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Biomedical subjects

W Becker

Publications and source records attributed to W Becker.

At least 271 records · Page 15Linked to original sources
PubMed1994

Molecular cloning of a protein serine/threonine phosphatase containing a putative regulatory tetratricopeptide repeat domain.

Two novel protein serine/threonine phosphatases were cloned from a rat fat cell library with probes generated by a polymerase chain reaction-based cloning approach. One of these cDNAs encoded a protein presumably representing the rat homologue of PPV from Drosophila (75% identity of amino acids). The other novel cDNA encoded a protein phosphatase of 499 amino acids and was designated PPT. Its catalytic domain contains motifs typical for protein phosphatases but is only distantly related with PP1, PP2A, and PP2B (38-42% identical amino acids). When expressed in Escherichia coli, the catalytic domain of PPT exhibited protein phosphatase activity (dephosphorylation of phosphorylase a) that was inhibitable by okadaic acid. As a unique feature among other members of this gene family, PPT has an amino-terminal extension of 200 amino acids harboring three tandemly arranged tetratricopeptide repeat (TPR) motifs. This domain has previously been found in other proteins involved in the regulation of RNA synthesis or mitosis. mRNA of PPT was predominantly found in brain and, in lower levels, in testis, but was nearly undetectable in spleen, lung, skeletal muscle, kidney, and liver. It is suggested that the TPR domain of PPT may be involved in the regulation of the function of this novel protein phosphatase.

3T3 Cells↗
PubMed1994

JIP60, a methyl jasmonate-induced ribosome-inactivating protein involved in plant stress reactions.

Plant tissues treated with the naturally occurring cyclopentanone compound methyl jasmonate or exposed to stress causing in planta jasmonate accumulation express distinctive proteins and, concomitantly, reduce the synthesis of most preexisting proteins. One of the recently identified jasmonate-induced proteins, designated JIP60, in barley is a ribosome-inactivating protein that cleaves polysomes of both animal and plant origin into their ribosomal subunits. By attacking foreign and self ribosomes, respectively, JIP60 appears to be both a defense protein and a potent regulator of protein synthesis in stressed plant tissues.

Acetates↗
PubMed1994

Substitution of conserved tyrosine residues in helix 4 (Y143) and 7 (Y293) affects the activity, but not IAPS-forskolin binding, of the glucose transporter GLUT4.

Six tyrosine residues (Y28, Y143, Y292, Y293, Y308, Y432(1)) which are conserved in all mammalian glucose transporters were substituted for phenylalanine by site-directed mutagenesis, and mutant glucose transporters were transiently expressed in COS-7 cells. Glucose transport activity as assessed by reconstitution of the solubilized transporters into lecithin liposomes was reduced by 70% in the mutant Y143F and appeared to be abolished in Y293F, but was not affected by substitution of Y28, Y292, Y308 and Y432. In contrast, covalent binding of the photolabel 125IAPS-forskolin was normal in all mutants. Stable expression of the mutants Y143F, Y293F, and Y292F in LTK cells yielded identical results. These data indicate that only two of the 6 conserved helical tyrosine residues, located in helices 4 and 7, are essential for full activity, but not for IAPS-forskolin binding of the GLUT4.

Affinity Labels↗
PubMed1994

Cloning of two novel ADP-ribosylation factor-like proteins and characterization of their differential expression in 3T3-L1 cells.

A polymerase chain reaction-based cloning approach was employed in order to identify ADP-ribosylation factors (ARF) in murine 3T3-L1 cells and to study their expression before and after differentiation of cells to the adipocyte-like phenotype. Partial sequences comprising the effector domains of ARF were amplified with degenerate primers and cloned. Five of these sequences were identified as murine homologues of known human ADP-ribosylation factors (ARF 1, 2, 4, 5, and 6). In addition, partial sequences of two previously unknown isoforms were found, and complete cDNA clones were isolated from a rat fat cell library and were sequenced. Both sequences harbor a putative myristoylation site in position 2, the known consensus sequences presumably involved in GTP binding and hydrolysis, and lack cysteine residues in the C terminus. Their amino acid sequences share a 56 and 41% identity, respectively, with human ARF 1. Based on a comparison with the known ARF isoforms, the first clone appears to represent the mammalian homologue of a known sequence from Drosophila (dARL 1, 79% identity) and was therefore designated rARL 1. The second clone resembled none of the known ARF-like proteins and was designated rARL 4. mRNA of ARL 4 was undetectable in the fibroblasts but abundant in the adipocyte-like phenotype, its expression starting on day 6 of the differentiation. In contrast, ARF 1, 2, and 5 were unaltered by differentiation of the 3T3-L1 cells; mRNA levels of ARF 6, and also of ARL 1 and ARF 4, were reduced after differentiation. It is suggested that the function of ARL 4 is related to the adipocyte-like phenotype of 3T3-L1 cells.

