All-in-fiber SESAM based comb oscillator with an intra-cavity electro-optic modulator for coherent high bandwidth stabilization.

We demonstrate the stabilization of an all-in-fiber polarization maintaining semi-conductor saturable absorber mirror (SESAM) mode locked frequency comb oscillator with an intra-cavity waveguide electro-optic phase modulator (EOM) to a narrow linewidth HeNe laser over 46 hours. The high feedback bandwidth of the EOM allows a coherent optical lock with an in-loop integrated phase noise of 1.12 rad (integrated from 10 Hz to 3 MHz) from the carrier signal. No piezo fiber stretcher was required to guarantee long-term stabilization, preventing mechanical degradation of the optical fibers and enabling a long lifetime of the oscillator. As an application a hybrid stabilization scheme is presented, where a comb tooth is phase locked to a longitudinal mode of the large ring laser "G" located at the Geodatic Observatory Wettzell. The hybrid stabilization scheme describes the optical lock of the frequency comb to the G laser and the simultaneous compensation of the ring laser frequency drift by comparing the comb repetition rate against an active H-maser reference. In this context the ring laser reached a fractional Allan deviation of 5 · 10-16 at an integration time of 16384 s.

The role of beta-catenin mutation and SOX9 expression in sex cord-stromal tumours of the testis.

The WHO classification of testis tumours includes the group of sex cord-stromal tumours. They are divided into several histological types, i.e. Leydig cell (LCT) and Sertoli cell tumours (SCT). Based on the physiological expression of ß-catenin in normal testis/Sertoli cells, it was previously shown that SCT can carry a ß-catenin mutation, causing a nuclear positivity for ß-catenin and cyclin D1. Furthermore, it could be shown that the stabilization of ß-catenin in Sertoli cells causes the loss of the Sertoli cell marker SOX9. We wanted to know whether the stabilization of ß-catenin in sex cord-stromal tumours influences SOX-9 expression and thus could be used in the diagnosis of sex cord-stromal tumours. Therefore, 53 cases of sex cord-stromal tumours and tumour-like lesions were investigated for their immunohistochemical expressions of ß-catenin, cyclin D1 and SOX9. In addition, mutation analyses of the ß-catenin gene (exon 3; CTNNB1) were performed. ß-catenin mutation in SCT results in nuclear ß-catenin and cyclin-D1 expressions on immunohistochemical analysis. The nuclear expression/stabilization of ß-catenin causes the loss of SOX9 in these tumours. In contrast, SOX9 is considerably expressed in non-mutated SCT as well as in Sertoli cells of non-neoplastic testes. In summary, immunohistochemical analyses of ß-catenin and SOX9 are useful to distinguish SCT from other sex cord-stromal tumours of the testis. Furthermore, the presence of SOX9 indicates that the cells of origin may be Sertoli cells.

[Leydig cell, Sertoli cell and adult granulosa cell tumors]. / Leydig-Zell-, Sertoli-Zell- und adulte Granulosazelltumoren.

According to the World Health Organization (WHO) classification Leydig cell tumors, Sertoli cell tumors and granulosa cell tumors of the testes belong to the group of sex cord-stromal tumors. These tumors most frequently occur sporadically but in rare cases can be associated with syndromes. These tumor entities show characteristic morphological changes, which in combination with specific immunohistochemical markers facilitate the diagnosis. Recent results of molecular pathological investigations, especially beta-catenin mutation analysis, allow a better categorization of these tumor entities.

[Non-seminomatous germ cell tumours]. / Nichtseminomatöse Keimzelltumoren.

The group of non-seminomatous germ cell tumors can be morphologically and therapeutically distinguished from the group of seminomas. The group of non-seminomatous germ cell tumors includes embryonal carcinoma, yolk sac tumor, choriocarcinoma and teratoma. All entities can occur rarely in pure form or much more commonly in mixed germ cell tumors consisting of more than one histological type. The non-seminomatous germ cell tumors are also characterized by the appearance of an isochromosome 12p, i(12p) and arise from a common precursor lesion called intratubular germ cell neoplasia of the unclassified type (ITGCNU). Various immunohistochemical markers are used to distinguish the different tumor components in addition to morphological characteristics.

