[New optical examination procedures for the diagnosis of skin diseases]. / Neue optische Untersuchungsverfahren für die Diagnostik von Hautkrankheiten.

BACKGROUND: Since the establishment of dermoscopy as a routine examination procedure in dermatology, the spectrum of noninvasive, optical devices has further expanded. In difficult-to-diagnose clinical cases, these systems may support dermatologists to arrive at a correct diagnosis without the need for a surgical biopsy. OBJECTIVE: To give an overview about technical background, indications and diagnostic performance regarding four new optical procedures: reflectance confocal microscopy, in vivo multiphoton tomography, dermatofluoroscopy, and systems based on image analysis by artificial intelligence (AI). MATERIALS AND METHODS: This article is based on a selective review of the literature, as well as the authors' personal experience from clinical studies relevant for market approval of the devices. RESULTS: In contrast to standard histopathological slides with vertical cross sections, reflectance confocal microscopy and in vivo multiphoton tomography allow for "optical biopsies" with horizontal cross sections. Dermatofluoroscopy and AI-based image analyzers provide a numerical score, which helps to correctly classify a skin lesion. The presented new optical procedures may be applied for the diagnosis of skin cancer as well as inflammatory skin diseases. CONCLUSION: The presented optical procedures provide valuable additional information that supports dermatologists in making the correct diagnosis. However, a surgical biopsy followed by dermatohistopathological examination remains the diagnostic gold standard in dermatology.

[Multiphoton tomography]. / Multiphotonentomographie.

In recent years, multiphoton tomography (MPT) and multiphoton microscopy have gained increasing importance as noninvasive examination techniques in dermatology. MPT imaging is based on the specific stimulation of biogenic fluorophores. The induction of second harmonic generation is also used for imaging of particular molecules. Additional fluorescence staining or fluorescence markers are not necessary-an important advantage for the in vivo examination of human skin. Multiphoton techniques are not only appropriate for clinical diagnostics but also for biomedical research. MPT provides an optical biopsy depth up to 200 µm with subcellular resolution depicting cellular and extracellular structures. In combination with fluorescence lifetime imaging, additional information about the microenvironment, the energetic state and the cellular metabolism can be obtained. This review presents recent developments of MPT for the in vivo evaluation of physiological and pathological changes of skin and diagnostics of dermal diseases.

The impact of psychiatric comorbidity on quality of life in patients undergoing herniated disc surgery.

BACKGROUND: Recent studies examined the role of psychiatric comorbidity in the process of rehabilitation in patients undergoing herniated disc surgery. These patients suffer from physical and psychosocial complaints or symptoms, which impact their everyday life negatively and the success of rehabilitation potentially. The objectives of this study are (1) to examine the quality of life (QoL) in disc surgery patients and to compare the findings with reference data from the general German population, and (2) to investigate the impact of psychiatric comorbidity on QoL of patients undergoing herniated disc surgery. METHODS: This study consists of 305 patients aged between 18 and 55 years who took part in face-to-face interviews during their hospital stay. Psychiatric comorbidity was assessed with the Composite International Diagnostic Interview (CIDI-DIA-X). By means of the 36-Item Short-Form Health Survey (SF-36), QoL was assessed in patients undergoing herniated disc surgery with and without psychiatric comorbidity. These findings were compared with the QoL of a representative sample of the general German population. RESULTS: Compared with the general population, QoL in patients with herniated disc surgery was lower in all domains of the SF-36. Psychiatric comorbidity impacts the QoL in patients with herniated disc surgery in all SF-36 domains except "physical function". The patients with psychiatric comorbidity showed significantly lower levels of QoL in the domains "bodily pain", "vitality", "social function", "role emotional", and "mental health". CONCLUSIONS: Psychiatric comorbidity has a substantial adverse effect on QoL in patients undergoing disc surgery. Therefore, it will be necessary to diagnose psychiatric comorbidities at an early stage and to include psychosocial interventions in the treatment of herniated disc patients aimed at improving deficits in psychosocial functioning and QoL.

