Circular test structure for the determination of piezoelectric constants of Sc x Al1-x N thin films applying Laser Doppler Vibrometry and FEM simulations.

Piezoelectric scandium aluminium nitride (Sc x Al1-x N) thin films offer a large potential for the application in micro electromechanical systems, as advantageous properties of pure AlN thin films are maintained, but combined with an increased piezoelectric actuation and sensing potential. Sc x Al1-x N thin films with x = 27% have been prepared by DC reactive magnetron sputtering to find optimized deposition parameters to maximize the piezoelectric constants d33 and d31. For the accurate and simultaneous measurement of these constants Laser Doppler Vibrometry has been applied and compared to finite element (FEM) simulations. The electrode design has been optimized to rotational symmetric structures enabling a 180° phase shifted excitation, so that a straight-forward comparison of experimental displacement curves with those obtained from FEM is feasible.

[Optimized assessment of the outcome in patients with radicular back pain of the lumbar spine. The modified NASS questionnaire]. / Bessere Verlaufsdokumentation bei Patienten mit lumboradikulären Schmerzsyndromen. Der modifizierte NASS-Fragebogen.

BACKGROUND: The purpose of the study was to present a reliable instrument with easy application to assess the outcome and improvement of therapy in patients with radicular symptoms of the lumbar spine. METHODS: Data from patients who underwent microdiscectomy because of lumbar radicular symptoms were collected and analyzed and interviews were performed using the well-known North American Spine Society (NASS) lumbar spine questionnaire (17 items) before and after the intervention. In addition patient data including comorbidities were collected. By calculating effect size (ES) and standardized response mean (SRM) for each item of the questionnaire, the questions with the highest change before and after the intervention could be selected. RESULTS: A total of 139 patients undergoing microdiscectomy for lumbar radicular symptoms due to a disc herniation were included in the analysis. Concerning the three dimensions pain, neurological symptoms and impairment of activities in daily life, the questions with best predictive value (high ES and SRM) were selected. According to their clinical relevance eight questions of the NASS questionnaire were finally selected for the short form. CONCLUSION: This short, significant and easy to use questionnaire is in our opinion a useful instrument to assess the course of patients with radicular back pain and especially to measure and monitor the outcome of therapeutic interventions, in addition to conventional clinical diagnostics and examinations. This novel instrument could be a useful tool for improving quality assurance in conventional and interventional pain management of these patients.

MAGE-C1/CT-7 expression in plasma cell myeloma: sub-cellular localization impacts on clinical outcome.

Plasma cell myelomas (PMs) have a poor prognosis. Cancer-testis (CT) antigens are immunogenic proteins, representing potential targets for tumor vaccination strategies. The expression of the CT antigens GAGE, MAGE-A4, MAGE-C1/CT-7, and NY-ESO-1 was investigated on paraffin-embedded bone marrow biopsies from 219 PM and 8 monoclonal gammopathy of undetermined significance (MGUS) patients. The frequency and prognostic impact of these CT antigens were compared with known morphological prognostic markers (i.e. Mib1 labeling index) and the presence of the translocations t(4;14)(p16.3; q32) and t(11;14)(q13;q32). We show that MAGE-C1/CT-7 is the most prevalent CT antigen, expressed in 57% of PMs in a high percentage of tumor cells. While MAGE-C1/CT-7 was absent in non-malignant plasma cells, plasma cells of patients with MGUS did express MAGE-C1/CT-7, but no other CT antigens. MAGE-C1/CT-7 was more frequently expressed in PMs with an elevated proliferation rate (Mib1 >10%) compared to PMs with a low proliferation rate (Mib1

Prostate stem cell antigen is a promising candidate for immunotherapy of advanced prostate cancer.

