Jörg Marienhagen

GLUT1 expression is increased in hepatocellular carcinoma and promotes tumorigenesis.

Accelerated glycolysis is one of the biochemical characteristics of cancer cells. The glucose transporter isoform 1 (GLUT1) gene encodes a key rate-limiting factor in glucose transport into cancer cells. However, its expression level and functional significance in hepatocellular cancer (HCC) are still disputed. Therefore, we aimed to analyze the expression and function of the GLUT1 gene in cases of HCC. We found significantly higher GLUT1 mRNA expression levels in HCC tissues and cell lines compared with primary human hepatocytes and matched nontumor tissue. Immunohistochemical analysis of a tissue microarray of 152 HCC cases revealed a significant correlation between Glut1 protein expression levels and a higher Ki-67 labeling index, advanced tumor stages, and poor differentiation. Accordingly, suppression of GLUT1 expression by siRNA significantly impaired both the growth and migratory potential of HCC cells. Furthermore, inhibition of GLUT1 expression reduced both glucose uptake and lactate secretion. Hypoxic conditions further increased GLUT1 expression levels in HCC cells, and this induction was dependent on the activation of the transcription factor hypoxia-inducible factor-1alpha. In summary, our findings suggest that increased GLUT1 expression levels in HCC cells functionally affect tumorigenicity, and thus, we propose GLUT1 as an innovative therapeutic target for this highly aggressive tumor.

Glucose Transporter 1 Gene Expression is Related to Thyroid Neoplasms with an Unfavorable Prognosis: An Immunohistochemical Study

PURPOSE An accelerated rate of glucose metabolism mediated by overexpression of key regulatory glycolytic enzymes and glucose transporters is among the most characteristic biochemical marker of malignant transformed cells. In thyroid neoplasms, however, an increased uptake of glucose [measured by 2-[18F]-fluoro-2-deoxy-D-glucose (FDG) and positron emission tomography (PET)] seems to be restricted to more aggressive and high-grade tumors, whereas tumors with favorable prognosis demonstrate no significant tracer uptake. We therefore studied the expression of glucose transporters in thyroid carcinomas with different grades of malignancy. METHODS Sections of formalin-fixed and paraffin-embedded tissue obtained from 45 patients with thyroid cancer (5 anaplastic, 20 papillary and 20 follicular tumors) were investigated. Polyclonal rabbit antiglucose transporter antibodies, reactive with glucose transporters 1-5 (GLUT1-5), were used after heat pretreatment of the sections. Staining was performed by the avidin-biotin conjugate immunoperoxidase reaction and evaluated semiquantitatively. RESULTS Expression of GLUT1 transporter on the cell membrane was closely related to the grade of malignancy in thyroid neoplasms (Fisher exact test p < 0.05). All anaplastic tumors showed a high level of GLUT1 expression in the cytoplasm and on the cell membrane. Positive membranous staining in differentiated tumors was detected predominantly in neoplasms with unfavorable prognosis, e.g., in widely invasive follicular or metastatic tumors, whereas low or no immunoreactivity could be seen in well-differentiated tumors or in normal thyroid epithelium. CONCLUSIONS These data indicate that overexpression of GLUT1 on the cell membrane of thyroid neoplasms is closely related to tumors demonstrating a more aggressive biological behavior. Therefore, determination of GLUT1 expression in thyroid cancer tissue may be a prognostic marker, and FDG-PET may be a helpful technique in identifying patients at a higher risk.

