FOXL2 Mutation Status in Sex Cord-stromal Tumors Cannot be Predicted by Morphology.

Granulosa-cell tumors (GCTs) are the most common type of malignant ovarian sex cord-stromal tumor (SCST). The histopathologic diagnosis of these tumors can be challenging. A recurrent somatic mutation of the forkhead box L2 (FOXL2) gene has been identified in adult GCT. In this retrospective single-center study of 44 SCST, a morphologic review together with analysis of FOXL2 C134W was evaluated in relation to tumor morphology. In addition, TERT promoter mutation testing was performed. Twelve of 36 cases got an altered diagnosis based on morphology alone. The overarching architectural growth pattern in 32/44 (72.7%) tumors was diffuse/solid with several tumors showing markedly heterogeneous architecture. In correlation to FOXL2 C134W mutation status, cytoplasmic color, and nuclear shape, differed between the FOXL2 C134W positive and FOXL2 C134 W negative groups, but these differences were not significant when comparing them separately. Nineteen of 44 cases underwent TERT promoter sequencing with a positive result in 3 cases; 2 adult GCTs and 1 cellular fibroma. Three patients developed a recurrence of which 2 were FOXL2 C134W positive adult GCTs and the third was an unclassified SCST. In conclusion, the morphologic and immunohistochemical diagnosis of different SCSTs is challenging and one cannot reliably identify FOXL2 mutation-positive tumors solely by morphologic features. Therefore, broad use of molecular analysis of the FOXL2 C134W mutation is suggested for SCSTs, and further studies are needed to evaluate the clinical outcome of these tumors as well as the diagnostic and prognostic implications of TERT promoter mutations.

Molecular detection and species identification of dermatophytes by SYBR-Green real-time PCR in-house methodology using hair samples obtained from dogs and cats.

The classical dermatophytes diagnosis is based on mycological culture and microscopy observation both human and animal hair, skin, and nail samples. The aim of this work was to develop the new in-house real-time PCR with pan-dematophyte reaction for detection and identification of the main dermatophytes directly from hair samples, providing a simple and rapid diagnosis of dermatophytosis in dogs and cats. An in-house SYBR-Green real-time PCR was designed and used for detecting a DNA fragment encoding chitin synthase 1 (CHS1). A total of 287 samples were processed by culture, microscopic examination with KOH 10%, and real-time PCR (qPCR) analysis. Melting curve analysis of the CHS1 fragment revealed to be reproducible, showing a single distinct peak for each species of dermatophyte, namely Trichophyton mentagrophytes, T. verrucosum, Microsporum canis, and Nannizzia gypsea (formerly M. gypseum). Then, out of the 287 clinically suspected cases of dermatophytosis, 50% were positive for dermatophytes by qPCR, 44% by mycological culture, and 25% by microscopic examination. Microsporum canis was identified in 117 samples tested by culture and 134 samples tested by qPCR, followed by N. gypsea in 5 samples (either tested by culture or qPCR) and T. mentagrophytes detected in 4 and 5 samples when tested by culture or qPCR, respectively. Overall, qPCR allowed the diagnosis of dermatophytosis in clinical samples. The results suggest this newly proposed in-house real-time PCR assay can be used as alternative diagnosis and rapid identification of dermatophytes frequently associated to clinical hair samples of dogs and cats.

The aim of this work was to develop a molecular detection strategy for dermatophytes by SYBR-Green real-time PCR of hair samples from animals. The melting curve analysis of the CHS1 fragment revealed to be reproducible, showing a single distinct peak for distinct dermatophyte species and allowed the diagnosis of dermatophytosis in dogs and cats caused mainly by Trichophyton mentagrophytes, Microsporum sp., and Nannizzia gypsea).

Precision Oncology of High-Grade Ovarian Cancer Defined through Targeted Sequencing.

