Detection of HHV-6 Virus in specimen of a ductal pancreatic adenocarcinoma with comparison in tumor and normal tissue.

AIMS: The association of human herpesvirus 6 (HHV-6) species with pancreatic cancer is controversially discussed. The aim of this study was to further investigate the postulated association and to identify the basis of HHV-6 DNA positivity reported for pancreatic cancer tissue. METHODS: All samples of patients with pancreatic cancer (cancer and surrounding tissue) were analyzed for presence of HHV-6 DNA by PCR and then selected cases by immunohistochemistry. RESULTS: Sixty eight per cent (68% = 52/77) of all patients were HHV-6 DNA positive in any of the samples, 49% (38/77) were positive in tumor tissue. Specimens of just one patient were HHV-6A DNA positive, all other patients were positive for HHV-6B. Immunohistochemical analysis of HHV-6 DNA positive samples did not reveal any specific HHV-6B protein positive tumor cell. In contrast, supposed immune cells presented intra- and peritumorally expressed HHV-6B-protein. The cause of presence of these cells in the tumor stroma is unknown, as of yet. CONCLUSIONS: HHV-6 DNA-positivity of pancreatic cancer tissue described by us and others is probably not due to the infection of pancreatic cells by HHV-6, but rather due to the migration of HHV-6 positive immune cells into the pancreas. Based on our data, we suppose that there is no direct evidence for HHV-6 as a causative agent of pancreatic cancer, but further in-depth studies (including investigation of immune status of patients) are necessary to make definitive conclusions.

[Scope of activities and research in pathology laboratories from 1920 to 1940]. / Tätigkeitsfelder und Forschungslandschaft der Pathologie in den Jahren 1920 bis 1940.

The ideal of a pathology institute consisted of a large autopsy department and a wide collection of specimens for the training of students and clinically active doctors. Additional departments supporting the autopsy department were a chemical and bacteriological laboratory. Other departments were experimental laboratories for biology, immunology, and cell research as well as an animal facility.The main pillar of the training was the understanding of the concept of disease and the development of disease. In this context, the final state of a disease was usually presented in the context of the autopsy. The special contribution of Rudolf Virchow was the additional consideration of cellular changes using microscopic preparations.In contrast, in England and America, clinical care on patients was carried out within the institutes in addition to autopsies. For this purpose, some institutes had their own wards with patient beds. The areas of research were accordingly different. The analysis of journal articles from 1920 to 1940 in two German-language journals, Virchows Archive and Naunyn-Schmiedebergs Archiv für experimentelle Pathologie und Pharmakologie (today: Naunyn-Schmiedeberg's Archives of Pharmacology), and two English-language journals, Journal ofPathology and Bacteriology and American Journal of Pathology, showed different scopes and numbers of publications.On the basis of these publications, it was found that the German journals published a huge variety of diseases, especially series of identical tumors and often the first description of the disease. The British Journal of Pathology published mainly infectious topics and numerous animal experiments. The American journal covered a very broad spectrum of publications, including many on clinically relevant histological techniques.

[Resection margins in soft tissue sarcomas]. / Resektionsgrenzen bei Weichteilsarkomen.

BACKGROUND: Soft tissue sarcomas (STS) are a rare and heterogeneous group of malignant tumors that arise from the mesenchymal tissue. STS can form anywhere in the human body, with the extremities being preferred sites of predilection. TREATMENT: A fundamental pillar of treatment is the surgical resection of soft tissue sarcomas. The goal is always an R0 resection with a safety margin. There is no consensus in the literature about the desired tumor-free resection margin. The decisive factors for these resection margins are histopathology, presence of anatomical barriers (capsule, tendon, fascia, cartilage, periosteum) and possibilities of (neo-) adjuvant therapy. DISCUSSION: References in the literature support the role of resection margins as a predictor of local recurrence. Regarding the role of resection margins in overall survival, available data is divergent. There are known prognostic factors that influence overall survival, such as histological subtype, tumor size, tumor grading, and presence of metastases. So far, several studies have attempted to quantify the margins of resection, but no consensus has been reached, and debates are ongoing. When analyzing all the results of the data in the literature, it seems appropriate to aim for a negative resection margin >1 mm including an anatomical border structure, if possible.

Reversal of dabigatran by intraosseous or intravenous idarucizumab in a porcine polytrauma model.