Immunotherapy of prostate cancer (CaP) may be a promising novel treatment option for the management of advanced CaP. However, the lack of suitable tumor antigens remains a major obstacle for the rational design of vaccines. To characterize potential CaP antigens, we determined the mRNA expression of the prostate-specific genes C1, C2, C5, PAGE-1, and prostate stem cell antigen (PSCA) in hormone-refractory CaP, benign prostatic hyperplasia, CaP cell lines, and CaP specimens. Among these gene products, only expression of PSCA appears to be retained in the majority of advanced CaP samples, as shown by reverse transcription-PCR analyses. Peptide fragments of PSCA presented in the context of major histocompatibility molecules could serve as recognition targets for CD8 T cells, provided these lymphocytes were not clonally deleted or peripherally tolerized. Our goal was to determine whether the human T-cell repertoire could recognize PSCA-derived peptide epitopes in the context of a common class I allele, HLA-A0201. Of nine peptides that, according to HLA-A0201 binding motifs, were candidate ligands of A0201 class I molecules, three peptides were able to stabilize HLA-A0201 molecules on the cell surface. One of the latter peptides, encompassing amino acid residues 14-22, was capable of generating a PSCA-specific T-cell response in a human lymphocyte culture from a patient with metastatic CaP. PSCA-specific CTLs recognized peptide-pulsed targets as well as three prostate carcinoma lines in cytotoxicity assays, indicating that this peptide could be endogenously processed. In conclusion, our findings establish PSCA as a potential target for antigen-specific, T cell-based immunotherapy of prostate carcinoma.

Chromosomal imbalances in noninvasive papillary bladder neoplasms (pTa).

Almost 70% of urinary bladder neoplasms present as low-grade papillary noninvasive tumors (stage pTa). To determine which genomic alterations can occur in pTa tumors of different grades and to evaluate the prognostic significance of chromosomal imbalances, we analyzed 113 pTa tumors (40 grade 1, 55 grade 2, 18 grade 3) by comparative genomic hybridization. pTaG1 (1.9 +/- 2.0) and pTaG2 (3.1 +/- 2.9) tumors had only few genomic alterations with 9q- (44%), 9p- (36%), and -Y (21%) being most prevalent. Neither the total number of aberrations nor any individual alteration was linked to the risk of recurrence in 95 pTaG1/G2 tumors with clinical follow-up information. pTaG3 tumors were characterized by a high number of alterations (7.7 +/- 4.5; P < 0.0001 for G3 versus G2). Several chromosomal imbalances that have previously been reported to be typical for invasive bladder neoplasms were significantly more frequent in pTaG3 than in pTaG2 tumors, including 2q-, 5p+, 5q-, 6q-, 8p-, 10q-, 18q-, and 20q+. A malfunction of genes at these loci may contribute to the development of high-grade urothelial neoplasias. However, there is no evidence for a direct role of these alterations for development of invasive tumor growth.

High-throughput tissue microarray analysis of 3p25 (RAF1) and 8p12 (FGFR1) copy number alterations in urinary bladder cancer.

Studies by comparative genomic hybridization revealed that the chromosomal regions 3p25 and 8p11-p12 are recurrently amplified in bladder cancer. To investigate the prevalence of DNA copy number alterations in these chromosomal regions and study their clinical significance, we used probes for the RAF1 (3p25) and FGFR1 (8p12) genes for fluorescence in situ hybridization. A tissue microarray containing 2317 tumors was analyzed. The analysis revealed RAF1 amplification in 4.0% and FGFR1 amplification in 3.4% of interpretable tumors. In addition, deletions were found at the 3p25 locus in 2.2% and at the 8p11-12 locus in 9.9% of interpretable tumors. Both amplifications and deletions of RAF1 and FGFR1 were significantly associated with high tumor grade (P < 0.0001), advanced stage (P < 0.0001), and poor survival (P < 0.05) if tumors of all of the stages where analyzed together. RAF1 amplifications were associated with subsequent tumor progression in pT1 carcinomas (P < 0.05). The marked differences in the frequency of all of the analyzed changes between pTa grade 1/grade 2 and pT1-4 carcinomas support the concept of these tumor groups representing different tumor entities.

Preferential antibody targeting to small lymphoma metastases in the absence of the primary tumour.

