Prognostic factors in renal-cell carcinoma: Immunohistochemical detection of p53 protein versus clinico-pathological parameters

Immunoreactivity forp53 protein was assessed in 100 cases of primary renal-cell carcinoma (RCC). The results were correlated with clinical survival data (follow-up 24 to 84 months: mean: 39 months) and with clinico-pathological parameters, including nuclear grade, tumour stage, cell type, tumour architecture and tumour diameter. In all, 32% of the tumours were p53-positive; there was no difference in survival between p53-positive and -negative cases. Similarly, p53 expression did not correlate with any of the clinico-pathological parameters mentioned. Nuclear grade (grade 1 + 2 vs. grade 3 + 4) had a striking impact on prognosis and so, to a lesser extent, did tumour stage and the occurrence of a spindle-cell component. The immunohistochemical detection of p53 in RCC is not of prognostic value. The estimation of nuclear grade, however is a major predictor of prognosis

Delineation of the healthy rabbit tonsil by immunohistochemistry - A short communication

Situated in the oral cavity, the rabbit palatine tonsils are part of the mucosal immune system and help to defend the body against foreign pathogens. Expressed as two oval protrusions in the wall of the oropharynx, the rabbit palatine tonsils are characterized by excretory ducts and trabeculae. We here compare paraffin embedded and cryosections of the healthy rabbit tonsils. This analysis centers on evaluating the differential outcomes resulting from the application of these fixation methodologies in conjunction with immunohistochemical assays targeting collagen I, collagen III, fibronectin, α-smooth muscle actin (α-SMA), and ki67. Subsequent recommendations are provided based on our findings. Furthermore, we demonstrate the advantage of an antigen retrieval step in immunohistochemical labeling of paraffin sections. Basic classical histological stainings as HE, GT and elastin were also performed. Comparison of different stainings and labelings was furthermore performed in serial sections, showing that adjacent to the excretory ducts, the tonsillar tissue was particularly composed of collagen I and fibronectin, while the vessel walls were predominantly α-SMA positive. Moreover, PAR-2 immunohistochemical staining was performed, where a small fraction of the cells found in the tonsillar connective tissue were PAR-2 positive (probably a subpopulation of mast cells), as well as the lumen of some excretory ducts and trabeculae. Collagen III on the other hand was only weakly expressed in the tonsils. Proliferating ki67 positive cells were rare. This endeavor serves to furnish the scientific community with reference imagery pertinent to researchers opting for the rabbit palatine tonsil model. The diversity of staining techniques employed herein establishes a foundational repository of images, primed for comparative analysis against pathological conditions. Furthermore, these images hold the potential to illustrate inter-species variations. For instance, they can be juxtaposed against murine or rodent tonsils, or even offer insights into the human context

Delineation of the healthy rabbit tongue by immunohistochemistry - A technical note

In the oral cavity the tongue is an important muscular organ that supports the swallowing of food and liquids. It is responsible for the sense of taste, based on the many different taste buds it contains. Research in the field of tongue diseases demands for suitable preclinical models. The healthy rabbit tongue may therefore serve as baseline and reference for the pathological situation. With this consideration, we covered the fixation and histological stainings as well as the immunohistochemical labelling of the healthy rabbit tongue. In this technical note, initial choice of the fixative is discussed, with a comparison of formalin fixation and subsequent paraffin embedding versus cryopreservation. Moreover, we delineate the effect of an antigen retrieval step for formalin fixation by several examples. Finally, we provide ECM markers collagen I, collagen III, fibronectin, α-SMA and elastin staining as well as ki67 for proliferative status and PAR-2 protein expression as a marker for inflammatory status and nociception in tongue sections, mainly from the tongue body. Technically, we found superiority of paraffin sections for collagen I, collagen III, fibronectin, ki67 and α-SMA labelling, for selected detections systems. As for ECM components, the lamina propria was very rich in collagen and fibronectin, while the muscular body of the tongue showed only collagen and fibronectin positive areas between the muscle fibers. Moreover, α-SMA was clearly expressed in the walls of arteries and veins. The inflammatory marker PAR-2 on the other hand was prominently expressed in the salivary glands and to some extent in the walls of the vessels. Particular PAR-2 expression was found in the excretory ducts of the tongue. This technical note has the aim to provide baseline images that can be used to compare the pathological state of the diseased rabbit tongue as well as for inter-species comparison, such as mouse or rat tongue. Finally, it can be used for the comparison with the human situation

Delineation of the healthy rabbit kidney by immunohistochemistry - A technical note.