Dual function of membrane-bound heat shock protein 70 (Hsp70), Bag-4, and Hsp40: protection against radiation-induced effects and target structure for natural killer cells

CX+/CX− and Colo+/Colo− tumor sublines with stable heat shock protein 70 (Hsp70) high and low membrane expression were generated by fluorescence activated cell sorting of the parental human colon (CX2) and pancreas (Colo357) carcinoma cell lines, using an Hsp70-specific antibody. Two-parameter flow cytometry revealed that Hsp70 colocalizes with Bag-4, also termed silencer of death domain, not only in the cytosol but also on the plasma membrane. After nonlethal γ-irradiation, the percentage of membrane-positive cells and the protein density of Hsp70 and Bag-4 were found to be strongly upregulated in carcinoma sublines with initially low expression levels (CX−, Colo−). Membrane expression of Hsp70 was also elevated in Bag-4 overexpressing HeLa cervix carcinoma cells when compared to neo-transfected cells. In response to γ-irradiation, neo-transfected HeLa cells behaved like Hsp70/Bag-4 low-expressing CX− and Colo−, and Bag-4-transfected HeLa cells like Hsp70/Bag-4 high-expressing carcinoma sublines CX+ and Colo+. Immunoprecipitation studies further confirmed colocalization of Hsp70 and Bag-4 but also point to an association of Hsp70 and Hsp40 on the plasma membrane of CX+ and Colo+ cells; on CX− and Colo− tumor sublines, Hsp40 was detectable in the absence of Hsp70 and Bag-4. Other co-chaperones including Hsp60 and Hsp90 were neither found on the cell surface of CX+/CX−, Colo+/Colo− nor on HeLa neo-/HeLa Bag-4-transfected tumor cells. Functionally, Hsp70/Bag-4 and Hsp70/Hsp40 membrane-positive tumor cells appeared to be better protected against radiation-induced effects, including G2/M arrest and growth inhibition, on the one hand. On the other hand, membrane-bound Hsp70, but neither Bag-4 nor Hsp40, served as a recognition site for the cytolytic attack mediated by natural killer cells.

Cardiovascular Risk Factors and Estimated Risk for CAD in a Randomized Trial Comparing Calcineurin Inhibitors in Renal Transplantation

Cardiovascular morbidity and mortality is high in patients following renal transplantation. The present analysis assessed major cardiovascular risk factors and estimated the risk of coronary artery disease in the largest present‐day comparative trial of tacrolimus vs. microemulsified cyclosporine A.

High-frequency repetitive transcranial magnetic stimulation in schizophrenia: a combined treatment and neuroimaging study.

BACKGROUND Repetitive transcranial magnetic stimulation (rTMS) of frontal brain regions is under study as a non-invasive method in the treatment of affective disorders. Recent publications provide increasing evidence that rTMS may be useful in treating schizophrenia. Results are most intriguing, demonstrating a reduction of negative symptoms following high-frequency rTMS. In this context, disentangling of negative and depressive symptoms is of the utmost importance when understanding specific rTMS effects on schizophrenic symptoms. METHOD Using a sham-controlled parallel design, 20 patients with schizophrenia were included in the study. Patients were treated with high-frequency 10 Hz rTMS over 10 days. Besides clinical ratings, ECD-SPECT (technetium-99 bicisate single photon emission computed tomography) imaging was performed before and after termination of rTMS treatment. RESULTS High-frequency rTMS leads to a significant reduction of negative symptoms combined with a trend for non-significant improvement of depressive symptoms in the active stimulated group as compared with the sham stimulated group. Additionally, a trend for worsening of positive symptoms was observed in the actively treated schizophrenic patients. In both groups no changes in regional cerebral blood flow could be detected by ECD-SPECT. CONCLUSIONS Beneficial effects of high-frequency rTMS on negative and depressive symptoms were found, together with a trend for worsening positive symptoms in schizophrenic patients.

