Patient-derived follicular lymphoma spheroids recapitulate lymph node signaling and immune profile uncovering galectin-9 as a novel immunotherapeutic target.

Follicular lymphoma (FL), the most common indolent non-Hodgkin lymphoma, constitutes a paradigm of immune tumor microenvironment (TME) contribution to disease onset, progression, and heterogenous clinical outcome. Here we present the first FL-Patient Derived Lymphoma Spheroid (FL-PDLS), including fundamental immune actors and features of TME in FL lymph nodes (LNs). FL-PDLS is organized in disc-shaped 3D structures composed of proliferating B and T cells, together with macrophages with an intermediate M1/M2 phenotype. FL-PDLS recapitulates the most relevant B-cell transcriptional pathways present in FL-LN (proliferation, epigenetic regulation, mTOR, adaptive immune system, among others). The T cell compartment in the FL-PDLS preserves CD4 subsets (follicular helper, regulatory, and follicular regulatory), also encompassing the spectrum of activation/exhaustion phenotypes in CD4 and CD8 populations. Moreover, this system is suitable for chemo and immunotherapy testing, recapitulating results obtained in the clinic. FL-PDLS allowed uncovering that soluble galectin-9 limits rituximab, rituximab, plus nivolumab/TIM-3 antitumoral activities. Blocking galectin-9 improves rituximab efficacy, highlighting galectin-9 as a novel immunotherapeutic target in FL. In conclusion, FL-PDLS maintains the crosstalk between malignant B cells and the immune LN-TME and constitutes a robust and multiplexed pre-clinical tool to perform drug screening in a patient-derived system, advancing toward personalized therapeutic approaches.

Enrichment of Immune Dysregulation Disorders in Adult Patients with Human Inborn Errors of Immunity.

Human inborn errors of immunity (IEI) comprise a group of diseases resulting from molecular variants that compromise innate and adaptive immunity. Clinical features of IEI patients are dominated by susceptibility to a spectrum of infectious diseases, as well as autoimmune, autoinflammatory, allergic, and malignant phenotypes that usually appear in childhood, which is when the diagnosis is typically made. However, some IEI patients are identified in adulthood due to symptomatic delay of the disease or other reasons that prevent the request for a molecular study. The application of next-generation sequencing (NGS) as a diagnostic technique has given rise to an ever-increasing identification of IEI-monogenic causes, thus improving the diagnostic yield and facilitating the possibility of personalized treatment. This work was a retrospective study of 173 adults with IEI suspicion that were sequenced between 2005 and 2023. Sanger, targeted gene-panel, and whole exome sequencing were used for molecular diagnosis. Disease-causing variants were identified in 44 of 173 (25.43%) patients. The clinical phenotype of these 44 patients was mostly related to infection susceptibility (63.64%). An enrichment of immune dysregulation diseases was found when cohorts with molecular diagnosis were compared to those without. Immune dysregulation disorders, group 4 from the International Union of Immunological Societies Expert Committee (IUIS), were the most prevalent among these adult patients. Immune dysregulation as a new item in the Jeffrey Model Foundation warning signs for adults significantly increases the sensitivity for the identification of patients with an IEI-producing molecular defect.

Patient-derived lymphoma spheroids integrating immune tumor microenvironment as preclinical follicular lymphoma models for personalized medicine.

