CD68 and CD83 immune populations in non-metastatic axillary lymph nodes are of prognostic value for the survival and relapse of breast cancer patients.

BACKGROUND: The foremost cause of death of breast cancer (BC) patients is metastasis, and the first site to which BC predominantly metastasizes is the axillary lymph node (ALN). Thus, ALN status is a key prognostic indicator at diagnosis. The immune system has an essential role in cancer progression and dissemination, so its evaluation in ALNs could have significant applications. In the present study we aimed to investigate the association of clinical-pathological and immune variables in the primary tumour and non-metastatic ALNs (ALNs-) of a cohort of luminal A and triple-negative BC (TNBC) patients with cancer-specific survival (CSS) and time to progression (TTP). METHODS: We analysed the differences in the variables between patients with different outcomes, created univariate and multivariate Cox regression models, validated them by bootstrapping and multiple imputation of missing data techniques, and used Kaplan-Meier survival curves for a 10-years follow-up. RESULTS: We found some clinical-pathological variables at diagnosis (tumour diameter, TNBC molecular profile and presence of ALN metastasis), and the levels of several immune markers in the two studied sites, to be associated with worse CSS and TTP. Nevertheless, only CD68 and CD83 in ALNs- were confirmed as independent prognostic factors for TTP. CONCLUSIONS: The study identified the importance of macrophage and dendritic cell markers as prognostic factors of relapse for BC. We highlight the importance of studying the immune response in ALNs-, which could be relevant to the prediction of BC patients' outcome.

Chemical composition, enantiomeric analysis and anticholinesterase activity of Lepechinia betonicifolia essential oil from Ecuador.

CONTEXT: Due to the interesting potential of essential oils (EO) against cholinesterases and their close relation in Alzheimer's disease, the EO of Lepechinia betonicifolia (Lam) Epling (Lamiaceae), a native shrub from Ecuador, was assessed. Chemical profiling and enantiomeric distribution were also recorded for the first time. OBJECTIVE: To analyse the chemical profile including the enantiomeric composition and anticholinesterase effect exerted by EO of L. betonicifolia. MATERIALS AND METHODS: The EO of L. betonicifolia fresh aerial parts was obtained by hydrodistillation in a Clevenger-type apparatus. Physical properties were determined according to standard norms. The chemical composition was determined by GC-MS and GC-FID. Enantioselective GC-MS analysis was carried out by using a capillary chiral column. Anticholinesterase effect was assessed by Ellman's method with acetylthiocoline as substrate and Ellman's reagent (DTNB) to detect its hydrolysis at 405 nm for 60 min. Donepezil was used as a reference drug. EO was dissolved in methanol to reach 10 mg/mL concentration and two more 10× dilutions were included. RESULTS: Thirty-nine constituents were identified corresponding to 97.55% of the total oil composition. The main components were ß-pinene (30.45%), sabinene (27.98%), α-pinene (4.97%), ß-phellandrene (4.79%), E-caryophyllene (4.44%) and limonene (3.84%). L. betonicifolia EO exerted a strong inhibitory effect over the AChE enzyme with an IC50 value of 74.97 ± 1.17 µg/mL. DISCUSSION AND CONCLUSIONS: Current chemical characterisation and anticholinesterase effect of EO of L. betonicifolia encourage us to propose this EO as a candidate for the preparation of functional foods or as adjuvant therapy for Alzheimer's disease.

Liquid Biopsy-Based Exo-oncomiRNAs Can Predict Prostate Cancer Aggressiveness.