Heat shock protein 70 (Hsp70) membrane expression on head-and-neck cancer biopsy—a target for natural killer (NK) cells☆

PURPOSE Heat shock protein 70 (Hsp70) was detected on the cell membrane of human tumor cell lines, but not on normal cells. Here we studied Hsp70 membrane expression as a target for natural killer (NK) cells on tumor material and control tissues of head-and-neck cancer patients. METHODS AND MATERIALS Membrane-bound Hsp70 was determined by flow cytometry on single-cell suspensions of tumors and the corresponding normal tissues of head-and-neck cancer patients. The cytolytic activity of NK cells against Hsp70-positive tumor cells was measured in a standard cytotoxicity assay. RESULTS In total, 54 of 74 primary tumors were found to be Hsp70 membrane-positive (73%); tongue/mouth, 21 of 24 (88%); oropharynx, 13 of 20 (65%); hypopharynx, 3 of 6 (50%); larynx, 8 of 11 (73%); trachea 1 of 2 (50%); esophagus, 4 of 5 (80%); lymph node metastases, 4 of 6 (67%). The corresponding control tissue was negative for membrane-bound Hsp70. Biopsies (6 of 6) of patients after in vivo gamma-irradiation (fractionated 5 x 2 Gy) were strongly Hsp70 membrane-positive. Irradiated, Hsp70-positive tumor cells are targets for Hsp70-peptide stimulated NK cells. CONCLUSION An irradiation-inducible, tumor-selective Hsp70 membrane localization provides a target structure for Hsp70-peptide stimulated human NK cells.

Patient information in radiooncology results of a patient survey.

Background: As a result of increased interest and public demand, providing patients with adequate information about radiooncology has become more and more difficult for the doctor. Insufficient patient information can not only cause anxiety for the patient, but can also lead to legal action against the physician. In order to gain a deepter insight into our clinical practice of providing patient information, we developed a special questionnaire. We describe our first experiences in using this questionnaire at our institute. Patients and Methods: We examined the amount of information and level of satisfaction, as well as the agreement of assessment between patient and physician after the provision of standard patient information before and at the end of radiotherapy. 51 consecutive patients were interviewed with a newly designed questionnaire. The first questioning with 13 items was carried out before radiotherapy and the second with ten items was done at the end of treatment. Sum scores for information and satisfaction were defined and agreement was measured by the weighted κ coefficient. Results: Global level of information and satisfaction was good, and a significant increase in information level and a significant decline in satisfaction were seen between questionnaire 1 and 2. Agreement between patient and physician was fair, for example intent of treatment resulted in a κ coefficient of 0.34, and poor for the doctors role with a κ coefficient of −0.002. Only 52% of the patients who received palliative radiotherapy rated correctly the non-curative intent of treatment, whereas 86% of the patients who received curative radiotherapy made a currect statement. Before radiotherapy, emotional state was often both negatively and positively assessed by the patients. Conclusion: Our short questionnaire is simple and easy to understand. It provides insights into patient information with respect to assessment of the information, satisfaction level, and agreement between doctor and patient. Therefore, it is suitable for use in the clinical routine. We found a high information and satisfaction score, but limited agreement between physician and patient. In the future, the questionnaire can be used as an aid to evaluate patient information in everyday practice and to train the communication skills of the physician. Further evaluation of the questionnaire is needed and, in particular, the aspect of patient information with palliative radiotherapy has to be improved.Hintergrund: Mit den gestiegenen Interessen und Ansprüchen der Öffentlichkeit ist auch die Patientenaufklärung in der Strahlentherapie für den Arzt mehr und mehr schwieriger geworden. Eine fehlerhafte Aufklärung kann für den Arzt juristische Konsequenzen haben oder beim Patienten Angst auslösen. Um einen genaueren Einblick in die klinische Routine unserer Patientenaufklärung zu erhalten, entwickelten wir einen speziellen Fragebogen. Erste Erfahrungen werden dargestellt, die wir in unserer Einrichtung sammelten. Patienten und Methoden: Das Niveau der Informiertheit, der Zufriedenheit und das Ausmaß der Übereinstimmung der Bewertung zwischen Patient und Arzt nach einer herkömmlichen Aufklärung vor Strahlentherapie und am Ende einer Strahlentherapie wurden geprüft. Dazu wurden 51 konsekutive Patienten mit einem neu entwickelten Fragebogen interviewt. Die erste Befragung mit 13 Items wurde vor der Strahlentherapie und die zweite mit zehn Items wurde am letzten Tag der Therapie durchgeführt. Summenscores der Informiertheit und Zufriedenheit wurden errechnet, und die Übereinstimmung wurde gemäß dem gewichteten κ-Koeffizienten gemessen. Resultate: Die globale Informiertheit und Zufriedenheit war gut; ein signifikanter Anstieg der Informiertheit und ein signifikanter Abfall der Zufriedenheit waren zwischen den Befragungen vor und am Ende der Bestrahlung zu beobachten. Die Übereinstimmung zwischen Patient und Arzt war passabel, z. B. in der Beurteilung der Behandlungsintention mit einem κ-Koeffizienten von 0,34 und gering z. B. in der Sicht der Arztrolle mit einem κ-Koeffizienten von −0,002. In der palliativen Situation gaben nur 52% der Patienten die nicht kurative Intention korrekt an, wohingegen 86% der kurativ behandelten Patienten die Behandlungsintention korrekt wiedergaben. Vor Strahlentherapie wurde die emotionale Befindlichkeit von den Patienten häufig sowohl negativ als auch positiv bewertet. Schlussfolgerung: Nach unserer Erfahrung ist der kurze Fragebogen einfach und gut verständlich. Er erlaubt Einblicke in die Aufklärung unter Bewertung der Informiertheit, Zufriedenheit und Übereinstimmung zwischen Arzt und Patient. Er ist deshalb für dien klinischen Einsatz geeignet. Wir fanden ein hohes Maß der Informiertheit und der Zufriedenheit, aber eine geringe Übereinstimmung in der Bewertung zwischen Arzt und Patient. Zukünftig kann der Fragebogen als Hilfsmittel eingesetzt werden, um die Patientenaufklärung in der klinischen Praxis zu evaluieren und die ärztliche Kommunikationsfähigkeit zu schulen. Der Fragebogen muss weiter verbessert werden, der Aufklärung über die palliative Strahlentherapie ist dabei besondere Beachtung zu schenken.