BACKGROUND: Follicular lymphoma (FL), the most common indolent non-Hodgkin's Lymphoma, is a heterogeneous disease and a paradigm of the contribution of immune tumor microenvironment to disease onset, progression, and therapy resistance. Patient-derived models are scarce and fail to reproduce immune phenotypes and therapeutic responses. METHODS: To capture disease heterogeneity and microenvironment cues, we developed a patient-derived lymphoma spheroid (FL-PDLS) model culturing FL cells from lymph nodes (LN) with an optimized cytokine cocktail that mimics LN stimuli and maintains tumor cell viability. RESULTS: FL-PDLS, mainly composed of tumor B cells (60% on average) and autologous T cells (13% CD4 and 3% CD8 on average, respectively), rapidly organizes into patient-specific three-dimensional (3D) structures of three different morphotypes according to 3D imaging analysis. RNAseq analysis indicates that FL-PDLS reproduces FL hallmarks with the overexpression of cell cycle, BCR, or mTOR signaling related gene sets. FL-PDLS also recapitulates the exhausted immune phenotype typical of FL-LN, including expression of BTLA, TIGIT, PD-1, TIM-3, CD39 and CD73 on CD3+ T cells. These features render FL-PDLS an amenable system for immunotherapy testing. With this aim, we demonstrate that the combination of obinutuzumab (anti-CD20) and nivolumab (anti-PD1) reduces tumor load in a significant proportion of FL-PDLS. Interestingly, B cell depletion inversely correlates with the percentage of CD8+ cells positive for PD-1 and TIM-3. CONCLUSIONS: In summary, FL-PDLS is a robust patient-derived 3D system that can be used as a tool to mimic FL pathology and to test novel immunotherapeutic approaches in a context of personalized medicine.

Prevalence and clinical significance in our setting of incidental uptake in the thyroid gland found on 18F-fluordeoxyglucose positron emission tomography-computed tomography (PET-CT).

INTRODUCTION: The expanding use of 18F-fluorodeoxyglucose (FDG) positron emission tomography-computed tomography (PET-CT) has resulted in an increased frequency of incidentally discovered areas of FDG uptake within the thyroid gland. In these incidentalomas, high malignancy rates are reported. The study aimed, on the one hand, to determine the prevalence in our setting of thyroid incidentalomas in patients with no previous history of thyroid cancer undergoing an FDG PET-CT as well as the risk of malignancy and, on the other hand, to evaluate the usefulness of the maximum standard uptake value (SUVmax) for detecting thyroid cancer. MATERIAL AND METHODS: The FDG PET-CT scans performed at our hospital between June 2013 and December 2020 were retrospectively reviewed. In those incidentalomas with sufficient additional investigation, a diagnosis of benign or malignant was established based on the complementary tests. RESULTS: From the 21,594 PET-CT scans performed, 398 (1.8%) patients had an incidental FDG uptake, either focal (n=324) or diffuse (n=74). Among incidentalomas with further investigation, the rate of malignancy was higher in patients with focal FDG uptake than in those with diffuse uptake (26.5% versus 4%, respectively, p<0.05). The SUVmax value was significantly lower in benign focal lesions (5.7 [range: 2.3-66] than in malignant ones 10.6 [range: 3.1-51.2]; p<0.05). Nearly a quarter of malignant diagnoses (23.3%) were related to potentially aggressive tumours. CONCLUSION: The high rate of malignant tumours found among PET-CT incidentalomas and the high proportion of aggressive tumours demonstrate the need for a standardised approach in the investigation of incidental focal FDG uptake in the thyroid gland.

A novel patient-derived 3D model recapitulates mantle cell lymphoma lymph node signaling, immune profile and in vivo ibrutinib responses.

Mantle cell lymphoma (MCL), a rare and aggressive B-cell non-Hodgkin lymphoma, mainly develops in the lymph node (LN) and creates a protective and immunosuppressive niche that facilitates tumor survival, proliferation and chemoresistance. To capture disease heterogeneity and tumor microenvironment (TME) cues, we have developed the first patient-derived MCL spheroids (MCL-PDLS) that recapitulate tumor oncogenic pathways and immune microenvironment in a multiplexed system that allows easy drug screening, including immunotherapies. MCL spheroids, integrated by tumor B cells, monocytes and autologous T-cells self-organize in disc-shaped structures, where B and T-cells maintain viability and proliferate, and monocytes differentiate into M2-like macrophages. RNA-seq analysis demonstrated that tumor cells recapitulate hallmarks of MCL-LN (proliferation, NF-kB and BCR), with T cells exhibiting an exhaustion profile (PD1, TIM-3 and TIGIT). MCL-PDLS reproduces in vivo responses to ibrutinib and demonstrates that combination of ibrutinib with nivolumab (anti-PD1) may be effective in ibrutinib-resistant cases by engaging an immune response with increased interferon gamma and granzyme B release. In conclusion, MCL-PDLS recapitulates specific MCL-LN features and in vivo responses to ibrutinib, representing a robust tool to study MCL interaction with the immune TME and to perform drug screening in a patient-derived system, advancing toward personalized therapeutic approaches.