Depression and anxiety in patients undergoing herniated disc surgery: relevant but underresearched - a systematic review.

BACKGROUND: An association between depression and anxiety and musculoskeletal disorders has been consistently reported in the past years. This article provides a systematic overview of the literature on the prevalence rates of depression and anxiety in patients undergoing surgery for a herniated disc. METHODS: A systematic literature search was conducted in the following electronic databases: PubMed, PsycINFO, Web of Science, Cochrane Library and PSYNDEXplus. The identified articles were evaluated for prevalence rates of depression and anxiety, methodological issues, change of depression and anxiety over time, and major findings on the impact of depression and anxiety on patients undergoing disc surgery. RESULTS: Fourteen studies were identified. Prevalence rates for depression and anxiety in patients undergoing disc surgery varied between 21.5% and 49.3% before and between 4.1% and 79.6% after disc surgery. The study designs, the use of assessment instruments and cut-off values varied greatly. Depression and anxiety decreased within the population of disc surgery patients over time. Depression and anxiety were found to have a great impact on the postoperative outcome of surgery, return to work, analgesia abuse, pain experience, and abnormal illness behaviour. CONCLUSIONS: Little research has been done to investigate depression and anxiety in patients undergoing surgery for a herniated disc. Evidently disc surgery patients are at higher risk of suffering from depression and anxiety than the general population. The review outlines the importance for clinicians to be more sensitive to psychological concerns in patients undergoing disc surgery. Psychological assessment and assistance from mental health professionals should be considered during the hospital stay and rehabilitation period, depending on local feasibility. Further investigations are necessary to examine whether the implementation of a multidisciplinary in-patient treatment program will improve postoperative outcome in patients undergoing intervertebral disc surgery.

[Determination of femoral head position with transinguinal ultrasound in DDH treatment]. / Der transinguinale Ultraschall zur Bestimmung der Hüftkopfzentrierung in der Behandlung der Hüftdysplasie und Hüftluxation.

AIM: Determination of the femoral head position after closed or open reduction and application of a spica cast is possible by X-ray, MRI, CT or transinguinal ultrasound. In this study we compared the efficacy of transinguinal ultrasound and radiography. Further options with transinguinal ultrasound such as the determination of soft tissue and intraoperative possibilities are also described. MATERIAL AND METHODS: In a first cohort of 25 patients with 33 affected hips ultrasound and radiography were compared. In a second cohort of 8 patients with 11 affected hips ultrasound and arthrography were compared. RESULTS: 32 radiographs proved to be not useful for the precise determination of the femoral head position. In all ultrasound images the criteria described by van Douveren et al. could be identified. All ultrasound images in the study were useful and gave reliable information with regard to the femoral head position. CONCLUSION: Consequently, standard radiographic documentation is no longer used as a standard in our clinic. MRI and CT are reserved for special cases. We recommend transinguinal ultrasound as a standard diagnostic method for determination of the femoral head position in hip spica casts. With a portable ultasound system, determination of the hip position using transinguinal ultrasound is immediately possible in the operating theatre.

[Thrombin induced tumour growth - pharmacological control]. / Thrombininduziertes Tumorwachstum - Pharmakologische Kontrolle.

The central enzyme of blood coagulation, the serine proteinase thrombin, is capable to modify the growth of tumour cells by interaction with protease activated receptors 1 and 4 of the tumour cells. Thrombin is permanently available in tumour micro environment; meizothrombin is generated from prothrombin at a tumour specific activation complex and can influence tumour cell growth via PAR-1 and 7-transdomain protein receptor signalling pathway, too. PEG-coupled direct thrombin inhibitors that possess special pharmacokinetic characteristics and that have been designed for long lasting efficacy in extracellular space, control serine proteinase activity in tumour micro environment and therefore they own a high potential anti-tumour efficacy. In xenographic tumour models this new substance class has shown a significant carcinostatic effect.

