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Biomedical subjects

A Berger

Publications and source records attributed to A Berger.

At least 55 records · Page 3Linked to original sources

Anandamide and diet: inclusion of dietary arachidonate and docosahexaenoate leads to increased brain levels of the corresponding N-acylethanolamines in piglets.

Endogenous ligands of cannabinoid receptors have been discovered recently and include some N-acylethanolamines (NAEs; e.g., N-arachidonoylethanolamine) and some 2-acylglycerols (e.g., sn-2-arachidonoylglycerol). Previously, we found these compounds to be active biologically when administered per os in large quantities to mice. In the present work, piglets were fed diets with and without 20:4n-6 and 22:6n-3 fatty acid precursors of NAEs, in levels similar to those found in porcine milk, during the first 18 days of life, and corresponding brain NAEs were assessed. In piglets fed diets containing 20:4n-6 and 22:6n-3, there were increases in several biologically active NAEs in brain homogenates-20:4n-6 NAE (4-fold), 20:5n-3 NAE (5-fold), and 22:5n-3 and 22:6n-3 NAE (9- to 10-fold). These results support a mechanism we propose for dietary long-chain polyunsaturated fatty acids influences on brain biochemistry with presumed functional sequelae. This paradigm will enable targeted investigations to determine whether and why specific populations such as infants, elderly, or persons suffering from certain clinical conditions may benefit from dietary long-chain polyunsaturated fatty acids.

Animals↗

Experience with split-course external beam irradiation +/- chemotherapy and integrated Ir-192 high-dose-rate brachytherapy in the treatment of primary carcinomas of the anal canal.

PURPOSE: The effect of the treatment of anal cancer by performing a high-dose-rate (HDR) brachytherapy boost during a short split between the external beam radiotherapy series (EBR) +/- chemotherapy was investigated. METHODS AND MATERIALS: Thirty-nine patients with anal canal cancers, stages T1-T4 N0-2 M0, were treated with split-course EBR (50-50.4 Gy) and a Iridium 192 ((192)Ir-) HDR boost (6 Gy) performed during the 1-2-week split. Patients who failed to achieve a complete tumor response received additional brachytherapy. Chemotherapy with 5-fluorouracil and mitomycin C was offered to patients with tumors > 3 cm and employed concomitantly on days 1-5 and day 1, respectively, of each EBR series. RESULTS: Follow-up ranged from 3 to 140 months (median 31). Median treatment duration was 56 days. The 3-year (5-year) actuarial rates of locoregional control (LRC) and disease-specific survival (DSS) were 81% (76%) and 80% (76%), respectively. The crude rate of anal preservation was 77% overall, and 97% in patients in whom LRC was achieved. Uncompromised anal function was recorded in 93% of these patients. The actuarial 3-year (5-year) rate of colostomy-free survival (CFS) was 78% (73%). There was a statistically significant difference in LRC and DSS according to stage, tumor size, and nodal status. Complications requiring surgical intervention occurred in 7.6% of patients. CONCLUSION: The integration of the HDR boost in a split-course EBR regimen +/- chemotherapy resulted in excellent sphincter function without an increase of severe complications and with rates of LRC, DSS, and CFS, which compare favorably with those reported in the literature.

Adult↗

HLA typing.

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Cytotoxicity Tests, Immunologic↗

Human fibulin-4: analysis of its biosynthetic processing and mRNA expression in normal and tumour tissues.