3T3 Cells↗
PubMed1994

Imaging rheumatoid arthritis joints with technetium-99m labelled specific anti-CD4- and non-specific monoclonal antibodies.

A direct comparison of the joint-imaging properties of inflammation-specific- and non-specific monoclonal antibodies (Mabs) was possible in a patient suffering from long-standing, severe rheumatoid arthritis (RA). This patient received an anti-CD4- and an anticarcinoembryonic antigen (anti-CEA) Mab, both labelled with technetium-99m, 9 days apart from each other. The anti-CD4 Mab was superior to the isotype-matched anti-CEA Mab in imaging inflamed joints. In the knee joint, the target-to-background ratio of the synovial membrane (SM) activity in comparison to that of adjacent large vessels was 1.22 (SM/muscle 1.55) for the anti-CD4 Mab and 0.53 (SM/muscle 0.92) for the anti-CEA Mab, in both cases 4 h after injection of the immunoglobulin. Since the CD4 antigen is present on the surface of T-helper lymphocytes and macrophages infiltrating the inflamed synovial membrane, imaging with the anti-CD4 Mab may allow more specific detection of inflammatory infiltrates in RA.

Aged↗
PubMed1994

Hirudinicidal activities of some natural molluscicides used in schistosomiasis control.

Experiments were conducted with (molluscicides) aridanin isolated from Tetrapleura tetraptera, aridan, an extract from T. tetraptera, endod, an extract from Phytolacca dodecandra and niclosamide on non-target aquatic organisms such as leech, hydra, tadpoles, anopheline mosquito larvae and brine shrimps and compared with their toxicity to the target snail. Biomphalaria glabrata. Aridanin, aridan, endod, and niclosamide produced rapid knockdown effects on B. glabrata at 0.04, 1.00, 30.00, and 40.00 ppm, respectively. All the molluscicides killed the leech, a pest of animals and man at molluscicidal concentrations. The hydra and tadpoles tested were sensitive to the molluscicides except aridanin but the shrimps and anopheline mosquito larvae were resistant to all the molluscicides.

Animals↗
PubMed1994

The use of monoclonal antibodies and antibody fragments in the imaging of infectious lesions.

The advantage of immunoscintigraphy over autologous leukocyte techniques in the imaging of infection is the simplicity of its use compared with techniques that require the isolation of autologous white blood cells. The advantage over 99mTc- or 111In-labeled nonspecific human immunoglobulin is the excellent target-to-background ratio, which results from an endogenous background subtraction with the whole antigranulocyte antibody (99mTc-anti-NCA-95; BW250/183) and a rapid clearance of the antibody using an antigranulocyte Fab'-fragment (99mTc-anti-NCA-90; IMMU-MN3). The whole antibody is helpful for imaging of soft tissue infections, peripheral bone, and abdominal infections. Problems may arise occasionally in regions with normal bone marrow that are caused by human antimouse antibody (HAMA) formation. An infection or the entire extent of inflammation can be proven or excluded only at 24 hours postinjection. In contrast, antigranulocyte Fab'-fragment may have its advantage in an earlier localization of infectious lesions (1 hour) owing to its smaller molecular size and its lower affinity for its epitope. This may be promising in the detection of spinal infections and in infections in regions with the presence of normal bone marrow. Also advantageous is the negligible HAMA response rate. Problems may arise in abdominal infections because of an intestinal excretion of activity as early as 4 hours postinjection in some cases but more usually at 24 hours.

Abdomen↗
PubMed1994

Structural modifications of monoclonal antibodies following direct versus indirect labelling with 99Tcm: does fragmentation really occur?

