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D Gerding

Publications and source records attributed to D Gerding.

11 recordsLinked to original sources

Multicenter typing comparison of sporadic and outbreak Clostridium difficile isolates from geographically diverse hospitals.

In a collaborative study by three laboratories, arbitrarily primed polymerase chain reaction (AP-PCR), HindIII restriction enzyme analysis (REA), and pulsed-field gel electrophoresis (PFGE) using SmaI were compared for typing of Clostridium difficile. The study included 30 isolates from nosocomial outbreaks in six geographically disparate hospitals and 15 isolates from sporadic cases of C. difficile diarrhea. REA distinguished a total of 23 types representing 10 groups; AP-PCR performed at Deaconess Hospital resolved 19 types; AP-PCR performed at the Centers for Disease Control resolved 15 types. Thirty isolates exhibited degradation of larger sized fragments during processing and therefore were nontypeable by PFGE; among the remaining 15 isolates, PFGE resolved 11 types. Outbreak isolates in five different hospitals represented REA group J and constituted a single AP-PCR strain. In summary, nosocomial outbreaks of C. difficile diarrhea in five hospitals were associated with a single genetic lineage as resolved by multiple strain typing systems.

Bacterial Typing Techniques↗

Increased LDL receptor mRNA expression in colon cancer is correlated with a rise in plasma cholesterol levels after curative surgery.

It is currently under debate whether the low serum cholesterol levels that are frequently observed in cancer patients represent a risk factor for/or, rather, are a consequence of the tumour. We postulate that malignant tumours are directly involved in an increased catabolism of cholesterol-rich low-density lipoprotein (LDL) particles. In a prospective study of 25 patients with colorectal carcinoma, we measured intraindividual shifts in serum cholesterol levels after surgery, and the expression of LDL-receptor mRNA in surgically removed specimens. A significant rise in plasma cholesterol levels was observed in patients 3 and 12 months after curative surgery, but not after non-curative surgery. In human colon carcinoma tissues LDL receptor mRNA expression, as determined by competitive reverse-transcriptase-polymerase-chain reaction, was found to be significantly increased when compared to tissues from the tumour-free margin (median values, 1.2 x 10(6) vs. 2.0 x 10(5) molecules/micrograms total cellular RNA, respectively, n = 17). The extent of LDL-receptor mRNA expression positively correlated to the percentage rise of plasma cholesterol levels 3 months (n = 7, r = 0.8763) and 12 months (n = 6, r = 0.9181) after curative surgery. This finding provides in vivo evidence that the tumour tissue itself contributes to decreased plasma cholesterol levels in patients suffering from colorectal carcinomas. It supports the hypothesis that low cholesterol levels in cancer patients are a consequence, and not the cause, of the malignancy.

Adult↗

Rapid quantitation of mRNA species in ethidium bromide-stained gels of competitive RT-PCR products.

A rapid method for quantifying the low-abundant mRNAs of the low density lipoprotein receptor and the 3-hydroxy-3-methylglutaryl coenzyme A reductase by competitive polymerase chain reaction is presented. This approach requires neither special labeling nor blotting procedures. For each analysis, a defined amount of total cellular RNA is co-reverse transcribed and co-amplified with a titration series of in vitro synthesized competitor RNA that carries an internal deletion. The equivalence point, which defines the amount of specific RNA in the sample, can be scored in ethidium bromide-stained agarose or polyacrylamide gels of the reaction products. As an example, responses to pravastatin, a competitive inhibitor of the HMG-CoA reductase, in a human tumor cell line were analyzed with this new technique. As a control, the expression of the unregulated gene, glyceraldehyde-3-phosphate dehydrogenase was measured in parallel using the same methodology. The results obtained were compared with those obtained by conventional Northern blotting.

Base Sequence↗

Effectiveness and tolerance to highly concentrated vs conventional TPN formulas.