BACKGROUND: Idarucizumab is licensed to reverse dabigatran in life-threatening haemorrhage. Establishment of venous access can be challenging, and the intraosseous (IO) route is a potentially life-saving alternative. In this study, we compared the efficacy and safety of IO or intravenous (i.v.) idarucizumab for dabigatran reversal in a porcine polytrauma model. METHODS: Male pigs (n=21) received oral dabigatran etexilate (30 mg kg-1 bid) for 3 days. On the 4th day, animals received dabigatran infusion and were randomised 1:1:1 to receive IO saline (control), i.v. idarucizumab (60 mg kg-1), or IO idarucizumab (60 mg kg-1), or animals were included in a sham group (n=7). Study treatment was administered after polytrauma and the animals were monitored for 240 min, or until death. Coagulation status was monitored by thromboelastometry, thromboelastography, and thrombin measurements. RESULTS: Total blood loss was lowest in sham animals [521 (52) ml, P<0.01 vs all other groups], and comparable in the two idarucizumab groups [IO: 1085 (102) ml vs i.v.: 1142 (125) ml], and highest in the control group [4065 (557) ml, P<0.001 vs all other groups]. Survival to 240 min was 100% in the sham group and both idarucizumab groups, and 14% in the control group. IO and i.v. idarucizumab promptly normalised global coagulation assays and thrombin generation. Thromboelastography showed a strong correlation between dabigatran concentrations and R-time (R2=0.90 and 0.89) in idarucizumab-treated animals. CONCLUSIONS: Intravenous and intraosseous idarucizumab were comparable for reversing dabigatran in a porcine trauma model. Dabigatran reversal could be monitored using fully automated thromboelastography.

[Fetal autopsies : Relic or still a gold standard?] / Fetale Obduktionen : Veraltetes Relikt oder nach wie vor ein Goldstandard?

BACKGROUND: Fetal autopsy rates are decreasing in Western countries although post-mortem examinations render important information for the parents concerning the cause of abortion and risk of recurrence in future pregnancies. OBJECTIVE: The intention of the presented study was to analyze the development of fetal autopsies in Germany during the last decade and to review accessible information obtained by fetal autopsy. MATERIAL AND METHODS: Reports of fetal autopsies conducted in two German university Institutes of pathology between 2005 and 2014 were evaluated retrospectively. Demographic data and the correlation between clinical diagnoses and autopsy findings were assessed. In addition, differences between spontaneous and induced cases of abortion and differences between the institutes were also documented. RESULTS: Overall, 428 fetal autopsies were performed, whereby the number of autopsies decreased by 24.2% during the study period. Of the examined fetuses 29.7% were induced abortions which as expected exhibited different malformations compared to cases of spontaneous abortion (p < 0.001). There was no evidence of a malformation or other cause of death in 27.1% of the cases and 95.7% of these abortions occurred spontaneously. A discrepancy between clinical and autopsy findings was evident in 6.8% of cases and 3.5% of the autopsy examinations revealed at least one additional malformation compared to the prenatal clinical data. CONCLUSION: Despite improvements in prenatal diagnostics, fetal autopsies remain an important diagnostic tool even today contributing additional information in a considerable number of cases potentially revising clinical diagnoses.

Immunotherapy with imiquimod and interferon alfa for metastasized Merkel cell carcinoma.

Merkel cell carcinoma (mcc) is a highly aggressive neuroendocrine tumour of the skin. Remission rates are high with chemotherapy in patients with metastasis, but without any improvement in overall survival. We present the case of a 90-year-old woman with facial mcc. After radiation and surgery, the mcc recurred with widespread cutaneous and regional lymph node metastases. The metastases were treated with weekly intralesional injections of 1-2×10(6) IU interferon alfa-2a, accompanied by topical imiquimod 5% cream 3 times weekly. After partial regression, subcutaneous pegylated interferon alfa-2b was added at a dose of 30 µg weekly, which was then increased to 50 µg weekly. At 4 months after the start of immunotherapy, all cutaneous metastases and the intralesionally treated lymph node metastases receded. Interruption or reduction of systemic interferon application resulted in locoregional relapses that were successfully treated with surgery or intralesional interferon injections. The patient remains alive 30 months after initiation of immunotherapy, suggesting that locally metastasized mcc might be able to be controlled with local and systemic immunotherapy.

Mtss1 is a critical epigenetically regulated tumor suppressor in CML.

