassays. Serum samples from Biobanc I3PT-CERCA were analyzed by Roche Elecsys® (N protein) and Vircell IgG (N and S proteins) immunoassays. Discordant samples were reanalyzed with other commercial immunoassays. Roche Elecsys® showed the positivity of 539 (15.2%) HCWs, 664 (18.7%) were found to be positive by Vircell IgG immunoassays, and 164 samples (4.6%) showed discrepant results. According to our SARS-CoV-2 infection criteria, 563 HCWs had SARS-CoV-2 infection. The Roche Elecsys® immunoassay has a sensitivity, specificity, accuracy, and concordance with the presence of infection of 94.7%, 99.8%, 99.3%, and 0.96, respectively. Similar results were observed in a validation cohort of vaccinated HCWs. We conclude that the Roche Elecsys® SARS-CoV-2 N protein immunoassay demonstrated good performance in diagnosing previous SARS-CoV-2 infection in a large cohort of HCWs.

Lymphocyte transformation assay for C neoformans antigen is not reliable for detecting cellular impairment in patients with neurocryptococcosis.

BACKGROUND: Cryptococcus neoformans causes meningitis and disseminated infection in healthy individuals, but more commonly in hosts with defective immune responses. Cell-mediated immunity is an important component of the immune response to a great variety of infections, including yeast infections. We aimed to evaluate a specific lymphocyte transformation assay to Cryptococcus neoformans in order to identify immunodeficiency associated to neurocryptococcosis (NCC) as primary cause of the mycosis. METHODS: Healthy volunteers, poultry growers, and HIV-seronegative patients with neurocryptococcosis were tested for cellular immune response. Cryptococcal meningitis was diagnosed by India ink staining of cerebrospinal fluid and cryptococcal antigen test (Immunomycol-Inc, SP, Brazil). Isolated peripheral blood mononuclear cells were stimulated with C. neoformans antigen, C. albicans antigen, and pokeweed mitogen. The amount of 3H-thymidine incorporated was assessed, and the results were expressed as stimulation index (SI) and log SI, sensitivity, specificity, and cut-off value (receiver operating characteristics curve). We applied unpaired Student t tests to compare data and considered significant differences for p<0.05. RESULTS: The lymphotoxin alpha showed a low capacity with all the stimuli for classifying patients as responders and non-responders. Lymphotoxin alpha stimulated by heated-killed antigen from patients with neurocryptococcosis was not affected by TCD4+ cell count, and the intensity of response did not correlate with the clinical evolution of neurocryptococcosis. CONCLUSION: Response to lymphocyte transformation assay should be analyzed based on a normal range and using more than one stimulator. The use of a cut-off value to classify patients with neurocryptococcosis is inadequate. Statistical analysis should be based on the log transformation of SI. A more purified antigen for evaluating specific response to C. neoformans is needed.

Insertion of short transmembrane helices by the Sec61 translocon.

The insertion efficiency of transmembrane (TM) helices by the Sec61 translocon depends on helix amino acid composition, the positions of the amino acids within the helix, and helix length. We have used an in vitro expression system to examine systematically the insertion efficiency of short polyleucine segments (L(n), n = 4 ... 12) flanked at either end by 4-residue sequences of the form XXPX-L(n)-XPXX with X = G, N, D, or K. Except for X = K, insertion efficiency (p) is <10% for n < 8, but rises steeply to 100% for n = 12. For X = K, p is already close to 100% for n = 10. A similar pattern is observed for synthetic peptides incorporated into oriented phospholipid bilayer arrays, consistent with the idea that recognition of TM segments by the translocon critically involves physical partitioning of nascent peptide chains into the lipid bilayer. Molecular dynamics simulations suggest that insertion efficiency is determined primarily by the energetic cost of distorting the bilayer in the vicinity of the TM helix. Very short lysine-flanked leucine segments can reduce the energetic cost by extensive hydrogen bonding with water and lipid phosphate groups (snorkeling) and by partial unfolding.

Evaluation of the communication of nursing students in the simulated teleconsultation: A cross-sectional study.