Prediction of Smoking Habits From Class-Imbalanced Saliva Microbiome Data Using Data Augmentation and Machine Learning.

Human microbiome research is moving from characterization and association studies to translational applications in medical research, clinical diagnostics, and others. One of these applications is the prediction of human traits, where machine learning (ML) methods are often employed, but face practical challenges. Class imbalance in available microbiome data is one of the major problems, which, if unaccounted for, leads to spurious prediction accuracies and limits the classifier's generalization. Here, we investigated the predictability of smoking habits from class-imbalanced saliva microbiome data by combining data augmentation techniques to account for class imbalance with ML methods for prediction. We collected publicly available saliva 16S rRNA gene sequencing data and smoking habit metadata demonstrating a serious class imbalance problem, i.e., 175 current vs. 1,070 non-current smokers. Three data augmentation techniques (synthetic minority over-sampling technique, adaptive synthetic, and tree-based associative data augmentation) were applied together with seven ML methods: logistic regression, k-nearest neighbors, support vector machine with linear and radial kernels, decision trees, random forest, and extreme gradient boosting. K-fold nested cross-validation was used with the different augmented data types and baseline non-augmented data to validate the prediction outcome. Combining data augmentation with ML generally outperformed baseline methods in our dataset. The final prediction model combined tree-based associative data augmentation and support vector machine with linear kernel, and achieved a classification performance expressed as Matthews correlation coefficient of 0.36 and AUC of 0.81. Our method successfully addresses the problem of class imbalance in microbiome data for reliable prediction of smoking habits.

Is perioperative brachytherapy effective in carcinoma of the tongue?

Purpose: To analyze the results of patients treated with perioperative interstitial brachytherapy (ISBT) in tongue carcinoma (TC). Material and methods: From April 2009 to May 2015, 43 squamous cell carcinoma consecutive patients diagnosed with TC were treated with limited partial glossectomy and perioperative ISBT, using high-dose-rate (HDR). Twenty- seven patients were treated by brachytherapy (BT), and sixteen received BT as a complement to subsequent external beam radiotherapy (EBRT) after results of lymph node dissection. Median age was 66 years. Distribution by stage, included 10 patients stage I, 14 stage II, 10 stage III, and 9 stage IV. Eighteen patients had negative margins, nineteen margin involvement, and in six cases, the margin was < 5 mm. Results: With a median follow-up of 54 months, LC at 3 and 5 years was 87% and 84%, respectively. LC was 95% at five years in patients with clear margins, and 75% with involved margins. LC in N0 patients treated with BT was 83% at 5 years, and in patients N+ with posterior EBRT treatment, LC was 86%. By tumor size, we found one local relapse in 13 cases T1, in 5 of 27 patients T2 was found, and no local relapse T3 with LC of 87%, 70%, and 100% respectively at five years. Regional control (RC) was 81% at 3 and 5 years. We found a metastasis-free survival of 91% at 3- and 5-year. Twenty-three patients have died, 11 of them due to other causes, with overall survival of 56% at three years and 53% at five years. Conclusions: Combined treatment with conservative surgery and ISBT shows similar results to radical surgery or RT alone, allowing a more patient-tailored approach, with good organ function preservation and cosmetic outcomes.

Next Generation Sequencing for Detecting Somatic FAS Mutations in Patients With Autoimmune Lymphoproliferative Syndrome.