The serine proteinase thrombin promotes migration of human renal carcinoma cells by a PKA-dependent mechanism.

In this study, we demonstrated that thrombin activates protein kinase C (PKC), mitogen activated protein kinases (MAP kinases), transcription factor nuclear factor-kappa B (NF-kappa B), and cAMP-dependent protein kinase (PKA) in the human renal carcinoma cell line A-498. In addition, it enhanced the migratory capacity, but had no effect on the proliferation of A-498 cells. The effect of thrombin on migration could be blocked by the PKA inhibitor H-89 but was not influenced by inhibition of PKC, MAP kinases or NF-kappa B. We concluded, that thrombin acts as a regulator on human A-498 renal carcinoma cell migration including PKA.

Evidence for a novel thrombopoietin signalling event: activation of protein kinase A in human megakaryoblastic CMK cells.

Thrombopoietin (TPO) plays a crucial role in megakaryocyte development. TPO signalling, which is mediated by its receptor Mpl, includes Janus kinase, (JAK) signal transducer and activator of transcription (STAT) and Shc/Ras/mitogen-activated protein kinase (MAPK) pathways. The precise nature of these signalling routes has not been clarified in detail up until now. We investigated the effect of TPO on activation of cAMP-dependent protein kinase (PKA) and its involvement in MAPK signalling in human megakaryoblastic leukaemia CMK cells. For estimation of PKA activity, phosphorylation of a PKA-specific peptide substrate was assayed in CMK cell lysates. Since activation of PKA is associated with translocation of its catalytic subunit alpha (C-PKA) into the cell nucleus, Western blot analysis of nuclear fractions with an anti-C-PKA antibody was additionally performed. The activation of TPO-induced MAPK activation and the effect of the PKA inhibitor H-89 was measured using immunoblotting with a monoclonal anti-pERK antibody. TPO enhanced cAMP and induced activation of PKA in CMK cells. In addition, H-89 partly blocked TPO-induced MAPK activation in CMK cells. Our results indicate a novel TPO-triggered signalling event, activation of the cAMP/PKA pathway in human megakaryoblastic CMK cells. This signal transduction route seems to be involved in TPO-induced MAPK signaling.

A novel PAR-1-type thrombin receptor signaling pathway: cyclic AMP-independent activation of PKA in SNB-19 glioblastoma cells.

Cellular effects of thrombin are mediated by members of a new subfamily of G protein-coupled receptors designated proteinase-activated receptors (PARs) with the prototype PAR-1. Investigation of PAR-1-induced signaling has been shown to be very important in clarifying thrombin's role in cell metabolism, differentiation, and growth. We evaluated connection of PAR-1 with the cAMP/PKA pathway in SNB-19 glioblastoma cells. Alpha-thrombin and the synthetic PAR-1 agonist SFLLRN stimulated PKA as shown by increased PKA activity and translocation of the catalytic PKA alpha subunits (PKA(cat)alpha) into the nucleus. However, no effect on cAMP could be observed. PKA(cat)alpha was found to be associated with nuclear factor-kappa B (NF-kappaB) p65 and its inhibitor protein IkappaB in SNB-19 cells. After PAR-1 stimulation, this association was markedly diminished. We conclude that PAR-1 mediates PKA activation without altering cAMP levels but includes NF-kappaB-associated PKA(cat)alpha in SNB-19 glioblastoma cells. This is the first evidence for a cAMP-independent PKA signaling by a G protein-coupled receptor.

Cloning, expression, and characterization of DNA polymerase I from the hyperthermophilic archaea Thermococcus fumicolans.

