RET Fusion Testing in Patients With NSCLC: The RETING Study.

Introduction: RET inhibitors with impressive overall response rates are now available for patients with NSCLC, yet the identification of RET fusions remains a difficult challenge. Most guidelines encourage the upfront use of next-generation sequencing (NGS), or alternatively, fluorescence in situ hybridization (FISH) or reverse transcriptase-polymerase chain reaction (RT-PCR) when NGS is not possible or available. Taken together, the suboptimal performance of single-analyte assays to detect RET fusions, although consistent with the notion of encouraging universal NGS, is currently widening some of the clinical practice gaps in the implementation of predictive biomarkers in patients with advanced NSCLC. Methods: This situation prompted us to evaluate several RET assays in a large multicenter cohort of RET fusion-positive NSCLC (n = 38) to obtain real-world data. In addition to RNA-based NGS (the criterion standard method), all positive specimens underwent break-apart RET FISH with two different assays and were also tested by an RT-PCR assay. Results: The most common RET partners were KIF5B (78.9%), followed by CCDC6 (15.8%). The two RET NGS-positive but FISH-negative samples contained a KIF5B(15)-RET(12) fusion. The three RET fusions not identified with RT-PCR were AKAP13(35)-RET(12), KIF5B(24)-RET(9) and KIF5B(24)-RET(11). All three false-negative RT-PCR cases were FISH-positive, exhibited a typical break-apart pattern, and contained a very high number of positive tumor cells with both FISH assays. Signet ring cells, psammoma bodies, and pleomorphic features were frequently observed (in 34.2%, 39.5%, and 39.5% of tumors, respectively). Conclusions: In-depth knowledge of the advantages and disadvantages of the different RET testing methodologies could help clinical and molecular tumor boards implement and maintain sensible algorithms for the rapid and effective detection of RET fusions in patients with NSCLC. The likelihood of RET false-negative results with both FISH and RT-PCR reinforces the need for upfront NGS in patients with NSCLC.

Differential Expression of MicroRNAs in Silent and Functioning Corticotroph Tumors.

The potential role of miRNAs in the silencing mechanisms of pituitary neuroendocrine tumors (PitNETs) has not been addressed. The aim of the present study was to evaluate the expression levels and the potential associated role of some miRNAs, pathways, and transcription factors in the silencing mechanisms of corticotroph tumors (CTs). Accordingly, the expression of miR-375, miR-383, miR-488, miR-200a and miR-103; of PKA, MAP3K8, MEK, MAPK3, NGFIB, NURR1, PITX1, and STAT3 were analyzed via qRT-PCR in 23 silent and 24 functioning CTs. miR-200a and miR-103 showed significantly higher expression in silent than in functioning CTs, even after eliminating the bias of tumor size, therefore enabling the differentiation between the two variants. Additionally, miR-383 correlated negatively with TBX19 in silent CTs, a transcription factor related with the processing of POMC that can participate in the silencing mechanisms of CTs. Finally, the gene expression levels of miR-488, miR-200a, and miR-103 were significantly higher in macroadenomas (functioning and silent) than in microadenomas. The evidence from this study indicates that miRNAs could be involved in the pathophysiology of CTs. The translational implications of these findings suggest that pharmacological treatments specifically targeting these miRNAs could become a promising therapeutic option for these patients.

Increased E2F1 mRNA and miR-17-5p Expression Is Correlated to Invasiveness and Proliferation of Pituitary Neuroendocrine Tumours.

