Banff Histopathological Consensus Criteria for Preimplantation Kidney Biopsies.

The Banff working group on preimplantation biopsy was established to develop consensus criteria (best practice guidelines) for the interpretation of preimplantation kidney biopsies. Digitally scanned slides were used (i) to evaluate interobserver variability of histopathologic findings, comparing frozen sections with formalin-fixed, paraffin-embedded tissue of wedge and needle core biopsies, and (ii) to correlate consensus histopathologic findings with graft outcome in a cohort of biopsies from international medical centers. Intraclass correlations (ICCs) and univariable and multivariable statistical analyses were performed. Good to fair reproducibility was observed in semiquantitative scores for percentage of glomerulosclerosis, arterial intimal fibrosis and interstitial fibrosis on frozen wedge biopsies. Evaluation of frozen wedge and core biopsies was comparable for number of glomeruli, but needle biopsies showed worse ICCs for glomerulosclerosis, interstitial fibrosis and tubular atrophy. A consensus evaluation form is provided to help standardize the reporting of histopathologic lesions in donor biopsies. It should be recognized that histologic parameters may not correlate with graft outcome in studies based on organs deemed to be acceptable after careful clinical assessment. Significant limitations remain in the assessment of implantation biopsies.

Hedgehog pathway overexpression in pancreatic cancer is abrogated by new-generation taxoid SB-T-1216.

The Hedgehog pathway is one of the major driver pathways in pancreatic ductal adenocarcinoma. This study investigated prognostic importance of Hedgehog signaling pathway in pancreatic cancer patients who underwent a radical resection. Tumors and adjacent non-neoplastic pancreatic tissues were obtained from 45 patients with histologically verified pancreatic cancer. The effect of experimental taxane chemotherapy on the expression of Hedgehog pathway was evaluated in vivo using a mouse xenograft model prepared using pancreatic cancer cell line Paca-44. Mice were treated by experimental Stony Brook Taxane SB-T-1216. The transcript profile of 34 Hedgehog pathway genes in patients and xenografts was assessed using quantitative PCR. The Hedgehog pathway was strongly overexpressed in pancreatic tumors and upregulation of SHH, IHH, HHAT and PTCH1 was associated with a trend toward decreased patient survival. No association of Hedgehog pathway expression with KRAS mutation status was found in tumors. Sonic hedgehog ligand was overexpressed, but all other downstream genes were downregulated by SB-T-1216 treatment in vivo. Suppression of HH pathway expression in vivo by taxane-based chemotherapy suggests a new mechanism of action for treatment of this aggressive tumor.

2016 Comprehensive Update of the Banff Working Group on Liver Allograft Pathology: Introduction of Antibody-Mediated Rejection.

The Banff Working Group on Liver Allograft Pathology reviewed and discussed literature evidence regarding antibody-mediated liver allograft rejection at the 11th (Paris, France, June 5-10, 2011), 12th (Comandatuba, Brazil, August 19-23, 2013), and 13th (Vancouver, British Columbia, Canada, October 5-10, 2015) meetings of the Banff Conference on Allograft Pathology. Discussion continued online. The primary goal was to introduce guidelines and consensus criteria for the diagnosis of liver allograft antibody-mediated rejection and provide a comprehensive update of all Banff Schema recommendations. Included are new recommendations for complement component 4d tissue staining and interpretation, staging liver allograft fibrosis, and findings related to immunosuppression minimization. In an effort to create a single reference document, previous unchanged criteria are also included.

[Histopathological differential diagnosis of primary liver tumors]. / Problematika diferenciální diagnostiky primárních nádoru jater.