In this study, the influence of direct and indirect 99Tcm-labelling on the molecular structural integrity of monoclonal antibodies and other immunoglobulin preparations was investigated. Molecular composition of antibody preparations [two IgG monoclonal antibodies, one F(ab')2 fragment (all directly labelled), one indirectly labelled polyclonal human immunoglobulin preparation] and of serum samples after antibody injection were studied using polyacrylamide gel electrophoresis (PAGE; non-reducing and reducing conditions) and gel filtration chromatography. With PAGE, depending on the conditions used, a variety of lower molecular weight products could be detected. When analysing the same antibody preparations by gel filtration chromatography, all complete antibody preparations appeared as homogenous proteins of IgG molecular weight (150 kD). In F(ab')2 fragments, some further fragmentation to Fab' was noticed. Neither in vitro nor in vivo (serum) evidence of smaller fragments could be detected by gel filtration, despite their presence in PAGE. We therefore conclude that through the reductive step of direct 99Tcm-labelling, interchain disulphide linkages are broken but the polypeptide chains of complete IgG remain associated by non-covalent linkages, whereas (F(ab')2 is fragmented further to form essentially Fab'. The protein-denaturating conditions of PAGE (even if performed non-reducingly) seem to produce artifacts, not representing the real in vivo condition. PAGE results should therefore be interpreted only with great care.

Antibodies, Monoclonal↗
PubMed1994

A comparison of demineralized freeze-dried bone and autologous bone to induce bone formation in human extraction sockets.

The purpose of this study was to test the bone-forming capacity of demineralized freeze-dried bone (DFDBA) and autologous bone grafts in extraction sockets. Seven paired sites were grafted with either DFDBA or autologous bone. The sites were reentered between 3 and 13 months for the purposes of obtaining biopsies of the grafted sites and to place endosseous implants. Biopsies from 6 of the 7 grafted sites were evaluated for new bone formation. DFDBA sites revealed the presence of dead particles of DFDBA with no evidence of bone formation on the surfaces of the implanted particles and no evidence of osteoclastic resorption of the bone particles. Biopsies from the 6 autologous sites revealed vascular channels with woven and lamellar bone. Some specimens had retained cortical, non-vital bone chips. These bone chips were undergoing active osteoclastic resorption. The results of this study questions the use of DFDBA as a bone inductive graft material.

Alveolar Process↗
PubMed1994

[Somatostatin receptor scintigraphy in neuroendocrine tumors exemplified by a patient with hepatic metastases of gastrinoma].

In a 66-year old woman, who suffered from recurrent melena, diarrhea and hematemesis with multiple untreatable gastric and duodenal ulcers, a markedly increased basal and secretin-stimulated gastrin level, clinically a Zollinger-Ellison syndrome was assumed. The conventional diagnostic procedures (esophago-gastro-duodenoscopy, colonoscopy, endosonography, ERCP, abdominal CT and small bowel enema) had failed to reveal the localisation of any gastrinoma. The thereupon performed scintigraphy with In-111-pentetreotide showed four somatostatin receptor expressing liver lesions: two of them could be detected at first site in the consecutively performed MR scans, another retrospectively bearing in mind the scintigraphic images. Today, the somatostatin receptor imaging seems to be a highly sensitive procedure for detecting and localizing hormonally active gastroenteropancreatic tumors. At the same time it is a method for in vivo evaluation of the somatostatin receptor status of localized GEP tumors, thus delivering a decisive diagnostic step for the evaluation of the effectiveness of a therapy with somatostatin analogues before such an expensive therapy is started.

Aged↗
PubMed1994

Localized ridge augmentation using absorbable pins and e-PTFE barrier membranes: a new surgical technique. Case reports.

This paper presents a new surgical technique to promote bone formation in localized alveolar ridge defects. The objective was to regenerate sufficient bone volume for implant placement. The technique is dependent on careful defect debridement and the use of absorbable orthopedic pins, which serve as tent poles and prevent the e-PTFE barrier membranes from collapsing into the defects. The three defects treated with this technique were completely resolved with new bone, and implants were successfully placed into the augmented ridges. Biopsies from the treated sites revealed new bone formation.

Adult↗
PubMed1994

[Liver transplantation in halothane-induced liver necrosis].