The DNA polymerase I gene of a newly described deep-sea hydrothermal vent Archaea species, Thermococcus fumicolans, from IFREMERS's collection of hyperthermophiles has been cloned in Escherichia coli. As in Thermococcus litoralis, the gene is split by two intervening sequences (IVS) encoding inteins inserted in sites A and C of family B DNA polymerases. The entire DNA polymerase gene, containing both inteins, was expressed at 30 degrees C in E. coli strain BL21(DE3)pLysS using the pARHS2 expression vector. The native polypeptide precursor of 170kDa was obtained, and intein splicing as well as ligation of the three exteins was observed in vitro after heat exposure. The recombinant enzyme was purified and some of its activities were characterized: polymerization, thermostability, exonuclease activities, and fidelity.

Meizothrombin, an intermediate of prothrombin activation, stimulates human glioblastoma cells by interaction with PAR-1-type thrombin receptors.

Thrombin induces well-characterized effects on normal and neoplastic brain cells by interaction with protease-activated receptor (PAR)-type thrombin receptors. However, nothing is known about the function of intermediate enzymes of prothrombin activation recently shown to evoke PAR-1-mediated signaling in smooth muscle cells. Therefore, we investigated the effect of recombinant human meizothrombin (rMT), one of thrombin's catalytically active precursor enzymes in the prothrombin cleavage cascade, on calcium mobilization in human SNB-19 glioblastoma cells. By using reverse-transcription polymerase chain reaction, immunofluorescence studies with a monoclonal anti-PAR-1 antibody and calcium measurements, SNB-19 cells were shown to express functional PAR-1-type thrombin receptors. PAR-1 is not only a receptor for thrombin in SNB-19 cells but was also activated by rMT very effectively. Under the conditions used in our experiments, SNB-19 cells stimulated with thrombin after rMT challenge were unable to elicit a new calcium response and vice versa. In addition, both rMT and thrombin induced no further calcium signal after that observed with the PAR-1-activating peptide SFLLRN. Therefore, rMT and thrombin seem to activate calcium signaling by similar mechanisms including PAR-1. Our results demonstrate rMT as a potent activator of PAR-1-type thrombin receptors in SNB-19 glioblastoma cells, suggesting a function of catalytically active thrombin precursor enzymes in cells of glial origin.

The two-receptor system PAR-1/PAR-4 mediates alpha-thrombin-induced [Ca(2+)](i) mobilization in human astrocytoma cells.

The proteinase-activated receptor 1 (PAR-1) was characterized as a functional receptor for thrombin in cells from different brain tumor entities. Whether PAR-1 alone accounts for thrombin-induced effects in human cancer cells, or whether other PAR contribute is unknown. We established primary cultures from two neurosurgically removed human astrocytomas and investigated intracellular signaling roles of PAR-1 and PAR-4 by estimating the effect of alpha-thrombin and PAR-activating peptides on [Ca(2+)](i) mobilization in single astrocytoma cells. alpha-Thrombin or the PAR-1-activating peptide SFLLRN induced a transient calcium mobilization. This suggests the involvement of PAR-1 in alpha-thrombin-induced calcium signaling in human astrocytoma cells. In addition, a second, PAR-4-dependent, mechanism exists. This was deduced from the findings that a further calcium signal could be observed in human astrocytoma cells stimulated with alpha-thrombin after SFLLRN and the PAR-4-activating peptide GYPGQV also induced a calcium response. In addition, the observation that trypsin, known to activate both PAR-2 and PAR-4, but not the specifically PAR-2-activating peptide SLIGRL induced calcium signaling is a further indication of functional PAR-4-type thrombin receptors in human astrocytoma cells. This is the first report demonstrating a signaling role for a dual thrombin receptor system in human tumor cells.

PAR 1-type thrombin receptors are involved in thrombin-induced calcium signaling in human meningioma cells.