Pre-clinical animal models are needed to investigate and study kidney injuries and diseases. The rabbit kidney model is frequently used because various important parameters can be assessed with it. For example, histology and immunohistochemistry are indispensable as tissue morphology and composition can be investigated qualitatively as well as quantitatively. Here, different histological and immunohistochemical stainings were performed in the rabbit healthy naïve kidney tissue. First, overnight formalin fixation followed by paraffin embedding and cryopreservation with a subsequent 10-minute formalin fixation prior to staining were compared. Cryosections showed a more pronounced staining pattern, with clear borders at low magnifications, but blurred borders at higher magnifications. Then, antigen retrieval (AR) for paraffin embedded sections resulted in more prominent corresponding signals compared to stainings without AR. Moreover, several advantages and disadvantages of chromogenic versus immunofluorescence stainings were considered. Chromogenic staining was advantageous compared to immunofluorescence for collagen I and III, and to a minor degree for fibronectin. Finally, distinct structures, such as the pelvis, the calices, the glomeruli and tubuli, were stained in serial sections with diverse immunohistochemical stainings in order to delineate their composition. The following stainings were performed: standard Haematoxylin&Eosin and Elastica van Gieson staining, collagen I, collagen III, fibronectin, α-SMA, ki-67 and protease-activated receptor-2 (PAR-2). While chromogenic stainings of collagen I and collagen III were particularly useful to depict kidney structures in paraffin sections compared with cryosections, cryosections immunofluorescently stained for α-SMA were superior to paraffin sections, particularly at higher magnifications. With regard to specific structures, we found renal vessel walls positive for fibronectin and α-SMA, while the Bowman's capsule was only positive for fibronectin and α-SMA showed only tiny spots. The mesangial cells of the glomeruli and the distal tubuli were PAR-2 positive, while the proximal tubuli were PAR-2 negative

Delineation of the healthy rabbit heart by immunohistochemistry - A technical note

Heart failure poses a big health problem and may result from obesity, smoking, alcohol and/or growing age. Studying pathological heart tissue demands accurate histological and immunohistochemical stainings in animal models, including chromogenic and fluorescent approaches. Moreover, a reliable set of healthy heart stainings and labeling are required, in order to provide a reference for the pathological situation. Heart and brain tissue of a healthy rabbit were collected, and different histological key steps were compared, such as paraffin embedding after formalin fixation versus cryopreservation; an antigen retrieval (AR) step in processing paraffin sections versus the same procedure without AR; or a chromogenic with a fluorescent detection system, respectively. Using serial sections, we stained the same morphological structure with classic approaches (HE, Masson Goldner Trichrome (GT) and Elastica van Gieson (EL)) and with different markers, including collagen I, collagen III, fibronectin, α-SMA, protease-activated receptor-2 (PAR-2) which is an inflammation-related marker, and ki67 for proliferating cells. Differences between conditions were quantitatively assessed by measuring the color intensity. Generally, cryosections exhibited a more prominent signal intensity in immunohistochemically labeled sections than in paraffin sections, but the strong staining was slurry, which sometimes impeded proper identification of morphological structures, particularly at higher magnifications. In addition, the advantage of an AR step was observed when compared to the condition without AR, where signal intensities were significantly lower. Different stainings of the heart arteries and the myocardium revealed a clear distribution of extracellular matrix components, with prominent collagen III in the artery wall, but an absence of collagen III in the myocardium. Moreover, paraffin-embedded sections provided more distinct structures compared to cryosections after collagen III, ki67, fibronectin, and α-SMA labeling. As for the Purkinje cells that were depicted in the heart and the cerebellum (Purkinje neurons), we found GT staining most suitable to depict them in the heart, while HE as well as EL staining was ideal to depict Purkinje neurons in the cerebellum. In sum, we provide useful reference images with different stainings for researchers using the rabbit heart or brain model. Such images can help to decide which of the immunohistochemical protocols are valuable to reach a specific aim. Recommendations are given for the best visualization of the target structures and specific (immunohistochemical) staining

Verificação da administração ilegal de estrógenos através de marcadores imunohistoquímicos na próstata de bovinos

The immunodetection of diverse cell markers was evaluated in prostatic samples from bullocks, and bullocks showing epithelial hyperplasia-metaplasia, with oestrogen-induced changes, and in experimental samples from bullocks inoculated with dietylstilbestrol (DES). Antigen-retrieval procedures allowed the use of tissues that had been fixed in formalin for long periods. Three tissue markers were chosen for the study: cytokeratins 13 and 16, vimentin and desmin. Monoclonal antibody K8.12 (specific for cytokeratins 13 and 16) stained basal cells and hyperplastic-metaplastic epithelium; monoclonal antivimentin, and desmin, allowed the definition of fibromuscular changes.A imunodeteccão de marcadores celulares foi avaliada em amostras prostáticas de bovinos com hiperplasia ou hiperplasia-metaplasia epiteliais, induzidas por estrógenos administrados ilegalmente e em próstatas de bovinos inoculados com dietilstilbestrol (DES). A técnica de recuperac ão antigênica permitiu o uso de tecidos fixados em formalina, por longos períodos. Foram utilizados os anticorpos monoclonais K8.12, anti-vimentina e anti-desmina para determinação de células basais coradas/epitélio hiperplásico-metaplásico, células do estroma e células musculares, respectivamente. As alterações tissulares observadas nos casos de campo e nos experimentais foram semelhantes, através do que se concluiu que houve administração ilegal de estrógenos. O teste imunohistoqu ímico com esses marca-dores específicos foi útil ao exame histológico da próstata, uma vez que a análise das imagens permite maior e melhor quantificação das altera- ções observadas. Os testes bioquí-micos, entretanto, são necess ários para uma avaliação mais precisa.Facultad de Ciencias Veterinaria