Targeting of spontaneous liver metastases of the ESb.MP murine lymphoma was achieved with anti-CD2 monoclonal antibody (MAb) 12-15A, which does not react with normal liver tissue. Using quantitative autoradiography on whole body sections of animals that had received a standard dose of 1.1 MBq of 125I-labelled monoclonal antibody, metastases accumulated up to > 90% of the injected dose per gram (id/g). The average uptake of primary tumour lesions was at a low level of 24 Bq/mg (corresponding to 2.2% id/g) because of highly non-uniform accumulation, while metastatic lesions were all above 50 Bq/mg. Uptake was particularly pronounced in animals tested after resection of the primary tumour: 85% of metastases showed levels above 300 Bq/mg, which was the upper limit of uptake in metastases of non-resected animals. These findings demonstrate the potential of the antibody approach with regard to attacking residual metastatic lesions after debulking.

Cutaneous follicular lymphoid hyperplasia with monotypic plasma cells. A clinicopathologic study of 18 patients.

Twenty cases of cutaneous follicular lymphoid hyperplasia with monotypic plasma cells are presented in a clinicopathologic study on 18 patients. The plaque-like or nodular lesions were solitary in 10 and multiple in eight patients. Immunohistochemistry showed well-defined B- and T-cell areas. Sheets of monotypic plasma cells occurred either interfollicularly or adjacent to the sclerotic stroma, with expression of IgG/kappa in 14 and IgG/lambda in six cases. In one patient with multiple lesions, one sample contained polyclonal plasma cells, whereas the other specimen showed light chain restriction. In another patient, disease recurred with a polytypic cutaneous plasma cell infiltrate. Polymerase chain reaction (PCR) revealed clonal immunoglobulin heavy chain gene rearrangements in eight of 13 cases, which was confirmed by Southern blot analysis in three samples. Clonal T-cell receptor chain gene rearrangements were not detected. Disease progression to overt malignant lymphoma did not occur within the follow-up period of up to 12 years, but recurrent disease was seen in three patients. Our data indicate that cutaneous lymphoid hyperplasia with monotypic plasma cells is a biologically distinct clinicopathological entity.

Melanoma targeting with a cocktail of monoclonal antibodies to distinct determinants of the human HMW-MAA.

The monoclonal antibodies (MoAbs) 149.53, 225.28, and 763.74 which recognize distinct and spatially distant determinants of the human high molecular weight-melanoma associated antigen (HMW-MAA) do not influence the binding of each other to cultured human melanoma cells. In vitro incubation of melanoma cells with a combination of the three 125I-labeled anti-HMW-MAA MoAbs results in a marked additive binding only when the MoAbs are used at saturating concentrations. Injection of the combination of the three 125I-labeled MoAbs (up to 300 micrograms per mouse) into human melanoma-bearing nude mice does not increase the amount of radioactivity specifically localized in melanoma lesions above the level observed upon injection of corresponding doses of individual MoAbs. These results may reflect the low concentration of MoAbs which reaches tumor lesions in vivo. Therefore, administration of combinations of MoAbs to distinct determinants of HMW-MAA may not increase the sensitivity of immunoscintigraphy to visualize lesions in patients with melanoma.

Polysomies but not Y chromosome losses have prognostic significance in pTa/pT1 urinary bladder cancer.

A disturbed cellular DNA content is of potential diagnostic and prognostic relevance in urinary bladder cancer. To evaluate the prognostic significance of individual chromosomal aberrations in superficial bladder cancer, specimens of 105 tumors (67 pTa, 38 pT1) were examined by fluorescence in situ hybridization (FISH). FISH allows quantitation of chromosomes on a cell by cell level. Centromere probes for the chromosomes Y, 1, and 17 were used. There was a strong association between polysomies of the chromosomes 1 (found in 46% of tumors) and 17 (40% of tumors, P < .0001). Polysomies (1 and 17) were significantly more frequent in pT1 than in pTa tumors (P < .0001 each). In pTa tumors, polysomies of both chromosomes were linked to a high risk of recurrences; polysomy 17 was associated with an increased risk of progression (P < .05 each). There was no significant association between polysomies and an unfavorable prognosis in pT1 carcinomas. Previous studies had suggested a prognostic role of Y losses in bladder cancer. However, Y losses were not linked to recurrences or tumor progression in pTa or pT1 tumors of 67 male patients. These data show that marked genetic differences exist between pTa and pT1 carcinomas. They also indicate that polysomies of different chromosomes may have prognostic relevance in pTa urinary bladder cancer.