Chronic myeloid leukemia (CML) is driven by malignant stem cells that can persist despite therapy. We have identified Metastasis suppressor 1 (Mtss1/MIM) to be downregulated in hematopoietic stem and progenitor cells from leukemic transgenic SCLtTA/Bcr-Abl mice and in patients with CML at diagnosis, and Mtss1 was restored when patients achieved complete remission. Forced expression of Mtss1 decreased clonogenic capacity and motility of murine myeloid progenitor cells and reduced tumor growth. Viral transduction of Mtss1 into lineage-depleted SCLtTA/Bcr-Abl bone marrow cells decreased leukemic cell burden in recipients, and leukemogenesis was reduced upon injection of Mtss1-overexpressing murine myeloid 32D cells. Tyrosine kinase inhibitor (TKI) therapy and reversion of Bcr-Abl expression increased Mtss1 expression but failed to restore it to control levels. CML patient samples revealed higher DNA methylation of specific Mtss1 promoter CpG sites that contain binding sites for Kaiso and Rest transcription factors. In summary, we identified a novel tumor suppressor in CML stem cells that is downregulated by both Bcr-Abl kinase-dependent and -independent mechanisms. Restored Mtss1 expression markedly inhibits primitive leukemic cell biology in vivo, providing a therapeutic rationale for the Bcr-Abl-Mtss1 axis to target TKI-resistant CML stem cells in patients.

Margins of oral leukoplakia: autofluorescence and histopathology.

Autofluorescence devices are widely used to examine oral lesions. The aim of this study was to see whether there were any signs of dysplasia, parakeratosis, or mucosal inflammation in the borders of homogeneous oral leukoplakia using autofluorescence, and we also compared clinically visible extensions with those detected by autofluorescence. Twenty patients with 26 homogeneous areas of oral leukoplakia were included in the study. After the clinically visible extensions of the lesion had been marked, we took a photograph through the autofluorescence device, which showed both borders in one picture. We then used photo-editing software to measure the size of the area of leukoplakia together with the area with loss of autofluorescence. We took 3 punch biopsy specimens: one from the leukoplakia, one 2.5mm from its marked borders, and one from healthy mucosa. Seventy-eight biopsy specimens were examined by an experienced pathologist, and 95% CI calculated to assess the amount of parakeratosis. Spearman's rank correlation was used to assess the association with mucosal inflammation. Ten areas of leukoplakia were surrounded by normal green autofluorescence, and 16 were consistent with loss of autofluorescence with a mean size of 66%, which exceeded the clinically visible size of the area of leukoplakia. We calculated that there was a strong association between these entities and their surrounding areas, with loss of autofluorescence for parakeratosis. Some leukoplakias showed clinically invisible extensions during histopathological examination and autofluorescence. The technique described enables clinicians to measure the extent of these lesions beyond their visible margins. We found no dysplasia, which emphasises that autofluorescence detects non-dysplastic lesions caused by mucosal inflammation and parakeratosis.

A 'waterfall' transfer-based workflow for improved quality of tissue microarray construction and processing in breast cancer research.

A major focus in cancer research is the identification of biomarkers for early diagnosis, therapy prediction and prognosis. Hereby, validation of target proteins on clinical samples is of high importance. Tissue microarrays (TMAs) represent an essential advancement for high-throughput analysis by assembling large numbers of tissue cores with high efficacy and comparability. However, limitations along TMA construction and processing exist. In our presented study, we had to overcome several obstacles in the construction and processing of high-density breast cancer TMAs to ensure good quality sections for further research. Exemplarily, 406 breast tissue cores from formalin-fixed and paraffin embedded samples of 245 patients were placed onto three recipient paraffin blocks. Sectioning was performed using a rotary microtome with a "waterfall" automated transfer system. Sections were stained by immunohistochemistry and immunofluorescence for nine proteins. The number and quality of cores after sectioning and staining was counted manually for each marker. In total, 97.1 % of all cores were available after sectioning, while further 96 % of the remaining cores were evaluable after staining. Thereby, normal tissue cores were more often lost compared to tumor tissue cores. Our workflow provides a robust method for manufacturing high-density breast cancer TMAs for subsequent IHC or IF staining without significant sample loss.

A 'telomere-associated secretory phenotype' cooperates with BCR-ABL to drive malignant proliferation of leukemic cells.

Telomere biology is frequently associated with disease evolution in human cancer and dysfunctional telomeres have been demonstrated to contribute to genetic instability. In BCR-ABL(+) chronic myeloid leukemia (CML), accelerated telomere shortening has been shown to correlate with leukemia progression, risk score and response to treatment. Here, we demonstrate that proliferation of murine CML-like bone marrow cells strongly depends on telomere maintenance. CML-like cells of telomerase knockout mice with critically short telomeres (CML-iG4) are growth retarded and proliferation is terminally stalled by a robust senescent cell cycle arrest. In sharp contrast, CML-like cells with pre-shortened, but not critically short telomere lengths (CML-G2) grew most rapidly and were found to express a specific 'telomere-associated secretory phenotype', comprising secretion of chemokines, interleukins and other growth factors, thereby potentiating oncogene-driven growth. Moreover, conditioned supernatant of CML-G2 cells markedly enhanced proliferation of CML-WT and pre-senescent CML-iG4 cells. Strikingly, a similar inflammatory mRNA expression pattern was found with disease progression from chronic phase to accelerated phase in CML patients. These findings demonstrate that telomere-induced senescence needs to be bypassed by leukemic cells in order to progress to blast crisis and provide a novel mechanism by which telomere shortening may contribute to disease evolution in CML.