INTRODUCTION: The COVID-19 pandemic presented universities with the challenge of virtualizing the teaching-learning process. Simulated teleconsultation has been used in undergraduate training, which allows nursing students to interact with simulated patients remotely. Studies have identified that distance imposes communication barriers on all elements-sender, receiver and message-and in both forms of transmission: verbal and nonverbal. OBJECTIVE: To describe the communication of nursing students in teleconsultation with simulated patients in the context of primary health care. METHODS: This was a descriptive, cross-sectional study of 92 fifth-year nursing students. The communication variable was measured with the Connect Identify Understand Agree Help scale. In the data analysis, the mean and standard deviation of the scores on the 29 items of the scale were determined, as were the mean values for the total scale and for the 3 domains of the scale. RESULTS: The items that presented an average of less than 1 were primarily those related to the Agree and Help to Act domain. The total mean was 1.15, and the means for the domains Connect, Identify and Understand Problems and Agree and Help to Act were 1.53, 0.90 and 1.28, respectively. A weakness in the exploration of the psychosocial context of the simulated patients was observed. CONCLUSIONS: The results of this study, which evaluated communication in the context of distance care, corroborate the evidence regarding communication in real or simulated face-to-face situations. Studies that compare communication in various teaching-learning contexts, whether real or virtual, face-to-face or at a distance, are recommended.

Membrane partitioning: "classical" and "nonclassical" hydrophobic effects.

The free energy of transfer of nonpolar solutes from water to lipid bilayers is often dominated by a large negative enthalpy rather than the large positive entropy expected from the hydrophobic effect. This common observation has led to the idea that membrane partitioning is driven by the "nonclassical" hydrophobic effect. We examined this phenomenon by characterizing the partitioning of the well-studied peptide melittin using isothermal titration calorimetry (ITC) and circular dichroism (CD). We studied the temperature dependence of the entropic (-TΔS) and enthalpic (ΔH) components of free energy (ΔG) of partitioning of melittin into lipid membranes made of various mixtures of zwitterionic and anionic lipids. We found significant variations of the entropic and enthalpic components with temperature, lipid composition and vesicle size but only small changes in ΔG (entropy-enthalpy compensation). The heat capacity associated with partitioning had a large negative value of about -0.5 kcal mol(-1) K(-1). This hallmark of the hydrophobic effect was found to be independent of lipid composition. The measured heat capacity values were used to calculate the hydrophobic-effect free energy ΔG (hΦ), which we found to dominate melittin partitioning regardless of lipid composition. In the case of anionic membranes, additional free energy comes from coulombic attraction, which is characterized by a small effective peptide charge due to the lack of additivity of hydrophobic and electrostatic interactions in membrane interfaces [Ladokhin and White J Mol Biol 309:543-552, 2001]. Our results suggest that there is no need for a special effect-the nonclassical hydrophobic effect-to describe partitioning into lipid bilayers.

Sudden Death without a Clear Cause after Comprehensive Investigation: An Example of Forensic Approach to Atypical/Uncertain Findings.

Sudden death (SD) is defined as the unexpected natural death occurred within an hour after the onset of symptoms or from the last moment the subject has been seen in a healthy condition. Brugada syndrome (BrS) is one of the most remarkable cardiac causes of SD among young people. We report the case of a 20-year-old man who suddenly died after reportedly having smoked cannabis. Autopsy, toxicology, and genetic testing were performed. Autopsy found a long and thick myocardial bridging (MB) at 2 cm from the beginning of the left anterior descending coronary artery. Furthermore, at the histopathological examination, fibrosis and disarray in myocardial area above the MB, fatty tissue in the right ventricle and fibrosis of the sino-atrial node area were found. Toxicology testing was inconclusive, while genetic testing found a rare missense variant of the TTN gene, classified as likely benign, and a variant of unknown significance in the SLMAP gene (a gene that can be associated with BrS). Hence, despite several atypical features were found, no inference on the cause of the death could be made under current evidence.

CD spectroscopy of peptides and proteins bound to large unilamellar vesicles.

Circular dichroism (CD) spectroscopy is an essential tool for determining the conformation of proteins and peptides in membranes. It can be particularly useful for measuring the free energy of partitioning of peptides into lipid vesicles. The belief is broadly held that such CD measurements can only be made using sonicated small unilamellar vesicles (SUVs) because light scattering associated with extruded large unilamellar vesicles (LUVs) is unacceptably high. We have examined this issue using several experimental approaches in which a chiral object (i.e., peptide or protein) is placed both on the membrane and outside the membrane. We show that accurate CD spectra can be collected in the presence of LUVs. This is important because SUVs, unlike LUVs, are metastable and consequently unsuitable for equilibrium thermodynamic measurements. Our data reveal that undistorted CD spectra of peptides can be measured at wavelengths above 200 nm in the presence of up to 3 mM LUVs and above 215 nm in the presence of up to 7 mM LUVs. We introduce a simple way of characterizing the effect on CD spectra of light scattering and absorption arising from suspensions of vesicles of any diameter. Using melittin as an example, we show that CD spectroscopy can be used to determine the fractional helical content of peptides in LUVs and to measure their free energy of partitioning of into LUVs.

