Clonal competition assays identify fitness signatures in cancer progression and resistance in multiple myeloma.

Multiple myeloma (MM) is a genetically heterogeneous disease and the management of relapses is one of the biggest clinical challenges. TP53 alterations are established high-risk markers and are included in the current disease staging criteria. KRAS is the most frequently mutated gene affecting around 20% of MM patients. Applying Clonal Competition Assays (CCA) by co-culturing color-labeled genetically modified cell models, we recently showed that mono- and biallelic alterations in TP53 transmit a fitness advantage to the cells. Here, we report a similar dynamic for two mutations in KRAS (G12A and A146T), providing a biological rationale for the high frequency of KRAS and TP53 alterations at MM relapse. Resistance mutations, on the other hand, did not endow MM cells with a general fitness advantage but rather presented a disadvantage compared to the wild-type. CUL4B KO and IKZF1 A152T transmit resistance against immunomodulatory agents, PSMB5 A20T to proteasome inhibition. However, MM cells harboring such lesions only outcompete the culture in the presence of the respective drug. To better prevent the selection of clones with the potential of inducing relapse, these results argue in favor of treatment-free breaks or a switch of the drug class given as maintenance therapy. In summary, the fitness benefit of TP53 and KRAS mutations was not treatment-related, unlike patient-derived drug resistance alterations that may only induce an advantage under treatment. CCAs are suitable models for the study of clonal evolution and competitive (dis)advantages conveyed by a specific genetic lesion of interest, and their dependence on external factors such as the treatment.

Identification of Immunoglobulin Gene Rearrangement Biomarkers in Multiple Myeloma through cfDNA-Based Liquid Biopsy Using tchDNA-Seq.

Multiple myeloma (MM) is a hematological malignancy characterized by the clonal proliferation of pathogenic CD138+ plasma cells (PPCs) in bone marrow (BM). Recent years have seen a significant increase in the treatment options for MM; however, most patients who achieve complete the response ultimately relapse. The earlier detection of tumor-related clonal DNA would thus be very beneficial for patients with MM and would enable timely therapeutic interventions to improve outcomes. Liquid biopsy of "cell-free DNA" (cfDNA) as a minimally invasive approach might be more effective than BM aspiration not only for the diagnosis but also for the detection of early recurrence. Most studies thus far have addressed the comparative quantification of patient-specific biomarkers in cfDNA with PPCs and BM samples, which have shown good correlations. However, there are limitations to this approach, such as the difficulty in obtaining enough circulating free tumor DNA to achieve sufficient sensitivity for the assessment of minimal residual disease. Herein, we summarize current data on methodologies to characterize MM, and we present evidence that targeted capture hybridization DNA sequencing (tchDNA-Seq) can provide robust biomarkers in cfDNA, including immunoglobulin (IG) rearrangements. We also show that detection can be improved by prior purification of the cfDNA. Overall, liquid biopsies of cfDNA to monitor IG rearrangements have the potential to provide important diagnostic, prognostic, and predictive information in patients with MM.

CNL and aCML should be considered as a single entity based on molecular profiles and outcomes.

Chronic neutrophilic leukemia (CNL) and atypical chronic myeloid leukemia (aCML) are rare myeloid disorders that are challenging with regard to diagnosis and clinical management. To study the similarities and differences between these disorders, we undertook a multicenter international study of one of the largest case series (CNL, n = 24; aCML, n = 37 cases, respectively), focusing on the clinical and mutational profiles (n = 53 with molecular data) of these diseases. We found no differences in clinical presentations or outcomes of both entities. As previously described, both CNL and aCML share a complex mutational profile with mutations in genes involved in epigenetic regulation, splicing, and signaling pathways. Apart from CSF3R, only EZH2 and TET2 were differentially mutated between them. The molecular profiles support the notion of CNL and aCML being a continuum of the same disease that may fit best within the myelodysplastic/myeloproliferative neoplasms. We identified 4 high-risk mutated genes, specifically CEBPA (ß = 2.26, hazard ratio [HR] = 9.54, P = .003), EZH2 (ß = 1.12, HR = 3.062, P = .009), NRAS (ß = 1.29, HR = 3.63, P = .048), and U2AF1 (ß = 1.75, HR = 5.74, P = .013) using multivariate analysis. Our findings underscore the relevance of molecular-risk classification in CNL/aCML as well as the importance of CSF3R mutations in these diseases.

NGS-Based Molecular Karyotyping of Multiple Myeloma: Results from the GEM12 Clinical Trial.

