Biomedical subjects
R Grobholz
Publications and source records attributed to R Grobholz.
Overexpression of a member of the pentraxin family (PTX3) in human soft tissue liposarcoma.
A unique feature of human soft tissue liposarcoma is a stable (12;16)(q13;p11) translocation observed mainly in myxoid and roundcell liposarcomas. This translocation results in FUS/CHOP fusion transcripts with a corresponding oncogenic protein. We hypothesised that genes downstream of FUS/CHOP might serve as attractive candidates for novel tumour associated antigens. Among a panel of analysed genes, only pentraxin related gene (PTX3) demonstrated high expression in liposarcomas as compared to normal tissues. The analysis of RNA and protein expression demonstrated concordant results. However, the level of RNA and protein overexpression did not correlate in all cases. Finally, PTX3 expression was not related to presence of a FUS/CHOP fusion transcript within the liposarcoma tissues. PTX3 has been associated with adipocyte differentiation and now, additionally, is characterised by a markedly increased expression in human soft tissue liposarcoma. This finding mandates further research efforts to clarify the exact role of PTX3 in liposarcoma oncogenesis.
Expression of transketolase TKTL1 predicts colon and urothelial cancer patient survival: Warburg effect reinterpreted.
Tumours ferment glucose to lactate even in the presence of oxygen (aerobic glycolysis; Warburg effect). The pentose phosphate pathway (PPP) allows glucose conversion to ribose for nucleic acid synthesis and glucose degradation to lactate. The nonoxidative part of the PPP is controlled by transketolase enzyme reactions. We have detected upregulation of a mutated transketolase transcript (TKTL1) in human malignancies, whereas transketolase (TKT) and transketolase-like-2 (TKTL2) transcripts were not upregulated. Strong TKTL1 protein expression was correlated to invasive colon and urothelial tumours and to poor patients outcome. TKTL1 encodes a transketolase with unusual enzymatic properties, which are likely to be caused by the internal deletion of conserved residues. We propose that TKTL1 upregulation in tumours leads to enhanced, oxygen-independent glucose usage and a lactate-based matrix degradation. As inhibition of transketolase enzyme reactions suppresses tumour growth and metastasis, TKTL1 could be the relevant target for novel anti-transketolase cancer therapies. We suggest an individualised cancer therapy based on the determination of metabolic changes in tumours that might enable the targeted inhibition of invasion and metastasis.
[Regressive changes after short-term neoadjuvant antihormonal therapy in prostatic carcinoma: the value of Gleason grading].
Although neoadjuvant, antihormonal therapy does not lead to an improvement in the outcome of prostatic carcinoma it is still used in the short-term in a subset of patients. Here we report the regressive changes due to this short-term treatment and analyse the impact on Gleason grading. The most frequent regressive changes in 82 tumors treated short-term were determined and quantified. The results were compared to a matched control group and also to the preoperative needle biopsies.A steep increase in regressive changes was observed within the first 4 weeks. After this point, changes increased only mildly. Within the first 2 weeks of treatment no significant changes compared to control tissue were present. Compared to the preoperative needle biopsies, pretreated tumors showed a significant upgrading. After 2 weeks of neoadjuvant antihormonal therapy, regressive changes are so great, that Gleason grading can no longer be recommended.
[Scrotal lymphangioma--a rare cause of acute scrotal pain in childhood].
INTRODUCTION: Scrotal lymphangioma is a rare differential diagnosis in boys with acute scrotal pain. CASE REPORT: A 5-year-old boy presented with acute scrotal pain and swelling. Physical examination demonstrated an unusual cystic scrotal mass with a normal testis. Ultrasound and MRI showed a complex septated cystic mass. The tumor was excised completely via an inguinal and scrotal approach. 2-year follow-up examinations with physical examination and ultrasound have shown no evidence of recurrence. CONCLUSIONS: Scrotal lymphangiomas are commonly misdiagnosed preoperatively. When ymphangioma is suspected, ultrasound and MRI imaging of the adjacent inguinal, perineal and pelvic region should be performed. To prevent recurrence, complete surgical excision is mandatory.
[Malignant mesothelioma of the testes].
