Qualitative and quantitative protease activity tests based on protein degradation in three-dimensional structures.

The pattern of the activity of proteases is related to distinct physiological states of living organisms. Often activity changes of a certain protease can be assigned to a specific disease. Hence, they are useful biomarkers and a simple and fast determination method of their activity could be a valuable tool for the efficient monitoring of numerous diseases. Here, two different methods for the qualitative and quantitative determination of protease activity are demonstrated using the model system of proteinase K. The first test system is based on a protein-modified and colored 3D silica structure that changes color when exposed to the enzyme. This method has also been used for the detection of matrix metallo-protease 2 (MMP2) with gelatine as protease substrate on the plates. The second detection system uses the decrease in the voltammetric signal of a cytochrome c/DNA multilayer electrode after incubation with a protease to quantitatively determine its proteolytic activity. While activities down to 0.15 U/ml can be detected with the first method, the second one provides detection limits of about 0.03U/ml (for proteinase K.) The functionality of both systems can be demonstrated and ways for further enhancement of sensitivity have been elucidated.

Early autologous and allogeneic peripheral blood stem cell transplantation for adult patients with acute B and T cell precursor neoplasms: a 12-year single center experience.

Adult acute lymphoblastic leukemia/lymphoma (ALL/LBL) is a rare and heterogeneous malignancy characterized by uncontrolled proliferation of B or T cell precursor cells. Here, we retrospectively analyzed the outcome of early autologous stem cell transplantation in standard-risk patients in first complete remission (n=24) and of allogeneic transplantation in high and highest risk, and relapsed/refractory patients (n=35). The 10-year overall survival after autologous transplantation was 45%. The 10-year overall survival after allogeneic transplantation was 58%. The cumulative incidence of relapse was 29% after allogeneic and 67% after autologous transplantation. The cumulative incidence of non-relapse mortality was 0% after autologous and 12% after allogeneic transplantation. This retrospective single center analysis in a limited number of standard-risk patients clearly demonstrates that early autologous transplantation in first complete remission leads to an acceptable long-term outcome with a short overall treatment duration of less than 6 months compared with more than 2 years with conventional chemotherapy. More sensitive and standardized methods to detect minimal residual disease (MRD) will further help to identify those patients more accurately who are most likely to benefit from such a short and intensive treatment strategy (i.e., MRD negative standard-risk patients) or those who require early targeted therapy (e.g., blinatumomab) in case of MRD positivity. Early allogeneic transplantation results in long-term survival/cure in nearly two-thirds of all high and highest risk, and relapsed/refractory patients.

Early Post-Operative Pancreatitis and Systemic Inflammatory Response Assessed by Serum Lipase and IL-6 Predict Pancreatic Fistula.

BACKGROUND: Post-operative pancreatic fistula (POPF) remains a critical complication after pancreatic resection. This prospective pilot study evaluates perioperative markers of pancreatitis and systemic inflammation to predict clinically relevant grade B/C-POPF (CR-POPF). METHODS: All patients undergoing pancreatic resection from December 2017 to April 2019 were prospectively enrolled. Surgical procedures and outcomes were correlated with perioperative blood markers. ROC analysis was performed to assess their predictive value for CR-POPF. Cut-offs were calculated with the Youden index. RESULTS: In total, 70 patients were analysed (43 pancreatoduodenectomies and 27 distal pancreatectomies). In-hospital/90-d mortality and morbidity were 5.7/7.1% (n = 4/n = 5) and 75.7% (n = 53). Major complications (Clavien-Dindo ≥ 3a) occurred in 28 (40.0%) patients, CR-POPF in 20 (28.6%) patients. Serum lipase (cut-off > 51U/L) and IL-6 (> 56.5 ng/l) on POD3 were significant predictors for CR-POPF (AUC = 0.799, 95%-CI 0.686-0.912 and AUC = 0.784, 95%-CI 0.668-0.900; combined AUC = 0.858, 95%-CI 0.758-0.958; all p < 0.001). Patients with both or one factor(s) above cut-off more frequently developed CR-POPF than cases without (100 vs. 50% vs. 7.5%, p < 0.001). This also applied for overall and severe complications (p = 0.013 and p = 0.009). CONCLUSIONS: Post-operative pancreatitis and inflammatory response are major determinants for development of POPF. A combination of serum lipase and IL-6 on POD3 is a highly significant early predictor of CR-POPF and overall complications, potentially guiding patient management. CLINICAL TRIAL REGISTRATION: The study protocol was registered at clinicaltrials.gov (NCT04294797).