Liquid biopsy-based biomarkers, including microRNAs packaged within extracellular vesicles, are promising tools for patient management. The cytokine tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is related to PCa progression and is found in the semen of patients with PCa. TWEAK can induce the transfer of exo-oncomiRNAs from tumor cells to body fluids, and this process might have utility in non-invasive PCa prognosis. We investigated TWEAK-regulated exo-microRNAs in semen and in post-digital rectal examination urine from patients with different degrees of PCa aggressiveness. We first identified 14 exo-oncomiRNAs regulated by TWEAK in PCa cells in vitro, and subsequently validated those using liquid biopsies from 97 patients with PCa. Exo-oncomiR-221-3p, -222-3p and -31-5p were significantly higher in the semen of high-risk patients than in low-risk peers, whereas exo-oncomiR-193-3p and -423-5p were significantly lower in paired samples of post-digital rectal examination urine. A panel of semen biomarkers comprising exo-oncomiR-221-3p, -222-3p and TWEAK was designed that could correctly classify 87.5% of patients with aggressive PCa, with 85.7% specificity and 76.9% sensitivity with an area under the curve of 0.857. We additionally found that TWEAK modulated two exo-oncomiR-221-3p targets, TCF12 and NLK. Overall, we show that liquid biopsy detection of TWEAK-regulated exo-oncomiRNAs can improve PCa prognosis prediction.

Differences in the Immune Response of the Nonmetastatic Axillary Lymph Nodes between Triple-Negative and Luminal A Breast Cancer Surrogate Subtypes.

Breast cancer (BC) comprises four immunohistochemical surrogate subtypes of which triple-negative breast cancer (TNBC) has the highest risk of mortality. Axillary lymph nodes (ALNs) are the regions where BC cells first establish before distant metastasis, and the presence of tumor cells in the ALN causes an immune tolerance profile that contrasts with that of the nonmetastatic ALN (ALN-). However, few studies have compared the immune components of the ALNs- in BC subtypes. The present study aimed to determine whether differences between immune populations in the primary tumor and ALNs- were associated with the luminal A or TNBC subtype. We evaluated a retrospective cohort of 144 patients using paraffin-embedded biopsies. The TNBC samples tended to have a higher histologic grade and proliferation index and had higher levels of immune markers compared with luminal A in primary tumors and ALNs-. Two methods for validating the multivariate analysis found that histologic grade, intratumoral S100 dendritic cells, and CD8 T lymphocytes and CD57 natural killer cells in the ALNs- were factors associated with TNBC, whereas CD83 dendritic cells in the ALNs- were associated with the luminal A subtype. In conclusion, we found that intratumoral regions and ALNs- of TNBC contained higher concentrations of markers related to immune tolerance than luminal A. This finding partially explains the worse prognosis of patients with TNBC.

Biofluid quantification of TWEAK/Fn14 axis in combination with a selected biomarker panel improves assessment of prostate cancer aggressiveness.

BACKGROUND: Conventional clinical biomarkers cannot accurately differentiate indolent from aggressive prostate cancer (PCa). We investigated the usefulness of a biomarker panel measured exclusively in biofluids for assessment of PCa aggressiveness. METHODS: We collected biofluid samples (plasma/serum/semen/post-prostatic massage urine) from 98 patients that had undergone radical prostatectomy. Clinical biochemistry was performed and several cytokines/chemokines including soluble(s) TWEAK, sFn14, sCD163, sCXCL5 and sCCL7 were quantified by ELISA in selected biofluids. Also, the expression of KLK2, KLK3, Fn14, CD163, CXCR2 and CCR3 was quantified by real-time PCR in semen cell sediment. Univariate, logistic regression, and receiver operating characteristic (ROC) analyses were used to assess the predictive ability of the selected biomarker panel in conjunction with clinical and metabolic variables for the evaluation of PCa aggressiveness. RESULTS: Total serum levels of prostate-specific antigen (PSA), semen levels of sTWEAK, fasting glycemia and mRNA levels of Fn14, KLK2, CXCR2 and CCR3 in semen cell sediment constituted a panel of markers that was significantly different between patients with less aggressive tumors [International Society of Urological Pathology (ISUP) grade I and II] and those with more aggressive tumors (ISUP grade III, IV and V). ROC curve analysis showed that this panel could be used to correctly classify tumor aggressiveness in 90.9% of patients. Area under the curve (AUC) analysis revealed that this combination was more accurate [AUC = 0.913 95% confidence interval (CI) 0.782-1] than a classical non-invasive selected clinical panel comprising age, tumor clinical stage (T-classification) and total serum PSA (AUC = 0.721 95% CI 0.613-0.830). CONCLUSIONS: TWEAK/Fn14 axis in combination with a selected non-invasive biomarker panel, including conventional clinical biochemistry, can improve the predictive power of serum PSA levels and could be used to classify PCa aggressiveness.