INTRODUCTION: Malignant mesothelioma of the tunica vaginalis is a very rare malignant tumour. The prevalence of mesothelioma is about 1 : 1 000 000: Only 1 % have their origin in the tunica vaginalis testis with a 5-year survival of less than 5 %. About 80 cases have been reported in the literature. In 41 % of these cases exposure to asbestos for many years was determines. CASE REPORT: We report on a 76-year-old patient who presented with an atypical hydrocele testis. Intraoperatively, thickened testicular walls and an crystalline lawn adjacent to the tunica vaginalis testis were found. CONCLUSIONS: Ultrasonography showing snow flurries together with cloudy hydrocele fluid and visible crystal particles and thickened testicular walls may help to identify a mesothelioma of the testicular walls. Intraoperative frozen section and a high inguinal orchiectomy is the operative method of choice.
A new technique of endoscopic full-thickness resection using a flexible stapler.
There is a significant need for a safe and easy technique for endoluminal endoscopic resection of gastrointestinal lesions, but such procedures are usually restricted to resection of the mucosal layer in order to preserve the integrity of the wall of the gastrointestinal tract. We present two patients, one with early cancer and one with a carcinoid tumor, who were treated by endoscopic full-thickness resection. We used a stapling device, consisting of a flexible shaft, which was positioned intraluminally, and a remote control. After pilot investigations in pigs and in human anatomical preparations, we performed a full-thickness resection of the gastric wall in these patients, using the flexible stapling device under gastroscopic control. Gastric wall specimens up to 4 cm x 4 cm in size were resected with the use of two to three stapler magazines. Both procedures followed an uneventful course and the patients made an uncomplicated recovery. Further studies are necessary to test the applicability of the technique in the management of other gastric neoplastic lesions.
Gene signatures of testicular seminoma with emphasis on expression of ets variant gene 4.
Gene expression patterns of testicular seminoma were analysed applying oligonucleotide microarrays in 40 specimens of different tumour stages (pT1, pT2, pT3) and in normal testes. Transcripts of maternally expressed 3 transcripts were expressed in seminoma without correlation with delta-like 1 homologue expression indicating an impaired imprinting status in seminoma. Interestingly, the transcripts of bromodomain-containing 2 and nuclear autoantigenic sperm protein associated with spermatogenesis were significantly upregulated in progressing tumour stages. Transcription factors TEA domain family member 4 and ETS variant gene 4 (ETV4), weakly expressed in normal testis, were strongly augmented during tumourigenesis. For ETV4 expression, a significant correlation with the increased expression of matrix metalloproteinase 2 and a disintegrin and metalloproteinase domain 15 was determined. The ETV4 protein was localised to nuclei of spermatogonia and revealed an intense staining in seminoma cells. Taken together, we characterised additional transcription factors and spermatogenesis-associated genes involved in the progression of seminoma.
[Relevance of the neuroendocrine differentiation in prostatic carcinoma].
BACKGROUND: NE tumor cells are present in virtually all prostatic adenocarcinomas. As they express no androgen receptor they are hormone independent. During anti-androgenic therapy their number increases. Their prognostic value is controversial. MATERIAL AND METHODS: In 233 patients with prostatic carcinoma the NE differentiation was determined in a hot spot (7.9 mm(2)) with maximum CgA positive cell density. A high NE differentiation (HNE) was defined by a least 30 NE tumor cells, while less means a low NE differentiation (LNE). In addition the occurrence of NE tumor cells was defined as solitary or clustered (> or =5 NE tumor cells in close proximity). RESULTS: In advanced and high grade tumors more and clustered NE tumor cells could be found than in low grade and organ confined tumors. Moreover HNE tumors and occurrence of NE clusters resulted in a significant shorter progression-free interval. CONCLUSIONS: Besides the quantity of NE differentiation the quality of the growth pattern of NE tumor cells is of relevance in prostatic carcinoma.
Expression and activity of matrix metalloproteinase-2 in calcific aortic stenosis.