Vascular Neurotoxicity Following Chemotherapy with Cisplatin, Ifosfamide, and Etoposide

OBJECTIVE To report a case of acute central nervous system (CNS) toxicity with multiple hemorrhages restricted to the corpus callosum associated with combination therapy of cisplatin, ifosfamide, and etoposide. CASE SUMMARY A 38-year-old white man with a testicular germ cell tumor received a cisplatin-based chemotherapy consisting of cisplatin 45 mg (20 mg/m 2 ), etoposide 570 mg (250 mg/m 2 ), and ifosfamide 4600 mg (2000 mg/m 2 ) given on 5 consecutive days during each course. After the first course of chemotherapy, the patient appeared to be neuropsychologically impaired with episodes of decreased alertness and features of a depressive syndrome. He became severely diminished in mental function, orientation, and psychomotor activity after a second course of treatment. In addition, he showed transient urinary incontinence. Motor and sensory deficits could not be detected. Magnetic resonance imaging demonstrated multiple hemorrhages restricted to the corpus callosum. An objective causality assessment revealed that an adverse drug reaction was probable. DISCUSSION Neurotoxicity has been associated with the administration of various antineoplastic agents. In particular, cisplatin and ifosfamide can cause both acute and delayed CNS toxicity. While ifosfamide neurotoxicity has been predominantly associated with neuropsychological impairment without evidence of structural abnormalities in neuroimaging studies, cisplatin has been shown to cause cerebrovascular complications. Various pathophysiologic conditions may contribute to these complications including thrombosis secondary to vascular endothelial injury or thromboembolic events. To our knowledge, as of December 2, 2003, vascular lesions restricted to the corpus callosum have not been reported as a complication of cisplatin- or ifosfamide-based chemotherapy. CONCLUSIONS Clinicians should be aware of the potential neurovascular adverse effects of cisplatin-based protocols. This is especially true in patients with subtle neurologic or neuropsychological symptoms. Chemotherapy-induced neurotoxicity should be considered in the differential diagnosis.

