Representative Tissue Samples Research Articles

The PD-L1/PD-1 axis expression on tumor-infiltrating immune cells and tumor cells in pediatric rhabdomyosarcoma

BackgroundActivation of immune checkpoints, e.g. PD-1/PD-L1 axis, in cancer microenvironment, enables evasion of host anti-cancer immune response and drives tumor progression. To date, there have been only a few studies analyzing PD-1/PD-L1 expression in pediatric malignancies. AimIn the current study, we aimed to assess PD-L1 and PD-1 expression in pediatric rhabdomyosarcoma (RMS) and to investigate their clinicopathological associations.Materials and methods: The study enrolled 31 children with RMS. Tissue microarrays with representative tumor tissue samples were stained with anti-PD-1 NAT105 clone (Ventana, Roche) and two different antibodies against PD-L1: SP142 (Ventana, Roche) and 22C3 (DAKO). Adequate positive controls were applied. Their expression was assessed in tumor-associated immune cells (TAICs) and in the tumor cells separately. ResultsWe did not detect any positive PD-L1 staining in analyzed tumors using SP142 antibody; however, in 11 cases (35.48%) its expression was revealed by means of 22C3 clone. The staining was restricted to TAICs in all cases, which no reaction in tumor cells. The 5-year relapse free survival (RFS) rate was significantly higher in PD-L1 positive cases (61.5% vs 25.0%, p = 0.024), but it most likely results from more frequent PD-L1 expression in low-stage RMS. PD-1 expression on TAICs was detected in 7 cases and did not influence the prognosis. ConclusionsWe found that PD-L1 expression on TAICs, as detected with the use of 22C3 clone but not SP142 antibody, tends to be associated with low-stage RMS in children. PD-1 expression on TAICs in RMS is neither associated with distinct clinical course nor with clinicopathological features.

Cryobiopsy in the Upper Urinary Tract: Preclinical Evaluation of a Novel Device

ObjectiveTo develop a novel device for cryobiopsy of the upper urinary tract (UUT) and to evaluate its feasibility in a standardized preclinical setting. Materials and MethodsFlexible cryoprobes (diameter 0.9 mm; cooling agent CO2) were developed and used to extract biopsies in porcine UUTs. Cryosamples obtained by ureterorenoscopy were systematically compared with biopsy specimens obtained with standard of care devices in terms of physical characteristics (deflection angle and irrigation flow rates) and histologic criteria (assessability). ResultsIrrigation flow rates were significantly higher with introduced BIGopsy (2.8 ± 0.1) compared with standard forceps (0.94 ± 0.06; P < .001) and cryoprobe (1.1 ± 0.1; P < .001). Angular deflection was significantly reduced by the inserted cryoprobe (130.7° ± 1.2° vs 166.9° ± 1.1° [BIGopsy] or 161.4° ± 1.9° [standard forceps]; both P < .001). Significantly larger UUT tissue samples were obtained by the cryoprobe (mean specimen area 7.5 ± 2.5 vs 4.6 ± 2.5 mm² [BIGopsy] or 1.4 ± 1.4 mm² [standard forceps]; both P < .001). No crush artifacts were observed in cryosamples. Superior histologic assessability scores were achieved in samples obtained by the cryoprobe (mean 2.8 ± 0.8) and BIGopsy (2.3 ± 1.9) when compared with standard forceps (0.4 ± 0.9; P < .001). ConclusionCryobiopsy in the UUT is feasible and represents a viable new option to improve the diagnostic accuracy of histopathologic evaluation. Larger and more representative tissue samples can be obtained using a cryoprobe and artifacts may be avoided. Further optimization of the probe will reduce possible restrictions of ureterorenoscopy handling when the device is inserted.

Assessment of palmar subcutaneous tissue vascularization in patients with Dupuytren's contracture.