Evaluation of Retinoblastoma and Ki-67 Immunostaining as Diagnostic Markers of Benign and Malignant Parathyroid Disease

RID="" ID="" Correspondence to: F. Farnebo, M.D., Ph.D.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/42410/1/268-23-1-68_23n1p68.pd

Zusammenhang zwischen Expression des Proteins CD44v6, Metastasierung und Überleben bei Patienten mit Plattenepithelkarzinomen des Oropharynx

Wir untersuchten in dieser retrospektiven Kohortenstudie die Expression des Glycoproteins CD44v6, das im Kopf-Hals-Bereich stark exprimiert wird, in Korrelation zu Metastasierung, Differenzierungsgrad und Überleben von 120 Patienten mit Plattenepithelkarzinomen des Oropharynx. Dabei wurde die Färbeintensität der zuvor immunhistochemisch behandelten Tumorschnitte ausgewertet. Mittels Kruskal-Wallis- und Whitney-Man-Test fanden wir keinen Zusammenhang zwischen den Tumoren ohne, mit geringem und ausgedehntem Lymphknotenbefall und der Antigenexpression (p=0,665), bzw. zwischen Differenzierungsgrad und Antigenexpression (p=0,235). Auch die Überlebenskurven nach Kaplan-Meier zeigten keinen signifikanten Unterschied im Überleben der Patienten mit starker bzw. schwacher Antigenexpression (p=0,397). CD44v6 ist somit kein Prognosefaktor für Oropharynxkarzinome und hat keinen Einfluss auf die Metastasierung

The cell biology of basal cell carcinoma. relationship to histology and clinical outcome.

Basal cell carcinomas presents with extremely diverse clinical and histological appearances and behaviour. Currently there is little understanding of the biological processes that determine these variations. In an attempt to understand these differences, this thesis evaluated some aspects of the cell biology of BCC both a prospective series and in archival specimens. A variety of measurements were assessed in combination with patient factors (age, presentation etc.) and medical factors (type and adequacy of treatment). The cell kinetics of BCC was studied in vivo following administration of bromodeoxyuridine, which was analysed by flow cytometry. The growth fraction (Ki-67 immunohistochemistry) and the contribution of cell loss to the overall tumour kinetics were studied by evaluating apoptosis (morphologically) and the bcl-2, bax and p53 protein expression, using immunohistochemistry, in both the prospective and archival specimens (including non recurrent, recurrent and horrifying BCCs). It was apparent that BCCs are highly proliferative tumours with a median Ts of 7.6 hours (range 5.0-14-6), Tpot 2.8 days (range 4.0-18.3 days), LI 14%, and Gf 32%. Cell production rates were related to the histological growth pattern with infiltrative and morpheic tumours having a higher Gf than the nodular tumours (p<0.01) and a shorter Tc and Tpot. Cell proliferation was not related to differentiation status. The median apoptotic index was 1% (range l%-5%) and in the absence of apoptotic rate measurements, it was difficult to equate the contribution of apoptosis to the paradox of the slow clinical growth of BCCs. However, the concept of a high apoptotic rate was not supported by bcl-2 and bax protein expression. 88% of BCCs expressed bcl-2 and 23% expressed bax. The relationship between p53 expression and apoptosis was unclear since there was no correlation of p53 with bax, bcl-2 or apoptosis. The apoptotic parameters displayed some relationship to the histological growth patterns. The infiltrative and morpheic tumours exhibited the least apoptosis and least bcl-2 expression (p=0.02), but p53 did not correlate with tumour histology. The contribution of biological factors in determining outcome (the development of recurrence or a horrifying tumour) in BCC are limited because patient factors (late presentation) and treatment factors are dominant. Incomplete excision was associated with recurrence and the development of a horrifying tumour when compared to non recurrent tumours (p<0.01). Primary radiotherapy was also associated with the development of a horrifying tumour (p<0.01). A novel treatment modality, the optomechanically flash scanned carbon dioxide laser, was evaluated to assess its ability to completely ablate BCCs. Complete ablation was associated with ablation depth (p<0.01) and tumour type (p=0.01). Superficial BCCs were most suitable for this modality but required lasering to the middle dermis or deeper for complete eradication. Identification of problem BCCs at an early stage still requires further research but this thesis highlights the need for further improvement in surgical treatment