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Biomedical subjects

M Krüger

Publications and source records attributed to M Krüger.

At least 19 recordsLinked to original sources

Removal of bacteria by filtration in planted and non-planted sand columns.

In order to diminish hygienic hazards from pathogens, the elimination of pathogenic bacteria in a pre-treatment step is important for the use of domestic wastewater for irrigation purposes. Therefore, we analysed the removal of bacteria in laboratory-scale model sand filters simulating vertical flow systems of constructed wetlands (CW). Sand-filled glass columns were planted with Juncus effusus or Phragmites australis and non-planted columns were used as controls. Processes of bacteria removal such as adsorption, lysis, and the biotic effects caused by plants, protozoa, and Bdellovibrio were studied with E. coli as a model bacterium. E. coli suspensions (10(8) cellsml(-1)) were trickled on the columns by intermittent loading under non-water-saturated operation conditions. In non-planted and sterilized sand columns, an initial removal of cells was observed in the range as expected by the adsorption capacity of the sand columns. After loading of the sand with cells, an increasing reduction of the cell concentrations by 3-4 orders of magnitude in the effluent was registered up to volumetric loads of more than 548mll(-1) day(-1) (230mm day(-1)). In planted columns, no higher levels of removal were observed. Predation by protozoa, which were found in concentrations up to 10(4)ml(-1) in the effluent, is considered to be the main reason for the elimination. However, Bdellovibrio bacteriovorus was also found in plaque-forming units of about 10(4)g(-1) sand, suggesting that this bacteriovorous bacterium plays an additional role in the removal process. In a second series of experiments, domestic wastewater was applied and removal efficiencies up to four orders of magnitude of the cfu's of coliform bacteria, faecal streptococci, and salmonellae were observed. Considering the transpiration of the plants, higher removal efficiencies were found in the planted variants. Protozoa and Bdellovibrio were detected in the domestic wastewater in varying concentrations, suggesting that predation and lysis were the major removal mechanisms.

Bacteria↗

[Retroperitoneal abscess in a man with severe necrotizing pancreatitis].

HISTORY AND ADMISSION FINDINGS: A 65-year-old man was admitted to the emergency unit of another hospital with severe acute abdominal pain. Medial laparotomy revealed a tear in the lower pole of the capsule of the enlarged spleen. Results of some laboratory tests became available only after the procedure: Lipase and amylase levels were markedly elevated at 5097 U/l (normal range < 100 U/l) and 1776 U/l (normal range < 60 U/l), respectively. These findings suggested acute pancreatitis as cause of the acute abdominal pain. Conservative treatment was initiated, but because of progressive clinical deterioration with fever, dyspnea and tachyarrhythmia he was transferred to our hospital for further treatment. INVESTIGATIONS, DIAGNOSIS AND TREATMENT: Chest X-ray revealed bilateral pleural effusion. Contrast-enhanced abdominal CT scan revealed an acute peripancreatic fluid collection and extensive pancreatic necrosis extending into the perirenal space, hilum of the spleen and transverse mesocolon. Despite the placement of multiple percutaneous pigtail catheters clinical improvement of the severe necrotizing pancreatitis was only temporary and repeated episodes of fever and septicaemia occured. Supported by a multidisciplinary consensus an endoscopic ultrasound (EUS)-guided transgastric drainage of the pancreatic abscess was performed with placement of two double-pigtail catheters and a nasocystic drain for irrigation. After this intervention the patient}s condition rapidly improved, inflammatory parameters normalized and the retroperitoneal abscess gradually resolved. CONCLUSION: A multidisciplinary consensus and modern interventional approaches such as EUS-guided endoscopic therapy are mandatory to successfully approach severe necrotizing pancreatitis and its complications.

Abdomen, Acute↗

Cholangioscopic characterization of dominant bile duct stenoses in patients with primary sclerosing cholangitis.