AIMTo investigate the structural and functional characteristics of palmar hypodermal tissue vascularization in Dupuytren’s contracture patients of different age groups.METHODSEighty-seven Dupuytren’s contracture patients underwent partial fasciectomy. Twenty-two of them were less than 55 years old (Y-group, n = 22); the others were 55 and older (O-group, n = 65). In surgically excised representative tissue samples, a histomorphometric analysis of the perforating arteries of the palmar aponeurosis and stereologic analysis of hypodermis vascularity were performed. The method of laser flowmetry estimated the microcirculation of the skin of the palm.RESULTSFrequency of cases with rapid development of contracture (less than 5 years) was 13.6% in the Y-group and 40% in the O-group, P < 0.05. The external and luminal diameters of perforating arteries in palmar fascia were decreased more severely in Y. The thickness of intima increased three times compared with healthy control, and the intima/media relation also increased, especially in O. Increased numerical and volumetric micro-vessel densities in hypodermis, percentage of large vessels (more than 12 μm in diameter), and percentage of vessels with signs of periadventitial inflammatory infiltration were noted in Y. The percentage of vessels with adventitial fibrosis was greater in O than in Y. Base capillary flow in Y was increased compared to healthy control subjects and to O, and peak capillary flow was increased in comparison with control.CONCLUSIONCompared to the O-group, Y-group patients exhibited more severe constrictive remodeling of palmar fascia perforating arteries supplying hypodermis but more expressed compensatory changes of its capillarization.

Diffuse reflectance spectroscopy of human liver tumor specimens - towards a tissue differentiating optical biopsy needle using light emitting diodes.

Significant numbers of liver biopsies fail to yield representative tissue samples. This study was conducted to evaluate the ability of LED-based diffuse reflectance spectroscopy to discriminate tumors from liver parenchyma. Ex vivo spectra were acquired from malignant lesions and liver parenchyma of 32 patients who underwent liver resection using a white light source and several LEDs. Integrated spectra of two combined LEDs with emission peaks at 470 nm and 515 nm were classified with 98.4% sensitivity and 99.2% specificity. The promising results could yield to a simple handheld and cost-efficient tool for real-time tissue differentiation implemented in a biopsy needle.

Abstract 3929: Heterogeneity in circulating tumor cells in blood samples of metastatic castration-resistant prostate cancer patient: comparison of isolation technique

Abstract Introduction/ObjectivesMetastatic castration-resistant prostate cancer (mCRPC) is a heterogen disease. Since most prostate cancer patients have a biopsy performed only at the time of diagnosis, representative tumor tissue sample giving real time information about the disease status is generally missing. CTC enumeration is a biomarker associated with clinical outcomes in patients with mCRPC. However, capturing these rare cells from whole blood is still a major challenge because they are extreme rare. Futhermore CTCs may exhibit a feature of epithelial-mesenchymal transition. First we assessed the method is best suited for the isolation of CTCs in mCRPC patients. Additionally was to evaluate the association between CTCs counts and ERG rearrangement in tumor samples. Materials &amp; Methods A tumor sample from metastatic site, along with blood samples were collected from mCRPC patients. Tumor samples were characterized of ERG rearrangement by fluorescence in situ hybridization. CTCs were detected using two methods. One was SreenCells system, CTC isolation based on size exclusion. The second method was the CellCollector which CTC captured using monoclonal specific antibody to EpCAM. This system can be used for CTC isolation ex vivo and in vivo. We used it here ex vivo. Captured CTCs were identified based on histological cell architecture by immunofluorescence staining using cytokeratin (8, 18, 19) and nuclear Hoechst staining positive as well as CD45 negative as criteria to exclude EpCam positive leucocytes Results We obtained duplicate blood samples from 10 mCRPC patient using SreenCells system and CellCollector isolation methods. Bothe methods detected CTCs in 8 of 10 blood samples, detection rate is 80 %. ScreenCells isolated in average of 22.6 CTCs (range of 0-80 CTCs). The CellCollector captured CTCs ex vivo in average of 3 CTCs (range of 0-6 CTCs). In a direct comparison of the ScreenCell system and the Cellcollector is significant different (p 0.0156) in the CTC counts. The CTC number of both systems is moderate correlated (r 0.5). All mCRPC tissue samples harbored an ERG rearrangement in range of 89 until 98 %. The CTC number of the ScreenCell system is good correlate with PSA level (r 0.776) and also with the ERG- rearrangement (r 0.5) of tissue samples. Conclusion Our results demonstrate that CTC isolation based on the physical properties has a higher sensitivity as the CTC isolation technique based on the biological cell properties in mCRPC patients. The size exclusion technique coupled with characterization of specific staining morphologies might be used to identify a heterogeneous CTC population which is always present in advanced cancer. It should be decided in dependence of the advanced cancer stage which CTC isolation technique can be used. The cost of the material and the hours of work are equal in both systems. Citation Format: Gerit Theil, Christine Weiss, Kersten Fischer, Andre Schumann, Paoloa Fornara. Heterogeneity in circulating tumor cells in blood samples of metastatic castration-resistant prostate cancer patient: comparison of isolation technique [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3929. doi:10.1158/1538-7445.AM2017-3929