Intervención participativa comunitaria: necesidades de salud de las personas mayores durante el período de la pandemia por COVID-19 / Community participatory intervention: health needs of older people during the COVID-19 pandemic period

Objetivo. Proponer una intervención participativa comunitaria según las necesidades de salud percibida por las personas mayores del Programa de Salud Cardiovascular de un centro de salud familiar y comunitaria de una comuna de Santiago de Chile durante el período de la pandemia de la COVID-19. Métodos. Se utilizó una metodología participativa de la comunidad, la cual se desarrolló en dos fases: diagnóstico participativo comunitario y desarrollo de la propuesta participativa de una intervención en salud comunitaria. Resultados. Desde el diagnóstico participativo comunitario, los participantes eligieron el desarrollo de un programa educativo fundamentado en las necesidades de salud, además de la entrega de material educativo sobre estilos de vida saludable y recomendaciones para tener un buen control de sus enfermedades. Conclusión. Desde el diagnóstico y planificación participativa, es posible intervenir en pro de las necesidades de las personas, familias y comunidades y capacitarlas en el manejo de su propia salud. (AU)

Aim. To propose a community participatory intervention according to the health needs perceived by elderly people in the Family and Community Health Centre Cardiovascular Health programme of a Santiago de Chile commune during the COVID-19 pandemic. Methods. A community participatory methodology was used, which was developed over two phases: community participatory diagnosis and development of a participatory proposal for a community health intervention. Results. From the community participatory diagnosis, participants opted for development of an educational programme based on health needs, in addition to the delivery of educational material related to healthy lifestyles and recommendations for robust control of their diseases. Conclusion. From diagnosis and participatory planning, it is possible to intervene in favour of the needs of individuals, families and communities and train them in managing their own health. (AU)

Folding amphipathic helices into membranes: amphiphilicity trumps hydrophobicity.

High amphiphilicity is a hallmark of interfacial helices in membrane proteins and membrane-active peptides, such as toxins and antimicrobial peptides. Although there is general agreement that amphiphilicity is important for membrane-interface binding, an unanswered question is its importance relative to simple hydrophobicity-driven partitioning. We have examined this fundamental question using measurements of the interfacial partitioning of a family of 17-residue amidated-acetylated peptides into both neutral and anionic lipid vesicles. Composed only of Ala, Leu, and Gln residues, the amino acid sequences of the peptides were varied to change peptide amphiphilicity without changing total hydrophobicity. We found that peptide helicity in water and interface increased linearly with hydrophobic moment, as did the favorable peptide partitioning free energy. This observation provides simple tools for designing amphipathic helical peptides. Finally, our results show that helical amphiphilicity is far more important for interfacial binding than simple hydrophobicity.

Liposome destabilization induced by synthetic lipopeptides corresponding to envelope and non-structural domains of GBV-C/HGV virus. Conformational requirements for leakage.

Liposomes have been used primarily as a model system for studying biological membranes. Numerous chemical, biochemical and biophysical methods have been used to elucidate the various aspects of the interaction between proteins or peptides and phospholipids. Having in mind the potential use of synthetic lipopeptides as antiviral therapies and aiming for a better understanding of the molecular interaction of the GBV-C/HGV with liposomes as model membranes, epitopes of GBV-C/HGV located at the E2 (99-118) and NS3(440-460) regions were selected. Peptides were modified at the N-terminus with acyl chains of different length (C(14) and C(16)) yielding the corresponding myristoil and palmytoil lipopeptides. The main aim of the present study was to get insight into the membrane-interacting properties of the above-described synthetic lipopeptides and to study their inhibition of the capacity of perturbing model membranes of fusion peptide of HIV-1 using fluorescence spectroscopy. In an attempt to establish a relationship between peptide membrane activity and structure, we use Circular Dichroism (CD) and Fourier-Transform Infrared Spectroscopy (FTIR).