[Sex cord gonadal stromal tumors]. / Tumoren des Gonadenstromas.

According to the World Health Organization (WHO) classification from 2004, sex cord gonadal stromal tumors are divided into Leydig cell tumors, Sertoli cell tumors, granulosa cell tumors, tumors of the thecoma-fibroma group, incompletely differentiated sex cord gonadal stromal tumors, mixed forms of sex cord gonadal stromal tumors and tumors containing both germ cell and sex cord gonadal stromal elements. These tumors can appear sporadically or in combination with hereditary syndromes. To diagnose these rare tumors the combination of characteristic morphological aspects and various immunohistochemical markers is useful. Latest investigations demonstrate the potential role of mutation analyses in the diagnosis of this heterogeneous group of tumors.

[Rare tumors and tumor-like lesions of the testis and paratesticular structures]. / Seltene Tumoren und tumorähnliche Läsionen des Hodens und der paratestikulären Region.

Tumors and tumor-like lesions of the testes and paratesticular structures are rare neoplasms often documented solely in case reports but are morphologically similar to their counterparts in other organ systems. According to the World Health Organization (WHO) classification, miscellaneous tumors of the testis, tumors of collecting ducts and rete testis, tumors of paratesticular structures are differentiated from mesenchymal tumors of the spermatic cord and testicular adnexa. In the differential diagnostics of a space-occupying mass in the testis or paratesticular region, tumor-like lesions should be considered because these lesions represent a large collection pot and occur more often than was originally assumed.

Switch of cadherin expression as a diagnostic tool for Leydig cell tumours.

Leydig cell tumours comprise about 3% of all testicular tumours. The pathogenesis of Leydig cell tumours is still poorly understood. We investigated testis with Leydig cell hyperplasia and Leydig cell tumours for their expression pattern of P- and N-cadherin. We could show a switch of expression of P- and N-cadherin in Leydig cell hyperplasia and Leydig cell tumours in comparison with normal Leydig cells. Cadherins could be established as a new immunohistochemical marker for this testicular tumour entity; their possible role in tumour genesis will be discussed.

The pluripotency factor LIN28 in monkey and human testes: a marker for spermatogonial stem cells?

Mammalian spermatogenesis is maintained by spermatogonial stem cells (SSCs). However, since evidentiary assays and unequivocal markers are still missing in non-human primates (NHPs) and man, the identity of primate SSCs is unknown. In contrast, in mice, germ cell transplantation studies have functionally demonstrated the presence of SSCs. LIN28 is an RNA-binding pluripotent stem cell factor, which is also strongly expressed in undifferentiated mouse spermatogonia. By contrast, two recent reports indicated that LIN28 is completely absent from adult human testes. Here, we analyzed LIN28 expression in marmoset monkey (Callithrix jacchus) and human testes during development and adulthood and compared it with that in mice. In the marmoset, LIN28 was strongly expressed in migratory primordial germ cells and gonocytes. Strikingly, we found a rare LIN28-positive subpopulation of spermatogonia also in adult marmoset testis. This was corroborated by western blotting and quantitative RT-PCR. Importantly, in contrast to previous publications, we found LIN28-positive spermatogonia also in normal adult human and additional adult NHP testes. Some seasonal breeders exhibit a degenerated (involuted) germinal epithelium consisting only of Sertoli cells and SSCs during their non-breeding season. The latter re-initiate spermatogenesis prior to the next breeding-season. Fully involuted testes from a seasonal hamster and NHP (Lemur catta) exhibited numerous LIN28-positive spermatogonia, indicating an SSC identity of the labeled cells. We conclude that LIN28 is differentially expressed in mouse and NHP spermatogonia and might be a marker for a rare SSC population in NHPs and man. Further characterization of the LIN28-positive population is required.

Developmental expression of the pluripotency factor sal-like protein 4 in the monkey, human and mouse testis: restriction to premeiotic germ cells.