Leiomyosarcoma of the Skin and Subcutaneous Tissue in a Nellore Cow

Background: Leyomiosarcomas are uncommon malignant smooth muscle tumors of hollow organs. Bovine leyomiosarcomas have been described in several sites, but smooth muscle tumors that arise from the skin are rare in domestic animals. These neoplasms may show variable histologic features depending on their grade of differentiation; therefore, diagnostic tools as special stain, electron microscopy and immunohistochemistry are essential for the definitive diagnosis. Due to the lack of reports of skin leyomiosarcomas in cattle, we described the occurrence of a leyomiosarcoma in the skin and subcutaneous tissue of a Nellore cow.Case: A 10-year-old Zebu Nellore cow was admitted at the Veterinary Diagnosis Center (CEDIVET) of Pará Federal University (UFPA), Brazil, to investigate a tumor on the hip. At clinical examination the cow showed a decreased appetite and weight loss. Due to the bad prognosis the veterinarian recommended euthanasia. Representative tissue samples were submitted for histopathological and immunohistochemistry evaluation at the Department of Pathology of the Federal Rural University of Rio de Janeiro (UFRRJ). The slides were stained by Hematoxylin and Eosin (HE), Masson’s trichrome and Van Gieson stains and submitted for immunohistochemistry with primary antibodies vimentin, troponin C, smooth muscle alpha actin and desmin. Grossly, the tumor appeared as a multilobular, ulcerated and firm 30 cm diameter mass with black and hemorrhagic areas on the surface. The mass was scattered throughout the subcutaneous tissue and infiltrated the adjacent muscle. On light microscopy, the neoplasm consisted of a non-encapsulated proliferation into the derma, composed of highly pleomorphic spindle to round cells arranged in broad interlacing fascicles and small islands. Metastatic lesions were found in the lungs and local lymph nodes. Masson’s trichrome stain revealed collagen production and Van Gieson stains was inconsistent. Immunohistochemically, the neoplastic cells expressed vimentin diffusely and smooth Alfa actin in 10% of the cells. Desmin and troponin C failed to express immunoreactivity.Discussion: The histological, histochemical and immunohistochemical features of the neoplasm were consistent with a leiyomiosarcoma of the skin and subcutaneous tissue. Leiomyosarcomas are uncommon in farm animals and most of them arise from the female genital tract, especially in the vagina and within the uterus. The majority of smooth muscle tumors of the skin are benign; however, we described here a neoplasm that displayed clear signs of malignancy. Due to the high grade of differentiation, Van-Gieson stain failed to determine the origin of the neoplasm. Many sarcomas have resembling histologic features and busually display interlacing bundles of spindle shaped cells, similar to the pattern described in the present tumor. Therefore, immunohistochemistry stain with vimetin and smooth alpha actin was essential for the definitive diagnosis. We identified metastatic lesions in the local lymph nodes and lungs. This tumor tends to develop slowly and metastasize late. These findings underline the importance of diagnostic tools, such as immunohistochemistry and special stains to guide the pathologist and to establish the diagnosis of bovine smooth muscle tumors. Consequently, an association of histochemical and immunohistochemical procedures is indispensable to guide the pathologist for the establishment of the diagnosis of bovine smooth muscle tumors.