Next-generation sequencing (NGS) has greatly improved our ability to detect the genomic aberrations occurring in multiple myeloma (MM); however, its transfer to routine clinical labs and its validation in clinical trials remains to be established. We designed a capture-based NGS targeted panel to identify, in a single assay, known genetic alterations for the prognostic stratification of MM. The NGS panel was designed for the simultaneous study of single nucleotide and copy number variations, insertions and deletions, chromosomal translocations and V(D)J rearrangements. The panel was validated using a cohort of 149 MM patients enrolled in the GEM2012MENOS65 clinical trial. The results showed great global accuracy, with positive and negative predictive values close to 90% when compared with available data from fluorescence in situ hybridization and whole-exome sequencing. While the treatments used in the clinical trial showed high efficacy, patients defined as high-risk by the panel had shorter progression-free survival (p = 0.0015). As expected, the mutational status of TP53 was significant in predicting patient outcomes (p = 0.021). The NGS panel also efficiently detected clonal IGH rearrangements in 81% of patients. In conclusion, molecular karyotyping using a targeted NGS panel can identify relevant prognostic chromosomal abnormalities and translocations for the clinical management of MM patients.

Molecular profiling of immunoglobulin heavy-chain gene rearrangements unveils new potential prognostic markers for multiple myeloma patients.

Multiple myeloma is a heterogeneous disease whose pathogenesis has not been completely elucidated. Although B-cell receptors play a crucial role in myeloma pathogenesis, the impact of clonal immunoglobulin heavy-chain features in the outcome has not been extensively explored. Here we present the characterization of complete heavy-chain gene rearrangements in 413 myeloma patients treated in Spanish trials, including 113 patients characterized by next-generation sequencing. Compared to the normal B-cell repertoire, gene selection was biased in myeloma, with significant overrepresentation of IGHV3, IGHD2 and IGHD3, as well as IGHJ4 gene groups. Hypermutation was high in our patients (median: 8.8%). Interestingly, regarding patients who are not candidates for transplantation, a high hypermutation rate (≥7%) and the use of IGHD2 and IGHD3 groups were associated with improved prognostic features and longer survival rates in the univariate analyses. Multivariate analysis revealed prolonged progression-free survival rates for patients using IGHD2/IGHD3 groups (HR: 0.552, 95% CI: 0.361-0.845, p = 0.006), as well as prolonged overall survival rates for patients with hypermutation ≥7% (HR: 0.291, 95% CI: 0.137-0.618, p = 0.001). Our results provide new insights into the molecular characterization of multiple myeloma, highlighting the need to evaluate some of these clonal rearrangement characteristics as new potential prognostic markers.

Facial mucocele and brain complications: diagnostic priorities.

We present the case of a male with a history of nasal polyposis underwent bilateral nasosinusal endoscopic surgery. He went to the emergency department because of having behavioral changes and left frontal headache. An emergency CT showed nasal-sinus polyposis and several nodular lesions with a characteristic "ring" enhancement and perilesional edema. These findings were compatible facial mucocele complicated with rupture of the bone wall of the left frontal sinus and frontal abscess. Urgent surgery was performed, with left frontal craniectomy and drainage of the abscesses. Mucoceles are benign slowly growing lesions which can associate important complications. The most frequent are abscesses and the invasion of neighboring structures. It is very important to remember that frontal mucoceles can cause intracranial invasion when there is an erosion of the internal osseous table.

Spectrum and functional validation of PSMB5 mutations in multiple myeloma.

Despite an increasing number of approved therapies, multiple myeloma (MM) remains an incurable disease and only a small number of patients achieve prolonged disease control. Some genes have been linked with response to commonly used anti-MM compounds, including immunomodulators (IMiDs) and proteasome inhibitors (PIs). In this manuscript, we demonstrate an increased incidence of acquired proteasomal subunit mutations in relapsed MM compared to newly diagnosed disease, underpinning a potential role of point mutations in the clonal evolution of MM. Furthermore, we are first to present and functionally characterize four somatic PSMB5 mutations from primary MM cells identified in a patient under prolonged proteasome inhibition, with three of them affecting the PI-binding pocket S1. We confirm resistance induction through missense mutations not only to Bortezomib, but also, in variable extent, to the next-generation PIs Carfilzomib and Ixazomib. In addition, a negative impact on the proteasome activity is assessed, providing a potential explanation for later therapy-induced eradication of the affected tumor subclones in this patient.