Tumor desmoide intraabdominal simulando una tumoración pancreática en una paciente con poliposis colónica familiar / Intra-abdominal desmoid tumor mimicking a pancreatic tumor in a patient with familial colonic polyposis

IntroducciónLa fibromatosis es una proliferación fibroblástica benigna con crecimiento infiltrativo local. Se clasifica en una forma superficial y en una forma profunda, también denominada tumor desmoide (TD). Esta última puede presentarse en forma esporádica o asociada a la poliposis adenomatosa familiar y síndrome de Gardner. La presentación pancreática es excepcional y sólo existen 8 casos descritos en la literatura médica.Observación clínicaMujer de 29 años con antecedentes de PCF y 2 lesiones pancreáticas. En la pieza quirúrgica se observaron 2 lesiones mal delimitadas en el páncreas con infiltración de órganos vecinos. Histológicamente, correspondían a una proliferación de estirpe mesenquimatosa de patrón fusocelular sin atipias citológicas, que se diagnosticaron de TD.DiscusiónLa etiología de la fibromatosis es desconocida. En pacientes con PCF la localización más habitual de los TD es intraabdominal siendo inusual la presentación pancreática. Esto plantea el diagnóstico diferencial con otras neoplasias del páncreas(AU)

IntroductionFibromatosis consists of a benign fibroblastic proliferation with local infiltrative growth. Two types are recognized: a superficial and a deep form, also known as desmoid tumor. The latter may occur sporadically or in association with familial adenomatous polyposis and Gardner's syndrome. Pancreatic presentation is exceptional and only eight cases have been described in the literature.Case reportWe report the case of a 29-year-old woman with a history of familial colonic polyposis and two pancreatic lesions. In the surgical specimen, two poorly defined pancreatic lesions were observed with infiltration of neighboring organs. Histologically, the lesions corresponded to mesenchymal proliferation with a fusocellular pattern without cytological atypica, which were diagnosed as desmoid tumors.DiscussionThe etiology of fibromatosis is unknown. In patients with familial colonic polyposis, the most common localization of desmoid tumor is intra-abdominal. Pancreatic presentation is unusual, requiring differential diagnosis with other pancreatic neoplasms(AU)

[Intra-abdominal desmoid tumor mimicking a pancreatic tumor in a patient with familial colonic polyposis]. / Tumor desmoide intraabdominal simulando una tumoración pancreática en una paciente con poliposis colónica familiar.

INTRODUCTION: Fibromatosis consists of a benign fibroblastic proliferation with local infiltrative growth. Two types are recognized: a superficial and a deep form, also known as desmoid tumor. The latter may occur sporadically or in association with familial adenomatous polyposis and Gardner's syndrome. Pancreatic presentation is exceptional and only eight cases have been described in the literature. CASE REPORT: We report the case of a 29-year-old woman with a history of familial colonic polyposis and two pancreatic lesions. In the surgical specimen, two poorly defined pancreatic lesions were observed with infiltration of neighboring organs. Histologically, the lesions corresponded to mesenchymal proliferation with a fusocellular pattern without cytological atypica, which were diagnosed as desmoid tumors. DISCUSSION: The etiology of fibromatosis is unknown. In patients with familial colonic polyposis, the most common localization of desmoid tumor is intra-abdominal. Pancreatic presentation is unusual, requiring differential diagnosis with other pancreatic neoplasms.