A rare case of metastatic hepatocellular carcinoma in a Himalayan black bear (Selenarctos thibetanus)

A 24-year-old female Himalayan black bear (Selenarctos thibetanus) was necropsied and gross lesions were observed in liver, kidney, heart, lung and spleen, from which representative tissue samples in 10% formalin were submitted for histopathology to the Department of Veterinary Pathology, GADUVAS, Ludhiana. Microscopically, the liver showed variable sized multiple tumorous foci surrounded by fibrous tissue stroma, with neoplastic cells having prominent nuclei, basophilic cytoplasm and marked hypercellularity. Lungs showed widespread variable sized randomly distributed metastatic foci, as well as, vascular tumor thrombo-embolism. On the basis of microscopic findings, the present case was diagnosed as hepatocellular carcinoma with widespread pulmonary metastases.

Metabolite mapping reveals severe widespread perturbation of multiple metabolic processes in Huntington's disease human brain

Huntington's disease (HD) is a genetically-mediated neurodegenerative disorder wherein the aetiological defect is a mutation in the Huntington's gene (HTT), which alters the structure of the huntingtin protein (Htt) through lengthening of its polyglutamine tract, thus initiating a cascade that ultimately leads to premature death. However, neurodegeneration typically manifests in HD only in middle age, and mechanisms linking the causative mutation to brain disease are poorly understood. Brain metabolism is severely perturbed in HD, and some studies have indicated a potential role for mutant Htt as a driver of these metabolic aberrations. Here, our objective was to determine the effects of HD on brain metabolism by measuring levels of polar metabolites in regions known to undergo varying degrees of damage. We performed gas-chromatography/mass spectrometry-based metabolomic analyses in a case-control study of eleven brain regions in short post-mortem-delay human tissue from nine well-characterized HD patients and nine matched controls. In each patient, we measured metabolite content in representative tissue-samples from eleven brain regions that display varying degrees of damage in HD, thus identifying the presence and abundance of 63 different metabolites from several molecular classes, including carbohydrates, amino acids, nucleosides, and neurotransmitters. Robust alterations in regional brain-metabolite abundances were observed in HD patients: these included changes in levels of small molecules that play important roles as intermediates in the tricarboxylic-acid and urea cycles, and amino-acid metabolism. Our findings point to widespread disruption of brain metabolism and indicate a complex phenotype beyond the gradient of neuropathologic damage observed in HD brain.

Spectral sensing for tissue diagnosis during lung biopsy procedures: The importance of an adequate internal reference and real-time feedback

ObjectivesDifficulties in obtaining a representative tissue sample are a major obstacle in timely selecting the optimal treatment for patients with lung cancer or other malignancies. Having a modality to provide needle guidance and confirm the biopsy site selection could be of great clinical benefit, especially when small masses are targeted. The objective of this study was to evaluate whether diffuse reflectance spectroscopy (DRS) at the tip of a core biopsy needle can be used for biopsy site confirmation in real time, thereby enabling optimized biopsy acquisition and improving diagnostic capability. Materials and methodsWe included a total of 23 patients undergoing a routine computed tomography (CT) guided transthoracic needle biopsy of a lesion suspected for lung cancer or metastatic disease. DRS measurements were acquired during needle insertion and clinically relevant parameters were extracted from the spectral data along the needle paths. Histopathology results were compared with the DRS data at the final measurement position. ResultsAnalysis of the collective data acquired from all enrolled subjects showed significant differences (p<0.01) for blood content, stO2, water content, and scattering amplitude. The identified spectral contrast matched the final pathology in 20 out of 22 clinical cases that could be used for analysis, which corresponds with an overall diagnostic performance of 91%. Three cases underlined the importance of adequate reference measurements and the need for real time diagnostic feedback. Continuous real time DRS measurements performed during a biopsy procedure in one patient provided clear information with respect to the variation in tissue and allowed identification of the tumour boundary. ConclusionsThe presented technology creates a basis for the design and clinical implementation of integrated fibre-optic tools for a variety of minimal invasive applications.