The most common primary hepatic malignancy is hepatocellular carcinoma, which constitutes 80-85% of all malignant epithelial neoplasms originating in the liver. The second most common primary hepatic malignancy is cholangiocellular carcinoma. In recent years, remarkable progress has been made in elucidating the molecular pathology of hepatic tumors. Advances in our understanding of molecular subtypes have led to a creation of a new classification system of hepatocellular adenomas, with important genotype-phenotype correlations and easy application to routine diagnostic practice. In the field of early hepatic neoplasia, a consensus on the definition of dysplastic nodules and early hepatocellular carcinoma has been reached. Immunohistochemical detection of glypican-3 and stromal invasion are used for the differential diagnosis. A lot of problems still exist in the category of mixed hepatocellular and cholangiocellular carcinoma. Although this category is recognized in the WHO classification, confusing terminology has been generated to describe tumors with morphological features of both HCC and CC. We further specify problems and pitfalls of the differential histopathological diagnosis of liver malignant tumors.

[Evaluation of safe resection margins in tumours of parenchymatous organs]. / Problematika stanovení bezpecných resekcních okraju u nádoru parenchymatózních orgánu

The definition of a safe surgical margin in tumours of parenchymatous organs has been the subject of frequent debates and a number of studies. This topic is not generally unified for parenchymatous organs and has many organ specifics. Moreover, there are still controversies in the diagnostic criteria and the definitions of positive resection margins, in indications of perioperative examinations of resection margins, and in the prognostic significance of margin involvement in different parenchymatous organs. The consensus in terminology and standardization of histopathological examination procedures is also very desirable across the diagnostic areas.

[Biliary (cholestatic) complications in the liver graft biopsy samples early after transplantation]. / Biliární (cholestatické) komplikace v bio-psiích jater casne po transplantaci.

During the first three months after a liver transplant, cholestasis of various type and degree represents 1 of the most frequent morphological findings in liver graft biopsies. The morphology of cholestasis is typical for all conditions with bile duct impairment but also for other conditions with more severe impairment of hepatocytes, including rejection and recurrence of viral hepatitides. Histological diagnosis represents the gold standard in addressing liver graft dysfunction causes, and in the majority of cases it allows for distinguishing between the main categories of diseases resulting in cholestasis. Usually a combination of various changes can be identified as a cause of the liver graft dysfunction early after transplantation. Therefore, the interpretation of limited morphological characteristics, which usually represent a certain type of tissue reaction, not the cause, is complicated. The close cooperation between the hepatologist and pathologist has become a necessary prerequisite for the best possible interpretation of the morphological conclusion.

Guidelines for the diagnosis of antibody-mediated rejection in pancreas allografts-updated Banff grading schema.

The first Banff proposal for the diagnosis of pancreas rejection (Am J Transplant 2008; 8: 237) dealt primarily with the diagnosis of acute T-cell-mediated rejection (ACMR), while only tentatively addressing issues pertaining to antibody-mediated rejection (AMR). This document presents comprehensive guidelines for the diagnosis of AMR, first proposed at the 10th Banff Conference on Allograft Pathology and refined by a broad-based multidisciplinary panel. Pancreatic AMR is best identified by a combination of serological and immunohistopathological findings consisting of (i) identification of circulating donor-specific antibodies, and histopathological data including (ii) morphological evidence of microvascular tissue injury and (iii) C4d staining in interacinar capillaries. Acute AMR is diagnosed conclusively if these three elements are present, whereas a diagnosis of suspicious for AMR is rendered if only two elements are identified. The identification of only one diagnostic element is not sufficient for the diagnosis of AMR but should prompt heightened clinical vigilance. AMR and ACMR may coexist, and should be recognized and graded independently. This proposal is based on our current knowledge of the pathogenesis of pancreas rejection and currently available tools for diagnosis. A systematized clinicopathological approach to AMR is essential for the development and assessment of much needed therapeutic interventions.

[How to improve the histopathological diagnosis of hepatocellular carcinoma in daily practice?]. / Jak v praxi zlepsit histopatologickou diagnostiku hepatocelulárního karcinomu.

Hepatocellular carcinoma (HCC) is one of the most common cancers in the world with an increasing incidence. Recently, an East-West consensus on the histopathologic criteria for the diagnosis of high-grade dysplastic nodules and early hepatocellular carcinoma was published. In addition to classical morphological criteria such as nucleocytoplasmic ratio, thickness of cell plates, mitotic index, and architectural disturbance, a new one--the stromal invasion--was recognized as a crucial criterion for the diagnosis of early HCC. Immunohistochemical detection of glypican--3 was shown as a specific marker for HCC that can be used to distinguish between the benign hepatocellular lesions and HCC.

