So-called "superficial" bladder tumors. Which classification in 2003? Part 1: Papillary tumors

The so-called superficial tumors of the urinary bladder three types of tumors: non infiltrating papillary tumors (pTa), tumors infiltrating the lamina propria (pT1) and carcinoma in-situ. Even if they all need conservative treatment, they belong to three distinct categories of tumors with their own recurrence and progression risks. All these tumors are evaluated according to TNM stages and morphological grades. The reproducibility of the grading has been recently discussed; the International Society of Urological Pathology (ISUP) and the World Health Organization (WHO) have proposed new criteria for new classifications in 1998 and 1999 respectively. These classifications have not been approved by most pathologists and they are discussed and commented here, including the recent point of view defending the refinement of the WHO 1973 classification for the grading. To understand well pT1 bladder carcinomas, the discussion of sub-staging (pT1a and pT1b) is exposed.     

Multiple cytokeratin-negative malignant tumors composed only of rhabdoid cells in the renal pelvis: a sarcomatoid urothelial carcinoma?

The author presents a unique case of multiple cytokeratin-negative malignant tumors consisting only of rhabdoid cells in the renal pelvis. A 54-year-old man complained of hematuria. A transurethral endoscopic examination revealed multiple papillary tumors, and transurethral resection of the bladder tumors was performed. Pathologically, they were ordinary papillary urothelial transitional cell carcinomas. Imaging modalities revealed multiple tumors of the right renal pelvis, and nephrectomy was performed. Grossly, three polypoid tumors measuring 2-4 cm were present in the pelvis. Histologically, they were composed only of malignant cells with rhabdoid features. There were no elements of transitional cell carcinoma. Immunohistochemically, the pelvic tumors were positive for vimentin and Ki-67 antigen (labeling=40%). They were negative for pancytokeratins (AE1/3, CAM5.2, KL-1 and polyclonal wide), 34βE12, cytokeratin (CK) 5/6, CK7, CK8, CK14, CK18, CK19, CK20, melanosome, EMA, CEA, desmin, S100 protein, α-smooth muscle actin, myoglobin, myogenin, CD34, p53 protein, p63, CD3, CD20, CD30, CD45, CD45RO, chromograin, synaptophysin, CD56, CD68, and KIT. NSE and PDGFRA were focally present, but this appeared nonspecific. Namely, the pelvic tumors expressed only vimentin. The author speculates that the pelvic multiple malignant “rhabdoid” tumors are not sarcomas but urothelial “rhabdoid” carcinoma with complete loss of CKs.     

Gastrointestinal stromal tumors (GISTs): CD117, DOG-1 and PKCθ expression. Is there any advantage in using several markers?

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the digestive tract. Expression of CD117, DOG1 and PKCθ was investigated immunohistochemically in a series of 99 paraffin-embedded GISTs in order to determine the sensitivity and diagnostic value of these markers. KIT exons 9, 11, 13 and 17 and PDGFRA exons 12 and 18 were amplified by PCR and sequenced. A total of 94/99 (94%) GISTs stained positive for CD117, 81/99 (82%) for PKCθ and 90/99 (91%) for DOG-1. A significant correlation was noted between CD117 and DOG-1 expression (p=0.0001). All three markers were expressed in 74% (73/99) of GISTs. Of the five CD117-negative cases, two were PKCθ-negative/DOG1-negative and had mutations in KIT exon 11. Two were PKCθ-positive/DOG1-positive and had mutations in PDGFRA (one each in exons 12 and 18), and one was DOG1-negative/PKCθ-positive, with a PDGFRA exon 18 mutation. The most sensitive marker was CD117, followed by DOG-1 and PKCθ. Although PKCθ was less sensitive, and its staining is more challenging and difficult to interpret, the use of this marker is highly recommended, particularly in CD117-negative/DOG-1-negative GISTs.     

BRAF mutation,TERT promoter mutation,and HER2 amplification in sporadic or neurofibromatosis-related neurofibromas and malignant peripheral nerve sheath tumors: do these molecules have a signature in malignant transformation?

Peripheral nerve sheath tumors may occur sporadically or related to neurofibromatosis (NF). Unless the mechanisms of tumorigenesis in NF related malignant peripheral nerve sheath tumors (MPNST) are better understood, it remained unclear in sporadic cases. We aimed to investigate the genetic route for malignancy in both individuals with NF-1 and sporadic ones to open a way for targeted therapies in the future. We investigated the role of HER2 with Dual ISH DNA Probe Cocktail test, BRAF mutation (exon 15) and TERT promoter mutation frequency with Sanger sequencing method in respectively 25 sporadic neurofibromas, 25 NF-1 related neurofibromas and 25 MPNST cases from two institutes. Categorical data were analyzed and summarized as frequency and percentage. Statistical analysis was done with SPSS v.22 statistical package, and the statistical significance level was considered as 0.05. We identified TERT promoter mutation only in one sporadic MPNST (4%) and no BRAF mutation in any case. HER2 amplification is found in 10/25 (40%) MPNST cases. No mutations or gene amplification detected in neurofibromas (p < 0.001). MPNSTs are sarcomas with poor prognosis and limited treatment options. TERT promoter mutations and HER2 amplification may play a putative role in therapeutic purposes.     

Chromogenic in situ hybridization for the detection of HER-2/neu gene amplification in breast cancer with an emphasis on tumors with borderline and low-level amplification: does it measure up to fluorescence in situ hybridization?