Light Microscopic, Ultrastructure Analysis and Functional Morphology of Cornu aspersum Spermatozoa Contained in the Frozen Hermaphroditic Duct / Análisis de Microscopía de Luz, Ultraestructura y Morfología Funcional de Espermatozoides de Cornu aspersum Contenidos en el Conducto Hermafrodita Congelado

SUMMARY: In this study we describe the functional morphology of Cornu aspersum (Helix aspersa), spermatozoa using light, scanning (SEM) and transmission electron (TEM) microscopies. The studies were performed with sperm located in the frozen hermaphroditic duct. Our results showed that the head presents an elongated conical shape slightly coiled in a corkscrew, with the nucleus partially covered by an acrosome, where an apical vesicle is located at the lateralized apex. This peculiar shape suggests the helical displacement movement of the spermatozoa. The head and the nucleus are slightly larger size compared to those of other gastropod species. The intermediate tract is surrounded by a mitochondrial complex and a glycogen helix. The glycogen helix is coiled helically along the intermediate tract, presenting at least five twists of glycogen helices. The complexity of both the mitochondrial complex and the glycogen helix suggests a high metabolic consumption considering the long period of time until fertilization occurs. Our findings on the detailed characterization of Cornu aspersum spermatozoa, obtained from a frozen hermaphroditic duct can contribute to a better understanding of the functional morphology of sperm and serve as a reference for future studies.

En este estudio describimos la morfología funcional de Cornu aspersum (Helix aspersa), espermatozoides utilizando microscopías de luz, barrido (SEM) y electrónica de transmisión (TEM). Los estudios se realizaron con espermatozoides localizados en el conducto hermafrodita congelado. Nuestros resultados mostraron que la cabeza presenta una forma cónica alargada ligeramente enrollada en un tirabuzón, con el núcleo parcialmente cubierto por un acrosoma, donde se ubica una vesícula apical en el ápice lateralizado. Esta peculiar forma sugiere el movimiento de desplazamiento helicoidal de los espermatozoides. La cabeza y el núcleo son de un tamaño ligeramente mayor en comparación con los de otras especies de gasterópodos. El tracto intermedio está rodeado por un complejo mitocondrial y una hélice de glucógeno. La hélice de glucógeno se enrolla helicoidalmente a lo largo del tracto intermedio, presentando al menos cinco giros de hélices de glucógeno. La complejidad tanto del complejo mitocondrial como de la hélice de glucógeno sugiere un alto consumo metabólico considerando el largo período de tiempo hasta que ocurre la fecundación. Nuestros hallazgos sobre la caracterización detallada de los espermatozoides de Cornu aspersum, obtenidos de un conducto hermafrodita congelado, pueden contribuir a una mejor comprensión de la morfología funcional de los espermatozoides y servir como referencia para futuros estudios.

Group Therapy for Patients with Adjustment Disorder in Primary Care.

A high percentage of patients have a diagnosis of adjustment disorder (AD) when they arrive at primary care (PC) appointments. However, most of them do not receive adequate psychological treatment. The present study's aim is to determine the efficacy of a group psychological treatment program in patients with AD. The sample consisted of patients with AD from two PC units in Valencia, from which two groups were randomly generated: A treatment group (n = 31) and a waiting-list group (n = 20), homogeneous in terms of socio-demographic and psychometric variables prior to treatment. Treatment consisted of eight one-hour group sessions held on a weekly basis; taking a cognitive-behavioral approach, they addressed aspects like controlling anxiety, cognitive restructuring, and coping techniques. The variables analyzed were: Psychopathology (Revised Symptom Inventory, SCL-90-R), health-related quality of life (Health Questionnaire, SF-12), and risk of suicidal behavior (Suicide Risk Scale). Means comparisons, ANCOVAs, and tests of effect size were performed. Statistically significant differences were observed in the variables, such that after intervention, the experimental group exhibited less anxious (F = 4.11, p =.048, η2 = .08) and depressive symptoms (F = 2.41, p =.029, η 2= .10) and higher quality of life related to physical (F = 7.17, p =.010, η2 = .13) and emotional health (F = 10.31, p =.002, η2 = .18). For the reasons above, we conclude that a comprehensive approach to emotional distress in PC, including group psychological interventions, is one solution for the demand for social services, and could provide savings on economic as well as human costs.

Measuring impulsivity in school-aged boys and examining its relationship with ADHD and ODD ratings.