Here, we report the identification of a human orthologue of fibulin-4, along with analysis of its biosynthetic processing and mRNA expression levels in normal and tumour tissues. Comparative sequence analysis of fibulin-4 cDNAs revealed apparent polymorphisms in the signal sequence that could account for previously reported inefficient secretion in fibulin-4 transfectants. In vitro translation of fibulin-4 mRNA revealed the presence of full-length and truncated polypeptides, the latter apparently generated from an alternative translation initiation site. Since this polypeptide failed to incorporate into endoplasmic reticulum membrane preparations, it was concluded that it lacked a signal sequence and thus could represent an intracellular form of fibulin-4. Using fluorescence in situ hybridisation analysis, the human fibulin-4 gene was localised to chromosome 11q13, this region being syntenic to portions of mouse chromosomes 7 and 19. Considering the fact that translocations, amplifications and other rearrangements of the 11q13 region are associated with a variety of human cancers, the expression of human fibulin-4 was evaluated in a series of colon tumours. Reverse transcription-polymerase chain reaction analysis of RNA from paired human colon tumour and adjacent normal tissue biopsies showed that a significant proportion of tumours had approximately 2-7-fold increases in the level of fibulin-4 mRNA expression. Taken together, results reported here suggest that an intracellular form of fibulin-4 protein may exist and that dysregulated expression of the fibulin-4 gene is associated with human colon tumourigenesis.

Amino Acid Sequence↗

A Schwann cell-seeded intrinsic framework and its satisfactory biocompatibility for a bioartificial nerve graft.

To optimize the internal environment of a collagen nerve tube, we designed a Schwann cell-seeded intrinsic framework and its biocompatibility was investigated. We fixed 6-0 polyglactin woven filaments (Vicryl) or polydioxanone monofilaments (PDS) on a silicone ring in a net fashion. It was coated with matrigel and then incubated with cultured newborn or adult Schwann cells. Furthermore, we implanted 1.5-cm-long filament-filled collagen tubes in a rat model. Using a live/dead fluorescent assay and electron microscopy, we found that adherent Schwann cells onto filaments remained viable and oriented longitudinally along filaments. The preliminary in vivo study indicated that a mild inflammatory reaction was present around the tube wall. However, nerve regeneration occurred around and between filaments. We concluded that the arrangement of Schwann cell columns onto filaments was achieved, mimicking Bünger bands. It was shown that the biomaterials did not impede nerve regeneration.

Animals↗

Hepatitis B virus markers in anti-HBc only positive individuals.

Isolated reactivity to hepatitis B virus (HBV) core antigen (anti-HBc) is observed relatively frequently in immunocompromised individuals, intravenous drug abusers (IVDA), and in the presence of HCV infection. The reason for the lack of HBsAg is not clear. The aim of the present study was to investigate which factors (genetic variability of S gene, low-level HBsAg, and immune complexes may be responsible for the failure of HBsAg detection with commercial HBsAg screening assays. Dilution series of two recombinant HBsAg escape mutants and dilutions of serum samples from chronic HBV carriers with multiple insertions in the a determinant and different HBsAg subtypes were tested with a highly sensitive assay that detects wild-type HBsAg (Elecsys HBsAg, Roche Diagnostics, Penzberg, Germany) and two assays that detect HBV wild-type and escape mutants (Murex HBsAg Version 3, Murex and Enzygnost HBsAg 5.0, Dade Behring, Marburg, Germany). Elecsys HBsAg showed in comparison to Murex HBsAg Version 3 and Enzygnost HBsAg 5.0 a reduced sensitivity for escape mutant detection. On the other hand, the best performance for HBsAg subtype detection was obtained with Elecsys HBsAg. In the second part of the study, a selected panel of isolated anti-HBc reactive (n = 104) serum samples (AxSYM Core) was submitted to testing by Elecsys HBsAg, Murex HBsAg Version 3, Enzygnost HBsAg 5.0, and HBsAg detection after immune complex dissociation (ICD) and anti-HBs determination with two different assays (AxSYM Ausab and Elecsys Anti-HBs). To assess the specificity of anti-HBc test results, all the samples were tested by a second anti-HBc assay (Elecsys Anti-HBc). Quantitative HBV DNA detection was undertaken with a commercially available HBV PCR assay (Amplicor HBV Monitor). HCV infection was present in 65.4% of anti-HBc only reactive individuals. Five AxSYM Core positive samples were negative by Elecsys Anti-HBc. Overall, 15 (14.4%) AxSYM Ausab negative samples gave positive results with Elecsys Anti-HBs (median value: 21 IU/ml). No low-level HBsAg carrier was detected among the isolated anti-HBc reactive individuals with Elecsys HBsAg. There was no evidence for the presence of immune complexes. Only one sample was repeatedly reactive by the Murex HBsAg, suggesting that the a mutant form of HBsAg was responsible for the isolated anti-HBc reactivity, however neutralisation assay was not interpretable and HBV DNA PCR was negative. Fifteen (14.4%) anti-HBc only positive individuals were HBV DNA carriers with concentrations ranging from 800 to more than >4,000,000 copies of viral DNA/ml. In conclusion, the most probable explanations for isolated anti-HBc reactivity in our study group are a possible interference of HBsAg synthesis by HCV infection (65.4%) and divergence of results of anti-HBs assays (14.4%). There is no evidence for the presence of low-level HBsAg carriers and immune complexes. HBsAg mutants cannot be excluded definitively by the test strategy used in the present evaluation.