Histochemical and Ultrastructural Studies on the thyroid gland ofPatiDucks(Anas platyrhynchos domesticus) of Assam

A total of 36 ducks were utilised in this study ranging in age from 2 weeks upto 40 weeks of age. For histochemical parameters, the thyroid glands were separated out of the thoracic cavity and the required representative tissue samples were preserved in deep freeze maintained at-80°C. For scanning electron microscopy the tissue samples were processed by SAIF, NEHU, Shillong. The colloid showed a strong PAS positive reaction in all the age groups. The activities of both acid and alkaline phosphatases were present in all the age groups. The scanning electron microscopy revealed the normal structure of the thyroid gland in all the age groups. Parenchyma of thyroid gland was composed of follicles which were separated from each other by interfollicular connective tissue made up of collagen and reticular fibers. The follicles were closely packed together and their shape varied from oval to polyhedral. Microvilli of the follicular cells were observed on the apical surface. The SEM also showed the thyroid artery penetrating the gland.

Extensive pancreatic heterotopia in distal esophagus mimicking esophageal malignancy

We report an unusual and rare case of a young adult patient with extensive symptomatic pancreatic heterotopia in the distal esophagus that appeared on imaging as a malignant tumor. After comprehensive preoperative workup, the soft tissue mass was treated surgically according to intraoperative histology that revealed its benign nature. A young male patient with worsening epigastric pain, gastroesophageal reflux, and weight loss presented in a university hospital for a comprehensive diagnostic workup. Apart from clinical examination and laboratory tests, he underwent gastroscopy, esophageal manometry, endoscopic ultrasound, barium meal study, computed tomography (CT), and positron emission tomography (PET-CT). Finally an interdisciplinary board recommended surgical removal of a mediastinal mass and final diagnosis was reached by intraoperative histology. The clinical examination and laboratory test were unremarkable, esophageal manometry suggested achalasia due to simultaneous spastic and retrograde contractions with high amplitude. Barium meal showed mild compliant stenosis of the distal esophagus, but gastroscopy found no explanation. CT and later also PET-CT revealed a large soft tissue mass with avid accumulation of 18F-fluorodeoxyglucose surrounding the distal esophagus. Endoscopic ultrasound was very limited and no representative tissue samples were obtained prior to surgery. The surgeon accessed the mediastinum through the left thoracotomy and found a noncompliant well-defined mass surrounding the distal esophagus. Intraoperative histology showed nothing but common structures of pancreatic tissue. A large soft-tissue mass of pancreatic heterotopia in the distal esophatus is extremely rare and preoperative workup inconclusive. Only surgical exploration and (intraoperative) histology are in a position to reveal its true nature.

Abstract 3474: Comparison of CTC capture efficiency of the CellCollector™ technology versus CellSearch® in prostate cancer patients at multiple time points