Calcific aortic stenosis is the main heart valve disease in the elderly, leading to massive focal calcification and thickening of the valve cusps. Matrix metalloproteinases (MMPs) are thought to contribute to this process. Therefore, the study assessed the expression of the gelatinases MMP-2 and MMP-9 and the endogenous tissue inhibitor of metalloproteinase (TIMP)-2 as well as the gelatinolytic activity in normal and stenotic valves. Human tricuspid aortic valves with and without calcific aortic stenosis were studied by immunohistochemistry for MMP-2, MMP-9 and TIMP-2. The gelatinolytic activity in native valve sections was assessed by gelatin in situ zymography with or without addition of the MMP activator p-aminophenymercuric acetate (APMA). Staining intensities for MMP-2 and TIMP-2 were elevated in stenotic valves as compared to controls. Minor staining of MMP-9 was present exclusively in stenotic valves. The morphologic distribution of gelatinolytic activity was comparable to the staining pattern of MMP-2, and since MMP-9 immunostaining demonstrated only a low number of positive cells, the observed gelatinolytic activity is likely due to MMP-2. Gelatinolytic activity was low in normal valves but significantly increased by the MMP activator APMA. In contrast, stenotic valves showed a strong basal gelatinolytic activity that could not be significantly enhanced by APMA suggesting that MMP-2 is present as a latent pro-enzyme in normal valves and activated in stenotic valves. Thus, MMP-2 might be involved in the matrix remodeling during calcific aortic stenosis.
Atraumatic intracutaneous skin closure with self-made fishing line suture compared to commercial thread.
OBJECTIVE: The aim of this study was to prove, whether the intracutaneous skin closure with self made fishing line suture is equivalent to commercial sutures. DESIGN: It was a randomised blinded animal study. SETTING: The study was performed in December 2002 at the Muhimbili University College of Health Sciences in Dar-es-salaam. SUBJECTS: Both German surgeons from Mannheim, and Tanzanian surgeons from Dar es salaam operated on nine one year old Tanzanian domestic sheep. INTERVENTIONS: 108 standardised cutaneous wounds on the backs of the animals were closed by intracutaneous sutures either with self produced fishing line suture or a commercial nylon suture (Ethilon). MAIN OUTCOME MEASURES: The clinical and histological outcomes, as well as the costs of this self-made fishing line suture, with a commercial nylon thread were evaluated and compared. RESULTS: There are no significant differences between the two sutures in histological or clinical findings, or in the reported ease of use by the surgeons. The cost of a self-produced atraumatic thread is US$ 0.12, less than one-twentieth of the cost of the commercial thread. CONCLUSION: Self-made fishing nylon suture has characteristics and properties in sheep skin wounds comparable to commercial nylon suture. The advantage of the commercial thread is the guaranteed quality assurance. It is discussed whether this quality assurance justifies the large price difference, and whether the self-produced thread should be recommended to surgeons in countries where the costs of surgical material often remains an obstacle for life saving operations.
[Mimicry of a liver abscess as hepatocellular carcinoma].
HISTORY: We report on a 82 year-old male who presented with chronic pain in the right upper abdomen, nausea and vomitus. An uncomplicated laparoscopic cholecystectomy occurred eight years and a hepatitis 60 years before. Clinical examination showed a normal abdominal status without palpable liver, spleen, enlarged lymph nodes or masses. He had no icteric skin and sclera. INVESTIGATIONS: The patient was afebrile without leukocytosis. The tumour markers CEA, CA 19 - 9 and AFP were negative. Ultrasound of the abdomen showed a normal homogeneous echotexture of the liver without signs of cholestasis or cirrhosis. In segment 6 a 3 cm large hypoechoic cystic formation with a central echogenic structure with acoustic shadowing was noted. Magnetic resonance revealed a 2.8 x 3.6 cm large lesion in the dorsal region of segment 6 of the liver, infiltrating the perihepatic fat. Pathologic lymph nodes and metastases were not found. DIAGNOSIS AND TREATMENT: At laparotomy, the tumour appeared to be malignant with invasion to the perirenal fat. Complete resection with negative margins included segment 6 of the liver and the adjacent fat (the kidney itself was not infiltrated). The cut specimen also showed signs of malignancy with central necrosis. However, histology of the resected specimen revealed a sterile liver abscess without malignancy, but with microscopic foreign bodies. The patient had an uneventful recovery without postoperative complications. CONCLUSIONS: This case report demonstrates the difficulty encountered in the differential diagnosis of inflammatory processes and malignant tumours. Chronic inflammatory changes may mimic solid neoplasms. Despite adequate magnetic resonance imaging and thorough intraoperative examination, the correct diagnosis was finally found by histology. The most likely genesis of the abscess are lost gallstones during cholecystectomy.