Conventional total parenteral nutrition regimens (TPN-C) involve concentrations of dextrose/protein which necessitate administration of 2.5-4 liters/day to meet target nutritional needs. Although this is frequently acceptable, certain clinical settings mandate a volume-restricted (VR) approach. This study compares a VR TPN regimen (TPN-VR) involving the use of 25% dextrose and 9.5% amino acid with D17.5 AA 5.0 (TPN-C). The two groups were compared for adequacy of nutritional delivery, balance, and tolerance. Twenty patients received the TPN-VR (Group 1) and 20 patients received TPN-C (Group 2). The groups were comparable in age, sex, injury severity, and APACHE 2 scores. Harris-Benedict (BEE) x 2 and 2 g/protein/kg of ideal body weight were delivered by the second day of TPN. A 27% reduction in administered fluid was achieved in Group 1 (P < 0.001). Metabolic cart data in both groups demonstrated that delivered calories exceeded REE. The average RQ in Group 1 was 0.84 and in Group 2 was 0.90 (P > 0.1). There was no significant difference between the two groups in nitrogen balance, mean serum bilirubin levels, PT and PTT, serum albumin levels, and triglycerides (P > 0.20). SGPT and alkaline phosphatase levels were significantly higher in Group 2 (P < 0.001). Group 2 received an average of 22% more carbohydrate than Group 1 and 45% required insulin compared to 25% in Group 1 (P < 0.01). In summary, TPN-VR is comparable to TPN-C in terms of effectiveness of delivery, nutritional balance, and tolerance.

Adult↗

Effect of continuous vs intermittent application of 3-OH-tamoxifen or tamoxifen on the proliferation of the human breast cancer cell line MCF-7 M1.

The antiproliferative potency of 5 x 10(-7) M tamoxifen (TAM) and 3-hydroxytamoxifen (3-OH-TAM) was investigated during continuous (8 days) or intermittent (2 h every 2nd or 3rd day, respectively) application to the oestrogen-receptor-positive, estradiol-sensitive human mammary carcinoma cell line MCF-7 M1, a variant of MCF-7 wild type. Growth modulation was evaluated in parallel by counting cells and by measuring DNA content. Continuous incubation resulted in a growth inhibition to 21.8 +/- 3.2% by 3-OH-TAM and to 39.5 +/- 4.8% by TAM when compared with control cultures defined as 100%. Intermittent addition induced a growth reduction to 23.0 +/- 2.1% by 3-OH-TAM and to 41.2 +/- 2.4% by TAM in relation to 100% controls. Addition of 3-OH-TAM for 2 h only at day 1 resulted in an inhibition to 70.3 +/- 3.2%, again in relation to 100% controls. When TAM was administered once for 2 h at day 1 it induced an inhibition to 79.0 +/- 4.9% at day 8. The in vitro results indicate that (a) at 5 x 10(-7) M 3-OH-TAM has a better antiproliferative effectiveness than TAM, (b) the intermittent application is as effective as continuous application (no significant difference), and (c) the addition once a week reveals only a slight growth reduction after 8 days of culture. Application of the long-living TAM results in continuously high serum concentrations, which have been shown to create resistant cell clones. Compared to TAM the 3-OH metabolite has a considerably shorter half-life and its application in vivo reveals rise and fall of its serum concentrations. Since the presented data demonstrate that 3-OH-TAM is more potent than TAM and that the intermittent application is as effective as the continuous form, interval therapy with 3-OH-TAM may slow down the process of acquiring resistance to antioestrogens.

Breast Neoplasms↗

Effectiveness of mitoxantrone on the proliferation of cell cultures derived from malignant mesenchymal tumors of human origin.

The antiproliferative potency of mitoxantrone (MITOX) has predominantly been established for epithelial and hematologic neoplasias. In this study the effectiveness of MITOX was investigated in vitro for 6 sarcomatous human cell lines derived from 2 synovial sarcomas, a malignant schwannoma, a malignant histiocytoma, a leiomyosarcoma, and a chondrosarcoma. The examination was performed using a proliferation assay with monolayer cell cultures. The effect of MITOX was compared with that of adriamycin (ADR) and cisplatin (CDDP). For each drug at least 3 concentrations were tested which covered the therapeutically achievable range, i.e., for MITOX 0.2-0.002 micrograms/ml, for ADR 0.5-0.005 micrograms/ml, and for CDDP 5.0-0.05 micrograms/ml. Test incubations were performed for 3 days. Antiproliferative potency of the cytostatic drugs was assessed by counting the number of cells at the start and the end of the test period with and without drug addition. Furthermore the dose inhibiting cell growth to 50% of controls (ID50) was determined for MITOX. For comparison 4 cell lines from carcinomatous lesions were included in the study. MITOX inhibited proliferation rates of 4 sarcomatous tumor cell lines more intensively than ADR, and was less effective in 2 cell lines. However, these differences were not significant. In all mesenchymal cell lines tested the antiproliferative potency of MITOX was more pronounced than that of CDDP. In carcinomatous cell lines the MITOX-induced growth inhibition was similar to that found in response to administration of ADR and CDDP confirming the described effect on epithelial tumors. The study suggests that MITOX possesses a growth inhibitory potency for malignant soft tissue tumors in vitro. From these data it may be worthwhile to initiate clinical trials testing the treatment of sarcomatous lesions with MITOX.