Percepción de usuarios de servicios de salud sobre la visibilidad social de los profesionales de enfermería durante la pandemia de COVID-19

Objetivo: Conocer la percepción de usuarios de los servicios de salud de tres comunas de Chile sobre la visibilidad social de los profesionales de enfermería durante el período de la pandemia de COVID-19. Material y Método: Estudio cualitativo descriptivo que utilizó la técnica del grupo focal. Se realizó un grupo focal con un muestreo de tipo intencionado conformado por siete participantes cuyos criterios de inclusión fueron ser usuarios del sistema de salud de Chile y mayor de 18 años. El reclutamiento de los participantes fue realizado por un afiche en redes sociales de la institución académica. La estrategia de análisis fue el de contenido. Resultados: Se encontraron tres temas y siete subtemas: la visibilidad de la profesión (imaginarios sociales, valoración de la labor, invisibilidad), primera línea (revaloración de los profesionales, seguridad y confianza) y género (profesión feminizada, invisibilidad). Conclusiones: Bajo el actual contexto de salubridad, la visibilidad de la enfermería se ha sostenido por situaciones que justamente son asociadas a la invisibilidad, salud mental, cargas laborales, cuestiones de género, entre otras. Se sugiere la realización de investigaciones que demuestren y difundan el importante rol de los profesionales de enfermería en la sociedad y promuevan su reconocimiento.

Objetivo: Conhecer a percepção de usuários dos serviços de saúde de três municípios do Chile sobre a visibilidade social dos profissionais de enfermagem durante o período da Pandemia da COVID-19. Material e Método: Estudo qualitativo descritivo que utilizou a técnica de grupo focal. Foi realizado um grupo focal com uma amostragem do tipo intencional, composta por sete participantes, cujos critérios de inclusão foram ser usuários do Sistema de Saúde do Chile e maiores de 18 anos. O recrutamento dos participantes foi realizado por um poster nas redes sociais da instituição acadêmica. A estratégia de análise foi de conteúdo. Resultados: Foram encontrados três temas e sete subtemas: a visibilidade da profissão (imaginários sociais, valorização do trabalho, invisibilidade), primeira linha (reavaliação dos profissionais, segurança e confiança) e gênero (profissão feminizada, invisibilidade). Conclusões: No contexto atual da saúde, a visibilidade da enfermagem tem sido sustentada por situações que se associam justamente à invisibilidade, saúde mental, carga de trabalho, questões de gênero, entre outras. Sugere-se a realização de pesquisas que demonstrem e divulguem o importante papel dos profissionais de enfermagem na sociedade e promovam seu reconhecimento.

Objective: To know the perception of users of health services in three communes of Chile regarding the social visibility of nursing professionals during the period of the COVID-19 Pandemic. Material and Method: Qualitative study with a descriptive design that used the focus group technique. A focus group was carried out with an intentional sampling, made up of seven participants whose inclusion criteria were: being users of the Chilean Health System and, older than 18 years. The recruitment of the participants was conducted by a poster in social networks of the academic institution. The analysis strategy was content. Results: Three themes and seven sub-themes were found. The themes were, the visibility of the profession (social imaginaries, value of work, invisibility), first line (revaluation of professionals, safety, and trust) and gender (feminized profession, invisibility). Conclusions: Under the current context of health, the visibility of nursing has been sustained by situations that are precisely associated with invisibility, mental health, workload, gender issues, among others. It is suggested that research be conducted to demonstrate and disseminate the significant role of nursing professionals in society and promote their recognition.

Case Report: Misleading Serological Diagnosis of Paracoccidioidomycosis in a Young Patient with the Acute Form Disease: Paracoccidioides brasiliensis or Paracoccidioides lutzii?

Negative results in serological routine screening of patients with microbiologically proven Paracoccidioidomycosis (PCM) are occasionally reported. Failure in detecting anti-Paracoccidioides antibodies has been ascribed to factors either related to serological techniques or to the status of the host immune reactivity. Recently, this issue has been renewed by the recognition that the Paracoccidioides genera comprises two species, lutzii and brasiliensis, which have distinct antigenic profiles and, therefore, may elicit different host antibody responses. We describe a patient with the acute form PCM due to Paracoccidioides brasiliensis with negative results on two reference centers' routine screening for P. brasiliensis antibodies, but positive results with Paracoccidioides lutzii antigens. The present case report suggests that antibodies elicited during P. brasiliensis infection recognize antigenic fractions shared by both species, highlighting the difficulties in distinguishing the two infections by means of the currently available routine serological assays.

