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Biomedical subjects

E Gross

Publications and source records attributed to E Gross.

At least 37 records · Page 2Linked to original sources
PubMed2000

Identification of specific BRCA1 and BRCA2 variants by DHPLC.

Denaturing high performance liquid chromatography (DHPLC) is generating increasing interest in clinical genetics as a reliable tool for the analysis of genetic alterations. In the work presented here our intentions were to optimize primer design and DHPLC analysis conditions for a qualitative detection of BRCA1 and BRCA2 variations. The BRCA1 and BRAC2 genes display a high proportion of polymorphisms. Sequencing efforts geared towards the distinction of tumor-related mutations and benign variants still remain time-consuming and expensive. DHPLC elution profiles, however, permit the correlation of a characteristic chromatographic profile with a specific sequence alteration. In this study we evaluate the sensitivity of DHPLC for the identification of unique polymorphisms, which are frequent in the Caucasian population, in lieu of sequence analysis. The complete BRCA1 gene and parts of BRCA2 were examined. In the case of BRCA1, 431 out of 432 heterozygotes were identified correctly. In addition, 18 new profiles were identified which had not been detected previously in our studies and which represented new mutations or rare polymorphisms. For BRCA2, 135 out of 137 simple sequence variants were classified correctly. In addition, six new profiles were identified which represented new mutations or rare polymorphisms.

BRCA2 Protein↗
PubMed2000

Ten novel BRCA1 and BRCA2 mutations in breast and/or ovarian cancer families from northern Germany.

Germline mutations in the BRCA1 and BRCA2 gene account for the majority of high-risk breast/ovarian cancer families. We have screened such families from Northern Germany by using DHPLC analysis and subsequent direct sequencing techniques. In ten families we identified six novel BRCA1 and 4 novel BRCA2 mutations comprising four frame shift mutations, one nonsense and one splice site mutation in the BRCA1 gene as well as three frameshift mutations and one nonsense mutation in the BRCA2 gene. Our analysis contributes to the further characterisation of the mutational spectrum of BRCA1 and BRCA2.

Adult↗
PubMed2000

Laparoscopic assisted surgery for Crohn's disease an initial experience and results.

The inflammatory process associated with Crohn's disease often makes dissection difficult, even in open surgery. The aim of this study was to assess the technical feasibility, safety and indication of laparoscopic assisted procedures performed in patients with Crohn's disease. Records of patients undergoing surgical operation for Crohn's disease from 1993 to 1998 at our hospital were reviewed. Intestinal resection was performed laparoscopically in 24 patients (LAP) and by open technique in 23 patients(OPEN). No significant differences existed as to age, gender, body-mass-index and previous surgery. In the laparoscopic group, seven operations (CON) were converted to open laparotomy (29%) because of large inflammatory mass and/or fistula. The mean intraoperative blood loss was significantly higher in CON- and OPEN-groups than in LAP-group (P < 0.01). Major complications occurred only in one patient who underwent laparoscopic assisted high anterior resection. Patients who underwent laparoscopic operation tolerated p.o. liquids sooner than patients who underwent open surgery (median: 2 vs. 5 day, P < 0.05). Compared with the CON- and OPEN-groups, patients in LAP-group had lower analgesic requirements (median: 3 vs. 6 and 5 day, P < 0.01). The median postoperative length of stay was significantly shorter in LAP-group than in OPEN-group (median: 11 vs. 14 day, P < 0.05). Our study showed that LAP is technically feasible for Crohn's disease. The preoperative correct diagnosis and selection of indications are very important, because the laparoscopic mobilization and resection may be difficult or impossible in patients with large fixed masses, multiple complex fistulas, or recurrent Crohn's disease.

Adult↗
PubMed2000

Large bowel obstruction: an unusual presentation of salmonella enterocolitis in infancy.

An infant with a large-bowel obstruction due to Salmonella B enterocolitis is presented. The clinical and radiologic findings were suggestive of Hirschsprung's disease with total colonic aganglionosis. Due to further deterioration, an ileostomy was performed. Pathologic examination disclosed ganglia in the colon. At laparatomy, 1 month later the colon, which was almost completely obliterated, was resected and an ileorectal anastomosis carried out. The patient remained a carrier of a multiple antibiotic-resistant group B Salmonella strain, and 2 months later died as a result of severe gastroenteritis.

Carrier State↗
PubMed2000

Compression anastomosis (AKA-2) in colorectal surgery: results in 442 consecutive patients.