Amino Acid Sequence↗

Simultaneous assessment of cholesterol absorption and synthesis in humans using on-line gas chromatography/ combustion and gas chromatography/pyrolysis/isotope-ratio mass spectrometry.

A number of dietary components and drugs are known to inhibit the absorption of dietary and biliary cholesterol, but at the same time can compensate by increasing cholesterol synthesis. It is, therefore, necessary to have a convenient and accurate method to assess both parameters simultaneously. Hence, we validated such a method in humans using on-line gas chromatography(GC)/combustion and GC/pyrolysis/isotope-ratio mass spectrometry (IRMS). Cholesterol absorption was measured using the ratio of [(13)C]cholesterol (injected intravenously) to [(18)O]cholesterol (administered orally). Simultaneously, cholesterol synthesis was measured using the deuterium incorporation method. Our methodology was applied to 12 mildly hypercholesterolemic men that were given a diet providing 2685 +/- 178 Kcal/day (mean +/- SD) and 255 +/- 8 mg cholesterol per day. Cholesterol fractional synthesis rates ranged from 5.0 to 10.5% pool/day and averaged 7.36% +/- 1.78% pool/day (668 +/- 133 mg/day). Cholesterol absorption ranged from 36.5-79.9% with an average value of 50.8 +/- 15.4%. These values are in agreement with already known data obtained with mildly hypercholesterolemic Caucasian males placed on a diet similar to the one used for this study. However, our combined IRMS method has the advantage over existing methods that it enables simultaneous measurement of cholesterol absorption and synthesis in humans, and is therefore an important research tool for studying the impact of dietary treatments on cholesterol parameters.

Cholesterol↗

Contribution of ICP-IDMS to the certification of antimony implanted in a silicon wafer--comparison with RBS and INAA results.

A thin-layer reference material for surface and near-surface analytical methods was produced and certified. The surface density of the implanted Sb layer was determined by Rutherford backscattering spectrometry (RBS), instrumental neutron activation analysis (INAA), and inductively coupled plasma isotope dilution mass spectrometry (ICP-IDMS) equipped with a multi-collector. The isotopic abundances of Sb (121Sb and 123Sb) were determined by multi-collector ICP-MS and INAA. ICP-IDMS measurements are discussed in detail in this paper. All methods produced values traceable to the SI and are accompanied by a complete uncertainty budget. The homogeneity of the material was measured with RBS. From these measurements the standard uncertainty due to possible inhomogeneities was estimated to be less than 0.78% for fractions of the area increments down to 0.75 mm2 in size. Excellent agreement between the results of the three different methods was found. For the surface density of implanted Sb atoms the unweighted mean value of the means of four data sets is 4.81 x 10(16) cm(-2) with an expanded uncertainty (coverage factor k = 2) of 0.09 x 10(16) cm(-2). For the isotope amount ratio R (121Sb/123Sb) the unweighted mean value of the means of two data sets is 1.435 with an expanded uncertainty (coverage factor k = 2) of 0.006.