Autoimmune lymphoproliferative syndrome (ALPS) is a primary immune regulatory disorder clinically defined by chronic and benign lymphoproliferation, autoimmunity and an increased risk of lymphoma due to a genetic defect in the FAS-FASL apoptotic pathway. Genetic defects associated with ALPS are germinal and somatic mutations in FAS gene, in addition to germinal mutations in FASLG, FADD, CASP8 and CASP10 genes. The accumulation of CD3+TCRαß+CD4-CD8- double negative T-cells (DNT) is a hallmark of the disease and 20-25% of ALPS patients show heterozygous somatic mutations restricted to DNT in the FAS gene (ALPS-sFAS patients). Nowadays, somatic mutations in the FAS gene are detected through Sanger sequencing in isolated DNT. In this study, we report an ALPS-sFAS patient fulfilling clinical and laboratory ALPS criteria, who was diagnosed through NGS with a targeted gene panel using DNA from whole blood. Data analysis was carried out with Torrent Suite Software and variant detection was performed by both germinal and somatic variant caller plugin. The somatic variant caller correctly detected other six ALPS-sFAS patients previously diagnosed in the authors' laboratories. In summary, this approach allows the detection of both germline and somatic mutations related to ALPS by NGS, avoiding the isolation of DNT as the first step. The reads of the somatic variants could be detected even in patients with DNT in the cut off limit. Thus, custom-designed NGS panel testing may be a faster and more reliable method for the diagnosis of new ALPS patients, including those with somatic FAS mutations (ALPS-sFAS).

3D Model Characterization by 2D and 3D Imaging in t(14;18)-Positive B-NHL: Perspectives for In Vitro Drug Screens in Follicular Lymphoma.

Follicular lymphoma (FL) is an indolent B cell lymphoproliferative disorder of transformed follicular center B cells, which accounts for 20-30 percent of all non-Hodgkin lymphoma (NHL) cases. Great advances have been made to identify the most relevant targets for precision therapy. However, no relevant models for in vitro studies have been developed or characterized in depth. To this purpose, we generated a 3D cell model from t(14;18)-positive B-NHL cell lines cultured in ultra-low attachment 96-well plates. Morphological features and cell growth behavior were evaluated by classical microscopy (2D imaging) and response to treatment with different drugs was evaluated by a high-content analysis system to determine the robustness of the model. We show that the ultra-low attachment (ULA) method allows the development of regular, spherical and viable ULA-multicellular aggregates of lymphoma cells (MALC). However, discrepancies in the results obtained after 2D imaging analyses on drug-treated ULA-MALC prompted us to develop 3D imaging and specific analyses. We show by using light sheet microscopy and specifically developed 3D imaging algorithms that 3D imaging and dedicated analyses are necessary to characterize morphological properties of 3D models and drug effects. This study proposes a new method, but also imaging tools and informatic solutions, developed for FL necessary for future preclinical studies.

Resistencia a múltiples antibióticos en serovariedades de Salmonella aisladas de muestras clínicas y alimentos / Resistance to multiple antibiotics in serovarities of Salmonella isolated from clinical samples and foods

Salmonella enterica es un patógeno transmitido por alimentos y agente etiológico de brotes alimentarios de gran impacto en la salud humana. El aumento de la resistencia bacteriana constituye una amenaza a la salud pública, la aparición de cepas de Salmonella con resistencia a múltiples antimicrobianos (MDR) fue descrita en humanos, alimentos y animales para consumo; por ello se considera muy importante conocer la situación epidemiológica local. El objetivo de este trabajo fue generar información sobre los serotipos circulantes, resistencia a los antibióticos y presencia de resistencia simultánea a múltiples fármacos en Salmonella provenientes de muestras clínicas humanas y muestras de alimentos en el periodo desde 2017 a 2019. Fueron analizadas un total de 668 cepas de Salmonella aisladas en los años 2017, 2018 y 2019 a partir de muestras clínicas humanas y de alimentos, en el Laboratorio Central de Salud Pública y/o remitidas por Laboratorios de la Red de Enteropatógenos. Se observaron serotipos muy diversos con prevalencia del serovar Heidelberg en alimentos y Typhimurium en muestras de humanos. Se encontró que el 45,4% de las cepas fueron sensibles a todos los antibióticos (ATB), el 35,6% fueron resistentes de 1 a 6 ATB y el 19% con sensibilidad intermedia; observándose mayor resistencia a Tetraciclina, Ác. Nalidíxico, Ampicilina y Nitrofurantoína, en menor grado se evidenció resistencia a cefalosporinas (C3ªG) y a ciprofloxacina. El 16.9% de las cepas presentaron resistencia múltiple (3 o más antibióticos) con 37 fenotipos distintos. Las serovariedades que presentaron mayor resistencia a los antimicrobianos fueron Heidelberg, Schwarzengrund y Typhimurium