BACKGROUND AND STUDY AIMS: Primary sclerosing cholangitis (PSC) is associated with the development of cholangiocarcinoma in up to 10 % of patients. Cholangiography or endoscopic tissue sampling does not reliably distinguish between cholangiocarcinoma and a benign dominant bile duct stenosis. The aim of the present study was to assess the value of cholangioscopy for distinguishing between benign and malignant dominant stenoses in PSC patients. PATIENTS AND METHODS: Fifty-three PSC patients with dominant bile duct stenoses were prospectively studied. Transpapillary cholangioscopy and endoscopic tissue sampling were carried out in addition to endoscopic retrograde cholangiography (ERC). The cholangiography and cholangioscopic findings were classified as malignant or benign by the investigators. A final diagnosis of malignant stenosis was based on positive histology and/or cytology, whereas a benign condition was assumed in cases of negative tissue sampling and uneventful extended clinical follow-up. RESULTS: Twelve PSC patients (23 %) had dominant bile duct stenoses caused by cholangiocarcinoma, whereas 41 of the 53 patients (77 %) had benign dominant bile duct stenoses. Cholangioscopy was significantly superior to ERC for detecting malignancy in terms of its sensitivity (92 % vs. 66 %; P = 0.25), specificity (93 % vs. 51 %; P < 0.001), accuracy (93 % vs. 55 %; P < 0.001), positive predictive value (79 % vs. 29 %; P < 0.001), and negative predictive value (97 % vs. 84 %; P < 0.001). Transpapillary cholangioscopy is more sensitive and specific for characterizing malignant bile duct stenosis in comparison with endoscopic brush cytology. CONCLUSIONS: Transpapillary cholangioscopy significantly increases the ability to distinguish between malignant and benign dominant bile duct stenoses in patients with PSC.

Adolescent↗

Polypyrimidine tract-binding protein (PTB) inhibits Hepatitis C virus internal ribosome entry site (HCV IRES)-mediated translation, but does not affect HCV replication.

Polypyrimidine tract-binding protein (PTB) has previously been shown to affect Hepatitis C virus (HCV) IRES-mediated translation. In the present study we investigated the functional role of PTB for HCV translation, replication and chronic HCV infection. Bicistronic HCV IRES reporter plasmids and two different subgenomic replicons (bicistronic: pHCVrep1bBB7 (s1179I); monocistronic: pFK1-389/hyg-ubi/NS3-3'/5.1) were used to analyze the effects of PTB. Following transfection of plasmids expressing PTB RNA in sense or antisense orientation, translational activity and HCV RNA were analyzed by luciferase assay, quantitative real-time RT-PCR and northern blot analysis. Additionally, in liver tissue (n = 53) intrahepatic PTB RNA levels were determined by quantitative real-time RT-PCR. Significant inhibition of HCV IRES activity up to 42.6% was observed upon PTB sense RNA expression for HCV IRES reporter plasmids, while translational activity was enhanced up to 63.8% for PTB antisense RNA expression. In the HCV replicons PTB did not affect replication and no correlation was found between intrahepatic PTB mRNA levels and serum HCV RNA or histological changes in liver tissue of HCV infected patients. Although PTB inhibits HCV IRES-mediated translation from bicistronic reporter constructs, data obtained from two subgenomic HCV replicons and liver tissue do not indicate a significant role of PTB for HCV replication and chronic HCV infection.

5' Untranslated Regions↗

Prediction algorithm for amino acid types with their secondary structure in proteins (PLATON) using chemical shifts.

The algorithm PLATON is able to assign sets of chemical shifts derived from a single residue to amino acid types with its secondary structure (amino acid species). A subsequent ranking procedure using optionally two different penalty functions yields predictions for possible amino acid species for the given set of chemical shifts. This was demonstrated in the case of the alpha-spectrin SH3 domain and applied to 9 further protein data sets taken from the BioMagRes database. A database consisting of reference chemical shift patterns (reference CSPs) was generated from assigned chemical shifts of proteins with known 3D-structure. This reference CSP database is used in our approach for extracting distributions of amino acid types with their most likely secondary structure elements (namely alpha-helix, beta-sheet, and coil) for single amino acids by comparison with query CSPs. Results obtained for the 10 investigated proteins indicates that the percentage of correct amino acid species in the first three positions in the ranking list, ranges from 71.4% to 93.2% for the more favorable penalty function. Where only the top result of the ranking list for these 10 proteins is considered, 36.5% to 83.1% of the amino acid species are correctly predicted. The main advantage of our approach, over other methods that rely on average chemical shift values is the ability to increase database content by incorporating newly derived CSPs, and therefore to improve PLATON's performance over time.

Algorithms↗

[Results of video-assisted thoracoscopic surgery for pneumothorax].