miR-17-5p and E2F1 have been described as deregulated in cancer, but they have scarcely been studied in pituitary neuroendocrine tumours (PitNETs). This study evaluates the relationship of E2F1 and miR-17-5p with the invasiveness and proliferation of PitNETs. In this cross-sectional descriptive study, we evaluated the expression of E2F1, MYC, and miR-17-5p by quantitative real time PCR analysis in 60 PitNETs: 29 gonadotroph (GT), 15 functioning somatotroph (ST), and 16 corticotroph (CT) tumours, of which 8 were silent (sCT). The clinical data were collected from the Spanish Molecular Register of Pituitary Adenomas (REMAH) database. We defined invasiveness according to the Knosp classification and proliferation according to a molecular expression of Ki-67 ≥ 2.59. E2F1 was more expressed in invasive than in non-invasive tumours in the whole series (p = 0.004) and in STs (p = 0.01). In addition, it was overexpressed in the silent subtypes (GTs and sCTs; all macroadenomas) and normoexpressed in the functioning ones (fCTs and STs; some microadenomas). miR-17-5p was more expressed in proliferative than in non-proliferative tumours (p = 0.041) in the whole series but not by subtypes. Conclusions: Our study suggests that in PitNETs, E2F1 could be a good biomarker of invasiveness, and miR-17-5p of proliferation, helping the clinical management of these tumours.

How Valuable Is the RT-qPCR of Pituitary-Specific Transcription Factors for Identifying Pituitary Neuroendocrine Tumor Subtypes According to the New WHO 2017 Criteria?

The classification of pituitary neuroendocrine tumors (PitNETs) subtypes continues generating interest. In 2017, the World Health Organization (WHO) proposed considering the immunohistochemical (IHC) analysis of pituitary-specific transcription factors (TF) for their typification. The present study targeted the quantification of pituitary-specific TF (TPIT, PIT-1, SF-1, GATA2, ESR1) gene expression by RT-qPCR to overcome the shortcomings of IHC and to complement it. We analyzed 251 tumors from our collection of PitNETs and performed additional IHC studies in a subset of 56 samples to analyze the concordance between gene and protein expression of the TF. The molecular and IHC studies allowed us to significantly reduce the percentage of null cell tumors in our series, most of which were reclassified as gonadotroph tumors. The concordance between the molecular and the immunohistochemical studies was good for tumors coming from the corticotroph and Pit-1 lineages but worsened for the rest of the tumors. Indeed, the RT-qPCR helped to improve the typification of plurihormonal Pit-1 and unusual tumors. Overall, our results suggest that the RT-qPCR of pituitary-specific TF and hormone genes could help pathologists, endocrinologists, and neurosurgeons to improve the management of patients with pituitary tumors.

Assessment of a New ROS1 Immunohistochemistry Clone (SP384) for the Identification of ROS1 Rearrangements in Patients with Non-Small Cell Lung Carcinoma: the ROSING Study.

INTRODUCTION: The ROS1 gene rearrangement has become an important biomarker in NSCLC. The College of American Pathologists/International Association for the Study of Lung Cancer/Association for Molecular Pathology testing guidelines support the use of ROS1 immunohistochemistry (IHC) as a screening test, followed by confirmation with fluorescence in situ hybridization (FISH) or a molecular test in all positive results. We have evaluated a novel anti-ROS1 IHC antibody (SP384) in a large multicenter series to obtain real-world data. METHODS: A total of 43 ROS1 FISH-positive and 193 ROS1 FISH-negative NSCLC samples were studied. All specimens were screened by using two antibodies (clone D4D6 from Cell Signaling Technology and clone SP384 from Ventana Medical Systems), and the different interpretation criteria were compared with break-apart FISH (Vysis). FISH-positive samples were also analyzed with next-generation sequencing (Oncomine Dx Target Test Panel, Thermo Fisher Scientific). RESULTS: An H-score of 150 or higher or the presence of at least 70% of tumor cells with an intensity of staining of 2+ or higher by the SP384 clone was the optimal cutoff value (both with 93% sensitivity and 100% specificity). The D4D6 clone showed similar results, with an H-score of at least 100 (91% sensitivity and 100% specificity). ROS1 expression in normal lung was more frequent with use of the SP384 clone (p < 0.0001). The ezrin gene (EZR)-ROS1 variant was associated with membranous staining and an isolated green signal FISH pattern (p = 0.001 and p = 0.017, respectively). CONCLUSIONS: The new SP384 ROS1 IHC clone showed excellent sensitivity without compromising specificity, so it is another excellent analytical option for the proposed testing algorithm.