Levels of CEACAM6 in Peripheral Blood Are Elevated in Patients with Plasma Cell Disorders: A Potential New Diagnostic Marker and a New Therapeutic Target?

INTRODUCTION: The prognosis of multiple myeloma is still unfavorable due to inherent characteristics of the disease and the often-delayed diagnosis due to widespread and unspecific symptoms such as back pain and fatigue. Therefore, a simple diagnostic blood test would be helpful to speed up the diagnostic procedure in such patients (pts.). Here, we evaluated the diagnostic value of plasma levels of carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) in the peripheral blood and bone marrow of pts. with plasma cell disorders and in healthy controls. MATERIALS AND METHODS: Immunoreactive CEACAM6 was determined in the peripheral blood and bone marrow (n = 95/100) of pts. with monoclonal gammopathy of unknown significance (MGUS: 28/37), newly diagnosed multiple myeloma (NDMM: 42/40), and relapsed/refractory multiple myeloma (RRMM: 25/23) by sandwich ELISA. RESULTS: Median CEACAM6 levels in the peripheral blood of pts. with plasma cell disorders were significantly higher than those of healthy controls (healthy controls: 15.2 pg/ml (12.1-17.1); MGUS: 19.0 pg/ml (16.4-22.5); NDMM: 18.0 pg/ml (13.4-21.2); and RRMM: 18.9 pg/ml (15.2-21.5); p < 0.001). Plasma levels of CEACAM6 discriminated healthy subjects from MGUS/NDMM pts. (AUC = 0.71, 95% CI: 0.6-0.8); i.e., a CEACAM6 level > 17.3 pg/ml has an 82% (95% CI: 70-90) predictive probability for the identification of MGUS or NDMM. Moreover, CEACAM6 levels in the bone marrow were significantly higher in RRMM pts. than in NDMM pts. (p = 0.04), suggesting a role of this molecule in disease progression. CONCLUSION: CEACAM6 plasma levels can noninvasively identify pts. with a plasma cell disorder and should be evaluated prospectively as a potential diagnostic marker. Moreover, due to high CEACAM6 levels in the bone marrow in RRMM pts., this adhesion molecule might be a therapeutic target in multiple myeloma pts.

Outcomes of pancreas retransplantation in patients with pancreas graft failure.

BACKGROUND: Pancreas retransplantation is still a controversial option after loss of a pancreatic graft. This article describes the experience of pancreas retransplantation at a high-volume centre. METHODS: This was a retrospective observational study of all pancreas retransplantations performed in a single centre between 1997 and 2013. Pancreatic graft loss was defined by the return to insulin dependence. Risk factors for graft loss as well as patient and graft survival were analysed using logistic and time-to-event regression models. RESULTS: Of 409 pancreas transplantations undertaken, 52 (12·7 per cent) were identified as pancreas retransplantations. After a median follow-up of 65·0 (range 0·8-174·3) months, 1- and 5-year graft survival rates were 79 and 69 per cent respectively, and 1- and 5-year patient survival rates were 96 and 89 per cent. During the entire follow-up, 22 grafts (42 per cent) were lost. Patient survival was not associated with any of the donor- or recipient-related factors investigated. Five-year graft survival was better after simultaneous kidney-pancreas retransplantation than pancreas retransplantation alone: 80 per cent (16 of 20) versus 63 per cent (20 of 32) (P = 0·226). Acute rejection (odds ratio 4·49, 95 per cent c.i. 1·59 to 12·68; P = 0·005) and early surgical complications (OR 3·29, 1·09 to 9·99, P = 0·035) were identified as factors with an independent negative effect on graft survival. CONCLUSION: Pancreas retransplantation may be considered for patients whose previous graft has failed.