Abstract Castration-Resistant Prostate Cancer (CRPC) is the second most common cancer cause of cancer-related death in men. Since most prostate cancer patients have a biopsy performed only at the time of diagnosis, representative tumor tissue sample giving real time information about the disease status is generally missing. The detection of circulating tumor cells (CTCs) in the blood of patients with CRPC might therefore, in addition to their prognostic value, serve as liquid biopsy, complementing or replacing prostate-specific antigen (PSA) determination in predicting and monitoring the response to different therapies. However, capturing these rare cells from whole blood is still a major challenge that needs significant improvement. Here we present the results of a comparison study, in which we compared the CellCollector™, a unique in vivo approach for the isolation of CTCs, with the CellSearch® system, the current standard. The study included 25 prostate cancer patients (15 with localized and 10 with metastasized prostate cancer (PCa-l and PCa-m)) and 29 subjects in the control group (24 men with benign prostate hypertrophy (BPH) and 5 women). At multiple time points of treatment, CTCs were enumerated (PCa-l: 42 applications; PCa-m: 29 applications). The CellCollector™, a medical wire coated with epithelial cell adhesion molecule (EpCAM) antibodies, was inserted in the cubital vein and incubated for 30 minutes. The captured CTCs were identified by immunofluorescence staining using cytokeratin (CK) and Hoechst positive as well as CD45 negative as criteria. For the CellSearch® measurements, a blood draw of 7.5 ml blood was performed. We found in 77.5 % (55/71) applications (PCa-l: 55.2 % (16/29); PCa-m: 88.1 % (37/42)) the cancer patient to be positive for CTCs using the CellCollector™. In contrast the CellSearch® system resulted only in 42.2 % (30/71) in the detection of CTCs (PCa-l: 17.2 % (5/29); PCa-m: 61.9 % (26/42)). The control groups showed in rare cases very low numbers of CK positive cells. In addition we found a correlation of the CTC levels detected by the CellCollector™ and the CellSearch® with the PSA level during treatment. In summary, our comparison study shows an improved sensitivity of the CellCollector™ compared to the current standard regarding the isolation of CTCs from prostate cancer patients and gives new insights in the value of CTCs for the monitoring of prostate cancer treatment. Citation Format: Gerit Theil, Kathrin Haubold, Paolo Fornara, Antje Stresemann, Arndt Schmitz, Thomas Krahn, Klaus Lücke. Comparison of CTC capture efficiency of the CellCollector™ technology versus CellSearch® in prostate cancer patients at multiple time points. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3474. doi:10.1158/1538-7445.AM2014-3474

Active surveillance of low risk prostate cancer

In Europe prostate cancer is one of the most common cancers among men. The diagnostics always include a control of the prostate-specific antigen (PSA) level and examination of a representative tissue sample from the prostate. With these findings it is possible to evaluate the degree of progression of the cancer and its prognosis. Several treatment options for localized prostate cancer are given by national and international guidelines including radical prostatectomy, percutaneous radiation therapy, or brachytherapy and surveillance of the cancer with optional treatment at a later stage. For the latter treatment option, known as active surveillance, strict criteria have to be met. The advantage of active surveillance is that only patients with progressive cancer are subjected to radical therapy. Patients with very slow or non-progressing cancer do not have to undergo therapy and thus do not have to suffer from the side effects. The basic idea behind active surveillance is that some cancers will not progress to a stage that requires treatment within the lifetime of the patient and therefore do not require treatment at all. Unfortunately the criteria for active surveillance are not definitive enough at the current time leading only to a delay in effective treatment for many patients. The surveillance strategy has without doubt a high significance among the treatment options for prostate cancer; however, at the current time it lacks reliable indicators for a certain prognosis. Therefore, patients must be informed in detail about the advantages and disadvantages of active surveillance.

The role of histopathology in forensic practice: an overview

The role of forensic histopathology in routine practice is to establish the cause of death in particular cases. This is achieved on the basis of microscopic analysis of representative cell and tissue samples taken from the major internal organs and from abnormal findings made at autopsy. A prerequisite of this is adherence to the quality standards set out for conventional histological/cytological staining and enzyme histochemical and immunohistochemical methods. The interpretation of histological findings is performed by taking into account macroscopic autopsy findings and information on previous history. Histological analysis may prompt postmortem biochemical and chemical-toxicological investigations. The results of histological analysis need to be classified by experts in the context of the available information and the need to withstand the scrutiny of other experts.

Irrigation and Debridement

Irrigation and Debridement

Expression of P57 Immunohistochemical Marker in Complete And Partial Hydatidiform Mole by Using Tissue Microarray Technique

