Role of Immunohistochemical Analysis of p16 and p53 in Vulvar Carcinoma

Tumor human papillomavirus (HPV) status is an important prognostic factor in vulvar cancer as indicated in the latest WHO classification of female genital tract tumors. Immunohistochemical detection of p16 is well established as a surrogate biomarker for tumor HPV association, including squamous cell carcinomas of the vulva. HPV-independent vulvar carcinomas are heterogeneous with 2 subcategories according to the TP53 mutation status. Therefore, the simultaneous use of p53 and p16 immunohistochemistry is recommended for accurate subclassification of vulvar squamous cell carcinomas. However, the role of molecular analytical tools, in particular RNA ISH and TP53 sequencing, is not so clear. This study aimed to investigate the performance of p53 and p16 immunohistochemistry for the diagnosis of vulvar carcinomas in comparison to TP53 mutation analysis and HPV RNA ISH. We analyzed 48 vulvar carcinomas in a tissue microarray format. Sensitivity and specificity for both methods, p16 (100% and 96%) and p53 (95% and 90%) immunohistochemistry for detection of HPV association as well as for TP53 mutations was high. Combining p16 and p53 immunohistochemistry we correctly classified all carcinomas in our series according to current WHO criteria. The sensitivity of HPV RNA ISH for the detection of HPV association was lower compared to p16 immunohistochemistry. Rare HPV-associated cases with TP53 mutation and HPV-independent tumors with p16 overexpression are discussed. In summary, the combined use of p16 and p53 immunohistochemistry for subclassification of vulvar carcinomas is justified in daily practice. Molecular tests should be restricted to rare cases with ambiguous clinicopathologic or immunohistochemical features

Abdominale Tuberkulose

Zusammenfassung: Die Tuberkulose (Tbc) zählt immer noch zu den 15 häufigsten Todesursachen weltweit. Die pulmonale Tbc stellt die klassische Manifestation dar, während die abdominale Tbc etwas aus dem Blickfeld verschwunden ist. Eine Tbc kann sich im ganzen Gastrointestinaltrakt (am häufigsten in der Ileozökalregion), im Peritoneum, in den abdominalen Lymphknoten oder in jedem soliden Organ des Abdomens manifestieren. Als "Chamäleon" kann sie u.a. eine Neoplasie oder eine entzündliche Darmerkrankung imitieren. Klinisch zeigen sich unspezifische Symptome (Fieber, Gewichtsverlust, Schmerzen) oder auch gastrointestinale Blutungen, Perforationen, Obstruktionen oder Aszites, jeweils abhängig vom Ort des Befalls. Die Diagnosesicherung erfolgt meist aus einer Gewebebiopsie oder gelegentlich mit einer Feinnadelpunktion mittels Nachweis von Mycobacterium tuberculosis in Kultur oder PCR. Die Therapie besteht in der Regel aus einer initialen Vierertherapie mit Isoniazid, Rifampicin, Pyrazinamid und Ethambutol, die bei Abwesenheit von Resistenzen auf eine zweimonatige Behandlung mit Isoniazid, Rifampicin und Pyrazinamid reduziert werden kann, gefolgt von 4Monaten Isoniazid und Rifampicin. Die Patienten müssen während der Therapie wegen medikamentösen Interaktionen und Nebenwirkungen sowie der Möglichkeit eines Immunrekonstitutionssyndroms eng begleitet werden. Zwingend ist in jedem Fall der Ausschluss einer offenen pulmonalen Tbc, da sich daraus krankenhaushygienische und epidemiologische Konsequenzen ergeben

Abdominale Tuberkulose

Die Tuberkulose (Tbc) zählt immer noch zu den 15 häufigsten Todesursachen weltweit. Die pulmonale Tbc stellt die klassische Manifestation dar, während die abdominale Tbc etwas aus dem Blickfeld verschwunden ist. Eine Tbc kann sich im ganzen Gastrointestinaltrakt (am häufigsten in der Ileozökalregion), im Peritoneum, in den abdominalen Lymphknoten oder in jedem soliden Organ des Abdomens manifestieren. Als „Chamäleon“ kann sie u. a. eine Neoplasie oder eine entzündliche Darmerkrankung imitieren. Klinisch zeigen sich unspezifische Symptome (Fieber, Gewichtsverlust, Schmerzen) oder auch gastrointestinale Blutungen, Perforationen, Obstruktionen oder Aszites, jeweils abhängig vom Ort des Befalls. Die Diagnosesicherung erfolgt meist aus einer Gewebebiopsie oder gelegentlich mit einer Feinnadelpunktion mittels Nachweis von Mycobacterium tuberculosis in Kultur oder PCR. Die Therapie besteht in der Regel aus einer initialen Vierertherapie mit Isoniazid, Rifampicin, Pyrazinamid und Ethambutol, die bei Abwesenheit von Resistenzen auf eine zweimonatige Behandlung mit Isoniazid, Rifampicin und Pyrazinamid reduziert werden kann, gefolgt von 4 Monaten Isoniazid und Rifampicin. Die Patienten müssen während der Therapie wegen medikamentösen Interaktionen und Nebenwirkungen sowie der Möglichkeit eines Immunrekonstitutionssyndroms eng begleitet werden. Zwingend ist in jedem Fall der Ausschluss einer offenen pulmonalen Tbc, da sich daraus krankenhaushygienische und epidemiologische Konsequenzen ergeben

Relevance of activated leukocyte cell adhesion molecule (ALCAM) in tumor tissue and sera of cervical cancer patients