Salmonella enterica is a foodborne pathogen and etiological agent of food outbreaks with a great impact on human health. The increase in bacterial resistance constitutes a threat to public health. The appearance of Salmonella strains with resistance to multiple antimicrobials (MDR) has already been described in humans, food and animals for consumption; for this reason, it is considered very important to know the local epidemiological situation. The target of this work was to generate information on circulating serotypes, antibiotic resistance and the presence of simultaneous resistance to multiple drugs in Salmonella from human clinical samples and food samples in the period from 2017 to 2019. A total of 668 Salmonella strains isolated in the years 2017, 2018 and 2019 were analyzed from human and food clinical samples, at the Central Public Health Laboratory and / or sent by Laboratories of the Enteropathogens Network. Very diverse serotypes were observed with prevalence of Heidelberg serovar in food and Typhimurium in human samples .It was found that 45,4% of the strains were sensitive to all antibiotics (ATB), 35,6% were resistant from 1 to 6 ATB and 19% with intermediate sensitivity; observing greater resistance to Tetracycline, Ác. Nalidixic, Ampicillin and Nitrofurantoin, to a lesser degree resistance to cephalosporins (C3ªG) and ciprofloxacin was evidenced. The 16.9% the strains presented multiple resistance (3 or more antibiotics) with 37 different phenotypes. The serovars with the highest antimicrobial resistance were Heidelberg, Schwarzengrund and Typhimurium

Follicular Lymphoma Microenvironment: An Intricate Network Ready for Therapeutic Intervention.

Follicular Lymphoma (FL), the most common indolent non-Hodgkin's B cell lymphoma, is a paradigm of the immune microenvironment's contribution to disease onset, progression, and heterogeneity. Over the last few years, state-of-the-art technologies, including whole-exome sequencing, single-cell RNA sequencing, and mass cytometry, have precisely dissected the specific cellular phenotypes present in the FL microenvironment network and their role in the disease. In this already complex picture, the presence of recurring mutations, including KMT2D, CREBBP, EZH2, and TNFRSF14, have a prominent contributory role, with some of them finely tuning this exquisite dependence of FL on its microenvironment. This precise characterization of the enemy (FL) and its allies (microenvironment) has paved the way for the development of novel therapies aimed at dismantling this contact network, weakening tumor cell support, and reactivating the host's immune response against the tumor. In this review, we will describe the main microenvironment actors, together with the current and future therapeutic approaches targeting them.

Curso masivo en línea "Salud sexual y reproductiva": cambios en conocimientos, deserción, reprobación y satisfacción / Massive online course "Sexual and reproductive health": changes in knowledge, desertion, disapproval and satisfaction

Resumen Objetivo: Presentar resultados globales de la deserción, reprobación y satisfacción de los usuarios del Curso Masivo en Línea Abierto (MOOC, por sus siglas en inglés), así como los cambios en conocimientos y prácticas del personal de salud inscrito al MOOC "Salud sexual y reproductiva y prevención del embarazo en adolescentes". Material y métodos: Se analizaron 45 549 registros y se ajustaron modelos de regresión logística para la deserción y reprobación del curso. Resultados: Finalizó el curso 57.8% de los usuarios, mientras que 15.1% reprobó. Los modelos logísticos muestran que la edad y el cargo ocupado se asocian tanto con la deserción como con la reprobación. Conclusiones: El MOOC es una herramienta útil para generar mejoras en conocimientos y competencias. Es necesario ampliar la cobertura a profesionales que tienen trato con población adolescente, con el fin de mejorar la salud sexual y reproductiva y prevenir embarazos en la adolescencia.

Abstract Objective: To present global results of the dropout and failure and satisfaction of the Massive Open Online Course (MOOC) users, as well as changes in knowledge and practices of personnel registered in "Sexual and Reproductive Health and Prevention of adolescent pregnancy". Materials and methods: Using 45 549 records, we adjust logistic regression models to find associations between variables of dropout and failure. Results: 57.8% of people completed the course and 15.1% failure. Logistic models show that age and position are associated with both dropout and failure. Conclusions: The MOOC is useful to improve knowledge and skills of health personnel. It is necessary to expand the coverage to professionals who deal with adolescent population, in order to improve sexual and reproductive health and prevent teenage pregnancies.