Back ground: Hydatidiform mole is an abnormal form of pregnancy divided in to two types; complete hydatidiform mole and partial hydatidiform mole. Detailed histopathologic examination remains to be the basis for the diagnosis of hydatidiform mole (HM). However, poor sampling, necrosis, and earlier uterine evacuation can lead to uncertainty in the diagnosis. Also, the criteria are subjective, resulting in considerable interobserver variability.The P57KIP2 gene is paternally imprinted and maternally expressed, and the presence of its protein product serves as an adjuvant marker for the nuclear maternal genome. Because a complete HM is the only type of conceptus lacking a maternal contribution, P57 KIP2 immunostaining is correspondingly absent, whereas it is present in partial HM and normal pregnancy. Aim of The Study: The purpose of this study is to evaluate the significance of the P57KIP2 immunohistochemical marker in the diagnosis of complete and partial hydatidiform mole , and to compare the P57KIP2 immunohistochemical marker results with that of ordinary Hematoxylin and Eosin slides histopathology results, in addition to evaluation of the Tissue Microarray Technique. Materials and Methods: Seventy cases of endometrial biopsies were obtained. Histologic evaluation of all cases was performed on routine sections stained with Hematoxylin and Eosin (H & E) and classified in complete and partial hydatidiform mole (30 each) and (10 cases) of normal product of pregnancy (abortion), and negative control slides were also used. These cases were collected during the period from December-2011 to Augest-2012. New technique is adopted in this research that is; Tissue microarray in which twelve small cores of representative tissue samples, each measure 3 mm in diameter sections from microarray block are cut using a microtome, mounted on a single microscope slide and then analyzed by staining with Hematoxylin and Eosin, then another section made for the immunohistochemical staining with P57 KIP2 antibody. Results: Regarding the expression of p57KIP2 IHC Marker in cases of PHM diagnosed previously by HE grade I, and they represented 13.3%. The correlation between the grades of the CHM and the results of the p57KIP2 IHC marker was statistically significant. Conclusion: P57KIP2 IHC marker is a useful adjunct, providing a definitive diagnosis of CHM. Errors in morphologic analysis resulted primarily in over diagnosis of PHM are more frequent than in CHM. There are significant differences in correct classification of hydatidiform mole between using P57KIP2 IHC marker and using H&E stain Cases in grade I of CHM were correlated significantly with the errors in morphological analysis resulted primarily in over diagnosis of CHM. Tissue Microarray is a relatively simple method can be used in pathology laboratories to decreased assay volume, time and cost.

Deadly outbreak of iron storage disease (ISD) in Italian birds of the family Turdidae.

ABSTRACTA widespread deadly outbreak occurred in captive birds belonging to the family Turdidae in Italy. The present study was performed on 46 dead birds coming from 3 small decoy-bird breeders in central Italy. Only Turdus pilaris, Turdus iliacus, Turdus philomelos and Turdus merula were affected. No other species of bird held by these breeders died. A change of diet before the hunting season was reported from all breeders. Full necropsy of the animals and histological investigations of representative tissue samples were performed. Microscopical examination showed marked iron deposits in liver samples. Bacteriological investigations and molecular analysis to exclude bacterial and viral diseases were carried out. Contamination of food pellet samples by mycotoxins and analysis to detect heavy metal contaminants in food pellet samples were considered. An interesting result was the high iron content found in food pellets. It was higher than that considered suitable for birds, especially for species susceptible to development iron storage disease (ISD). Taken together, the results suggested an outbreak of ISD caused by the high iron content of food given to the birds before the hunting season. The high mortality recorded only in species belonging to the family Turdidae suggests a genetic predisposition in the affected birds.

Concurrent outbreak of pasteurellosis and aflatoxicosis in Kuttanad ducks

One hundred and forty eight ducks of age group of 12–14 months out of a total population of two hundred and fifty ducks died of concurrent outbreak of pasteurellosis and aflatoxicosis in an organized farm. Tissue samples were collected for histopathology and bacterial isolation. Hepatic tumours were observed in six ducks. Heart blood smear and impression smears from various visceral organs were taken and bipolar stained Pasteurella organisms were detected by Leishman staining. Feed samples were collected for aflatoxin estimation and found to be 500 ppb. Pasteurella multocida organism was isolated from representative tissue samples collected. Characteristic pathological alterations corresponding to aflatoxicosis and pasteurellosis were found in different visceral organs.