Seven different laboratory measures of impulsivity were administered to a group of 165 school-aged boys. Parents' and teachers' ratings of Attention Deficit and Hyperactivity Disorder and Oppositional/Defiant Disorder were also obtained. Factor analyses of impulsivity measures revealed the existence of a strong Inhibitory Control Factor including measures derived from Stop Task, the Continuous Performance Test, the Matching Familiar Figures Test, and the Circle Tracing Task. Other forms of impulsivity like resistance to interference, the Wisconsin Card Sorting Test and efficiency in the DRL Task loaded on a second independent factor. The Inhibitory Control factor was correlated with ADHD ratings, whereas the second factor was slightly related to the presence of ODD symptoms. Discussion is focused on the relevance of inhibitory control in impulsivity and ADHD research.

Human papillomavirus infection in HIV-1 infected women in Catalonia (Spain): implications for prevention of cervical cancer.

BACKGROUND: High-risk human Papillomavirus infection is a necessary factor for cervical squamous intraepithelial lesions and invasive cervical cancer. In HIV-1-infected women, HPV infection is more prevalent and a higher risk of cervical cancer has been identified. We aimed to calculate the prevalence of infection by HR-HPV, determine the factors associated with this infection and abnormal cytology findings and to describe the history of cervical cancer screening in HIV-1-infected women. METHODS: We enrolled 479 HIV-1-infected women from the PISCIS cohort. Each patient underwent a gynecological check-up, PAP smear, HPV AND Hybrid capture, HPV genotyping, and colposcopy and biopsy, if necessary. We applied questionnaires to obtain information on sociodemographic, behavioral, clinical, and cervical screening variables. We present a cross-sectional analysis. RESULTS: Median age was 42 years. The prevalence of HR-HPV infection was 33.2% and that of high-grade squamous intraepithelial lesions (HSIL) was 3.8%. The most common genotypes were 16(23%), 53(20.3%), and 52(16.2%). The factor associated with HR-HPV infection was age <30 years (odds ratio[OR],2.5; 95%confidence interval[CI],1.1-5.6). The factors associated with the presence of HSIL or low-grade squamous intraepithelial lesions (LSIL) were CD4T-lymphocyte count <200 cells/mm(3) versus >500 cells/mm(3) (OR,8.4; 95%CI,3.7-19.2), HIV-1 viral load >10,000 copies/mL versus <400 copies/mL (OR,2.1; 95%CI,1.0-4.4), and use of oral contraceptives (OR,2.0; 95%CI,1.0-3.9). Sixty percent of HIV-1-infected women had had one Pap smear within the last 2 years. CONCLUSIONS: The high prevalence of HPV infection and cervical lesions in the HIV-1-infected population in Catalonia, as well as the low coverage and frequency of screening in this group, means that better preventive efforts are necessary and should include vaccination against HPV, better accessibility to screening programs, training of health care professionals, and specific health education for HIV-1-infected women.

Gastric Adenocarcinoma: Is Computed Tomography (CT) Useful in Preoperative Staging?

BACKGROUND AND PURPOSE: Although multiple studies testing the accuracy of CT in the preoperative staging of gastric adenocarcinoma have been carried out, their results are controversial. Whilst some authors claim that CT is an accurate method for preoperatively staging gastric cancer, others have advocated the contrary. Because of this discrepancy we have retrospectively reviewed preoperative CT findings compared with histopathological results in patients with gastric adenocarcinoma. PATIENTS AND METHODS: Seventy-two patients diagnosed with gastric cancer who underwent potentially curative surgery and preoperative staging CT of quality were included in the study. The size, gastric wall thickening, presence of lymphadenopathy, adjacent organ invasion and location of the gastric mass was recorded. Early tumors (T1 and T2) and more advanced tumors (T3 and T4) were grouped together. CT staging was correlated with the final histopathological stage (TNM). The global results were expressed as sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). RESULTS: Seventy-two cases were included with fifty-five being male and a median age of 67 years (range 33-91). CT correctly identified the location of the tumor in 56 (53% antropyloric, 18% subcardial). Median time from CT scan to surgery was fourteen days (range 2-49). In T detection: T1/T2 and T3/T4 with sensitivity of 70% and 61%. Lymph node involvement: Sensitivity 49%. Overstaged in 47% Understaged in 75%. Specificity of 53%. Nine patients with colon-mesocolon (5 patients) and pancreas (4 patients) invasion. Sensitivity 44% and specificity 96%. CONCLUSION: Spiral CT is not an accurate method in predicting preoperative stages in gastric cancer.