SALL4 (sal-like protein 4) is a pluripotency transcription factor, which is highly expressed in embryonic stem (ES) cells and which is essential for mouse preimplantation development. In adult mouse organs, Sall4 mRNA is highly expressed in the testis and ovary, while there is only little or no expression in other organs. There is also a high expression of SALL4 in human testicular germ cell tumors. However, there is as yet no detailed analysis of SALL4 expression during mammalian testicular development. We analyzed SALL4 expression in ES cells, preimplantation embryos, and the developing and adult testis of a nonhuman primate (NHP) species, the common marmoset monkey (Callithrix jacchus). Immunofluorescence revealed SALL4 in the nuclei of marmoset ES cells and preimplantation embryos. Marmoset SALL4 isoform analysis in ES cells and newborn and adult testis by RT- PCR and Western blotting showed two different isoforms, SALL4-A and SALL4-B. Immunohistochemistry localized this transcription factor to the nuclei of primordial germ cells and most gonocytes in the prenatal and early postnatal marmoset testis. In the pubertal and adult testis SALL4 was present in undifferentiated spermatogonia. In the developing and adult human and mouse testis SALL4 expression mimicked the pattern in the marmoset. Adult testes from additional NHP species, the treeshrew, the cat and the dog also exhibited SALL4 in undifferentiated spermatogonia, indicating a conserved expression in the mammalian testis. Taking into account the importance of SALL4 for mouse development, we conclude that SALL4 may play an important role during mammalian germ cell development and is involved in the regulation of spermatogonial proliferation in the adult testis.

Expression and function of the vitamin D receptor in malignant germ cell tumour of the testis.

Testicular germ cell tumours (TGCTs) are the most common malignancy in young men aged 18-35 years. They are clinically and histologically subdivided into seminomas and non-seminomas. 1,25-Dihydroxyvitamin D (1,25(OH)(2)D(3)) is the active form of vitamin D and exerts its actions via a specific intracellular vitamin D receptor (VDR). Several investigations in the recent years have revealed, in addition to a physiological occurrence of the VDR in various tissues, VDR expression in different human malignancies. Furthermore, 1,25(OH)(2)D(3) plays an important role in the regulation of cell proliferation and differentiation. In different normal and malignant cell types, antiproliferative and pro-differentiating effects of 1,25(OH)(2)D(3) are described. We investigated whether TGCT express the VDR, wether differences exist between the histological subtypes and if vitamin D has a function on the proliferation of tumour cells. Furthermore, we investigated the potential function of the vitamin D-regulated genes nuclear receptor co-repressor 1(NCOR1), nuclear receptor co-repressor 2 (NCOR2), thyroid receptor interacting protein 15 (TRIP15), Growth Arrest and DNA Damage (GADD45), MAP kinase-activated protein kinase 2 (MAPKAPK2), Cytochrome P450, family 24, subfamily A, polypeptide 1 (CYP24A1) and Cytochrome P450, family 27, subfamily B, polypeptide 1 (CYP27B1) in the pathogenesis of TGCT. We demonstrate, for the first time, that primary TGCT as well as TGCT cell lines, express VDR mRNA and protein. Vitamin D and VDR may play a role in the pathogenesis of TGCTs. Furthermore, vitamin D inhibits proliferation of TGCT cell-lines, potentially via an increase in expression of GADD45. Our data suggest that vitamin D could play a role in antitumour therapy.

Leupaxin acts as a mediator in prostate carcinoma progression through deregulation of p120catenin expression.

Recently, we could show that the focal adhesion protein leupaxin (LPXN) is expressed in human prostate carcinomas (PCa) and induces invasiveness of androgen-independent PCa cells. In this study we show that LPXN enhanced the progression of existing PCa in vivo by breeding transgenic mice with prostate-specific LPXN expression and TRAMP mice (transgenic adenocarcinoma of mouse prostate). Double transgenic LPXN/TRAMP mice showed a significant increase in poorly differentiated PCa and distant metastases as compared with control TRAMP mice. Additional studies on primary PCa cells generated from both transgenic backgrounds confirmed the connection regarding LPXN overexpression and increased motility and invasiveness of PCa cells. One mediator of LPXN-induced invasion was found to be the cell-cell adhesion protein p120catenin (p120CTN). Both in vitro and in vivo experiments revealed that p120CTN expression negatively correlates with LPXN expression, followed by a redistribution of beta-catenin. Downregulation of LPXN using small interfering RNAs (siRNAs) resulted in a membranous localization of beta-catenin, whereas strong nuclear accumulation of beta-catenin was observed in p120CTN knockdown cells leading to enhanced transcription of the beta-catenin target gene matrix metalloprotease-7. In conclusion, the present results indicate that LPXN enhances the progression of PCa through downregulation of p120CTN expression and that LPXN could function as a marker for aggressive PCa in the future.