Silent somatotropinomas.

Silent somatotroph pituitary neuroendocrine tumors (or silent growth hormone pituitary neuroendocrine tumors, SGH-PitNET) are neoplasias with positive immunostaining for growth hormone (GH), in patients with no signs and symptoms of acromegaly nor biochemical evidence of GH hypersecretion. From a clinical stand-point they are considered and managed as non-functioning pituitary tumors, since they usually come to evidence due to mass-effects (headache, visual impairment, hypopituitarism) or as asymptomatic pituitary incidentalomas. SGH-PitNET have deserved little attention in the medical literature, and no specific guidelines exist regarding their management. However, identification of a particular tumor lineage through immunostaining patterns of non-functioning pituitary tumors may determine postoperative medical therapy in the near future. This review updates the current knowledge about the epidemiologic, clinical, pathological and molecular characteristics of this particular type of pituitary tumors.

Is it time to consider the expression of specific-pituitary hormone genes when typifying pituitary tumours?

The aim of the present study is to check whether we can replicate, in an independent series, previous results showing that the molecular study of pituitary-specific gene expression complements the inmunohistochemical identification of pituitary neuroendocrine tumours. We selected 112 patients (51 (46.4%) women; mean age 51.4±16 years; 102 macroadenomas (91.9%), 9 microadenomas (8.1%)) with complete clinical, radiological, immunohistochemical and molecular data from our data set of pituitary neuroendocrine tumours. Patients were different from those previously studied. We measured the expression of the pituitary-specific hormone genes and type 1 corticotrophin-releasing hormone and arginine vasopressin 1b receptors, by quantitative real-time polymerase chain reaction using TaqMan probes. Afterwards, we identified the different pituitary neuroendocrine tumour subtypes following the 2017 World Health Organization classification of pituitary tumours, calculating the concordance between their molecular and immuhistochemical identification. The concordance between molecular and immunohistochemical identification of functioning pituitary neuroendocrine tumours with the clinical diagnosis was globally similar to the previous series, where the SYBR Green technique was used instead of TaqMan probes. Our results also corroborated the poor correlation between molecular and immunohistochemical detection of the silent pituitary neuroendocrine tumour variants. This discrepancy was more remarkable in lactotroph, null-cell and plurihormonal pituitary neuroendocrine tumours. In conclusion, this study validates the results previously published by our group, highlighting a complementary role for the molecular study of the pituitary-specific hormone genes in the typification of pituitary neuroendocrine tumours subtypes.

PAM50 breast cancer subtyping by RT-qPCR and concordance with standard clinical molecular markers.

BACKGROUND: Many methodologies have been used in research to identify the "intrinsic" subtypes of breast cancer commonly known as Luminal A, Luminal B, HER2-Enriched (HER2-E) and Basal-like. The PAM50 gene set is often used for gene expression-based subtyping; however, surrogate subtyping using panels of immunohistochemical (IHC) markers are still widely used clinically. Discrepancies between these methods may lead to different treatment decisions. METHODS: We used the PAM50 RT-qPCR assay to expression profile 814 tumors from the GEICAM/9906 phase III clinical trial that enrolled women with locally advanced primary invasive breast cancer. All samples were scored at a single site by IHC for estrogen receptor (ER), progesterone receptor (PR), and Her2/neu (HER2) protein expression. Equivocal HER2 cases were confirmed by chromogenic in situ hybridization (CISH). Single gene scores by IHC/CISH were compared with RT-qPCR continuous gene expression values and "intrinsic" subtype assignment by the PAM50. High, medium, and low expression for ESR1, PGR, ERBB2, and proliferation were selected using quartile cut-points from the continuous RT-qPCR data across the PAM50 subtype assignments. RESULTS: ESR1, PGR, and ERBB2 gene expression had high agreement with established binary IHC cut-points (area under the curve (AUC) ≥ 0.9). Estrogen receptor positivity by IHC was strongly associated with Luminal (A and B) subtypes (92%), but only 75% of ER negative tumors were classified into the HER2-E and Basal-like subtypes. Luminal A tumors more frequently expressed PR than Luminal B (94% vs 74%) and Luminal A tumors were less likely to have high proliferation (11% vs 77%). Seventy-seven percent (30/39) of ER-/HER2+ tumors by IHC were classified as the HER2-E subtype. Triple negative tumors were mainly comprised of Basal-like (57%) and HER2-E (30%) subtypes. Single gene scoring for ESR1, PGR, and ERBB2 was more prognostic than the corresponding IHC markers as shown in a multivariate analysis. CONCLUSIONS: The standard immunohistochemical panel for breast cancer (ER, PR, and HER2) does not adequately identify the PAM50 gene expression subtypes. Although there is high agreement between biomarker scoring by protein immunohistochemistry and gene expression, the gene expression determinations for ESR1 and ERBB2 status was more prognostic.