Quantitative error analysis for computer assisted navigation: a feasibility study.

PURPOSE: The benefit of computer-assisted navigation depends on the registration process, at which patient features are correlated to some preoperative imagery. The operator-induced uncertainty in localizing patient features-the user localization error (ULE)-is unknown and most likely dominating the application accuracy. This initial feasibility study aims at providing first data for ULE with a research navigation system. METHODS: Active optical navigation was done in CT-images of a plastic skull, an anatomic specimen (both with implanted fiducials), and a volunteer with anatomical landmarks exclusively. Each object was registered ten times with 3, 5, 7, and 9 registration points. Measurements were taken at 10 (anatomic specimen and volunteer) and 11 targets (plastic skull). The active NDI Polaris system was used under ideal working conditions (tracking accuracy 0.23 mm root-mean-square, RMS; probe tip calibration was 0.18 mm RMS). Variances of tracking along the principal directions were measured as 0.18 mm(2), 0.32 mm(2), and 0.42 mm(2). ULE was calculated from predicted application accuracy with isotropic and anisotropic models and from experimental variances, respectively. RESULTS: The ULE was determined from the variances as 0.45 mm (plastic skull), 0.60 mm (anatomic specimen), and 4.96 mm (volunteer). The predicted application accuracy did not yield consistent values for the ULE. CONCLUSIONS: Quantitative data of application accuracy could be tested against prediction models with iso- and anisotropic noise models and revealed some discrepancies. This could potentially be due to the facts that navigation and one prediction model wrongly assume isotropic noise (tracking is anisotropic), while the anisotropic noise prediction model assumes an anisotropic registration strategy (registration is isotropic in typical navigation systems). The ULE data are presumably the first quantitative values for the precision of localizing anatomical landmarks and implanted fiducials. Submillimetric localization is possible for implanted screws; anatomic landmarks are not suitable for high-precision clinical navigation.

Mena, a new available marker in tumors of salivary glands?

Mena (mammalian Ena) is an actin regulatory protein involved in cell motility and adhesion. Based on its potential role in malignant transformation revealed in other organs, we analyzed the Mena expression in normal salivary glands (SG) and salivary tumors. Mena expression was determined in normal SG (n=10) and also benign (n=20) and malignant (n=35) lesions of SG. For the immunohistochemical staining we used the anti-Mena antibody. All normal SG and the benign lesions (10 pleomorphic adenomas, 10 Warthin's tumors) were Mena negative. Salivary duct carcinomas (n=5), carcinomas in pleomorphic adenoma (n=5), acinic cell carcinomas (n=5), squamous cell carcinomas (n=10) and high-grade mucoepidermoid carcinomas (n=2) were positive. The lymphomas (n=5) and low-grade mucoepidermoid carcinomas (n=1) were Mena negative. In one case the lymphoblastic cells stained positive for Mena. Some of the endothelial cells, in the peritumoral vessels, were Mena positive. To the best of our knowledge, this is the first study in the literature about Mena expression in salivary tumors. Our study suggests that Mena protein seems to play a role in malignant transformation and its intensity is correlated with the type and grade of tumor and also with vascular invasion. Its positivity in endothelial cells may suggest its potential role in tumor angiogenesis.

Development of a (PQQ)-GDH-anode based on MWCNT-modified gold and its application in a glucose/O2-biofuel cell.

In this study a biofuel cell anode is developed on the basis of multi-walled carbon nanotubes (MWCNTs). Recombinant pyrroloquinoline quinone (PQQ) dependent glucose dehydrogenase (GDH) is covalently coupled to a PQQ-layer which is adsorbed onto thiol-modified MWCNTs at a gold electrode. In the presence of glucose a catalytic current starts at a potential of -80 mV vs. Ag/AgCl, 1M KCl. Under substrate saturation current densities of 170-200 µA/cm2 can be achieved. The operation is based on mediated electron transfer of the enzyme. This (PQQ)-GDH-MWCNT-electrode is combined with a MWCNT-modified electrode to which bilirubin oxidase (BOD) is covalently coupled. The resulting membrane-free biofuel cell has an open cell potential of 600 mV and can achieve a power density in the range of 23 µW/cm2.