Thrombin is known to play a role as regulator in tumor spreading and tumor growth. Proteinase-activated receptor 1 (PAR 1)-type thrombin receptors were identified in different cancer cells including human glioblastoma cells. Thus a function of PAR 1 in brain tumors may be suggested. In this study, the presence of PAR 1-type thrombin receptors was investigated in primary cell cultures established from operated human meningiomas from two 59- and 79-year-old women. Characterization of PAR 1 on binding level was performed using immunofluorescence studies with the monoclonal anti-PAR 1 antibody Mab 61-1 directed against a domain in the NH2-terminus of PAR 1. These binding sites constitute functional thrombin receptors that are involved in thrombin-induced signaling in human meningioma cells as demonstrated by investigation of alpha-thrombin- and PAR 1-activating hexapeptide (TRAP-6)-induced [Ca2+]i mobilization. To our knowledge, this is the first report demonstrating thrombin-induced intracellular signaling in human meningioma cells mediated by the PAR 1-type thrombin receptor.

Presence of the proteinase-activated receptor-2 (PAR-2) in human brain tumor cells--trypsin- and SLIGRL-induced calcium response in primary cultured meningiomas.

This study focused on proteinase-activated receptor-2 (PAR-2) in primary cultured human meningioma cells. Stimulation of these cells with the serine proteinase trypsin resulted in a dose-dependent transient calcium response. Since the specific PAR-2 agonist peptide SLIGRL also induced [Ca2+]i mobilization in human meningioma cells and successive application of SLIGRL and trypsin elicited no new calcium signal we conclude that trypsin-induced calcium signaling is mediated by PAR-2 in human meningioma cells. To our knowledge, this is the first report describing functional PAR-2-type receptors in human brain tumor cells.

Equations for obtaining melting points for the ternary system ethylene glycol/sodium chloride/water and their application to cryopreservation.

The present study describes the H(2)O-NaCl-ethylene glycol ternary system by using a differential scanning calorimeter to measure melting points (T(m)) of four different ratios (R) of ethylene glycol to NaCl and then devising equations to fit the experimental measurements. Ultimately an equation is derived which characterizes the liquidus surface above the eutectic for any R value in the system. This study focuses on ethylene glycol in part because of recent evidence indicating it may be less toxic to pancreatic islets than Me(2)SO, which is currently used routinely for islet cryopreservation. The resulting physical data and previously determined information regarding the osmotic characteristics of canine pancreatic islets are combined in a mathematical model to describe the volumetric response to equilibrium-rate freezing in varying initial concentrations of ethylene glycol.

Protein kinase C is involved in cholecystokinin octapeptide-induced proliferative action in rat glioma C6 cells.

Chotecystoknin octapeptide (CCK-8) has been shown to stimulate DNA synthesis in rat glioma C6 cells by activation of CCKB type receptors. However, the signalling pathways contributing to this proliferative action in C6 cells have not been investigated thus far. This study demonstrated that stimulation of rat glioma C6 cells with CCK-8S resulted in activation of protein kinase C isozymes betaI, betaII, gamma and zeta. The participation of protein kinase C in the CCK-8S-induced effect on C6 cell growth was demonstrated by measurement of [3H]thymidine incorporation and estimation of cell number. The data indicate that CCK-8S stimulates growth in rat glioma C6 cells by a protein kinase C-dependent mechanism.

[Ultrasound diagnosis of isolated aplasia of the septum pellucidum]. / Sonographische Diagnose der isolierten Aplasie des Septum pellucidum.

Absence of the septum pellucidum may occur as an isolated malformation. Some of the common paediatric ultrasound and radiology textbooks mention this anomaly. However, only 12 case reports have been published so far. Absent septum pellucidum is also seen in combination with other cerebral malformations like septo-optic pituitary dysplasia (SOPD, de Morsier), agenesis of corpus callosum, holoprosencephaly etc. More often it is seen as a secondary injury due to an increasing internal hydrocephalus. Occurrence of this aplasia and its significance are often underestimated. At a paediatric centre in Germany the anomaly is expected to occur in 1: 1,000 to 1:3,000 cerebral ultrasound examinations. After a review of the literature four cases are presented. All of them were observed by ultrasound during the neonatal period. Using ultrasonography diagnosis of the isolated absence of septum pellucidum can be easily performed by the experienced physician, and will usually be of eminent relevance for the patient.