BACKGROUND: We examined whether molecular characterization of high-grade epithelial ovarian cancer can inform the diagnosis and/or identify potential actionable targets. METHODS: All of the consecutively sequenced high-grade ovarian tumours with consent between 2014 until 2019 were included. A total of 274 tumours underwent next generation sequencing using a targeted panel. RESULTS: Patients with high-grade ovarian epithelial cancer were consented to prospective molecular characterization. Clinical information was extracted from their medical record. Tumour DNA was subjected to sequencing, and selected patients received PARP inhibitor therapy. CONCLUSIONS: Tumours from 274 women were sequenced, including high-grade serous carcinoma (n = 252), clear cell carcinoma (n = 4), carcinosarcoma (n = 9), endometrioid carcinoma (n = 3), undifferentiated carcinoma (n = 1), and mixed tumours (n = 5). Genomic profiling did not influence histologic diagnosis. Mutations were identified in TP53, BRCA1, BRCA2, as well as additional homologous recombination repair pathway genes BARD1, ATR, CHEK2, PALB2, RAD51D, RAD50, SLX4, FANCA, RAD51C, and RAD54L. In addition, mutations in PTEN and CDKN2A were identified. Several somatic mutations with implications for germline testing were identified, including RMI1, STK11, and CDH1. Germline testing identified 16 previously unknown BRCA1/2 carriers. Finally, 20 patients were treated with the PARP inhibitor olaparib based on the sequencing results.

MRI-Guided, Noninvasive Delivery of Magneto-Electric Drug Nanocarriers to the Brain in a Nonhuman Primate.

A magnetically guided brain delivery method previously demonstrated in mice has not yet been translated for clinical applications due to the mismatch of available static magnet dimensions in relation to the human brain size and shape. To develop a human-compatible methodology, we explored magnetic resonance imaging (MRI) as a tool for the delivery of magneto-electric nanoparticles (MENPs) into the brain of a baboon, as a proof-of-concept study. MRI brain image analysis showed a reduction in T2* value at the basal ganglia, hemisphere, and vertex, thereby confirming successful MENP delivery to the brain. The observation of well-integrated morphologically healthy tissues and no blood toxicity over the study duration confirmed the biocompatibility of MENPs and the delivery procedure. Outcomes of this research present MRI-assisted delivery of MENPs to the brain as a safe and noninvasive method in larger species such as baboons and one step closer to human translation. This MENP-based nanomedicine delivery method can be used for clinical application in order to investigate effective central nervous system (CNS) therapies.

Anxiety, hospital fears and conduct and behavioral alterations during pediatric hospitalization.

INTRODUCTION: A hospital admission is an experience capable of generating emotional and behavioral alterations at any age. This study pretends to analyze the response of anxiety, fears and/or behavioral alterations in pediatric patients exposed to a conventional non-surgical hospital admission and the existing relationship between these responses and certain modulating variables. Metodology. Design of cohorts. Data collection was carried out in three stages (M1: at admission; M2: at discharge; M3: 2 weeks after discharge), on a 30 patient sample between the ages of 6 to 15 years and 30 caregivers. A comparison was made on the mean of the repeated measurements (Student t) of the respond variables and their correlation (Pearson’s Coefficient Correlation) with modulating variables. RESULTS: The results of the intra-subject analysis showed significance in terms of anxiety levels state in patients in M1 versus M3 (t=3.93, p<.0001, d=0.69) and the magnitude of the total behavioral alterations registered in M1 versus M3 (t=-5.02, p<.0001, d=0.60). It was observed that a significant relationship between modulating variables of patients (anxiety risk) and of the caregiver (anxiety character state, strategy of confrontation) and the variables of response of the anxiety and behavioral alterations of the patient. CONCLUSIONS: Exposure of a conventional non-surgical hospital admission may have negative consequences at an emotional and behavioral level in children, present far beyond the hospital admission. Certain variables, from the patient and the caregiver, are psychological vulnerability factors before a hospitalization process.