The pluripotency transcription factor Krüppel-like factor 4 is strongly expressed in intratubular germ cell neoplasia unclassified and seminoma.

Germ cell tumors of the testis are the most frequent tumors in men between 20 and 40 years. Their most common subtype is the seminoma, which arises like the embryonal carcinoma from an intratubular germ cell neoplasia unclassified (IGCNU), i.e. fetal germ cells that escaped from the control of the developing testicular stem cell niche, eventually leading to a fully developed seminoma (or embryonal carcinoma). The molecular causes for the development of an IGCNU are still unknown. However, IGCNU cells share the expression of several factors with primordial germ cells and gonocytes and, interestingly, also with pluripotent embryonic stem (ES) cells and induced pluripotent stem (iPS) cells. One factor playing important roles in both iPS and ES cells is the transcription factor Krüppel-like factor 4 (KLF4). This study examined KLF4 expression data from 179 human testicular samples including normal controls and seminoma, deposited in Gene Expression Omnibus repository for microarray data at the National Centre for Biotechnology Information. Immunohistochemistry was used to detect KLF4 protein expression in IGCNU (n = 6), seminoma (n = 14) and fetal human testes (n = 14). Microarray data from three independent sources suggest higher mRNA expression in seminoma than in normal testis. Normal spermatogonia, which are the stem cells of spermatogenesis, controlled by their stem cell niche, do not express KLF4. In contrast, IGCNU and seminoma cells strongly express KLF4. In conclusion, this finding suggests that KLF4 may be an important factor for the maintenance of the developmental and the tumorigenic potential of IGCNU as well as for the malignancy of seminoma.

Intracellular ABC transporter A3 confers multidrug resistance in leukemia cells by lysosomal drug sequestration.

Multidrug resistance (MDR) seriously limits the efficacy of chemotherapy in patients with cancer and leukemia. Active transport across membranes is essential for such cellular drug resistance, largely provided by ATP-binding cassette (ABC) transport proteins. Intracellular drug sequestration contributes to MDR; however, a genuine intracellular ABC transport protein with MDR function has not yet been identified. Analyzing the intrinsic drug efflux capacity of leukemic stem cells, we found the ABC transporter A3 (ABCA3) to be expressed consistently in acute myeloid leukemia (AML) samples. Greater expression of ABCA3 is associated with unfavorable treatment outcome, and in vitro, elevated expression induces resistance toward a broad spectrum of cytostatic agents. ABCA3 remains localized within the limiting membranes of lysosomes and multivesicular bodies, in which cytostatics are efficiently sequestered. In addition to AML, we also detected ABCA3 in a panel of lymphohematopoietic tissues and transformed cell lines. In conclusion, we identified subcellular drug sequestration mediated by the genuinely intracellular ABCA3 as being a clinically relevant mechanism of intrinsic MDR.

[Leiomyoma of the urethra - cause of an obstruction misdiagnosed as hereditary urethral stricture in a young man]. / Leiomyom der Urethra - Ursache einer als angeborene Harnröhrenenge fehlgedeuteten Striktur beim jungen Mann.

Leiomyomas are benign neoplasms arising from smooth muscle cells. We describe the case of a 17-year-old boy admitted with progressive severe obstructive voiding symptoms. Retrograde urethrography showed a bulbous urethral stricture which was resected with primary urethral anastomosis. Histopathological examination confirmed the very rare case of a leiomyoma of the urethra. In patients with urethral stricture, leiomyoma should be included in the diagnostic considerations.

[A patient with Churg-Strauss syndrome presenting as Guillain-Barré syndrome]. / Churg-Strauss-Syndrom mit dem klinischen Bild eines Guillain-Barré-Syndroms.