Abstract Background An altered expression of the activated leukocyte cell adhesion molecule (ALCAM) is associated with cancer progression in various cancer types. In some cancers ALCAM has a prognostic value or is predictive for the benefit of therapeutic interventions. To date there are no data on the role of ALCAM in cervical cancer available. Methods In this study, ALCAM expression was analysed by immunohistochemistry (IHC) in tissue samples of 233 patients with cervical cancer, among them 178 with complete follow-up information. In addition, soluble (s-)ALCAM was measured in sera of a subset of the included patients (n = 55) by enzyme-linked immunosorbent assay (ELISA). Results ALCAM overexpression was detected (immunoreactive score (IRS) 2-12) in 58.4% of the cervical cancer samples. The normal ectocervical or endocervical epithelium showed no ALCAM reactivity. In untreated patients, ALCAM overexpression in tumor tissue tended to be associated with shorter cancer-specific survival (CSS) and disease-free survival (DFS). Patients, whose tumor samples showed ALCAM overexpression receiving a cytotoxic therapy like radiotherapy or chemoradiation, however, had a favourable prognosis compared to those patients, whose cancers showed no or minimal ALCAM staining. This effect was particularly apparent in patients receiving chemoradiation where the CSS was significantly longer in patients with ALCAM-positive tumors (p = 0.038; cumulative incidence rates at 96 months 8%, 95% CI 0%-23%, and 26%, CI 3%-43% in ALCAM-positive and ALCAM-negative cases, respectively). Median preoperative s-ALCAM concentration in sera from tumor patients was 27.6 ng/ml (range 17.5-55.1 ng/ml, mean 28.9 ng/ml), serum levels did not correlate with intratumoral ALCAM expression. Conclusions The data of our retrospective study suggest that the prognostic value of ALCAM expression in cervical carcinoma might be therapy-dependent, and that ALCAM might function as a predictive marker for the response to chemoradiation. This should be confirmed in further, prospective studies. </jats:sec

Optimization of a Novel Peptide Ligand Targeting Human Carbonic Anhydrase IX

BACKGROUND: Carbonic anhydrase IX (CA IX) is a hypoxia-regulated transmembrane protein over-expressed in various types of human cancer. Recently, a new peptide with affinity for human carbonic anhydrase IX (CaIX-P1) was identified using the phage display technology. Aim of the present study is to characterize the binding site in the sequence of CaIX-P1, in order to optimize the binding and metabolic properties and use it for targeting purposes. METHODOLOGY/PRINCIPAL FINDINGS: Various fragments of CaIX-P1 were synthesized on solid support using Fmoc chemistry. Alanine scanning was performed for identification of the amino acids crucial for target binding. Derivatives with increased binding affinity were radiolabeled and in vitro studies were carried out on the CA IX positive human renal cell carcinoma cell line SKRC 52 and the CA IX negative human pancreatic carcinoma cell line BxPC3. Metabolic stability was investigated in cell culture medium and human serum. Organ distribution and planar scintigraphy studies were performed in Balb/c nu/nu mice carrying subcutaneously transplanted SKRC 52 tumors. The results of our studies clearly identified amino acids that are important for target binding. Among various fragments and derivatives the ligand CaIX-P1-4-10 (NHVPLSPy) was found to possess increased binding potential in SKRC 52 cells, whereas no binding capacity for BxPC3 cells was observed. Binding of radiolabeled CaIX-P1-4-10 on CA IX positive cells could be inhibited by both the unlabeled and the native CaIX-P1 peptide but not by control peptides. Stability experiments indicated the degradation site in the sequence of CaIX-P1-4-10. Biodistribution studies showed a higher in vivo accumulation in the tumor than in most healthy tissues. CONCLUSIONS: Our data reveal modifications in the sequence of the CA IX affine ligand CaIX-P1 that might be favorable for improvement of target affinity and metabolic stability, which are necessary prior to the use of the ligand in clinical approaches

Table 2: Anti-tumor agents for targeting hypoxia-induced CA IX for therapy and diagnosis.

The expression of carbonic anhydrase (CA) IX is up-regulated in many types of solid tumors in humans under hypoxic and acidic microenvironment. Inhibition of CA IX enzymatic activity with selective inhibitors, antibodies or labeled probes has been shown to reverse the acidic environment of solid tumors and reduce the tumor growth establishing the significant role of CA IX in tumorigenesis. Thus, the development of potent antitumor drugs targeting CA IX with minimal toxic effects is important for the target-specific tumor therapy. Recently, several promising antitumor agents against CA IX have been developed to treat certain types of cancers in combination with radiation and chemotherapy. Here we review the inhibition of CA IX by small molecule compounds and monoclonal antibodies. The methods of enzymatic assays, biophysical methods, animal models including zebrafish and Xenopus oocytes, and techniques of diagnostic imaging to detect hypoxic tumors using CA IX-targeted conjugates are discussed with the aim to overview the recent progress related to novel therapeutic agents that target CA IX in hypoxic tumors

Examinations on possibilities to applicate a tourque measurement system at the power take-off of a tractor

Die vorliegende Arbeit befasst sich mit der Untersuchung von Messsystemen, die die Messung eines Drehmoments in einem Schleppergetriebe ermöglichen sollen. Dazu wurde zunächst die vorangegangene Bachelorarbeit kurz vorgestellt und ausgewertet. An-schließend wurden die infrage kommenden Messprinzipien noch einmal erklärt. Für die geeigneten Messsysteme wurden, nach der Erstellung einer Anforderungsliste an die Konstruktion, Entwürfe in mehreren Varianten erstellt. Nach der konstruktiven Erstellung mit einem 3D-Programm, wurde für beide Systeme eine technische und wirtschaftliche Bewertung erstellt. Anhand der angestellten Untersuchungen erfolgte im Anschluss ein Vergleich der beiden Varianten. Zum Schluss wurden die Ergebnisse der Arbeit noch ein-mal zusammengefasst und ein Ausblick für die weitere Vorgehensweise gegeben