We compared chromogenic in situ hybridization (CISH) with fluorescence in situ hybridization (FISH) for assessing HER-2/neu gene amplification using tissue microarrays (TMAs) made from formalin-fixed, paraffin-embedded tissue blocks from 113 cases of invasive breast carcinoma. TMAs were created using 0.6-mm tissue cores with 4 sampled cores per tumor. For both assays, a HER-2/chromosome 17 signal ratio of 2.0 or more was considered positive for gene amplification. The average ratio of cores from the same tumor was used for determination of gene amplification status of that particular tumor Of 113 cases, 102 were tested successfully by both assays. The results were concordant in 100.0% of cases (63 amplified; 39 nonamplified). All 22 cases of borderline (ratio, 2.0-2.5) or low-level (ratio, 2.6-3.9) amplification by FISH also showed HER-2 gene amplification by CISH. CISH is as sensitive as FISH in detecting borderline and low-level HER-2 amplification. Reliable recognition of the invasive carcinoma area by light microscopy and preservation of the test slides are added advantages of CISH. CISH performs as well as FISH in the analysis of HER-2 gene amplification in breast cancer and might have advantages in certain situations.     

Reflektorische Beeinflussung der Atmung durch Änderung der CO2-Konzentration in den Lungenalveolen bzw. Lungencapillaren: Ein bisher unbekannter Atemreflex?

Zusammenfassung Untersuchungen an acht Hunden mit Bronchospirometrie und einseitiger Pulmonalisblockade ergaben außer einer Verkleinerung des Atemvolumens auf der nichtdurchbluteten Lungenseite eine Verminderung der Atemfrequenz. CO₂-Zugabe auf der nichtdurchbluteten Seite führte zur Aufhebung der Volumenverkleinerung und zur Erhöhung der Atemfrequenz. Nach Vagotomie blieb die Atemvolumenverminderung, die auf lokale Bronchiolenconstriction durch CO₂-Mangel zurückzuführen ist, erhalten, während die Atemfrequenz keine Änderungen mehr zeigte. Wir glauben daraus schließen zu können, daß es sich um einen bisher unbekannten Atemreflex handelt, der von der CO₂-Konzentration in den Alveolen bzw. Lungencapillaren abhängig ist und über den afferenten Vagus vermittelt wird: Stärkere CO₂-Verminderung in der Lunge führt zur Verlangsamung der Atmung, CO₂-Erhöhung zur Beschleunigung. Eine Vermittlung der CO₂-Änderungen an das Atemzentrum auf dem Blutwege ist bei der Versuchsanordnung nicht möglich und wurde auch blutgasanalytisch ausgeschlossen. Einseitige Stenosen ohne Aufhebung des Blutdurchflusses auf derselben Seite führen zu keiner Änderung der Atemfrequenz.     

Immune alterations in three mouse strains following 2-deoxy- -glucose administration

Using 2-deoxy- -glucose (2-DG)-induced stress, our laboratory has developed studies to define stress effects on immune responses. Here, we report effects of increasing doses of 2-DG on the immune response of BALB/c, C57BL/6 and BDF₁ mice 2 h after three injections of 0 to 2000 mg/kg of 2-DG. Female 4- to 5-week-old mice were euthanized and blood and spleens were collected. A suspension of partially purified mature T splenocytes was obtained by negative selection using J11.d2 antibodies. Glucose and corticosterone levels were measured in the plasma of each mouse. Splenocyte and mature T splenocyte suspensions were tested in in vitro proliferation assays with or without concanavalin A. Splenocytes were analyzed for the following cell-surface markers: CD3, TCR α/β, CD4, CD8 and major histocompatibility complex (MHC) Class II. Significant increases in blood glucose levels were observed in C57BL/6 and BALB/c strains with the highest 2-DG dose (p<0.05). Corticosterone levels were higher in BDF₁ mice and C57BL/6 mice following the administration of 1000 and 2000 mg/kg of 2-DG, respectively (p<0.01). In vitro proliferation of mature T splenocytes in the presence of concanavalin A was decreased in BDF₁ (p<0.05) but not in BALB/c and C57BL/6 mice. In addition, in BDF₁ mice the decrease was highly correlated with an increase of CD3+ and TCR α/β+ cells in the spleen. These results demonstrated high variability in the response of different mouse strains to 2-DG-induced stress.     

Enzyme Replacement Therapy in Mucopolysaccharidosis I: Altered Distribution and Targeting of α- -Iduronidase in Immunized Rats

Enzyme replacement therapy (ERT) has been developed and trialed for the treatment of human lysosomal storage disorder patients. The viability of ERT for the treatment of these severe multiple pathology disorders has subsequently been established. However, in both animal model studies and human clinical trials, some individuals have been shown to develop an immune response to the replacement protein. This potential complication for treatment has been investigated by the infusion of recombinant human α- -iduronidase (rh-α- -iduronidase) into nonimmune and immunized rats to simulate mucopolysaccharidosis type I ERT in the presence of different levels of antibody. In rats with high antibody titers to rh-α- -iduronidase (titer 1,024,000) there was evidence of altered organ distribution and subcellular targeting when compared to either lower titer immunized rats (titers less than 64,000) or nonimmune rats (titers 512–1024). In addition, hypersensitivity reactions were observed for high titer rats (titer 1,024,000) during rh-α- -iduronidase infusion, but not for the other two treatment groups. A rat with an antibody titer of 64,000 had only minor changes in subcellular targeting and organ distribution when infused with rh-α- -iduronidase. This implied that a high level of antibody was required to effect changes in α- -iduronidase enzyme targeting and distribution. Notably, in the high titer rats, the antibody produced appeared to increase the tissue and subcellular level of rh-α- -iduronidase specific activity. This suggested that antibody production may not always result in an adverse effect on ERT.