CME: Papilläres Mikrokarzinom und papilläres Karzinom der Schilddrüse ≤1cm: Modifizierte Definition der WHO und therapeutisches ‧Dilemma

Aims: Major controversies exist regarding the treatment of papillary microcarcinoma of the thyroid (PMC). Prior to 2003 PMC was defined by the WHO as a papillary carcinoma of 1cm or less in diameter. In 2004 that definition changed, with the new classification requiring that the tumour also must be found incidentally. Patients, methods: In this study we reviewed the clinical records of 67 patients with papillary tumours of the thyroid ≤1 cm, taking into account the new WHO definition (54 pts. with incidentally found PMC, median age: 53 years, 13 pts. with suspicion of thyroid neoplasm before resection, median age: 38 years). Clinical presentation, surgical treatment, further therapy and follow-up are presented. Results: Median tumour size was 7 mm in both groups (1.10 mm). Multicentric tumours were found in 15 pts. (22%), 8 had more than one PMC on the same side, and 7 displayed PMC bilaterally. Eleven (16%) of the primary tumors had metastatic involvement of regional lymph nodes at the time of initial surgery or during follow-up. Two patients showed distant metastases. No correlation between tumour size and multifocality or the presence of lymph node metastases could be seen. The gender of patients was the only significant independent variable for all patients; age and lymph node involvement was significantly different between incidentally and non-incidentally found PMC. Conclusions: Despite the majority of patients with PMC having an excellent outcome, there are also cases showing an unfavorable course. Currently no predictive parameter exists to anticipate the course and long-term outcome for an individual patient. Until this problem is solved, each patient should have the option to decide for him or herself whether to be treated similarly or differently than for conventional thyroid cancer.

Homozygosity for the 168His variant of the minor histocompatibility antigen HA-1 is associated with reduced risk of primary Sjögren's syndrome

The genes for the human ATP‐binding cassette (ABC) transporter ABCA7 and the minor histocompatibility antigen HA‐1 are juxtaposed in close proximity on chromosome 19p13.3. The multispan transmembrane protein ABCA7 contains an extracellular domain that is recognized by antisera from patients with Sjögrens syndrome (“Sjögren‐epitope”). Recent work from our laboratory demonstrating the involvement of ABCA7 in cellular ceramide and phosphatidylserine export suggests a role for this transporter in programmed cell death. In HA‐1, a protein of unknown function, a His/Arg polymorphism (His168Arg), which constitutes the immunologic target for HA‐1‐specific cytotoxic T cells, has been causatively linked to graft‐versus‐host disease after allogeneic stem cell transplantation. Because these findings suggest a potential implication of ABCA7 and HA‐1 in immune processes, we tested the hypothesis that allelic variants in both genes are associated with autoimmune disorders. We identified a total of 31 exonic single‐nucleotide polymorphisms (SNP) in the ABCA7/HA‐1 gene complex, nine of which represent non‐synonymous nucleotide alterations. Genotypes of ABCA7 and HA‐1 SNP were determined in three distinct Caucasian populations of patients with primary Sjögrens syndrome and ethnically matched controls. Comparison of allele frequencies between these groups revealed that the incidence of the HA‐1 168His allele is significantly lower in Sjögrens syndrome patients than in controls (p<0.003). In contrast, the frequencies of all ABCA7 allelic variants and additional HA‐1 polymorphisms were similar in patients and controls. In cohorts of patients with systemic lupus erythematosus, rheumatoid arthritis and multiple sclerosis, no significant differences in the frequencies of ABCA7 and HA‐1 allelic variants were observed relative to controls. Our results suggest that the HA‐1 168His variant is associated with reduced susceptibility to primary Sjögrens syndrome.