INTRODUCTION: Video-assisted thoracoscopic surgery has been used in the treatment of pneumothorax since 1990. There is still no general agreement regarding the procedure to obliterate the pleural space or regarding the indication for wedge resection if no blebs or bullae can be found. PATIENTS AND METHODS: Clinical records referring to 100 video- assisted thoracoscopic operations for pneumothorax, performed in Berlin-Buch between 1998 and 2001, were retrospectively evaluated with regard to the rate of recurrences and postoperative complications in connection with the operative procedure. There were 70 male and 28 female patients aged from 16 to 78 years. The mean age was 34 years. The statistical significance was evaluated by the McNemar-test. RESULTS: 74 patients showed a regular postoperative course. Severe postoperative complications occurred in 9 patients: re-operation via thoracotomy n=5 (postoperative bleeding n=2, recurrence n=2, persistent air-leak n =1); re-thoracoscopy n=3 (postoperative bleeding n=2, recurrence n=1), pleural empyema n=2. The complication rate was significantly higher (p < 0.001) in patients with underlying pulmonary diseases (secondary spontaneous pneumothorax) or thoracic surgery in their history. Depending on the intraoperative situation, the cause of pneumothorax and the patient's general condition the pleural space was obliterated by the following procedures: apical parietal pleurectomy (n=85), electro-pleurodesis alone (n= 7), electro-pleurodesis and pleural abrasion (n=3), talcum poudrage (n=2). The rate of major complications was lower (p < 0.001) when pleurectomy was performed 8.2 % (7/85) compared to those operations performed without pleurectomy 13.3 % (2/15). The rate of re-operations for recurrences/persistent air leaks or empyemas following persistent air leaks was significantly lower (p < 0.001) when wedge resection 3.75 % (3/80) had been performed compared with operations without wedge resection 10 % (2/20). DISCUSSION: Apical parietal pleurectomy and wedge resection of blebs or bullae are effective methods to prevent recurrences. Considering the specific complications of parietal pleurectomy alternative methods to obliterate the pleural space should be used depending on the intraoperative situation. Among other factors underlying pulmonary diseases or thoracic surgery in the patient's history strongly influence the postoperative outcome independently of the operative procedure.

Adolescent↗

Luminal salmonella endotoxin affects epithelial and mast cell function in the proximal colon of pigs.

BACKGROUND: Salmonellosis and systemic endotoxaemia affect intestinal function. However, little is known about the functional importance of luminal Salmonella (S.) endotoxin during intestinal infection. METHODS: Pigs were either given or not given lipopolysaccharide (LPS, 30 mg day(-1)) of S. Typhimurium DT-104 orally for 14 days. Blood samples were taken weekly. After slaughter (day 14), epithelia of the proximal colon were investigated in Ussing chambers. Bacterial translocations to lung, liver, spleen and several lymph nodes were determined by culture. RESULTS: Endotoxin feeding increased plasma C-reactive protein (CRP) and histamine levels without evoking clinical signs. Postmortem, proximal colonic epithelia of LPS-treated animals showed both a decreased histamine release after mast cell stimulation with A23187 and a smaller increase in short-circuit current after A23187 application. Addition of the nitric oxide donor, sodium nitroprusside (SNP), also elicited lower increases in short-circuit current in the proximal colon of endotoxin-treated pigs. Endotoxin pre-feeding decreased colonic ion conductance, although mannitol and histamine fluxes were high in some epithelia of this group. Luminal Salmonella endotoxin increased bacterial translocation to proximal jejunal lymph nodes. LPS applied to colonic epithelia in vitro had no electrophysiological effects. CONCLUSIONS: Luminal endotoxin elicits an acute phase response and affects intestinal electrolyte transport and mast cell function. Furthermore, LPS induces epithelial spots of increased mannitol permeability that could be identical to spots of enhanced bacterial translocation.

Acute-Phase Reaction↗

Force kinetics and individual sarcomere dynamics in cardiac myofibrils after rapid ca(2+) changes.