Efficacy of magnetic thermoablation using SPIO in the treatment of osteoid osteoma in a bovine model compared to radiofrequency and microwave ablation.

PURPOSE: To evaluate heating efficacy of superparamagnetic iron oxide nanoparticles (SPIO) for electromagnetic ablation (EMA) of osteoid osteoma (OO) using an ex vivo model compared to radiofrequency ablation (RFA) and microwave ablation (MWA). METHODS: A model for OO using sliced bovine tibia and sliced muscle tissue was developed. A bone cavity filled with either a mixture of SPIO and agarose or pure agarose (control group) was established. EMA was performed using an experimental system, RFA and MWA using clinically approved systems, and the ablation protocols recommended by the vendor. For temperature measurements, fiberoptic temperature probes were inserted inside the cavity, on the outside of the periosteum, and at a 5 mm distance to the periosteum. RESULTS: Maximum temperatures with or without SPIO in the nidus were as follows: EMA: 79.9 ± 2.5/22.3 ± 0.7 °C; RFA: 95.1 ± 1.8/98.6 ± 0.9 °C; MWA: 85.1 ± 10.8/83.4 ± 9.62 °C. In RFA with or without SPIO significantly higher temperatures were achieved in the nidus compared to all other groups (p < 0.05). In MWA significantly higher temperatures were observed in the 5 mm distance to the periosteum compared to EMA and RFA with or without SPIO (p < 0.05). In MWA temperature decrease between nidus and the 5 mm distance to the periosteum was significantly lower than in RFA with or without SPIO (p < 0.0001). In MWA without SPIO temperature decrease was significantly lower than in the EMA group (p < 0.05). CONCLUSION: In the experimental setting, ablation of OO is safe and effective using EMA. It is less invasive than RFA and MWA, and it theoretically allows repeated treatments without repeated punctures. In comparison, the highest temperatures in the nidus are reached using RFA.

A rare case of massive carpal osteoblastoma requiring complex reconstructive surgery.

An osteoblastoma is a rare, commonly benign, osteoid-producing neoplasm of the bone with an incidence of 2% of all primary bone tumours. We present a case of a 54-year-old patient with persisting carpal pain and massive swelling of the hand for a period of 4 years. Incision biopsies revealed the histopathological finding of a carpal osteoblastoma. After complete tumour excision, including the carpal and, in parts, metacarpal bones, reconstructive surgery was performed with a free osteocutaneous iliac crest flap to obtain a natural hand contour and the best possible hand function. Follow-up revealed improvement of the hand function in terms of flexion, extension and strength without discomfort or further pain. Thus, ongoing carpal pain should lead to an intensive search with further diagnostic measures such as magnetic resonance imaging (MRI) scan as well as biopsies, if necessary, to obtain the correct diagnosis.

[Bone biopsy needles: mechanical properties, needle design and specimen quality]. / Knochen-Biopsiesysteme: mechanische Eigenschaften, Nadeldesign und Probenqualität.

PURPOSE: To quantitatively analyze differences in mechanical properties, needle design including signs of wear, subjective handling and specimen quality of bone biopsy needles. MATERIALS AND METHODS: In this study 19 different bone biopsy systems (total 38; 2 /type) were examined. With each biopsy needle five consecutive samples were obtained from vertebral bodies of swine. During puncture a force-torques sensor measured the mechanical properties and subjective handling was assessed. Before and after each biopsy the needles were investigated using a profile projector and signs of wear were recorded. Afterwards, a pathologist semi-quantitatively examined the specimen regarding sample quality. The overall evaluation considered mechanical properties, needle wear, subjective handling and sample quality. Differences were assessed for statistical significance using ANOVA and t-test. RESULTS: Needle diameter (p = 0.003) as well as needle design (p = 0.008) affect the mechanical properties significantly. Franseen design is significantly superior to other needle designs. Besides, length reduction recorded by the profile projector, as a quality criterion showed notable distinctions in between the needle designs. CONCLUSION: Bone biopsy needles vary significantly in performance. Needle design has an important influence on mechanical properties, handling and specimen quality. Detailed knowledge of those parameters would improve selecting the appropriate bone biopsy needle.