Journal Article↗

Organotypical engineering of differentiated composite-skin equivalents of human keratinocytes in a collagen-GAG matrix (INTEGRA Artificial Skin) in a perfusion culture system.

BACKGROUND: The production of autologous composite skin equivalents for the treatment of full-thickness skin defects in burns is time consuming and costly because of laboratory procedures which have to be performed manually. In the present study keratinocytes were seeded into INTEGRA Artificial Skin and placed in a perfusion culture system in order to evaluate the possibility of producing composite grafts in an automated system with the aim of establishing a cost-effective method of industrial production. METHODS: Composite grafts of INTEGRA and human keratinocytes were raised in perfusion culture and grafted onto athymic mice to evaluate their potential to reconstitute a full-thickness skin substitute in vivo compared to grafts from standard stagnant cultures. RESULTS: Cultured composites from perfusion cultures showed no significant histological differences compared to those from stagnant cultures; however, a tendency of improved cell growth and a more surface-oriented localization was observed. Cell proliferation and surface-bound differentiation were not impaired by the use of carbonate-independent buffering (HEPES), which is necessary for perfusion culture. The composite grafts from perfusion culture exhibited identical wound adherence and complete healing and histologically represented a multi-layered, keratinizing human epidermis. CONCLUSION: Engineering of differentiated composite skin equivalents is possible in a perfusion culture system, which offers technical and procedural and possibly even biological advantages compared to standard stagnant culture methods. The development of automated perfusion culture systems for the production of composite grafts in sizes required clinically (scale-up) will be the next step in the cost-effective engineering of large-scale composite skin equivalents.

Animals↗

Evaluation of the new LCx HIV RNA quantitative assay: comparison with the Cobas Amplicor HIV Monitor assay.

In the present study we evaluated the performance of the new LCx HIV RNA quantitative kit (Abbott Laboratories, Delkenheim, Germany) for the quantitative detection of HIV-1 RNA in human plasma in comparison to the Cobas Amplicor HIV-1 Monitor assay (Roche Diagnostics, Branchburg, N.J.), including samples containing a variety of HIV-1 subtypes. LCx and Cobas were compared using archived EDTA plasma samples collected from HIV-infected patients. Considering the lower limit of the linear range of 50 copies/ml, the detection rate of the LCx was 139 out of 174 (79.9%) versus 131 out of 174 (75.3%) of the Cobas. Overall agreement was 95.4% (166/174) at a cut-off of 50 copies. LCx and Cobas results on clinical samples were found linearly associated (r2 = 0.900) and strongly correlated (r = 0.949). The mean viral load in the 174 frozen patient samples was 3.25 log10 copies/ml by LCx compared to 2.71 log10 copies/ml by Cobas. Considering only samples with a viral load > or =50 copies/ml, the average difference was -0.132 log copy/ml. Using a panel consisting of 9 plasma samples spiked with 9 different HIV-1 cultured isolates (A-H, and O) LCx detected the 9 subtypes with a high degree of precision, i.e., 9-33% coefficient of variation. As expected, the Cobas failed to detect the group O isolates. The results of the remaining samples showed a higher degree of variation (when testing four replicates of the subtype panel) than the LCx of 14.2-40.3%. Nevertheless, the results were comparable with the LCx data.

DNA Ligases↗

[Esophagectomy for squamous cell carcinoma of the esophagus isolated or associated with head and neck cancer: long-term survival].