Abstract P1-08-06: Low tumor CD68 mRNA content (intratumoral macrophages) is predictive for benefit from adjuvant trastuzumab in HER2-positive breast cancer: An analysis of the FinHER trial

Abstract Background: Tumor immune cell infiltrates influence prognosis of node negative breast cancer (BC), and intratumoral B-cells and T-cells are of importance for achieving response to chemotherapy in triple negative BC. The role of tumor infiltrating macrophages remains unclear, but they might promote tumorigenesis. We investigated the prognostic value of CD68 mRNA levels within the luminal, HER2-positive and triple negative subtypes in the FinHer trial patient population, and evaluated their predictive value on survival outcomes achieved with adjuvant trastuzumab and chemotherapy in early breast cancer. Methods: RNA was extracted from formalin-fixed paraffin-embedded (FFPE) tumor tissue of 917 (90.8%) patients out of the 1010 patients who participated in the FinHer trial. Intratumoral macrophage infiltration was assessed by measuring breast tumor CD68 mRNA content using RT-qPCR from representative FFPE tissue samples. Breast cancer molecular subtypes (luminal, HER2 positive and triple-negative) were approximated using immunohistochemistry (IHC) and central CISH testing data obtained from the FinHer trial datafile. Prognostic significance of CD68 on distant disease-free survival (DDFS) was assessed using Kaplan-Meyer analysis and log-rank test. Results: Tumor CD68 mRNA expression was normally distributed with median expression of 33.56 (dCt). CD68 mRNA levels correlated weakly with estrogen receptor (ER) mRNA expression and ER protein levels (r = 0.11 and r = 0.15, respectively; p&amp;lt;0.0001), but not with tumor HER2 mRNA level or IHC/CISH status (r = -0.06, p = 0.12; and r = 0.01, p = 0.92, respectively). The median tumor CD68 mRNA content was not prognostic for DDFS in the subset of luminal cancers (ER+, HER2-; n = 533, 5-year DDFS 88% versus 88%; p = 0.92), but a lower than the median CD68 level tended to be associated with favorable DDFS in triple negative cancer (n = 143; 81% vs. 70%; p = 0.11) and was significantly associated with favorable DDFS in HER2-positive cancer (n = 191; 86% vs. 67%; p = 0.001). In the subset of HER2-positive cancer with tumor CD68 mRNA expression level lower than the median, patients treated with trastuzumab benefitted from addition of trastuzumab to chemotherapy (5-year DDFS 93% with trastuzumab vs. 79% without trastuzumab; p = 0.02), whereas no benefit from trastuzumab was observed when tumor CD68 content was higher than the median (67% vs. 68%; p = 0.92). Conclusions: The study validates tumor CD68 concentration as a prognostic biomarker in HER2-positive early breast cancer. Patients with HER2-positive cancer and few tumor macrophages (low tumor CD68 mRNA content) benefitted from addition of trastuzumab to chemotherapy, whereas patients with HER2-positive BC with high tumor macrophage content derived no benefit from adjuvant trastuzumab. Trastuzumab may be effective only for macrophage-poor HER2-positive cancers that are prone to antibody-dependent cellular cytotoxicity (ADCC), whereas it may have little efficacy for cancers that progress despite or due to high intratumoral macrophage content that interferes with ADCC. Other agents, such as T-DM1, might work better than trastuzumab in the subset of women who have HER2-positive BC with a high tumor macrophage content. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-06.

Characterization of B‐cell subpopulations in patients with chronic rhinosinusitis

Recent research suggest that B and plasma cells may play an important role in the pathogenesis of chronic rhinosinusitis with nasal polyposis (CRSwNP). The purpose of this study was to subcharacterize the B cell response in the sinus mucosa of control and CRS patients. Representative tissue samples and peripheral blood samples were obtained from controls, CRS without nasal polyps (CRSsNP) and CRSwNP. Using single-cell suspension flow cytometry these samples were analyzed for overall and stage-specific B and plasma cell percentages. Both atopic and nonatopic CRSwNP patients showed an increase in local numbers of naive, active, and memory B cells compared to controls. CRSsNP patients only showed local elevations of naive B cells. Plasma cells were only significantly elevated in the sinus tissue of atopic CRSwNP patients. These local tissue increases did not correlate with increased numbers of circulating B cells. This study provides further evidence of an important role of B cells in CRSwNP patients. The local increase appears to be independent of a systemic response.