Does surgical technique influence post-tonsillectomy haemorrhage? Our experience.

INTRODUCTION AND OBJECTIVES: Tonsillectomy represents one of the main surgical procedures for the otolaryngologist, with haemorrhage being the most common postoperative. The objective of this study was to determine the post-tonsillectomy haemorrhage rate, and relate the surgical technique, diagnosis and patient age. METHODS: This was a retrospective study, from April 2012 to January 2014, covering 429 patients. We used the following surgical-dissection techniques: cold, Colorado needle and monopolar forceps. Haemostasis was carried out in every case with monopolar forceps and gauze compression. RESULTS: Post-tonsillectomy haemorrhage rate was 6.99%. According to the surgical technique used, with Colorado needle dissection, the bleeding rate we found was 7.07%; with monopolar forceps dissection, the rate was 20.4%; and with cold dissection, 2.9%. Of all haemorrhages, only 9 (2.09%) needed reintervention, of which 40% were performed with monopolar forceps dissection. The group with the largest bleeding rate was that of more than 14 years old. The diagnosis most associated with bleeding was peritonsillar abscess. CONCLUSION: The lowest bleeding rate was found with cold dissection and monopolar forceps haemostasis (2.09%). Consequently, based on our experience and the results obtained in the study, we consider that surgical technique does influence post-tonsillectomy haemorrhage.

Anthelmintic effect of carob pods and sainfoin hay when fed to lambs after experimental trickle infections with Haemonchus contortus and Trichostrongylus colubriformis.

The aim of the study was to compare the in vivo anthelmintic activity of sainfoin hay (Onobrychis viciifolia) and carob pod meal (Ceratonia siliqua) against gastrointestinal nematodes. Seven days before infection, 64 naive lambs were assigned to four different groups: Group S received sainfoin hay and group CAR was fed with carob pods. The remaining lambs received lucerne hay (Medicago sativa) and were assigned to positive (non-treated, NT) and negative (treated, T) control groups (treatment with albendazole). On day 0, lambs were artificially trickle infected for 6 weeks, with a mixture of infective larvae of Haemonchus contortus and Trichostrongylus colubriformis. Parasitological and pathophysiological parameters were measured repeatedly during the 2-month study. Compared to the NT group, decreases in egg excretion were observed in the CAR and S groups with significant differences only found for sainfoin (p < 0.05). At necropsy, group S showed decreases in the total worm numbers of both nematode species with significant differences for H. contortus. In contrast, no differences were noticed for the CAR group. Compared to the NT group, lower values for fecundity of female H. contortus were found in the S and CAR groups, however differences were non-significant. No differences in body weight gains were found between groups. Consistent results were found showing significantly higher packed cell volume (PCV) values in the T and S groups compared to NT and CAR groups. Overall, these results confirm a positive effect associated with the feeding of lambs with tanniniferous resources on host resilience (PCV values) and against gastrointestinal parasitic nematodes by affecting some biological traits of worm populations (e.g. eggs per gram of faeces and worm numbers). However, the anthelmintic effects differed between the two tannin-containing resources, which might be associated with the quantity and/or quality of secondary metabolites (condensed tannins and/or other polyphenols).

[Methodology for evaluation of malignancy screening in surgical thyroid gland disease]. / Evaluación metodológica para detección de malignidad en patología quirúrgica tiroidea.