STUDY AIM: Esophageal squamous cell carcinomas are frequently associated with head and neck cancers. This retrospective study was conducted to compare the long-term outcome of the patients with a double cancer and of the patients with a solitary esophageal cancer after curative management. PATIENTS AND METHODS: From 1989 to 1999, 114 patients with an esophageal carcinoma were included in the study. Among them, 52 had an associated head and neck cancer (metachronous: n = 17 and synchronous: n = 35). Curative treatment was achieved in all patients. The patients were divided in "solitary" and "associated" group. RESULTS: Age, sex distribution, tumor location and histological findings were similar in the two groups. The esophageal resection was an esopharyngolaryngectomy (n = 13), a subtotal esophagectomy with cervical anastomosis (n = 92) and a Lewis-Santy esophagectomy with thoracic anastomosis (n = 9). Operative mortality (8 versus 7.7%), anastomotic leaks rate (14.5 versus 21%) and pneumonia rate (21 versus 9.6%) were not significantly different in the two groups. The mean hospital stay was 27 days. The mean follow-up was 85 +/- 50 months. Five-year survival rates were not significantly different in the two groups (p = 0.6411). In univariate survival analysis the only significant predictive factors were the depth of esophageal tumor invasion (p = 0.0002) and node involvement (p = 0.0373). The presence of head and neck cancer did not affect survival after esophagectomy. CONCLUSION: With an aggressive therapeutic plan, the survival of patients with an esophageal cancer associated to head and neck cancer was similar to the survival of patients with an isolated esophageal carcinoma. Long term esophageal survey seems to be useful to detect more superficial esophageal carcinomas in patients with head and neck cancer.

Aged↗

A treadmill ergometer for three-dimensional ground reaction forces measurement during walking.

This report describes new treadmill ergometer designed to measure the vertical and horizontal ground reaction forces produced by the left and right legs during walking. It was validated by static and dynamic tests. Non-linearity was from 0.2% (left vertical force) to 1.4% (right antero-posterior force). The resonance frequency was from 219 (right vertical direction) to 58 Hz (left medio-lateral direction). A calibration "leg", an air jack in series with a strain gauge, was developed and used to produce force signals comparable to those obtained during human locomotion. The mean differences between the force measured by the calibration leg and treadmill ergometer at 5 km h(-1) were 3.7 N (0.7%) for the left side and 6.5 N (1.2%) for the right. Measurements obtained during human walking showed that the treadmill ergometer has considerable potential for analysing human gait.

Calibration↗

Human cytomegalovirus infection: diagnostic potential of recombinant antigens for cytomegalovirus antibody detection.

Recombinant antigen-based enzyme immunoassays (EIAs) for the detection of human cytomegalovirus (HCMV) specific antibody are believed to yield a higher sensitivity and specificity than virus lysate EIAs. The aim of the present study was to evaluate the accuracy of newly established HCMV assays (Copalis CMV Multiplex, Sorin; Cobas Core CMV IgG and IgM EIAs, Roche Diagnostics; Anti-HCMV recombinant IgG, gB-IgG, IgM and IgA, Biotest; and ETI-CYTOK-G PLUS and M reverse PLUS, Sorin) based on recombinant antigens and/or virus lysate for laboratory diagnosis of HCMV infection. For the assessment of sensitivity, follow-up samples from patients suffering from active HCMV infection were tested. Testing a large number of potentially interfering samples challenged the specificity of the assays. There was no statistically significant difference in the performance of HCMV IgG assays. The results were more heterogeneous for the detection of serological markers of active infection (HCMV IgM, HCMV IgA and anti-CM2). The sensitivities of the different assays ranged between 40.5 and 71.4%. A variable number (17.8-1.7%) of false-positive results were obtained among potentially interfering serum samples. Two of the recombinant antigen based assays showed a high degree of interference with EBV VCA-IgM-positive sera. The best performance was achieved with ETI-CYTOK-M reverse PLUS since it combined the highest sensitivity with specificity. Commercially available assays based on recombinant antigens showed, overall, a poorer performance than the virus lysate EIA.

Antibodies, Viral↗

Primary adenocarcinoma of the rectovaginal septum: a case report and literature review.

The authors report the third case of primary adenocarcinoma of the rectovaginal septum without associated endometriosis and discuss the pathogenesis of this tumour. Some of the tumour cells were stained with OC 125 antibody which recognises epithelium of coelomic origin; adenocarcinoma of the rectovaginal septum may arise directly from embryological Müllerian remnants.

Adenocarcinoma↗