[Fibrosing cholestatic hepatitis--disease not only of transplanted patients. A report of eight cases]. / Fibrotizující cholestatická hepatitida--onemocneni nejen transplantovaných pacientu.

Fibrosing cholestatic hepatitis (FCH) is a rapidly progressive, sometimes fatal form of hepatitis B or C in patients who are under immunosuppressive treatment. This condition was originally described in hepatitis B virus-infected recipients after a liver transplantation. It is characterized clinically by cholestatic hepatic dysfunction, and pathologically by marked hepatocyte swelling, cholestasis, periportal peritrabecular fibrosis, and only mild inflammation. Here we present 8 patients with hepatitis B and C related FCH. Three patients developed FCH after liver transplantation, two of them died due to hepatic failure. One recipient of a kidney/pancreas transplant developed "de novo" hepatitis C with features of FCH. He underwent antiviral treatment and survived with good liver function, unfortunately both of his grafts failed. Four patients suffered from a reactivation of their respective hepatitis B infections after chemotherapy treated hematological malignancy. Three of them needed an urgent liver transplantation and survived with good liver function and with a remission of their hematological diseases. The last patient died due to liver failure. Although FCH is a rare variant of viral hepatitis, it should be emphasized that prompt diagnosis is important for the management of patients.

Glomus tumor of the stomach: a case report and review of the literature.

Glomus tumor is a benign soft tissue neoplasm which commonly affects the subungual region of the fingers. But the tumors can also arise in the other sites such as the antrum of the stomach. We are reporting a case of a glomus tumor of the stomach in a 71-year-old female patient who presented with dyspepsia. The tumor was confined to the lamina muscularis propria, it consisted of round cells with small uniform nuclei, which surrounded thin walled blood vessels. Immunohistochemistry revealed the tumor to be positive for smooth muscle actin, vimentin, calponin, h-caldesmon and negative for c-KIT, S-100, CD34, CD99, synaptophysin, chromogranin, desmin and EMA. The proliferation marker Ki-67 was positive in less than 5% of tumor cell nuclei. Glomus tumors are usually benign but malignant cases have been published. Criteria for the malignant potential of gastric glomus tumors remain poorly defined.

Hepatocyte growth factor and antibodies to HLA and MICA antigens in heart transplant recipients.

Recent unconfirmed literature data suggest that elevated concentrations of the multifunctional cytokine hepatocyte growth factor (HGF) might be a marker of increased incidence of acute rejection after organ transplantation. The aim of this study was to test the hypothesis that HGF levels may correlate with the rejection and/or with the production of HLA and MHC Class I chain-related antigens A (MICA) specific antibodies. Sixty-three heart transplant recipients were included into the study. Hundred and eighty-five endomyocardial biopsies (EMB) obtained up to 6 months after transplantation were retrospectively analyzed for signs of cellular (CR) and antibody-mediated rejection (AMR). Pre- and post-transplant sera were tested for HGF concentrations and antibodies to HLA class I, class II and MICA antigens by xMap technology (Luminex). Pre-transplant HGF did not correlate with the incidence of CR or AMR. However, higher HGF concentrations correlated significantly with HLA antibody production before and after transplantation (P = 0.006 and P < 0.0001 respectively). Patients with both HLA class I and class II antibodies before transplantation had significantly lower AMR-free survival. Furthermore, recipients with pre-transplant donor-specific antibodies (DSA) had significantly lower AMR-free survival (50%) than recipients without pre-transplant HLA antibodies (90%) and patients with antibodies not specific to donor antigens (92%) (P = 0.005). Post-transplant MICA antibodies tended to be more frequent in patients with AMR (P = 0.063). In conclusion, elevated HGF concentrations in our study were not associated with the incidence of CR and/or AMR but with the presence of HLA-specific antibodies. Testing for DSA before heart transplantation by Luminex may be helpful for the identification of patients with increased risk of AMR.