A MeSH based intelligent search intermediary for Consumer Health Information Systems.

GIN Austria (Gesundheitsinformationsnetz Austria) offers patients and consumers reliable medical knowledge about diseases, wellness and disease management in an easy understandable way and enables them to quick and incessant access to informations about the Austrian health system and Austrian health organizations. To achieve full customer (patients, citizens) satisfaction to find relevant information we propose a concept of a vector-model oriented IR-Tool based on the controlled vocabulary of the MeSH Thesaurus (german version). By this approach users who are often not used to scientific terms and expressions are supported to build up their own query with MeSH Main Headings. In a second step broader and narrower Main Headings are added to the query vector by the system. For this calculation an adapted version of the Floyd-Warshall algorithm for directed, azyclic graphs is used. The tool is part of the GIN Search Modul, which will ease gathering health information from different heterogenous internet datasources.

A multilingual medical thesaurus browser for patients and medical content managers.

This paper introduces a user-friendly browser interface which integrates multilingual search and browsing functionalities within medical thesauri via the internet. The tool is being developed as part of the GIN Austria Patient Information System and is based on an adapted datamodel of the MeSH thesaurus. A prototype offers the possibility to build up queries and export lists of MeSH main headings collected during browsing the relevant MeSH trees. The thesaurus browser can be used both by patients and citizens to build queries based on a controlled vocabulary to match them with existing documents within GIN and by medical information managers to find out appropriate keywords for interactive tagging or indexing of medical contents. A key component of this tool is the flexible choice of different languages of the MeSH datasource as well as of the user interface. Both can be changed independently at any point during a session. Another central aspect is the use of the UMLS Metathesaurus in combination with localized Thesaurus versions due to existing international character set problems.

Internalins from the human pathogen Listeria monocytogenes combine three distinct folds into a contiguous internalin domain.

Listeria monocytogenes is an opportunistic, food-borne human and animal pathogen. Host cell invasion requires the action of the internalins A (InlA) and B (InlB), which are members of a family of listerial cell-surface proteins. Common to these proteins are three distinctive N-terminal domains that have been shown to direct host cell-specific invasion for InlA and InlB. Here, we present the high-resolution crystal structures of these domains present in InlB and InlH, and show that they constitute a single "internalin domain". In this internalin domain, a central LRR region is flanked contiguously by a truncated EF-hand-like cap and an immunoglobulin (Ig)-like fold. The extended beta-sheet, resulting from the distinctive fusion of the LRR and the Ig-like folds, constitutes an adaptable concave interaction surface, which we propose is responsible for the specific recognition of the host cellular binding partners during infection.

A MeSH based intelligent search intermediary for consumer health information systems.

GIN AUSTRIA (Gesundheitsinformationsnetz AUSTRIA) offers patients and consumers reliable medical knowledge about diseases, wellness and disease management in an easy understandable way and enables them to quick and incessant access to informations about the Austrian health system and Austrian health organizations. To achieve full customer (patients, citizens) satisfaction to find relevant information we propose a concept of a vector-model oriented IR-Tool based on the controlled vocabulary of the MeSH Thesaurus (german version). By this approach users who are often not used to scientific terms and expressions are supported to build up their own query with MeSH Main Headings. In a second step broader and narrower Main Headings are added to the query-vector by the system. The tool is part of the GIN Search Modul, which will ease gathering health information from different heterogenous internet datasources.

Bonding porcelain laminate veneer provisional restorations: An experimental study.

STATEMENT OF PROBLEM: Minimal tooth preparation is required for porcelain laminate veneers, but an interim restoration is anticipated by patients to protect their teeth against thermal insult, chemical irritation, and to provide esthetics. Cement remaining after the removal of the provisional restoration can impair the etching quality of the tooth surface and the fit and final bonding of the porcelain laminate veneer. PURPOSE: This in vitro study inspected the tooth surface for remaining debris from cement after removal of a provisional restoration. MATERIAL AND METHODS: Seventy-two extracted natural anterior teeth were prepared for porcelain laminate veneers. For half of the teeth, the smear layer was removed before luting provisional restorations. Veneer provisional restorations were fabricated and luted to teeth with 3 bonding methods: temporary eugenol-free cement, spot etching combined with dual-curing luting cement, and polyurethane adhesive combined with dual-curing luting cement. After removal of provisional restorations 1 week later, the tooth surface was examined for residual luting material with an SEM. RESULTS: Traces of cement debris were found on provisionally prepared teeth for all 3 material methods. CONCLUSION: The use of polyurethane adhesive combined with the dual-curing cement revealed significantly less teeth with debris than the other methods. This difference was less distinct when the smear layer was removed.