Overall survival prediction in metastatic castration-resistant prostate cancer treated with radium-223

ABSTRACT Objective Radium-223(223Ra) is indicated for patients (p) with metastatic castration resistant prostate cancer (mCRCP). Objectives The aim of this study was to evaluate the role of baseline clinical variables associated with overall survival (OS) and toxicity of 223Ra. Its purpose was to identify the factors that can predict a better response to treatment and provide information regarding the most appropriate time for the application of 223Ra. Materials and Methods Prospective study in 40p with mCRPC treated with 223Ra. End points were OS, progression-free survival and time to progression. The follow-up parameters were: doses received, hemoglobin (Hb), absolute neutrophil count (ANC), platelet count (PC), prostate specific antigen (PSA), alkaline phosphatase (ALP), Visual Analogue Scale for pain, Eastern Cooperative Oncology Group (ECOG) and WHO's Cancer Pain Ladder. The use of other treatments was also evaluated. Results Median OS was 17.1 months(mo) (CI95%6.5-27.7); 26/40p received complete treatment of 223Ra, without reaching a median OS and 14p received incomplete treatment with a median OS 13.6mo(CI95%1.6-25.6). Median follow-up was 11.2mo (range:1.3-45.2). The univariate analysis showed that factors as VAS, ECOG, Hb and ALP values were independently associated with OS. First line treatment with 223Ra was started in 11/40p, while 19p had been heavily pre-treated and 13p received concomitant treatment. Conclusions 223Ra therapy require an adequate selection of patients to obtain the greatest clinical benefit. Low basal Hb, hight basal ALP, bone marrow involvement and an altered ECOG were the main factors that decreased OS in our patients. 223Ra should be considered relatively early in the course of treatment.

Genetic analysis, in silico prediction, and family segregation in long QT syndrome.

The heritable cardiovascular disorder long QT syndrome (LQTS), characterized by prolongation of the QT interval on electrocardiogram, carries a high risk of sudden cardiac death. We sought to add new data to the existing knowledge of genetic mutations contributing to LQTS to both expand our understanding of its genetic basis and assess the value of genetic testing in clinical decision-making. Direct sequencing of the five major contributing genes, KCNQ1, KCNH2, SCN5A, KCNE1, and KCNE2, was performed in a cohort of 115 non-related LQTS patients. Pathogenicity of the variants was analyzed using family segregation, allele frequency from public databases, conservation analysis, and Condel and Provean in silico predictors. Phenotype-genotype correlations were analyzed statistically. Sequencing identified 36 previously described and 18 novel mutations. In 51.3% of the index cases, mutations were found, mostly in KCNQ1, KCNH2, and SCN5A; 5.2% of cases had multiple mutations. Pathogenicity analysis revealed 39 mutations as likely pathogenic, 12 as VUS, and 3 as non-pathogenic. Clinical analysis revealed that 75.6% of patients with QTc≥500 ms were genetically confirmed. Our results support the use of genetic testing of KCNQ1, KCNH2, and SCN5A as part of the diagnosis of LQTS and to help identify relatives at risk of SCD. Further, the genetic tools appear more valuable as disease severity increases. However, the identification of genetic variations in the clinical investigation of single patients using bioinformatic tools can produce erroneous conclusions regarding pathogenicity. Therefore segregation studies are key to determining causality.

Evaluation of tests for antibody response in the follow-up of patients with acute and chronic forms of paracoccidioidomycosis.

Several serological tests have been used successfully in the diagnosis of paracoccidioidomycosis (PCM). In contrast, data about the use of these tests in the follow-up of PCM patients have been heterogeneous. In this study, serum samples from 43 PCM patients with different clinical forms were analysed by counter-immuno-electrophoresis (CIE), complement fixation (CF) and ELISA before treatment. With CIE and ELISA, the chronic unifocal form showed significantly lower antibody levels compared with chronic multifocal and acute forms. Acute form patients had significantly higher titres than patients with multifocal disease by CIE but not by ELISA. No significant differences were observed with CF. Twenty-seven of these patients were followed-up for 2 years and showed a decline in antibody levels by all three tests, paralleling clinical improvement. However, only patients with unifocal disease cleared their antibodies after 1 year of treatment as analysed by CF and ELISA and after 2 years by CIE, suggesting that these patients may need shorter courses of therapy. Patients with the other clinical form of the disease needed > or =2 years of therapy to clear their antibodies. Sera from a further five patients who presented with a relapse were analysed. At the time of relapse all showed increases in antibody levels by CIE and ELISA, but only three showed increases by CF tests. Therefore, CIE and ELISA demonstrated a better clinical correlation than CF, probably reflecting the fungal burden of PCM patients more accurately.