Leflunomide derivate FK 778 in accelerated renal injury in transgenic rat.

Renal ischaemia/reperfusion (I/R) injury and hypertension represent major alloantigen-independent risk factors contributing to the development of chronic allograft nephropathy. In a model of accelerated major histocompatibility complex-independent renal injury, we evaluated the effect of leflunomide derivate - FK778 - on the progression of accelerated nephropathy. Thirty-six uninephrectomized hypertensive transgenic (m-REN-2)-27 rats received a clip on renal pedicle for 45 minutes. Animals were treated with FK778 3 mg/kg/day (I/R 3 mg, N = 12), 10 mg/kg/day (I/R 10 mg, N = 12) or placebo (N = 12) via gavage for 16 weeks. Eighteen animals were sham-operated and treated with FK778 3 mg/kg/day (sham 3 mg, N = 6), 10 mg/kg/day (sham 10 mg, N = 6) or were untreated (sham, N = 6). Proteinuria and blood pressure were evaluated throughout and the kidneys were harvested for morphological and immunohistochemical analysis at the end of the experiment. At week 16, rats with I/R injury and FK778 treatment had lower proteinuria compared with placebo-treated rats (I/R 3 mg: 48.42 +/- 26.16, I/R 10 mg 27.28 +/- 21.86 vs. Placebo: 70.13 +/- 50.19 mg/day, P < 0.05). The untreated sham group exhibited lower proteinuria compared with FK778-treated sham groups (Sham 3 mg: 24.23 +/- 10.89; Sham 10 mg: 17.37 +/- 4.13; Sham: 14.23 +/- 1.18) There was no difference in glomerulosclerosis and interstitial fibrosis among the treated groups. In the untreated animals the rate of interstitial fibrosis decline reached statistical significance (Placebo vs. Sham: 1.125 +/- 0.641 % vs. 0.250 +/- 0.500 %, P < 0.05). There was higher CD5+ leukocyte infiltration in the placebotreated group. FK778-treated rats displayed amelioration of some changes induced by the I/R injury. Our observation also suggests potential nephrotoxicity of FK778.

[Autoimmune pancreatitis with biliary tree and liver involvement as a part of IgG4-related autoimmune sclerosing disease. Case report]. / Autoimunní pankreatitida s postizením zlucovodu a jater jako soucást IgG4 pozitivního onemocnení (IgG4-related autoimmune sclerosing disease). Kazuistika.

We report a rare case of autoimmune pancreatitis associated with sclerosing cholangitis, kidney, and salivary glands involvement. Abdominal ultrasound and CT scan showed enlarged pancreas, multiple nodular mass in the liver and kidney. ERCP showed features compatible with primary sclerosing cholangitis. The working clinical diagnosis considered malignant tumor with liver metastases. Histological examination of the liver biopsy sample revealed inflammatory process with numerous IgG4 positive plasma cells. The diagnostic conclusion of IgG4-related autoimmune sclerosing disease was drawn. The serum IgG level was elevated. Treatment with steroids improved the clinical course, all masses in the liver and kidneys disappeared, and laboratory tests were normalized. Now, 4 years after diagnosis the patient is free of all symptoms.

Allograft hepatitis after liver transplantation for epithelioid haemangioendothelioma.

Primary hepatic epitheloid heamangioendothelioma (EHE) is a rare neoplasm of vascular origin. We present a retrospective study of 6 patients with EHE treated by liver transplantation that were monitored clinically for more than 3 years and had protocol biopsy samples taken at 1, 3, 5, 7, and 10 years posttransplant. None of the patients suffered from any form of viral or autoimmune hepatitis before or after the transplantation. Two patients had lung metastases detected by preoperative imaging. All tumours showed factor VIII, CD31, and CD34 strong positive staining. In 5 of the 6 transplant recipients the protocol graft biopsies showed chronic non-specific hepatitis with slowly progressive periportal fibrosis that appeared during the 3rd post-transplant year. The septal fibrosis was diagnosed in the 6th and 10th year after transplantation. Liver tests did not reflect either the presence or the degree of inflammation or fibrosis and have remained normal. In retrospect, we consider that our recipients most probably developed alloantigen dependent inflammatory and fibrotic damage to their liver grafts. All six recipients are still alive for a median survival time of 95.1 month (range 44 months to 132 months), with good graft function, and without recurrence of the tumor. The lung metastases in 2 of the 6 patients have remained unchanged for 10 and 12 years retrospectively.