Adjuvant docetaxel for high-risk, node-negative breast cancer.

BACKGROUND: A regimen of docetaxel, doxorubicin, and cyclophosphamide (TAC) is superior to a regimen of fluorouracil, doxorubicin, and cyclophosphamide (FAC) when used as adjuvant therapy in women with node-positive breast cancer. The value of taxanes in the treatment of node-negative disease has not been determined. METHODS: We randomly assigned 1060 women with axillary-node-negative breast cancer and at least one high-risk factor for recurrence (according to the 1998 St. Gallen criteria) to treatment with TAC or FAC every 3 weeks for six cycles after surgery. The primary end point was disease-free survival after at least 5 years of follow-up. Secondary end points included overall survival and toxicity. RESULTS: At a median follow-up of 77 months, the proportion of patients alive and disease-free was higher among the 539 women in the TAC group (87.8%) than among the 521 women in the FAC group (81.8%), representing a 32% reduction in the risk of recurrence with TAC (hazard ratio, 0.68; 95% confidence interval [CI], 0.49 to 0.93; P=0.01 by the log-rank test). This benefit was consistent, regardless of hormone-receptor status, menopausal status, or number of high-risk factors. The difference in survival rates (TAC, 95.2%; FAC, 93.5%) was not significant (hazard ratio, 0.76; 95% CI, 0.45 to 1.26); however, the number of events was small (TAC, 26; FAC, 34). Rates of grade 3 or 4 adverse events were 28.2% with TAC and 17.0% with FAC (P<0.001). Toxicity associated with TAC was diminished when primary prophylaxis with granulocyte colony-stimulating factor was provided. CONCLUSIONS: As compared with adjuvant FAC, adjuvant TAC improved the rate of disease-free survival among women with high-risk, node-negative breast cancer. (Funded by GEICAM and Sanofi-Aventis; ClinicalTrials.gov number, NCT00121992.).

The cavitated accessory uterine mass: a Müllerian anomaly in women with an otherwise normal uterus.

OBJECTIVE: To present clinical cases of women who had an accessory and cavitated noncommunicating uterine mass with functioning endometrium associated with a normal uterus, suggestive of a new type of Müllerian anomaly. METHODS: We report on five institutional cases: four cases of cavitated accessory uterine mass and a case of true adenomyoma. A review of the literature was performed by looking for these terms and others related in MEDLINE. RESULTS: Including ours, there are 18 cases in the literature showing an accessory cystic cavity lined by endometrioid epithelium with an otherwise normal uterus. Another 11 cases only partially fulfilled the inclusion criteria. All of the first cases were in young women presenting with severe dysmenorrhea (n=4). Generally, the tumor was located in the anterior wall of the uterus at the level of insertion of the round ligament. It presents a certain similarity with the cavitated true adenomyomas observed in older women in whom the endometrial lining of the cystic cavity is generally absent. For differential diagnosis with cavitated noncommunicating rudimentary uterine horns, hysterosalpingography showing a normal eutopic uterine cavity is decisive. CONCLUSION: Noncommunicating accessory uterine cavities and isolated cystic adenomyomas correspond to the same pathology: cavitated accessory uterine mass associated with an otherwise normal uterus. They present problems of differential diagnosis with true cavitated adenomyomas and cavitated rudimentary uterine horns. Accessory uterine mass could be caused by duplication and persistence of ductal Müllerian tissue in a critical area at the attachment level of the round ligament, possibly related to a gubernaculum dysfunction. LEVEL OF EVIDENCE: III.