Multiorgan histiocytosis after B-cell acute lymphoblastic leukemia.

The association of multiorgan histiocytosis after acute lymphoblastic leukemias is very rare as most cases are localized forms of Langerhans cell histiocytosis (LCH). We report on an 18-year-old man diagnosed with B-cell acute lymphoblastic leukemia (B-ALL) with p16 deletion (9p21). He was treated with induction chemotherapy using the Spanish PETHEMA group protocol and achieved complete remission. Three months after the diagnosis of B-ALL, he developed a severe multiorgan histiocytosis that is clinically suggestive of LCH but lacked typical immunohistochemical features of LCH and indeterminate cell histiocytosis: CD1a was strongly positive, CD68 and S-100 protein were moderately positive, and langerin was negative. The drugs of the first-line treatment recommended for LCH had been part of the chemotherapy of B-ALL that the patient had received. Therefore, we prescribed the second-line treatment for LCH (cytarabine and 2'-chlorodeoxyadenosine), and he achieved partial remission. The patient died during the aplasia induced by the third cycle of chemotherapy from pneumonia. We could not demonstrate the transdifferentiation of tumoral lymphocytes into histiocytes, using p16 deletion (9p21) as a marker, because these cells did not share the mutation. Neither could we study immunoglobulin-H rearrangement as we had exhausted all the tissue samples. In the medical literature, there are a few reported cases of T-cell acute lymphoblastic leukemia followed by disseminated LCH and just 1 case of B-ALL followed by localized LCH affecting the bones. Therefore, our patient may be the first published case of B-ALL followed by histiocytosis, which had 2 singularities: it was multiorgan and the immunohistochemistry was not typical of LCH.

Effects of retinoic acid on the neural crest-controlled organs of fetal rats.

Prenatal exposure of rat embryos to retinoic acid induces severe malformations involving various organs. The mechanisms of this embryopathy are known only in part. This study describes the malformations of the neural crest-derived organs in this model and shows that many of them fit into the pattern of disturbed neural crest organogenic control. Pregnant rats were exposed to either all-trans retinoic acid (125 mg/kg; n=17) or vehicle ( n=10) on E10. Fetuses were recovered on E21 and external and internal malformations were sought. The craniofacial area, the trachea, parathyroids, thymus, thyroid, heart, great vessels, and adrenals were examined. In contrast with normal controls, 100% of retinoic acid animals had craniofacial, 94% anorectal, 90% limb, and 55% neural tube defects. The thymus was absent or ectopic in 76%, the parathyroids were absent or single in 88%, and the thyroid was abnormal in 41%. There were neural crest-type (outflow tract and/or pharyngeal aortic arch defects) cardiovascular malformations in 90% and the adrenals were absent in 52%. Interestingly, 9 of 11 (88%) animals with neural tube defects had absent adrenal glands. This association was significant ( p<0.01) by Fisher exact test. Among the complex mechanisms of retinoic acid teratogenesis, severe disturbances of the neural crest pathway play a leading role. The simultaneous development of neural tube defects and adrenal agenesis suggests common pathogenic pathways.

Effects of vitamin A on malformations of neural-crest-controlled organs induced by nitrofen in rats.