Monoclonal antibody accumulation in experimental and spontaneous lung metastases of human malignant melanoma in rodents.

Monoclonal antibody (mAb) uptake in metastatic lung lesions was depicted and evaluated by digital autoradiography. The models examined were experimental metastases of a human melanoma in nude mice and spontaneous metastases of human melanoma in immunocompromised young rats. By comparing uptake patterns in local (s.c.) tumours and in lung processes of various sizes it was found that patterns were essentially similar in both types of malignant tissue. From the point of view of visualization, however, the high blood content of lung tissue resulted in high background and low contrast. This could be overcome by the use of rapidly cleared antibody fragments.

Primary gastric apoptosis-rich T-cell lymphoma co-expressing CD4, CD8, and cytotoxic molecules.

In contrast to primary gastric lymphomas of B-cell type, little is known about primary gastric T-cell lymphomas. We describe three cases with remarkably similar features: diffuse growth, epitheliotropism, medium too large cell size, high apoptotic rates, and a CD3+, CD4+, CD8+, CD45RO+ immunophenotype. Clonal TCRgamma gene rearrangement was shown in two cases. Epstein-Barr virus infection was excluded in two cases. Taking advantage of fresh-frozen material, we analyzed two cases further, revealing CD5-, CD16+, CD56-, CD57-, CD25+, CD30+, CD103 (alphaEbeta7)+, bcl-2 protein+, CD95+, CD95 ligand(L)-. CD95L, however, was detected in histiocytic and fibroblastoid by stander cells. The lymphomas expressed granzyme B, perforin, and the TIA-1 antigen in various combinations. All three cases had a very unfavorable clinical course characterized by local recurrence and/or dissemination to other epithelial sites, leading to death within 6-12 months after the initial diagnosis despite surgery and aggressive antineoplastic treatment. These data suggest a novel variant of peripheral T-cell lymphoma operationally characterized as primary gastric, apoptosis-rich, CD103+, EBV-, T-cell lymphoma co-expressing CD4, CD8, CD16 and cytotoxic molecules.

T-cell-rich B-cell non-Hodgkin's lymphoma mimicking Hodgkin's disease.

We report on a patient with recurrent T-cell-rich B-cell lymphoma (TCRBCL), initially misdiagnosed as a lymphocyte-rich Hodgkin's disease. This case exemplifies the diagnostic problems of TCRBCL and the need for immunophenotypic analysis to differentiate TCRBCL from Hodgkin's disease, nodular paragranuloma and peripheral T-cell lymphoma. A rather unusual aspect is the long disease-free interval between the excision of the node in and the late relapse in 1996. The significance of the abundant T-cell infiltration in this B-cell neoplasm will be discussed and the concepts concerning antitumor response will be reviewed. Based on epidemiological data and the clinical behaviour TCRBCL does not seem to represent a distinctive pathological entity.

Identification of plasmacytoid T cells in lymphoid hyperplasia of the skin.

Well-defined clusters of plasmacytoid T cells were identified in two cases of lymphoid hyperplasia of the skin. With the use of paraffin-embedded and frozen sections and 46 different antibodies, the antigenic profile of plasmacytoid T cells was obtained. Corresponding to plasmacytoid T cells in reactive and neoplastic lymph node conditions, plasmacytoid T cells of the skin were devoid of most T-cell-restricted differentiation antigens CD2, CD3, CD5, and CD8 while expressing monocyte/macrophage-related antigens recognized by Ki-M6 and Ki-M7. In addition, plasmacytoid T cells were positive for CD4, CD45, HLA class II antigens (HLA-DR, Leu-10, MB3, LN2, LN3) and the transferrin receptor. Furthermore, they reacted with anti-Leu-8, MB1, Ki-B3 (CD45R), and MT1. Thus plasma-cytoid T cells are considered to be mononuclear cells with monocyte/macrophage differentiation rather than T lymphocytes. In addition, our findings substantiate the concept that plasmacytoid T cells may function as a hitherto unrecognized subgroup of antigen-presenting cells in the skin.