BACKGROUND: The morbidity and mortality associated with colorectal surgery results partly from anastomotic leakage. Animal experiments have shown some advantages of sutureless anastomosis over conventional colorectal anastomosis. Compression anastomosis follows the same biological principles as sutureless anastomosis. METHODS: The compression anastomosis AKA-2 was evaluated in a prospective consecutive audit of 442 patients between September 1989 and August 1998. RESULTS: Anastomoses were performed in 372 elective and 70 emergency situations. The indication for operation was colorectal cancer (56.3 per cent) and diverticulitis (23.5 per cent). A defunctioning colostomy was performed in 110 patients (24.9 per cent). Fourteen patients died (3.2 per cent). Death was related to anastomotic complications in three patients (0.7 per cent). Twenty-four patients (5.4 per cent) developed intra-abdominal complications. There were 11 symptomatic (2.5 per cent) and six asymptomatic (1.4 per cent) leakages. Anastomoses that were more than 10 cm from the anal verge leaked in seven (2.4 per cent) of 291 cases, while anastomoses between 5 and 10 cm leaked in three (2.6 per cent) of 116 cases and those less than 5 cm from the anal verge leaked in one (3 per cent) of 35 cases. CONCLUSION: The low incidence of anastomotic complications demonstrates good biological healing of compression anastomoses. The compression anastomosis AKA-2 is safe in both high and low anterior resection and can therefore be recommended for use in colorectal surgery.

Adult↗
PubMed2000

Cell lines from the same cervical carcinoma but with different radiosensitivities exhibit different cDNA microarray patterns of gene expression.

Combining chemotherapy with radiotherapy has improved the cure rate among patients with cancers of the cervix. Although one-half to two-thirds of the patients can be cured by radiation alone, such patients cannot be identified at present and must therefore suffer the burden of chemotherapy. Our long-range goal is to identify those cervical cancers that are radiosensitive and could be cured by radiotherapy alone. The advent of methods that permit the simultaneous analysis of expression patterns of thousands of genes, make it feasible to attempt to identify the molecular events related to radiosensitivity and the associated regulatory pathways. We hypothesize that the sensitivity of tumor cells to ionizing radiation (IR) is determined by the level of expression of specific genes that may be identified with the aid of cDNA microarrays. As the first step in testing this hypothesis, we determined the gene expression differences between two cell lines exhibiting different degrees of radiosensitivity. These were derived from the same tumor prior to treatment from a patient with squamous cell carcinoma of the cervix. The mRNA from these cells was subjected to cDNA analysis on a microarray of 5,776 known genes and ESTs. The expression of 52 genes of the total of 5,776 was elevated (maximum 4.1 fold) in the radioresistant cells as compared to the radiosensitive cells. Ten of the 52 sequences are known genes while 42 are ESTs. Conversely, the expression of 18 genes was elevated in the sensitive cells as compared to the resistant cells. Seven of these 18 are known genes while eleven are ESTs. Among the genes expressed differentially between the resistant and sensitive cells were several known to be associated with response to IR and many more genes and ESTs that had not previously been reported to be related to radiosensitivity. The genes that showed the greatest overexpression in the radioresistant cell line were metal-regulatory transcription factor-1, cytochrome P450 CYP1B1, adenomatosis polyposis coli, translation elongation factor-1, cytochrome-c oxidase, whereas in the sensitive cell line, transcription factor NF-kappa-B, metalloproteinase inhibitor-1 precursor, superoxide dismutase-2, insulin-like growth factor binding protein-3, guanine nucleotide-binding protein and transforming growth factor beta-induced protein were overexpressed.

Cell Survival↗
PubMed2000

The pivotal role of deep vein thrombophlebitis in the development of acute disseminated staphylococcal disease in children.

Deep vein thrombophlebitis (DVT) and septic pulmonary emboli (PE) are rare in children. The association of DVT and acute disseminated staphylococcal disease (DSD) during childhood has not been previously reported. We report 3 children who developed a triad of DVT, septic PE, and acute osteomyelitis with Staphylococcus aureus cultured from blood and bone. One child succumbed, while 2 survived following prolonged, morbid hospitalizations. The rapid clinical deterioration observed in these patients might be caused by the aggressiveness of staphylococcal infection combined with an ongoing showering of septic emboli from the ileo-femoral DVT. We suggest that infected DVT with septic PE had a pivotal role in the development of DSD in these children. The presence of this triad should prompt aggressive treatment with the appropriate antibiotics, anticoagulation, surgical drainage, and assisted ventilation when indicated.