Ansiedad, miedos hospitalarios y alteraciones conductuales en la hospitalización infantil / Anxiety, hospital fears and conduct and behavioral alterations during pediatric hospitalization

Introducción. El ingreso hospitalario es una experiencia susceptible de generar alteraciones emocionales o conductuales a cualquier edad. Este estudio pretende analizar la respuesta de ansiedad, miedos y/o alteraciones conductuales en pacientes pediátricos expuestos a un ingreso hospitalario convencional no quirúrgico y las relaciones existentes entre dichas respuestas y determinadas variables moduladoras. Metodología. Diseño de cohortes. La recogida de datos se realizó en tres momentos (M1: al ingreso; M2: al alta; M3: a las dos semanas del alta), sobre una muestra de 30 pacientes de 6 a 15 años y 30 cuidadores. Se realizó una comparación de medias de medidas repetidas (t de Student) de las variables de respuesta y su correlación (Coeficiente correlación de Pearson) con variables moduladoras. Resultados. Los resultados del análisis intrasujeto muestran significación en cuanto a los niveles de ansiedad estado del paciente en M1 versus M3 (t=3.93, p<0.0001, d=0.69) y la magnitud de alteraciones conductuales totales registradas en M1 versus M3 (t=-5.02, p<0.0001, d=0.60). Se objetivó relación significativa positiva entre variables moduladoras del paciente (ansiedad rasgo) y del cuidador (ansiedad rasgo-estado, estrategias de afrontamiento) y las variables de respuesta de ansiedad y alteraciones conductuales del paciente. Conclusiones. La exposición a una hospitalización convencional no quirúrgica puede producir consecuencias negativas a nivel emocional y conductual en el niño, presentes más allá del ingreso hospitalario. Determinadas variables, del paciente y del cuidador, son factores de vulnerabilidad psicológica ante el proceso de hospitalización

Introduction. A hospital admission is an experience capable of generating emotional and behavioral alterations at any age. This study pretends to analyze the response of anxiety, fears and/or behavioral alterations in pediatric patients exposed to a conventional non-surgical hospital admission and the existing relationship between these responses and certain modulating variables. Metodology. Design of cohorts. Data collection was carried out in three stages (M1: at admission; M2: at discharge; M3: 2 weeks after discharge), on a 30 patient sample between the ages of 6 to 15 years and 30 caregivers. A comparison was made on the mean of the repeated measurements (Student t) of the respond variables and their correlation (Pearson's Coefficient Correlation) with modulating variables. Results. The results of the intra-subject analysis showed significance in terms of anxiety levels state in patients in M1 versus M3 (t=3.93, p<.0001, d=0.69) and the magnitude of the total behavioral alterations registered in M1 versus M3 (t=-5.02, p<.0001, d=0.60). It was observed that a significant relationship between modulating variables of patients (anxiety risk) and of the caregiver (anxiety character state, constrategy of confrontation) and the variables of response of the anxiety and behavioral alterations of the patient. Conclusions. Exposure of a conventional non-surgical hospital admission may have negative consequences at an emotional and behavioral level in children, present far beyond the hospital admission. Certain variables, from the patient and the caregiver, are psychological vulnerability factors before a hospitalization process

Cationic intermediates in the Pd-catalyzed negishi coupling. kinetic and density functional theory study of alternative transmetalation pathways in the Me-Me coupling of ZnMe2 and trans-[PdMeCl(PMePh2)2].

The complexity of the transmetalation step in a Pd-catalyzed Negishi reaction has been investigated by combining experiment and theoretical calculations. The reaction between trans-[PdMeCl(PMePh(2))(2)] and ZnMe(2) in THF as solvent was analyzed. The results reveal some unexpected and relevant mechanistic aspects not observed for ZnMeCl as nucleophile. The operative reaction mechanism is not the same when the reaction is carried out in the presence or in the absence of an excess of phosphine in the medium. In the absence of added phosphine an ionic intermediate with THF as ligand ([PdMe(PMePh(2))(2)(THF)](+)) opens ionic transmetalation pathways. In contrast, an excess of phosphine retards the reaction because of the formation of a very stable cationic complex with three phosphines ([PdMe(PMePh(2))(3)](+)) that sequesters the catalyst. These ionic intermediates had never been observed or proposed in palladium Negishi systems and warn on the possible detrimental effect of an excess of good ligand (as PMePh(2)) for the process. In contrast, the ionic pathways via cationic complexes with one solvent (or a weak ligand) can be noticeably faster and provide a more rapid reaction than the concerted pathways via neutral intermediates. Theoretical calculations on the real molecules reproduce well the experimental rate trends observed for the different mechanistic pathways.