A 74-year-old man presented in a pulmonary clinic with symmetrically ascending tetraparesis. Physical and neurophysiological examinations suggested Guillain-Barré syndrome. The patient was treated with an initial course of 7s immunoglobulins without success. His state worsened until he was unable to walk. Severe eosinophilia (41%) was later noted in the differential white blood cell count. Combined with the onset of asthma-like symptoms, this prompted us to suspect Churg-Strauss syndrome. Despite treatment with high-dose corticoids, the palsy did not improve. It was only after immunosuppression with cyclophosphamide that the patient began to recover. The subgroup of necrotising vasculitides must be considered as differential diagnosis of rapidly progressive, symmetrical neuropathy with ascending course. Early identification and treatment are essential, since early immunosuppressive therapy is often successful, whereas delayed initiation of treatment may lead to a fatal outcome.

[Molecular principles of alternative treatment approaches for hormone-refractory prostate cancer]. / Molekulare Grundlagen alternativer Therapieansätze für das hormonrefraktäre Prostatakarzinom.

Prostate cancer is more frequently diagnosed in men from Western countries than from Asian societies. Therefore, nutritional factors such as phyto-oestrogens from soya are considered to cause this prostate cancer prevention effect. As there is no curative therapy for hormone-refractory prostate cancer, new strategies are in demand which might include phyto-oestrogens or inhibitors of histone deacetylases. Both approaches have in common the potential to reduce the aberrant androgen receptor and IGF receptor signalling. Furthermore, invasiveness and acquired survival strategies of tumours can be diminished. Reduced tumour cell proliferation and PSA secretion coincide with altered gene expression in the aforementioned processes. In addition, selective knock-down of genes by RNA interference afforded functional analyses regarding impact and succession of expression events involved in the beneficial effects caused by phyto-oestrogens and histone deacetylase inhibitors.

Expression and function of protein phosphatase PP2A in malignant testicular germ cell tumours.

Testicular germ cell tumours (TGCT) represent the most common malignancy in young males. We reported previously that two prototype members of the mitogen-activated protein kinase (MAPK) family, the MAPK ERK kinase (MEK) and extracellular signal-regulated kinase (ERK), are inactive in malignant testicular germ cells and become active after drug stimulation, leading to apoptosis of tumour cells. In this study, we asked whether the protein phosphatase PP2A, a known inhibitor of the MEK-ERK pathway, participates in the proliferation and/or apoptosis of primary TGCT (n = 48) as well as two TGCT cell lines (NTERA and NCCIT). Quantitative RT-PCR, immunohistochemistry, western blot analyses and phosphatase assay indicate that primary TGCT as well as TGCT cell lines express PP2A and that PP2A is active in TGCT cell lines. The inhibition of PP2A by application of two PP2A inhibitors, cantharidic acid (CA) and okadaic acid (OA), results in a significant increase in caspase-3-mediated apoptosis of TGCT cell lines. Thereby, PP2A inhibition was accompanied by phosphorylation and activation of MEK and ERK. Functional assays using the MEK inhibitor PD98059 demonstrated that the phosphorylation of MEK and ERK was required for the induction of caspase-3-mediated apoptosis of malignant germ cells. Thus, our data suggest that inhibition of PP2A mediates its apoptosis-inducing effect on TGCT through activation of the MEK-ERK signalling pathway that leads to caspase-3-mediated apoptosis of tumour cells. In addition our results support previous observations that PP2A exerts an anti-apoptotic effect on malignant tumour cells.

[Epithelioma cuniculatum plantare in the region of a skin scar due to mechanical trauma. Insurance law aspects of scar carcinomas]. / Carcinoma cuniculatum plantare im Bereich einer mechanisch-traumatisch verursachten Hautnarbe. Versicherungsrechtliche Aspekte des Narbenkarzinoms.

We herein report a case of a highly differentiated verrucous squamous cell carcinoma of the sole of the left forefoot ("epithelioma" or "carcinoma cuniculatum plantare") that had--as a specific feature--developed within a preexisting long-standing skin scar due to an earlier occupational crush injury. Because of expansive tumor growth extending to the metatarsal bones and penetrating the tissue between the tendons, an ultrashort amputation of the hindfoot was performed according to the method of Chopart. The pathogenetic mechanisms possibly underlying the formation of scar carcinomas are discussed. For judgement of legal liability insurance questions, the criteria indicating a causal relationship between traumatic scars and cancer development are presented.