[Hemodynamic and metabolic characterization of orthotopic rat prostate carcinomas using dynamic MRI and proton magnetic resonance spectroscopy].
AIM: The aim of this study was the noninvasive characterization of prostate carcinoma orthotopically implanted in rats using Gd-DTPA-assisted dynamic MRI (dMRI) and proton magnetic resonance spectroscopy ((1)H-MRS). MATERIAL AND METHODS: After surgical exposure of the prostate, Dunning R3327 orthotopic prostate carcinoma was induced by injecting cells of the MAT-LyLu subline. Six rats were examined 5 and 14 days after tumor induction with dMRI and (1)H-MRS at 1.5 T. Six tumor-free rats served as controls. Using an open two-compartment model, the parameters A (amplitude) and k(ep) (exchange rate constants) were calculated from the signal time curves of the dMRI. The relative signal intensities (Cho/Cr) of the resonances of choline (Cho) and the creatine-phosphocreatine complex (Cr) were computed from the MR spectra. RESULTS: Already after 5 days, the tumors in the prostate could be clearly identified based on the decrease in signal intensity to T2w and increase of A and k(ep). High Cho/Cr levels and resonances of two lipid fractions (Lip(1) at 0.8-1.5 ppm and Lip(2) at 2.0-2.2 ppm) were observed by MRS in the highly necrotic tumors. CONCLUSIONS: The orthotopic rat prostate carcinoma model resembles human prostate carcinoma in regard to MR morphology, dMRI, and (1)H-MRS. The noninvasive characterization of perfusion and metabolism makes a comparative examination of different treatment modalities possible.
[Detection of prostate carcinomas with T1-weighted dynamic contrast-enhanced MRI. Value of two-compartment model].
AIM: The suitability of dynamic parameters of the two-compartment model for detecting prostate carcinomas and its correlation with tumor microvascular density were evaluated. METHODS: The study included 43 patients with biopsy-proven prostate carcinoma: 28 were examined by 1.0-T MRI (Turbo-FLASH) and 15 by 1.5-T MRI (FLASH) with infusion of 0.1 mmol/kg Gd-DTPA. Signal time curves were parametrized with an open two-compartment model in amplitude and exchange rate constants (k(ep)). The microvascular density of resected prostate carcinomas was determined. RESULTS: The microvascular density in the tumors was significantly higher than in the adjacent healthy prostate tissue and correlated in both sequences with k(ep). Prostate carcinomas of the peripheral zone were demarcated by amplitude and k(ep). In the Turbo-FLASH sequence there was a significant difference between the tumor tissue and healthy peripheral zone in terms of k(ep) and in the FLASH sequence in terms of amplitude. CONCLUSION: Prostate carcinomas can be visualized with dynamic T1-weighted MR sequences using a two-compartment model. Moreover, the parameter k(ep) reveals the microvascular density in the tumor and can thus provide valuable clinical information for characterizing the tumors.
Bilateral meningiomatous lesions of the spinal accessory nerves.
BACKGROUND: Meningiomas arising from cranial nerves with no dural attachment are exceedingly rare. The authors present a patient with bilateral meningiomatous lesions originating symmetrically from both spinal accessory nerves. CASE REPORT: A 61-year old woman presented with a one-year history of spinal ataxia and minimal left-sided motor impairment. Magnetic resonance imaging demonstrated two extrinsic lesions dorsolaterally of the medulla. Surgical exposure via a midline suboccipital approach with C1 laminectomy revealed the lesions arising from the spinal accessory nerves and in direct contact with the vertebral arteries. Histological investigation showed hypocellular fibrous lesions with proliferating meningothelial cells, psammoma bodies and immunoreactivity for vimentin, S-100 protein and epithelial membrane antigen. INTERPRETATION: To the authors' knowledge this is the first report of intradural tumours of the spinal accessory nerves not derived from Schwann cells and the first report of bilateral intracranial meningiomatous lesions without dural attachment.
[Fibromyxoid pseudosarcoma of the urinary bladder. Case report and review of the literature of a rare disease].