Transcranial magnetic stimulation excites the labyrinthine segment of the facial nerve: an intraoperative electrophysiological study in man.

The site where transcranial magnetic stimulation (magStim) depolarizes the facial nerve was investigated in 6 patients who underwent surgery of the cerebellopontine angle (CPA). The facial nerve was stimulated (1) magnetically prior to craniotomy, (2) electrically near the brainstem (elREZ), (3) at the exit from the CPA into the facial canal (elPorus), and (4) in the stylomastoid fossa (elStylo). The range of latency differences (delta) of compound muscle action potentials (CMAPs) recorded from the ipsilateral mentalis muscle were as follows: delta elREZ-magStim: +0.5 to +1.1 ms (P less than or equal to 0.03, Wilcoxon test); delta elPorus-magStim: +0.2 to +0.5 ms (P less than or equal to 0.03); delta elStylo-magStim: +0.8 to +1.0 ms (P less than or equal to 0.03). On the basis of anatomical data and a facial nerve conduction velocity of 33-46 m/s in these patients, it was concluded that transcranial magnetic stimuli depolarized the facial nerve at a location 10-15 mm distal to its entrance into the facial canal. This corresponds to the end of the labyrinthine segment of the facial nerve, i.e. the transit zone where the nerve ceases to be surrounded by cerebrospinal fluid (CSF) with its high electrical conductivity and enters the high-resistance tissue of the petrous bone.

The excitation site of the trigeminal nerve to transcranial magnetic stimulation varies and lies proximal or distal to the foramen ovale: an intraoperative electrophysiological study in man.

The excitation site of the trigeminal nerve using transcranial magnetic stimulation (magStim) was analyzed in 5 patients in whom the trigeminal nerve was surgically exposed in the posterior fossa during microvascular decompression of the facial nerve for hemifacial spasm. The trigeminal nerve was stimulated (1) magnetically immediately prior to craniotomy, and (2) electrically near the root exit zone (elREZ) of the nerve from the brainstem. Mean latency differences (delta) of masseter compound muscle action potentials (CMAPs) (delta elREZ minus magStim) were 0.7 (range: +0.3 to +1.3) ms (P less than or equal to 0.05, Wilcoxon-test). From these results, an analysis of anatomical data, and using a trigeminal nerve conduction velocity (NCV) of 50 m/s as reported in the literature, the following conclusions were drawn: the excitation site to magStim (1) is variable among individuals, (2) is located 3.4 (1.6-6.5) cm distal to the trigeminal REZ, and (3) which corresponds to segments of the nerve that are located either within or outside the cerebrospinal fluid (CSF), either proximal or distal to the foramen ovale. These findings are in contrast to those we obtained in a previous study of the facial nerve in which the excitation site was found to be constant among subjects and restricted to the location on the nerve where it exists the high conductivity CSF to enter the high-resistance petrous bone.

The effect of ketamine anesthetic induction on muscle responses to transcranial magnetic cortex stimulation studied in man.

