Immunohistological study of the density and distribution of human penile neural tissue: gradient hypothesis.

Immunohistological patterns of density and distribution of neural tissue in the human penis, including the prepuce, are not fully characterized, and effects of circumcision (partial or total removal of the penile prepuce) on penile sexual sensation are controversial. This study analyzed extra- and intracavernosal innervation patterns on the main penile axes using formalin-fixed, paraffin-embedded human adult and fetal penile tissues, single- and double-staining immunohistochemistry and a variety of neural and non-neural markers, with a special emphasis on the prepuce and potential sexual effects of circumcision. Immunohistochemical profiles of neural structures were determined and the most detailed immunohistological characterizations to date of preputial nerve supply are provided. The penile prepuce has a highly organized, dense, afferent innervation pattern that is manifest early in fetal development. Autonomically, it receives noradrenergic sympathetic and nitrergic parasympathetic innervation. Cholinergic nerves are also present. We observed cutaneous and subcutaneous neural density distribution biases across our specimens towards the ventral prepuce, including a region corresponding in the adult anatomical position (penis erect) to the distal third of the ventral penile aspect. We also describe a concept of innervation gradients across the longitudinal and transverse penile axes. Results are discussed in relation to the specialized literature. An argument is made that neuroanatomic substrates underlying unusual permanent penile sensory disturbances post-circumcision are related to heightened neural levels in the distal third of the ventral penile aspect, which could potentially be compromised by deep incisions during circumcision.

IgG4-related disease diagnosed in a paratesticular pseudotumor simulating malignancy, in a patient with retroperitoneal fibrosis.

IgG4 related disease is a term used to describe a fibroinflammatory condition characterized by storiform fibrosis, inflammation with a dense lymphoplasmocytic infiltrate rich in plasma cells expressing IgG4, and often, if not always, raised serum levels of IgG4. We report a case of a patient with a past history of retroperitoneal fibrosis presenting with a swollen left testicle, who underwent an orchidectomy due to suspicion of malignancy. The surgical specimen revealed a paratesticular pseudotumor with histopathological and immunohistochemical characteristics of IgG4 related disease. To the best of our knowledge, just nine such cases have previously been reported, of which only three were manifestations of systemic disease, as in the present case. Whilst it is important to recognize the clinical and radiological features of this entity, histopathology is often essential in order to confirm the diagnosis.

WISP-2 modulates the induction of inflammatory mediators and cartilage catabolism in chondrocytes.

Wnt-1 inducible signaling pathway protein 2 (WISP-2/CCN5) is a recently identified adipokine that has been described as an important mediator of canonical Wnt activation in adipogenic precursor cells. In osteoarthritis (OA), the most common form of arthritis, chondrocytes exhibit aberrant and increased production of pro-inflammatory mediators and matrix degrading enzymes such as IL-1ß and MMP-13. Although recent evidence suggests a role for Wnt signaling in OA physiopathology, little is known about the involvement of WISP-2 in cartilage degradation. In the present study, we determined the expression of WISP-2 in healthy and OA human chondrocytes. WISP-2 expression is modulated along chondrocyte differentiation and downregulated at the onset of hypertrophy by inflammatory mediators. We also investigated the effect of WISP-2 on cartilage catabolism and performed WISP-2 loss-of-function experiments using RNA interference technology in human T/C-28a2 immortalized chondrocytes. We demonstrated that recombinant human WISP-2 protein reduced IL-1ß-mediated chondrocyte catabolism, that IL-1ß and WNT/b-catenin signaling pathways are involved in rhWISP-2 protein and IL-1ß effects in human chondrocytes, and that WISP-2 has a regulatory role in attenuating the catabolic effects of IL-1ß in chondrocytes. Gene silencing of WISP-2 increased the induction of the catabolic markers MMP-13 and ADAMTS-5 and the inflammatory mediators IL-6 and IL-8 triggered by IL-1ß in human primary OA chondrocytes in a Wnt/ß-catenin dependent manner. In conclusion, here we have shown for the first time that WISP-2 may have relevant roles in modulating the turnover of extracellular matrix in the cartilage and that its downregulation may detrimentally alter the inflammatory environment in OA cartilage. We also proved the participation of Wnt/ß-catenin signaling pathway in these processes. Thus, targeting WISP-2 might represent a potential therapeutical approach for degenerative and/or inflammatory diseases of musculoskeletal system, such as osteoarthritis.