INTRODUCTION: We review a methodology for detection of malignancy in thyroid gland surgery, comparing clinical exploration, ultrasonography, gammagraphy, fine-needle aspiration (FNA) and extemporaneous biopsy with the definitive pathological results. MATERIAL AND METHODS: We carried out a retrospective study on a sample of 433 (N=433) patients who had been intervened due to thyroid gland disease at the same ENT centre between 1999 and 2004. We collected information focusing on the characteristics of cervical exploration, ultrasonography, gammagraphy, FNA, extemporaneous biopsy and definitive pathological results. RESULTS: With N=433, the male/female ratio was approximately 1/9 (m/f 1/9), the average age was 45.5 years, with a range between 13 and 87 years, and with 20.3% of referred family history of thyroid gland disease; the cervical exploration, ultrasonography ang gammagraphy were not good tests for the screening of malignancy; FNA and extemporaneous biopsy offered 74% and 81% of sensitivity (Se), and 73% and 92% of specificity (Sp) in the detection of malignancy. The specific sensitivities towards papillary carcinoma of FNA and extemporaneous biopsy were 81% and 88%, whilst for follicular carcinoma they were 66% and 62.5%, respectively. CONCLUSIONS: FNA has good sensitivity in clinical suspicion of thyroid papillary carcinoma, although it should be complemented by extemporaneous biopsy due to its better specificity. In the case of follicular carcinoma there is a tendency towards conservative treatment, without carrying out extemporaneous biopsy previously and with reintervention in case of definitive malignancy.

[Laryngeal paraganglioma: diagnosis and treatment. A propos of a case]. / Paraganglioma laríngeo, diagnóstico y tratamiento. A propósito de un caso.

Paragangliomas of the larynx (LPG) are unusual neuroendocrine tumors that are seen as a vascular submucosal mass, with a female predilection. We add another case of LPG to the 77 previously reported in the literature. We present the clinical, radiologic, and surgical features of a supraglottic LPG seen in a 40-year-old woman with a 8-month history of slowly progressive hoarseness. The tumor was removed approaching a lateral thyrotomy. The patient remains disease free 18 months after surgery. Preoperative CT, angiography, and embolization are useful in making the diagnosis and reducing perisurgical bleeding of LPG before surgical intervention. Complete excision through an external mucosa-sparing approach is the treatment of choice.

m.6267G>A: a recurrent mutation in the human mitochondrial DNA that reduces cytochrome c oxidase activity and is associated with tumors.

Complete sequencing of the mitochondrial genome of 13 cell lines derived from a variety of human cancers revealed nine novel mitochondrial DNA (mtDNA) variations. One of them, m.6267G>A, is a recurrent mutation that introduces the Ala122Thr substitution in the mitochondrially encoded cytochrome c oxidase I (MT-CO1): p.MT-CO1: Ala122Thr (GenBank: NP_536845.1). Biochemical analysis of the original cell lines and the transmitochondrial cybrids generated by transferring mitochondrial DNAs to a common nuclear background, indicate that cytochrome c oxidase (COX) activity, respiration, and growth in galactose are impaired by the m.6267G>A mutation. This mutation, found twice in the cancer cell lines included in this study, has been also encountered in one out of 63 breast cancer samples, one out of 64 colon cancer samples, one out of 260 prostate cancer samples, and in one out of 15 pancreatic cancer cell lines. In all instances the m.6267G>A mutation was associated to different mtDNA haplogroups. These findings, contrast with the extremely low frequency of the m.6267G>A mutation in the normal population (1:2264) and its apparent absence in other pathologies, strongly suggesting that the m.6267G>A missense mutation is a recurrent mutation specifically associated with cancer.

Trichinella spiralis secretes a homologue of prosaposin.

Infective larvae and adult stage Trichinella spiralis secrete a protein homologous to prosaposin, the precursor of sphingolipid activator proteins (saposins) A-D originally defined in vertebrates. The protein contains four saposin domains, with the six cysteine residues which form the three intramolecular disulphide bonds in close register in each case. It differs substantially from vertebrate prosaposins in the N-terminal prodomain, the region separating saposins A and B, and completely lacks the C-terminal domain which has been demonstrated to be essential for lysosomal targetting in these organisms. The protein is secreted in unprocessed form with an estimated mass of 56 kDa, and contains a single N-linked glycan which is bound by the monoclonal antibody NIM-M1, characteristic of the TSL-1 antigens which are capped by tyvelose (3,6-dideoxy-D-arabinohexose). Immuno-electron microscopy localised the protein to membrane-bound vesicles and more complex multi-lamellar organelles in diverse tissues including the hypodermis, intestine and stichosomes, although it was absent from the dense-core secretory granules typical of the latter. Possible functions of a secreted prosaposin are discussed.