Lymphangitic carcinomatosis as a cause of malignant transient pleural transudate.

Although it is generally accepted that a malignant transient pleural transudate may appear during the early stages of lymphatic obstruction, cases demonstrating such probability are rare in literature. A 67-year-old woman was admitted to hospital because a lymphangitic carcinomatosis and a transudative infrapulmonary pleural effusion with a cytology positive for adenocarcinoma. One month later the effusion keeps being positive for adenocarcinoma but exudative in character. Lymphatic obstruction appears as the cause of the initial transudative characteristics of the pleural effusion.

Exon 20 PIK3CA mutations decreases survival in aggressive (HER-2 positive) breast carcinomas.

PIK3CA mutations at 9 and 20 exons were studied in a series of 56 selected aggressive breast carcinomas (BC): 27 with Her-2 over-expression and negativity for estrogen receptors (ER) and progesterone receptors (PR), and 29 "triple negative" BC (negative for ER, PR and Her-2). Also, immunohistochemical studies of p53, ki-67, Her-1 (EGFR), pIGF-1R, PTEN, p110alpha, and pAkt were performed. Six mutations in exon 20 PIK3CA were identified among the 27 Her-2 positive BC, whereas only one exon 9 PIK3CA mutation was detected in a triple negative tumor (p = 0.035). Furthermore, PIK3CA mutations were associated with p110alpha over-expression (p = 0.001). Overall survival was shorter in cases with PIK3CA mutations (p = 0.015 in all series; and p = 0.041 for Her-2+ tumors), although multivariate analyses did not show statistical differences. No statistical significance was related with disease-free survival. Exon 20 PIK3CA mutations are relatively frequent in Her-2+ tumors and shorten survival, whereas neither exons 9 and 20 mutations seem related with "triple negative" breast carcinomas.

[Idiopathic bronchiolocentric interstitial pneumonia: a new idiopathic interstitial pneumonia]. / Neumonía intersticial bronquiolocéntrica idiopática: una nueva neumonía intersticial idiopática.

Idiopathic interstitial pneumonias represent a diverse group of lung diseases with diffuse effects on the lung parenchyma. In 2002, the American Thoracic Society/European Respiratory Society consensus classification unified the descriptions of the different entities encompassed by idiopathic interstitial pneumonias. Despite this broad consensus there are still some entities without a clear definition and others, such as idiopathic bronchiolocentric interstitial pneumonia, that were only later described. We present the case and outcome of a woman diagnosed with idiopathic bronchiolocentric interstitial pneumonia by lung biopsy.

Idiopathic organizing pneumonia: a relapsing disease. 19 years of experience in a hospital setting.