Pseudosarcomatous fibromyxoidtumors (PSF) of the bladder are rare, benign tumors consisting of myofibroblastoid bladder wall proliferations of unknown etiology. Misinterpretation as a sarcoma is common and due to infiltrative growth and cellular pleomorphism and may leaed to false radical treatment. We present a 27 year old man with PSF presenting with microhematuria. Results of clinical, radiological and histopathological examinations and therapy are presented. A. review of the literature data of the to our knowledge 73 cases that have been reported in the literature is given.
Fas ligand expression in Hodgkin lymphoma.
Because previous investigations suggested involvement of the Fas ligand (FasL) in the selection process in the follicular dendritic cell (FDC)-associated cell cluster of the germinal center, we investigated the expression of FasL in Hodgkin lymphoma (HL) on protein and RNA level, while considering the Epstein-Barr virus status of the Hodgkin and Reed-Sternberg (HRS) cells. Tumor tissue from 47 patients with classic HL (32 nodular sclerosis [NS], 11 mixed cellularity [MC], and 4 lymphocyte-rich [LR]) was analyzed by immunohistochemistry for FasL, Fas, CD21, and CD23 and by Western blotting for FasL. FasL mRNA was detected by an exon 4-specific oligonucleotide and Epstein-Barr virus infection by in situ hybridization for Epstein-Barr virus early RNAs (EBER). Western blotting showed soluble and membrane-bound forms of FasL. Immunohistochemistry showed FasL expression in virtually all HRS of 94% of NS cases and 82% of MC cases. FasL expression did not correlate with the Epstein-Barr virus status of the HRS. Low FasL protein expression was found in some HRS of LR cases. FasL mRNA was detected in 39% of NS, 46% of MC, and 33% of LR cases. Seventy percent to 90% of the HRS cells expressed Fas. CD21 immunohistochemistry showed disrupted FDC networks in the tumor tissue with reduced and virtually absent expression of CD23 and FasL. These observations suggest that FasL expression in HRS cells and the absence of FasL in the FDC cluster represent a disturbed microenvironment that may be involved in the pathogenesis of HL.
Expression of MAGE antigens and analysis of the inflammatory T-cell infiltrate in human seminoma.
The MAGE gene family encodes antigens that are recognized by cytotoxic T-cells. The expression of MAGE antigens has been linked to tumor stage, and MAGE peptides are under investigation as possible vaccines. Seminomas are tumors that are typically accompanied by a heavy inflammatory infiltrate, but have not been studied with regard to their MAGE antigen expression and its correlation with the inflammatory infiltrate. We investigated, therefore, MAGE protein expression, the amount of cytotoxic T-cells, clonality of the lymphocytic infiltrate, apoptotic activity and occurrence of necrosis. Specimens of 27 patients with classical seminoma were examined by immunohistochemistry for CD4, CD8, CD56, CD45R0, beta2-microglobulin and HLA-DR. MAGE expression was detected with the monoclonal antibody 57B, reactive with MAGE-1, -3, -4, -6 and -12. Clonality of the inflammatory infiltrate was examined by multiplex polymerase chain reaction (PCR) analysis of the T-cell receptor rearrangement. Apoptotic cells were detected by DNA nick-end labeling of fragmented DNA, and the apoptotic index was determined semi-quantitatively. Expression of 57B was found in 19 (70%) of 27 seminomas. In all cases, more than 70% of T-cells expressed CD45R0. In four cases, a predominant infiltration of CD8-positive cytotoxic T-cells (CD4/CD8 ratio < 1) was present. However, 15 seminomas showed a CD4/CD8 ratio > 1. In all cases, infiltration of CD56-positive natural killer cells was only focal. HLA-DR expression was not detectable in tumor tissue; beta2-microglobulin was only focal in three cases. Analysis of the T-cell clonality revealed a polyclonal population. The apoptotic index was not significantly different in 57B-positive seminomas (4.15%) compared with 57B negative seminomas (3.80%). Also, no correlation between the 57B expression and the occurrence of necrosis was found. MAGE antigens are homogeneously expressed in most seminomas, but their presence does not appear to represent a dominant epitope responsible for the lymphocytic infiltrate.