[Tissue arrays: Two simple and economical methods for manual construction.] / Matrices tisulares: presentación de dos métodos sencillos y económicos de elaboración manual.

OBJECTIVES: Tissue array (TA) technologyis widely used as a method for the in situ investigation oftissue markers in cancer studies. A limitation of this techniqueis the high price of tissue arrayers. We describetwo easy and non-expensive manual methods, that haveproduced small and medium format arrays. MATERIAL AND METHODS: 16 TAs were manuallyconstructed from conventional paraffin blocks using twodifferent techniques. For the first method, a 16G Tru-Cutneedle whose bevel edge had been cut, was used tomake the holes in the donor blocks (80 cases) and thereceptor ones (resulting in 2 TAs each one with 55 casesand two with 25 cases). In the second technique, a 4mm-diameter punch for cutaneous biopsies was appliedto the donor blocks (obtaining 210 cylinders from 108blocks) and to the receptor ones (12 TAs). Hematoxylin-eosin, immunohistochemical and in situ hybridizationstains were performed on sections from these TAs. RESULTS: The tissue loss rate in the sections obtainedfrom the TAs constructed with the first method was26.5%, but as two cylinders were included from eachcase, at least one of them was retained. There was notany loss of tissue in the sections from the TAs constructedwith the second method. The results of all of the stainsperformed were successful. CONCLUSIONS: These two manual methods of elaborationof TAs result rather simple and they are economical.The tissue loss rate is significant in the first methodbut it can be compensated embedding more than onecylinder from each donor block. There was not anyproblem in the sectioning of the TAs constructed with thesecond method.

OBJETIVOS: La tecnología de matricestisulares (MTs) se ha implantado como método de trabajohabitual en la investigación de marcadores tisularesrelacionados con el cáncer. Su inconveniente esla necesidad de contar con un dispositivo especial deprecio elevado. Presentamos dos métodos manuales,económicos, que han sido válidos para construir MTsde pequeño y mediano formato.MATERIAL Y MÉTODOS: Se han elaborado 16 MTs deforma manual a partir de bloques convencionales deparafina, mediante dos técnicas diferentes. En la primerase utilizó una aguja Tru-Cut 16G a la que se cortó el bisel, para realizar los orificios en los bloques donantes(80 casos) y en los receptores (resultando dos MTs de55 casos y dos de 25 casos). Para la segunda técnicase utilizó un dispositivo "punch" para biopsias cutáneas,de 4 mm de diámetro, que se aplicó a los bloques donantes(obteniendo 210 cilindros de 108 bloques) y alos receptores (12 MTs). En las secciones de las MTs obtenidasse realizaron tinciones de hematoxilina-eosina,inmunohistoquímica (IHQ) e hibridación in situ. RESULTADOS: La tasa de pérdida de material en las seccionesobtenidas con el primer método fue del 26,5%,pero al haberse incluído dos cilindros de cada caso, almenos uno de ellos se conservó. En las MTs obtenidascon el método de punch biopsia no hubo pérdidas detejido. Los resultados de todas las tinciones realizadasfueron óptimos. CONCLUSIONES: Estos dos métodos manuales de elaboraciónde MTs resultan relativamente sencillos y soneconómicos. La tasa de pérdida de tejido es sólo significativaen el primero de los métodos pero se puedecompensar incluyendo varios cilindros de cada bloquedonante. En el segundo método no han existido problemasdestacables en la fase de microtomía.

Matrices tisulares: presentación de dos métodos sencillos y económicos de elaboración manual / Tissue arrays: Two simple and economical methods for manual construction