Adolescent↗

Clostridium difficile plasmid isolation as an epidemiologic tool.

A large hospital outbreak of Clostridium difficile diarrhea at the Minneapolis Veterans Administration Medical Center (MVAMC) was studied by plasmid profile typing. Plasmids were obtained from 30 (37%) of 82 clinical isolates from MVAMC patients and 10 (67%) of 15 non-MVAMC isolates. While bacteriophage plus bacteriocin typing and polyacrylamide gel electrophoresis (PAGE) plus bacterial agglutination typing proved more universally applicable, plasmid profiles may be useful for tracing isolated epidemic outbreaks, reinfections and relapses caused by plasmid-bearing strains.

Bacteriocins↗

Establishment of primary cell cultures: experiences with 155 cell strains.

Cell culture systems allow the examination of cell populations in a functional state. To simulate in vivo conditions as closely as possible freshly established cell strains are superior to permanent cell lines. Different aspects for the establishment of primary cell cultures obtained from various tissues are compared: Disintegration, culture media supplemented with basal additions, special supplements (growth factors, hormones), and attachment factors. The proliferation rates of the attained cell strains were evaluated by determination of cell doubling times. Procedures for how to obtain a relatively high plating efficiency (approx. 70% in our series of 219 attempts) of primary growth in vitro are described: (1) Mechanical disintegration is superior to enzymatic digestion. If mechanical treatment alone did not produce a sufficient number of viable cells, additional digestion with collagenase/dispase revealed a higher number of proliferating primary cultures than with trypsin. (2) Proliferation of cell cultures from normal and tumorous tissues of epithelial origin was superior in Leibovitz L 15 medium (58 of 87 (67%) cases). Cultures from mesenchymal tissues and tumors were found to have shortest cell doubling times in MEM and RPMI 1640 (16 of 23 (70%) cases). The media were supplemented with the basal additions indicated. (3) In approx. 30% of the cases special supplements like growth factors or hormones increased cell replication, although they were almost always not essential for cell growth. (4) Attachment factors only rarely contributed to the initiation of primary monolayer cultures. The application of various culture conditions does not lead to a protocol optimal for all tissues, for all probes of the same type of tumor, or for all tumor specimens of unique differentiation.

Cell Adhesion↗

Antigen-induced experimental septic arthritis in rabbits after intraarticular injection of Staphylococcus aureus.

With the Dumonde-Glynn model of antigen-induced arthritis, a rabbit model was developed to examine the histopathologic differences between normal and arthritic joints in the same animal infected by intraarticular injections of Staphylococcus aureus. Microscopic examination of whole joint sections and a quantitative histopathologic scale were used to compare changes in all the articular components of 17 normal and 17 arthritic joints infected for less than two weeks. The histological changes were more severe in infected arthritic joints than in infected normal joints (mean +/- SD total histology score, 13.8 +/- 2.4 and 9.3 +/- 4.0, respectively; P less than .001). In infected arthritic joints, subsynovial abscesses extended into subchondral bone via the pannus of chronic synovitis at articular margins and intraarticular attachments of cruciate ligaments, rather than by initial cartilage destruction and direct extension into subchondral bone.

Abscess↗

Use of technetium-99m methylene diphosphonate and gallium-67 citrate scans after intraarticular injection of Staphylococcus aureus into knee joints of rabbits with chronic antigen-induced arthritis.

Numerous clinical studies have questioned the ability of radionuclide scans to differentiate septic from aseptic joint inflammation. A clinical study may not be able to document an underlying disease process or duration of infection and, thus, may make conclusions about the accuracy of scan interpretations open to debate. In this study, the Dumonde-Glynn model of antigen-induced arthritis in rabbits was used as the experimental model to study technetium and gallium scans in Staphylococcus aureus infection of arthritic and normal joints. Gallium scans were negative in normal rabbits, usually negative in antigen-induced arthritis, but positive in septic arthritis. The bone scan was usually negative in early infection but positive in late septic arthritis, a finding reflecting greater penetration of bacteria into subchondral bone because of the underlying inflammatory process.

Animals↗