Diagnosis and follow-up of veno-occlusive disease of the liver by use of Doppler ultrasound. A case report.

The case report demonstrates that Doppler ultrasound is a very useful tool for the differential diagnosis and monitoring of veno-occlusive disease of the liver after chemotherapy.

Tissue expansion in pig skin--a histochemical approach.

In the present study, a porcine model for controlled skin expansion was investigated to improve our understanding of epidermal and vascular responses following stretching. The model is of outstanding importance not only for the clinical use of tissue expansion but provides interesting data for skin physiology and oncology, too. Thirteen out of 15 animals, who underwent silicone tissue expander implantation showed good clinical results. In all of them, skin biopsies were taken at the end of a controlled tissue expansion procedure (final expander volumes 350 or 500 ccm): one tissue specimen was obtained from the centre of the expanded skin area and a second from the neighbouring but nonexpanded skin. The tissue specimens were immediately frozen in liquid nitrogen and processed to 4 microns thick acetone-fixed frozen sections. Lectin histochemistry and immunohistology were performed using the following techniques: direct and indirect immunofluorescence technique (DIFT, IIFT), immunoperoxidase technique (POX) with either 3,3'-diamino-benzidine (DAB) or 3-amino-9-ethyl-carbazole (AEC). The histochemical findings were supplemented by measurements of the number of vital epidermal cell layers, the epidermal thickness (microns), and the papillary vascular count per visual field. There was a significant diminuation of the vascular count (mean +/- S.D. = 55.0% +/- 12.5%; U-test: p less than 5%). By immunohistochemistry, a loss of the basal cell reactivity for the following antibodies was noted: ACAM (against calmodulin), K 8.12 (against keratins 13 +/- 16) and A51-B/H4 (against keratins 8, 14, 18). There was a remarkable increase of filaggrin expression in the uppermost spinal cell layers in expanded skin, which was most pronounced in those specimens with the shortest interval to the last fluid injection into the expander. We gained no evidence for alterations of the expression of suprabasal epidermal keratins, lectin binding sites (UEA I, PNA, ConA, WGA), and vascular lectin- and immunoreactivity due to tissue expansion. The subdermal capsule, which had formed around the silicone expander, was strongly vimentin-reactive. In conclusion, controlled tissue expansion is capable to change the basal cell phenotype--a feature which is shared with a number of conditions with increased proliferative activity and with the epidermis covering different skin tumours. The regular expression of suprabasal keratins and epidermal lectin binding sites provides evidence for a normal epidermal cell differentiation. Furthermore, the porcine skin is a reliable model for studying physiology and pathophysiology of human skin.

[Experimental studies on tissue expansion in reconstructive surgery]. / Experimentelle Untersuchungen zur Gewebeexpansion in der Wiederherstellungschirurgie.

In the present study, a porcine model for controlled tissue expansion was investigated under the conditions or formation of extreme enlarged randomized flaps. The circulatory disturbances and necrotizing processes to be expected were evaluated immediately after flap formation by intravenous injection of fluorescein dye to predict the surviving area. Lectin histochemistry and immunohistology were performed using either the direct and indirect immunofluorescence technique (DIFT, IIFT) or immunoperoxidase techniques (POX). Over all, the expanded randomized flaps healed without complications in all animals. The acute randomized flaps showed clear necroses already after 10 days. In the thoracic area they averaged 60.3% (SD = 8.17%) and in the pelvic area 47.32% (SD = 14.11%) of the flap. The use of the fluorescein dye test in the present experiment demonstrated that the flap surviving area is significantly greater than the flap staining area. The regular expression of suprabasal keratins and epidermal lectin binding sites provides evidence for a normal epidermal cell differentiation.