Kinetics of force development and relaxation after rapid application and removal of Ca(2+) were measured by atomic force cantilevers on subcellular bundles of myofibrils prepared from guinea pig left ventricles. Changes in the structure of individual sarcomeres were simultaneously recorded by video microscopy. Upon Ca(2+) application, force developed with an exponential rate constant k(ACT) almost identical to k(TR), the rate constant of force redevelopment measured during steady-state Ca(2+) activation; this indicates that k(ACT) reflects isometric cross-bridge turnover kinetics. The kinetics of force relaxation after sudden Ca(2+) removal were markedly biphasic. An initial slow linear decline (rate constant k(LIN)) lasting for a time t(LIN) was abruptly followed by an ~20 times faster exponential decay (rate constant k(REL)). k(LIN) is similar to k(TR) measured at low activating [Ca(2+)], indicating that k(LIN) reflects isometric cross-bridge turnover kinetics under relaxed-like conditions (see also. Biophys. J. 83:2142-2151). Video microscopy revealed the following: invariably at t(LIN) a single sarcomere suddenly lengthened and returned to a relaxed-type structure. Originating from this sarcomere, structural relaxation propagated from one sarcomere to the next. Propagated sarcomeric relaxation, along with effects of stretch and P(i) on relaxation kinetics, supports an intersarcomeric chemomechanical coupling mechanism for rapid striated muscle relaxation in which cross-bridges conserve chemical energy by strain-induced rebinding of P(i).

Animals↗

C-SPACE (cleavage-specific amplification of cDNA ends): a novel method of ribozyme-mediated gene identification.

A hairpin ribozyme, RzCR2A, directed against position 323 of the hepatitis C virus 5'-untranslated region (HCV 5'-UTR) was used to establish and validate a novel method for the detection of cellular target molecules for hairpin ribozymes, termed C-SPACE (cleavage-specific amplification of cDNA ends). For C-SPACE, HeLa mRNA containing the transcript of interest was subjected to in vitro cleavage by RzCR2A in parallel with a control ribozyme, followed by reverse transcription using a modified SMART cDNA amplification method and cleavage-specific PCR analysis. C-SPACE allowed identification of the RzCR2A target transcript from a mixture containing the entire cellular mRNA while only requiring knowledge of the ribozyme binding sequence for amplification. In a similar approach, C-SPACE was used successfully to identify human 20S proteasome alpha-subunit PSMA7 mRNA as the cellular target RNA of Rz3'X, a ribozyme originally designed to cleave the negative strand HCV 3'-UTR. Rz3'X was found to substantially inhibit HCV internal ribosome entry site (IRES) activity and PSMA7 was subsequently confirmed to be involved in HCV IRES-mediated translation. Thereby, C-SPACE was validated as a powerful tool to rapidly identify unknown target RNAs recognized and cleaved by hairpin ribozymes.

5' Untranslated Regions↗

Freehand ultrasound elastography of breast lesions: clinical results.

We developed a freehand method for ultrasound elastography, which can be applied during a routine sonographic examination with off-line calculation of strain images (elastograms). Forty-eight patients with 53 breast lesions were examined and, after biopsy or operation, histologic reports were available for all lesions. The correlation coefficient of time delay estimates was used as a quality criterion for the subsequent calculation of elastograms. Beyond the qualitative evaluation of elastograms, we suggested a semiquantitative approach. For that purpose, the elastogram of each lesion was normalized to an overall strain of 1% (i.e., the average strain in the image was set to 1%). After normalization, we determined mean strain values inside and outside of each lesion, respectively. Defining solid lesions as benign and malignant lesions except for fibrous mastopathy, we found significant difference in strain between solid lesions and their surrounding tissue. However, that result must not be misunderstood to suggest that it was possible to distinguish benign from malignant lesions in general. Still, we address the potential of ultrasound elastography to improve the detection and localization of breast lesions as well as their differential diagnosis. Besides, we developed a freehand applicator for further studies, which guarantees a homogeneous axial compression regardless of the experience of the examiner.

Adult↗

FGFs control the patterning of the inner ear but are not able to induce the full ear program.

FGF2 or FGF8 applied ectopically, close to the developing otic placode enhances transcription of a subset of ear marker genes such as Nkx5-1, SOHo1 and Pax2. Other ear expressed genes (Dlx5 and BMP4) are not up-regulated by FGFs. Ectopic FGFs lead to an increase in size of the vestibulo-cochlear ganglion. This phenotypic change is due to an increased recruitment of epithelial cells to the neuronal fate rather than to an enhanced proliferation. We also observed an induction of additional, vesicle-like structures upon ectopic FGF treatment, but this induction never led to enrolment of a full ear program. We further demonstrate that FGF8 is expressed in two separate, short waves, first at the otic placode stage and later at the vesicle stage. Both activities correspond to critical morphogenetic events in ear development. We propose that FGF8 is an important regulator of otocyst patterning.