Vitamin A (vit A) alleviates the effects of nitrofen in exposed rat pups. The present study examines the effects of early exposure to vitamin A on the neural-crest-related cardiovascular, thymic, parathyroid, and thyroid malformations previously reported in the rat model of congenital diaphragmatic hernia (CDH). Pregnant rats were exposed on gestational day 9.5 to 100 mg 2-4-dichlorophenyl-p-nitrophenyl ether (nitrofen) alone or followed by 15,000 IU vit A. Controls were treated only with oil or oil + vit A. The fetuses were recovered near term and diaphragmatic, lung, heart, and thymic malformations were sought after dissection. The parathyroids and thyroid were histologically investigated. The hearts were also examined for protein, DNA, and proportion of proliferating cells. None of the control fetuses had malformations, whereas 41% of nitrofen and 27% of nitrofen + vit A fetuses had CDH. Anomalies of the heart outflow tract and pharyngeal arteries were seen in 64% and 43%, respectively, in both groups. Heart and thymic hypoplasia, which were severe in the nitrofen group with significant decreases of total DNA and percent proliferating cells, were significantly improved in the nitrofen + vit A group. The hypoplastic thymus was malformed in 53% and 27% of fetuses, respectively, and the parathyroids were abnormal in 48% and 35%, respectively. Only minimal anomalies of the thyroid were found. The significant improvement of heart and thymic hypoplasia associated with vit A was not seen for the other variables studied, but there was a trend in this direction for all of them. Vit A definitely improved heart hypoplasia induced by nitrofen by stimulating myogenesis. It also improved thymic hypoplasia, but had limited beneficial effects on malformations of the cardiac outflow tract and pharyngeal derivatives that accompany CDH in rats exposed to nitrofen.

Effects of early embryonal exposure to dexamethasone on malformations of neural-crest derivatives induced by nitrofen in rats.

Prenatal corticosteroids reverse to some extent lung and heart hypoplasia in nitrofen-exposed rat pups. The present study examines the effects of early exposure to dexamethasone on the neural crest-related malformations of the cardiovascular system, thymus, parathyroids, and thyroid observed in this model. Pregnant rats were exposed on gestational day 9.5 to either 100 mg 2-4-dichlorophenyl-p-nitrophenyl ether (nitrofen) alone or followed on days 10.5 and 11.5 by 0.4 mg/kg dexamethasone (dexa) i.p. Controls were treated with either oil alone or oil+dexa alone. The fetuses were recovered near term and diaphragmatic, lung, heart, and thymic malformations were sought after dissection. The parathyroids and thyroid were histologically investigated. Control fetuses had no malformations whereas 68% of nitrofen and 65% of nitrofen + dexa fetuses had congenital diaphragmatic hernias (CDH). Heart-outflow tract and pharyngeal artery anomalies were seen in 62% and 61%, respectively in both groups. Heart hypoplasia, which was severe in the nitrofen group, was fully reversed in nitrofen+dexa pups. In contrast, thymic hypoplasia was of similar severity in both groups. The hypoplastic thymus was malformed in 29% and 39%, the parathyroids in 50% and 41%, and the thyroid in 25% and 16% of fetuses, respectively. These differences were not significant. Early exposure to dexa in rat fetuses previously treated with nitrofen thus does not produce any benefit on the incidence or severity of malformations of the cardiac outflow tract and pharyngeal derivatives that accompany CDH in rats exposed to nitrofen. However, even administered so early, this medication prevents heart hypoplasia, suggesting a favorable effect on early heart organogenesis.

Enfermedad de Hodgkin. Papel etiopatogénico del virus de Epstein-Barr en las comarcas de Tarragona / Hodgkin's disease. Etiopathogenic role of EBV in Tarragona (Spain)

Fundamento: Se pretende determinar la proporción de enfermedad de Hodgkin (EH) que expresa el virus de Epstein-Barr (VEB) en nuestro medio. Pacientes y métodos: Se ha realizado un estudio retrospectivo sobre 49 casos de EH usando la técnica inmunohistoquímica LMP-1 y la técnica de hibridación in situ para EBER-1. Resultados: Un 40,8 por ciento (20/49) de los casos expresaba VEB (EBER-1 y/o LMP-1 positivos). Este porcentaje fue significativamente mayor en EH diagnosticadas a pacientes mayores de 55 años y no hubo diferencias por sexo, aunque fue mayor, pero no de forma significativa, en el subtipo histológico de EH de celularidad mixta. Conclusiones: El VEB se asocia a un 40,8 por ciento de las EH en las comarcas de Tarragona (AU)