The risk for developing acute liver failure after halothane exposition was calculated between 1:8,000 and 1:36,000. The case report given on a 22 year old man with halothane-induced hepatic failure is unusual, because the typical risk factors as age over 40, female sex, obesity, and previous exposure to halothane were not present. Two days after exposure to halothane the patient suffered acute liver failure with severe coagulopathy (factor V = 5% activity), and encephalopathy grade IV complicated by renal failure and respiratory insufficiency. Maximal increases of enzymes in blood were AST 3900 U/L, ALT 2570 U/L, LDH 10600 U/L. After six days the patient underwent liver transplantation with complete anuria and instable circulation. Explanted liver showed massive necrosis (70% of parenchyma) and fatty changes. The liver transplant had immediately a good function and renal failure resolved within three days. In the follow-up of 3 1/2 years the patient suffered no further complications. Culturing the patient's lymphocytes in the lymphocyte transformation test a strong reaction could be detected with a stimulatory index of 20. Maximal proliferation was observed when lymphocytes were incubated with plasma metabolites of a volunteer drawn 120 minutes after anesthesia with halothane was started.

Adult↗
PubMed1994

Detection of soft-tissue infections and osteomyelitis using a technetium-99m-labeled anti-granulocyte monoclonal antibody fragment.

UNLABELLED: Imaging of osteomyelitis and soft-tissue infections can be problematic with currently available agents; bone scans are often false-positive. Indium-111-oxine and 99mTc-HMPAO white blood cell (WBC) scans require ex vivo handling of blood with potential exposure to infectious agents, and 99mTc-antigranulocyte (IgG1) antibodies need 24 hr for final diagnosis. METHODS: We investigated the use of 99mTc-murine anti-granulocyte monoclonal Fab' fragment in 20 patients with suspected osteomyelitis of soft-tissue infections. All patients also had 99mTc bone scans and 111In-oxine or 99mTc-HMPAO white blood cell scans. The final diagnosis was confirmed by culture, biopsy, surgery, follow-up, x-rays, CT or MRI. In vitro studies performed on granulocytes demonstrated no effect on their function when the anti-granulocyte monoclonal antibody fragment was added. RESULTS: Sensitivity, specificity and diagnostic accuracy to detect infection was 88%, 75% and 80%, respectively. All lesions could be detected as early as 1 hr after injection of the antibody fragment. In comparison, WBC scanning had values of 86%, 78% and 81%, respectively. Some lesions could only be detected 24 hr following the injection of labeled WBCs. LeukoScan had three false-positives and WBC scanning had two false-positives. CONCLUSIONS: Immunoscintigraphy with 99mTc-NCA-90 Fab' fragments offers rapid localization of foci, rapid and simple use, a negligible HAMA response rate, no effect on granulocyte function and an accuracy comparable to WBC scanning.

Adult↗
PubMed1993

Use of immunoscintigraphy in the diagnosis of fever of unknown origin.

Fever of unknown origin (FUO) has been defined as an elevation in temperature (38 degrees C) for at least 2-3 weeks despite intensive investigation. The value of immunoscintigraphy with the technetium-99m-labelled anti-granulocyte antibody anti-NCA-95 (BW 250/183, IgG1) was studied retrospectively in 34 consecutive patients with FUO. Every effort was made to confirm a diagnosis, including methods such as ultrasonography, computed tomography, magnetic resonance imaging, bacteriological tests, surgical intervention and clinical follow-up. In 58.8% of the patients, an infectious cause for the fever was found, in 30.2% of the patients, a benign or malignant haematological disease, pancreatitis or thyrotoxicosis was found. No cause for fever could be found in 11%. The overall diagnostic sensitivity and specificity of immunoscintigraphy for infection were 40% and 92% respectively. The positive predictive value was calculated to be 88% and the negative predictive value was calculated to be 52%. False-negative scans were especially noted in patients with endocarditis, pneumonia and small brain abscesses, where the lesions did not exceed a diameter of 0.5 cm. If patients with endocarditis were excluded, the imaging sensitivity and specificity were increased to 57% and 95%. This study demonstrates that 99mTc-anti-NCA-95 scanning is able to localize infectious causes of FUO, other than endocarditis.

Adult↗