Animals↗

[Surgical treatment of malignant thoracic schwannomas].

UNLABELLED: The objective of this study is to evaluate the results after surgical treatment of malignant tumors arising from the peripheral nerves of the thorax under consideration of adjuvant therapy modalities. PATIENTS AND METHODS: Between 1988 and 1998, 9 patients (6 males, 3 females) underwent surgical treatment for MTNSO and 35 pts. for benign neurogenic tumors. The mean age in patients with malignant tumors was 45 years (range, 25 to 73 years). 3 pts. with MTNSO (33.3%) had neurofibromatosis (von Recklinghausen's syndrome) compared to 8.6% (3/35) in patients with benign neurogenic tumors. RESULTS: In patients with MTNSO partial chest wall resections (n = 4) including sternectomy (n = 1), lung resections (n = 2), paravertebral (n = 1) and mediastinal tumor resection (n = 1) and palliative resection of pleural recurrence (n = 1) were performed. Radical resection was achieved in 5 pts. (55.5%). There was no postoperative mortality. 3 patients (33.3%) had postoperative complications: wound infection (n = 2) and wound dehiscence due to fall with consequent pleural infection (n = 1). Adjuvant therapy was performed in two pts. (adjuvant radiotherapy/chemotherapy for metastatic disease n = 1; adjuvant chemotherapy/adjuvant radiotherapy after resection of recurrent tumor n = 1). Early recurrence is documented in 2 pts. (after 3 and 6 months). Two pts. are alive and free of disease at three years, and the patient after sternectomy with recurrent disease at 20 months. Three pts. died 8, 9 and 26 months after the primary surgical procedure. The first postoperative examination (at three months) in the remaining patient showed no evidence for recurrent disease. CONCLUSION: Patients with MTNSO have an unfavourable prognosis and local recurrence is frequent even after radical surgical therapy. Therefore an adjuvant treatment in these patients may be justified, even if the value of these therapy modalities is not proved yet. A tumor-free long-term survival especially after complete surgical resection is possible in selected cases.

Adult↗

Involvement of proteasome alpha-subunit PSMA7 in hepatitis C virus internal ribosome entry site-mediated translation.

Ribozymes are small catalytic RNA molecules that can be engineered to enzymatically cleave RNA transcripts in a sequence-specific fashion and thereby inhibit expression and function of the corresponding gene product. With their simple structures and site-specific cleavage activity, they have been exploited as potential therapeutic agents in a variety of human disorders, including hepatitis C virus (HCV) infection. We have designed a hairpin ribozyme (Rz3'X) targeting the HCV minus-strand replication intermediate at position 40 within the 3'X tail. Surprisingly, Rz3'X was found to induce ganciclovir (GCV)-resistant colonies in a bicistronic cellular reporter system with HCV internal ribosome entry site (IRES)-dependent translation of herpes simplex virus thymidine kinase (TK). Rz3'X-transduced GCV-resistant HeLa reporter cells showed substantially reduced IRES-mediated HCV core protein translation compared with control vector-transduced cells. Since these reporter systems do not contain the HCV 3'X tail sequences, the results indicate that Rz3'X probably exerted an inhibitory effect on HCV IRES activity fortuitously through another gene target. A novel technique of ribozyme cleavage-based target gene identification (cleavage-specific amplification of cDNA ends) (M. Krüger, C. Beger, P. J. Welch, J. R. Barber, and F. Wong-Staal, Nucleic Acids Res. 29:e94, 2001) revealed that human 20S proteasome alpha-subunit PSMA7 mRNA was a target RNA recognized and cleaved by Rz3'X. We then showed that additional ribozymes directed against PSMA7 RNA inhibited HCV IRES activity in two assay systems: GCV resistance in the HeLa IRES TK reporter cell system and a transient transfection assay performed with a bicistronic Renilla-HCV IRES-firefly luciferase reporter in Huh7 cells. In contrast, ribozymes were inactive against IRES of encephalomyocarditis virus and human rhinovirus. Additionally, proteasome inhibitor MG132 exerted a dose-dependent inhibitory effect on HCV IRES-mediated translation but not on cap-dependent translation. These data suggest a principal role for PSMA7 in regulating HCV IRES activity, a function essential for HCV replication.

Antiviral Agents↗