BACKGROUND: Although organizing pneumonia (OP) is a common pathological finding, studies including a substantial number of patients with idiopathic forms from a unique center and a long follow-up are rare. OBJECTIVES: To determine patients with cryptogenic forms of organizing pneumonia (COP), in order to characterize their clinical course, to identify predictive factors for relapse and to assess their effect on outcome. METHODS: For a 19-year period, all histopathological reports from a community teaching hospital were reviewed, and OP was found in 210 lung specimens belonging to 197 patients. RESULTS: Thirty-three (17%) patients presented cryptogenic forms and 32 of them (97%) responded to steroid therapy. At follow-up, 14 patients presented no relapses (no-relapse group, NR) and 18 (56%) presented relapses (relapsing group, RG) that resolved with ulterior treatment. Multifocal opacities on chest X-ray (RG 83% vs. NR 36%, p = 0.02) appeared to be a predictor for relapse. Patients with relapses showed a shorter time span to chest X-ray normalization (RG 8 +/- 8 weeks vs. NR 13 +/- 9 weeks, p = 0.09) that became significant in patients with 3 or more relapses (multiple-relapse group, MR, 4 +/- 2 weeks vs. NR 13 +/- 9 weeks, p < 0.04). Although the initial prednisone dose was similar in patients with relapsing forms, its maintenance was shorter than in patients without relapses, showing a trend to significance (RG 4 +/- 3 weeks, NR 7 +/- 6 weeks, p = 0.09). Lower levels of lactate dehydrogenase and gamma-glutamyltransferase, although always within the normal range, were found in patients with relapsing forms. CONCLUSION: COP is a specific but infrequent form of OP with a good response to steroid therapy. Relapses are frequent and typical characteristics of COP which resolved with ulterior treatment. Multifocal opacities on chest X-ray and a shorter maintenance of the initial steroid dose may increase the risk of relapse.

Respiratory failure secondary to organizing pneumonia in a patient with membranoproliferative glomerulonephritis and lung haemorrhage.

We report the case of a woman with membranoproliferative glomerulonephritis type I who presented with acute respiratory failure secondary to alveolar haemorrhage. The persistence of the respiratory failure, once the alveolar haemorrhage had ceased, was apparently due to diffuse alveolar damage in organization. The recognition of this proliferative reaction through the performance of transbronchial biopsy allowed the initiation of a correct treatment with which a favourable response was obtained.

Niveles de contaminación por ruido en una de las principales avenidas de la ciudad de México / Noise pollution levels in a large street in Mexico city

Se reportan los resultados de la medición de los niveles de contaminación por ruido, encontrados en seis horarios a lo largo de las 24 hr del día, en una de las principales avenidad de la ciudad de México en el año de 1993. Las mediciones se efectuaron mediante un sonómetro a las: 4.00, 8.00, 11.00, 15.00, 19.00 y 23.00 hr. El nivel de ruido en el percentil 10 (NR10) en los horarios estudiados fué de 80.3, 104.8, 100.9, 103.8, 95.6 y 95.2 dB "A" a las 4.00, 8.00, 11.00, 15.00, 19.00 y 23.00 hrs. respectivamente. El nivel de ruido en el percentil 50 (NR50) en los horarios estudiados fué de 67.7, 90.5, 87.5, 90.5, 86.5, y 77.2 dB "A" respectivamente. El nivel de ruido en el percentil 90 (NR90) fué de 60.2, 85.6, 82.1, 86.8, 84.7 y 73.9 dB "A". La correlación entre el ruido y la cantidad de automóviles circulantes fué de r=0.86. estos datos muestran que los niveles de ruido en calles de denso tráfico en la ciudad de México, se mantienen por arriba de la norma considerad como saludable (70 dB), a lo largo del día asociados al tráfico automotor

Validación del uso de juguetes sonoros mexicanos en la detección temprana de hipoacusias profundas bilaterales / Validation of the use of mexican sonorous toys for the early detection of profound bilateral hering loss

Se informa de los resultados de la validación de una prueba de rastreo de la audición en niños de OVol. 38 No. 4 1993 tres juguetes sonoros (matraca, timbre y palo de lluvia), de manufactura artesanal mexicana en dos poblaciones, una de ellas aleatoria y otra de población abierta. La sensibilidad fue de 88 por ciento y su especificidad fue de 90 por ciento. Se compararon los resultados diagnósticos de la prueba de juguetes sonoros contra la de potenciales provocados auditivos del tallo cerebral, no encontrándose diferencias significativas para identificar sorderas severas a produndas