[Systemic amyloidoses in renal biopsy samples]. / Systémové amyloidózy v biopsiích ledvin.

UNLABELLED: The kidneys are one of the most frequent sites of amyloid deposition during systemic AL, AA, and several hereditary amyloidoses. Distinguishing between different forms of amyloids is clinically important because of their different treatment. MATERIAL AND METHODS: We present a 5-year retrospective study of amyloidoses diagnosed in renal biopsy samples. The classification of amyloidosis was made by immunofluorescence and immunohistochemical staining with antibodies to kappa and lambda immunoglobulin light chains, and for serum amyloid A protein. RESULTS: From January 2003 to December 2007, 996 renal biopsy samples from one centre were evaluated. Amyloidosis was diagnosed in 62 samples (6.2%); 33 (53.2%) were classified as AL and 25 (40.3%) as AA amyloidosis. Four cases have remained unclassified. We did not identify any difference in the distribution of deposits among cases with AL and AA amyloidosis, respectively. The majority of patients underwent the renal biopsy due to severe proteinuria or nephrotic syndrome. Three patients had very low proteinuria, less than 0.5 g/day. Diagnosis of amyloidosis was suspected by nephrologists in 48 patients (77.4%). CONCLUSION: Diagnosis of amyloidosis involves detection of amyloid deposits and classification of the amyloid form, which represent the basic step for appropriate therapy. Altogether it is not an easy task for pathologists, and with the emergence of markedly different treatments depending on the specific type of amyloid, the precise typing is of increasing clinical importance and should be performed with great care. Immunofluorescence can be very useful in daily practice for classification of the type of amyloidosis.

[Pancreatic resection for metastatic renal cell carcinoma]. / Resekce pankreatu pro metastázy karcinomu ledviny.

BACKGROUNDS: Late metastases of renal cell carcinoma (RCC) are quite common. However, metastases in the pancreas are rare. Between 2004-2008 the Department of transplantation surgery of the institute of clinical and experimental medicine performed 87 pancreatic resections for tumour. From this, metastasis of RCC was histologically verified in four cases.The aim of this study was to summarize in the form of brief case reports our experience with the surgical treatment of pancreatic metastasis of RCC. OBSERVATION: The interval from nephrectomy to the occurrence of pancreatic metastasis was 10, 11, 15 and 16 years. All patients were examined to exclude metastatic generalization. Surgical treatment was: one total pancreatectomy, two subtotal pancreatectomies and one caudal resection. Two patients had solitary pancreatic metastasis, one had two metastases and one had multiple metastatic lesions. No complications were observed in the postoperative period. All patients are living with survival time of 7, 23, 26 and 52 months. None of them has signs of recurrence of the primary disease. CONCLUSION: The follow up in patients with a history of RCC should be lifelong. Considering the low response of RCC and its metastases to oncological treatment, pancreatic resection is a safe method with a low rate of complications in patients with RCC metastases limited only to the pancreas and detected in time.

[Transarterial chemoembolization in hepatocellular carcinoma]. / Transarteriální chemoembolizace u hepatocelulárního karcinomu jater.