GIN AUSTRIA. Assuring quality and relevance on Internet-health-informations for patients.

GIN AUSTRIA (Gesundheitsinformationsnetz AUSTRIA) offers patients and consumers reliable medical knowledge about diseases, wellness and disease management in an understandable way and enables them to quick and incessant access to informations about the Austrian health system and Austrian health organizations. To improve the quality of the database and to achieve full customer (patients, citizens) satisfaction a systematic approach for implementing total quality management is also applied. Focusing the attention on understanding and responding to customer needs, systematic and continuous improving of the IS and total involvement of all participants are the three core TQM principles at this project. The second focus of the project is the development and the implementation (prototype) of a medical dictionary or rather medical thesaurus as interface for patients, who are not used to scientific terms and expressions. This interface is based on the controlled vocabulary of the MeSH-Thesaurus (german version).

[Actinomycosis with abdominal manifestation]. / Abdominális manifesztációjú actinomycosis.

During the last 4 years the authors observed 3 actinomycotic cases with abdominal localization. The disease was found in all the three cases to be localized to those parts of the intestinal tract where stasis of fecal contents may occur, i. e. to the appendix, to Meckel's diverticle and to the left colonic flexure. The relatively rare incidence and preoperative diagnostic difficulties make publication of these cases worthwhile. Based on own experience and literary data they describe natural history, clinical picture, histology and treatment of the disease, respectively.

[Epidemiological profile of dental caries and treatment needs]. / Epidemiologisches Profil der Zahnkaries und Behandlungsanforderungen.

In a population survey 2,263 patients were examined for their caries status and treatment needs. The DMF/T-Index and the individual relative risk for caries were calculated. Adolescents and young adults showed a high caries risk and required most of all necessary restorations. But especially older adults and seniors over 55 years presented a high caries risk, required more extensive restorations and more public dental health care at all.

[Relation between functional-immunological and clinical parameters in juvenile periodontal diseases]. / Beziehungen zwischen funktionell-immunologischen und klinischen Parametern vei Periodontalerkrankungen von Jugendlichen.

In 57 Moscow adolescents aged 15 to 19 years gingival bleeding and periodontal pocketing was recorded. In addition to this functional parameters of Rheoperiodontography and Polarography were documented and immunological investigations were carried out. The clinical parameters of the test group showed a higher level of periodontal diseases compared with a random selected control group. A correlation between clinical and functional parameters in early periodontal destruction was not detected. This study demonstrates the importance of prevention of periodontal diseases in adolescents.

[The frequency of Chlamydia detection in gynecologic and obstetrical patients and newborns]. / Zur Häufigkeit des Chlamydiennachweises bei gynäkologischen und geburtshilflichen Patienten sowie Neugeborenen.

Chlamydia trachomatis is said to play an important role in the pathogenesis of gynecologic, perinatal and neonatal infectious morbidity. Chlamydia trachomatis is responsible for several infectious syndromes in non-pregnant women, mothers and their offspring. We investigated the incidence of Chlamydia trachomatis in 353 non-pregnant and pregnant women without complaints and in 50 newborn. Chlamydia trachomatis was isolated from 6% of genital carcinoma, about 20% of different gynecological diseases and pregnant women within the first and third trimenon, 35% of puerperas and 10% of newborns. The prevalence of Chlamydia trachomatis antibody occurred in 34 to 55% of the non-pregnant and pregnant women and in 38% of the newborns. We propose a screening for Chlamydia trachomatis infection in high-risk patients and in pregnancy and a treatment of infected individuals.