In man, an anesthetic agent that induces surgical anesthesia with minimal influence on descending pyramidal tract activity remains to be found. Anesthesia with ketamine allows recording of stable compound muscle action potentials (CMAPs) to single transcranial magnetic stimulations of the motor cortex (CortStim) in monkeys. This report describes the findings in 5 patients, where CMAPs to CortStim were recorded from the right hypothenar during anesthesia induction with ketamine. The agent was injected intravenously every 90 s in 6 steps of 0.5 mg up to a maximum of 3 mg/kg body weight (BW). Surgical anesthesia was achieved after ketamine injection of 1.5 (n = 4 patients) or 2.0 mg/kg BW (n = 1). In the five individuals tested, CMAP amplitudes and latencies (mean; range) were 2.6 (1.6-5.8) mV and 22.8 (20.4-24.6) ms before induction, and 1.6 (0.3-4.7) mV and 23.5 (21.7-24.5) ms after administration of the maximum dose. The paired differences (mean +/- 1 S.D.) were 0.8 +/- 0.6 mV and 1.0 +/- 0.8 ms and were statistically not significant (n = 5, P = 0.1, Wilcoxon-test). With ketamine as a single anesthetic induction agent CMAPs to single CortStim remain easily recordable even in dosages higher than those necessary to induce surgical anesthesia. All other previously tested anesthetic agents suppress CMAPs to CortStim as soon as the patient is unconscious.

Intraoperative motor and sensory monitoring of the cauda equina.

A set of standard techniques to monitor the motor and sensory function of the cauda equina is proposed for surgery in the lumbosacral spinal canal for the release of a tethered cord or the removal of a neoplasm. Continuous loudspeaker-controlled recording of electromyographic activity in four leg muscles of both sides supplied the surgeon with immediate feedback on injury to any of the motor roots from the second lumbar to the fourth sacral segment. Continuous recording of tibial nerve somatosensory evoked potentials yielded information about the functional state of parts of the lumbosacral sensory pathways. Motor roots could be identified by electrical stimulation in the operating field with bipolar stimulation forceps and recording of compound muscle action potentials from the leg muscles. Sensory nerve roots could be identified by nerve root somatosensory evoked potentials recorded from the scalp after the electrical stimulation of the exposed nerve. This set-up is a combination of previously developed monitoring techniques and provides the surgeon with functional information: 1) continuous feedback on the state of the endangered motor and sensory function of the cauda equina; and 2) rapid anatomical identification of nerve roots and their distinction from fibrous or neoplastic structures.

Orthodromic (intra/extracranial) neurography to monitor facial nerve function intraoperatively.

This report introduces the technique of orthodromic neurography for monitoring of facial nerve function during operation in the cerebellopontine angle. By stimulation of the intracisternal segment of the facial nerve, a compound nerve action potential with amplitudes of 15 to 480 microV can be recorded extracranially from the nerve near the stylomastoid foramen after 0.95 to 2.27 ms. Usually there is no need for signal averaging, and the method is independent of the effect of muscle relaxants. With the use of the same electrophysiological equipment as for evoked potential neuromonitoring, immediate and repeated localization of the facial nerve and its discrimination from the trigeminal and the lower cranial nerves during nerve preparation within the tumor capsule is possible.

Influence of some anesthetic agents on muscle responses to transcranial magnetic cortex stimulation: a pilot study in humans.

The effects of some commonly used intravenous and inhalational anesthetic agents on the motor evoked responses to transcranial magnetic cortex stimulation were assessed in 17 subjects. Compound motor action potentials (CMAPs) of the abductor digiti minimi muscle were recorded. Baseline values (12 stimulations/subject) were established before anesthesia was induced with a single agent administered in steps up to a maximal dose (MaxDose). Cortical stimulation was performed and depth of anesthesia was assessed according to Guedel after each dose. A common feature was the marked intra- and interindividual variability of baseline values of CMAPs in those patients not premedicated, those premedicated, and the anesthetized patients. The average amplitude of CMAPs was related to the depth of anesthesia in a given subject, whereas onset latencies of CMAPs did not systematically change. CMAPs were markedly reduced or abolished after administration of potent sedative drugs such as midazolam (MaxDose, 0.4 mg/kg body weight), pentothal (MaxDose, 8 mg/kg), propofol (MaxDose, 2 mg/kg), and isoflurane (MaxDose, 1.9 and 3.7 vol %), as soon as patients reached Stage II and Stage III anesthesia. When fentanyl (MaxDose, 8 micrograms/kg) or nitrous oxide (MaxDose, 79%) was used, the subjects reached Stages I and II, but not Stage III. With these drugs, reliable recording of CMAPs was possible even with the maximal administered dose.