INTRODUCTION: Hepatocelullar carcinoma (HCC) is the fifth most common cancer in the world. It mostly occurs in patients with cirrhosis. In the Czech Republic, about 250 new cases are reported per year. Surgery, i.e. liver resection or transplantation, as the only potentially curable method is possible in 15-20% of them. For the rest, palliative treatment is indicated. This includes ablative methods (radiofrequency ablation, alcoholization), transarterial chemoembolization (TACE), systemic chemotherapy or biological treatment by sorafenib. TACE is method of choice in patients unsuitable for surgery and ablative treatment. Another indication is embolization of HCC before liver transplantation to prevent tumour progression. In combination with other methods, down staging of the tumour and curable treatment afterward is possible. AIMS: To assess the outcome of transarterial chemoembolisation in patients with hepatocellular carcinoma. METHODS: Between 2004-2008 we performed 30 TACE. Of that number, 28 TACE were performed in 20 patients with HCC. We super selectively catheterized the tumour via arteria femoralis and used Doxorubicin with Lipiodol as embolic material. In follow up, we carried out laboratory studies and CT. RESULTS: We have not noticed any major complications. Post-embolization syndrome with fever, nausea and right upper quadrant pain occurred after 10 TACE (33%). One-, two- and three years survival of the patients was 53%, 40% a 20%. CONCLUSION: TACE is safe method prolonging patients' survival with unresectable HCC. For the correct treatment of HCC, its concentration to cancer centres and the cooperation between multiple specialists is necessary.

[Experimental small intestine transplantation]. / Transplantace tenkého streva v experimentu.

AIM: Transplantation of the small intestine is a standard treatment method in patients with small intestinal failures. The aim of this study was to master the surgical technique, optimalize immunosuppression regimes, diagnose acute cellular graft rejection based on cellular and humoral indicators. METHODS: The authors performed a total of 43 transplantation procedures in pigs. The first, surgical part of the experiment was aimed at mastering two principal methods of vascular anastomosis- firstly, connecting the graft with mesenteric vessels (Group n1 = 18) and secondly, connecting the graft with the aorta and the inferior vena cava (Group n2 = 25). The second part of the experiment included assessment of rejection changes in various immunosuppression regimes. Only animals who did not die because of a technical failure of the procedure or due to internal reasons (n = 24) were assessed. The study animals were assigned to four groups (A (n = 3)--autotransplantation, without immunosuppresion; B (n = 7) and C (n = 8)--allotransplantation with immunosuppression using tacrolimus, resp. in a combination with sirolimus; D (n = 6)--allotransplantation without immunosuppression. Rejection was diagnosed based on histological examination of the grafts@ biopsy samples. Plasmatic citruline was used as a non-invasive humoral indicator of the graft impairment. RESULTS: Procedural complications were observed in 12 (67%) study animals from the first group, and in 3 (12%) animals from the second group. In the assessment of rejection changes, the longest survival was observed with autotransplantations, the shortest survival period was shown with allotransplantations without immunosuppression. No significant survival differences were demonstrated between the both treated groups. (p < 0.05). Group C showed lower rates of cellular rejections, compared to Group B and D. CONCLUSION: During the experiment, the authors managed to master the graft collection, as well as the transplantation technique. Lower rates of surgical complications were observed when the graft was supplied by the central vascular system. No significant differencies were observed between the tacrolimus monoterapy regimen and the combination therapy with sirolimus. Histological examination is the golden standard for the cellular rejection diagnostics. Plasmatic citruline has no signifiance in the rejection assessment.

The influence of purification of carp collagen used in a novel composite graft with sandwich construction of the wall on its biological properties and graft patency rates.

We compared graft outcome between two types of a novel composite three-layer carp-collagen-coated vascular graft in low-flow conditions in a sheep model. Collagen in group A underwent more cycles of purification than in group B in order to increase the ratio between collagen and residual fat. The grafts were implanted end-to-side in both carotid arteries in sheep (14 grafts in 7 sheep in group A, 18 grafts in 9 sheep in group B) and artificially stenosed on the right side. The flow in the grafts in group A decreased from 297±118 ml/min to 158±159 ml/min (p=0.041) after placement of the artificial stenosis in group A, and from 330±164ml/min to 97±29 ml/min (p=0.0052) in group B (p=0.27 between the groups). From the five surviving animals in group A, both grafts occluded in one animal 3 and 14 days after implantation. In group B, from the six surviving animals, only one graft on the left side remained patent (p=0.0017). Histology showed degradation of the intimal layer in the center with endothelization from the periphery in group A and formation of thick fibrous intimal layer in group B. We conclude that the ratio between collagen and lipid content in the novel three-layer graft plays a critical role in its patency and structural changes in vivo.