OBJETIVOS: La tecnología de matrices tisulares (MTs) se ha implantado como método de trabajo habitual en la investigación de marcadores tisulares relacionados con el cáncer. Su inconveniente es la necesidad de contar con un dispositivo especial de precio elevado. Presentamos dos métodos manuales, económicos, que han sido válidos para construir MTs de pequeño y mediano formato. MATERIAL Y MÉTODOS: Se han elaborado 16 MTs de forma manual a partir de bloques convencionales de parafina, mediante dos técnicas diferentes. En la primera se utilizó una aguja Tru-Cut 16G a la que se cortó el bisel, para realizar los orificios en los bloques donantes (80 casos) y en los receptores (resultando dos MTs de 55 casos y dos de 25 casos). Para la segunda técnica se utilizó un dispositivo "punch" para biopsias cutáneas, de 4 mm de diámetro, que se aplicó a los bloques donantes (obteniendo 210 cilindros de 108 bloques) y a los receptores (12 MTs). En las secciones de las MTs obtenidas se realizaron tinciones de hematoxilina-eosina, inmunohistoquímica (IHQ) e hibridación in situ. RESULTADOS: La tasa de pérdida de material en las secciones obtenidas con el primer método fue del 26,5%, pero al haberse incluído dos cilindros de cada caso, al menos uno de ellos se conservó. En las MTs obtenidas con el método de punch biopsia no hubo pérdidas de tejido. Los resultados de todas las tinciones realizadas fueron óptimos. CONCLUSIONES: Estos dos métodos manuales de elaboración de MTs resultan relativamente sencillos y son económicos. La tasa de pérdida de tejido es sólo significativa en el primero de los métodos pero se puede compensar incluyendo varios cilindros de cada bloque donante. En el segundo método no han existido problemas destacables en la fase de microtomía

OBJECTIVES: Tissue array (TA) technology is widely used as a method for the in situ investigation of tissue markers in cancer studies. A limitation of this technique is the high price of tissue arrayers. We describe two easy and non-expensive manual methods, that haveproduced small and medium format arrays. MATERIAL AND METHODS: 16 TAs were manually constructed from conventional paraffin blocks using two different techniques. For the first method, a 16G Tru-Cut needle whose bevel edge had been cut, was used to make the holes in the donor blocks (80 cases) and the receptor ones (resulting in 2 TAs each one with 55 cases and two with 25 cases). In the second technique, a 4 mm-diameter punch for cutaneous biopsies was applied to the donor blocks (obtaining 210 cylinders from 108 blocks) and to the receptor ones (12 TAs). Hematoxylin-eosin, immunohistochemical and in situ hybridization stains were performed on sections from these TAs. RESULTS: The tissue loss rate in the sections obtained from the TAs constructed with the first method was 26.5%, but as two cylinders were included from each case, at least one of them was retained. There was not any loss of tissue in the sections from the TAs constructed with the second method. The results of all of the stains performed were successful. CONCLUSIONS: These two manual methods of elaboration of TAs result rather simple and they are economical. The tissue loss rate is significant in the first method but it can be compensated embedding more than one cylinder from each donor block. There was not any problem in the sectioning of the TAs constructed with the second method

Breast cancer subtype discrimination using standardized 4-IHC and digital image analysis.

Breast cancer is a heterogeneous disease. Surrogate classification of intrinsic subtypes of invasive carcinomas by combined immunohistochemistry for estrogen receptor (ER), progesterone receptor (PR), HER2, and Ki67 (4-IHC) has increased steadily since the 2011 St Gallen symposium, due to its rapid subtyping of tumors at a reasonable cost. An important step in improving 4-IHC reproducibility and reliability will be to provide reference values from the routine use of standardized 4-IHC followed by image analysis. The aims of the current study were (1) to analyze invasive breast carcinomas using standardized 4-IHC and quantitative image analysis and (2) to compare the results obtained in the classification of biological subtypes using current Ki67 and PR threshold values proposed by different authors to sub-classifying the luminal A-like and the luminal B-like (HER2-negative) subtypes. Five hundred twenty-one tumors were analyzed by standardized immunohistochemistry, with automatic image analysis, and HER2 FISH technique. Positivity for ER was found in 82.7% and for PR in 70.1% of cases. Using the Allred scoring system, hormone receptor results showed a bimodal distribution, particularly for ER. HER2 positivity was found in 15.7% of cases, and the mean Ki67 score was 32.3%. Using the most recently proposed surrogate definitions for the classification of luminal breast cancer subtypes, the percentages of different subtypes that we found were similar to those published with genomic platforms: 40.7% luminal A-like, 32.4% luminal B-like/HER2-negative, 9.8% luminal B-like/HER2-positive, 6.0% HER2-positive, and 11.1% triple negative. Standardized 4-IHC with automatic image analysis constitutes a low-cost method for surrogate definitions of biological subtypes of breast cancer that delivers accurate results in a day.