Acute Disease↗
PubMed1999

Angiographic anatomy of the inferior right atrial isthmus in patients with and without history of common atrial flutter.

BACKGROUND: Although most ablative procedures undertaken for common atrial flutter target the inferior right atrial isthmus, comparative studies of the morphology of this area are lacking. Our study examines its angiographic anatomy, making correlations with postmortem specimens, to provide a better understanding of the anatomic substrate of this arrhythmia. METHODS AND RESULTS: The gross morphological features and dimensions of the area between the orifice of the inferior caval vein and the attachment of the septal leaflet of the tricuspid valve were determined from angiograms made in 23 patients with documented atrial flutter and 30 control subjects. For comparison, we studied 20 normal heart specimens. When viewed in right anterior oblique projection, 2 morphologically distinct areas were identified. In the specimens, the inferior isthmus measured a mean length of 30+/-4 mm, not significantly different from the dimensions obtained from angiograms of control subjects. The mean length of the isthmus, however, was greater in patients with common atrial flutter than those without (37+/-8 versus 28+/-6 mm). Patients with atrial flutter and structural heart disease had an even longer isthmus than those with flutter alone (39. 6+/-8 versus 33+/-7 mm). Compared with those without flutter, the atrial diameter was also larger in patients with flutter (57.6+/-9 versus 48.5+/-6 mm). Reevaluation carried out at follow-up 10+/-2 months after ablation did not show any reduction in atrial size, although contractility improved. CONCLUSIONS: The inferior isthmus and right atrium in patients with common atrial flutter were significantly larger than those in a control population.

Adult↗
PubMed1999

A highly sensitive, fast, and economical technique for mutation analysis in hereditary breast and ovarian cancers.

Mutation analysis of complex genes without hotspots for sequence variations, such as BRCA1, is time-consuming and expensive. Of all currently available methods, direct sequencing has the highest sensitivity, but also the highest costs. Other techniques, such as SSCP, DGGE, and PTT, are more economical but, depending on the experience of the investigator, have at best a sensitivity of 90%. We investigated in a prospective study the feasibility and accuracy of the DHPLC technique. We present the application of the DHPLC protocol for BRCA1 mutation detection on a HPLC device from Bio-Tek Kontron Instruments (Neufahrn, Germany). DNA from 46 women with hereditary breast and ovarian cancer undergoing genetic testing for BRCA1 mutations were tested. Of 1,518 amplicons analyzed by DHPLC, corresponding to 33 fragments spanning the entire BRCA1 gene, 626 were also directly sequenced. The comparison demonstrated that DHPLC detected all alterations found by direct sequencing. No false-positive signals were seen in cases of homozygous sequences. Further, no false-negative results were ever obtained in women with mutations or polymorphisms, or both. In cases of known genetic variations, the nature of the alterations could be predicted by DHPLC. We also compared different separation matrices. Up to about 500 injections, no significant differences in sensitivity could be observed between poly(styrene divinylbenzene) and end-capped silica based columns. However, after more than 500 injections, the resolution of hetero- from homoduplex deteriorated rapidly on silica columns.

BRCA1 Protein↗
PubMed1999

Successful postexposure rabies prophylaxis after erroneous starting treatment.

BACKGROUND: Postexposure prophylaxis, adequately applied after exposure to a rabid animal, is highly effective in prevention of human disease. Deviations from the recommended vaccination postexposure treatment protocol have been associated with vaccination failure and human mortality. We investigated an incident in which seven Israel Defense Forces soldiers were bitten by a rabid fox and initially treated not in accordance with the recommended vaccination protocol. METHODS: The soldiers received modified anti-rabies postexposure prophylaxis, including a higher dosage of both the active and the passive vaccines. The humoral antibody response was monitored subsequently. RESULTS: All soldiers showed a satisfactory increase (above 0.5 UE/ml by ELISA) in serum anti-rabies antibody titers. None developed the disease more than a year after follow-up. CONCLUSIONS: Strict adherence to the treatment guidelines following an injury by a rabid animal is of utmost importance. We suggest possible compensatory management after a potentially lethal deviation from protocol.

Administration, Topical↗
PubMed1999

Laparoscopic treatment of colonic perforations related to colonoscopy.

BACKGROUND: Colonic perforations associated with colonoscopy are rare but major complications. Conservative treatment is less invasive than major surgery, but any case of failure leads to more extensive surgical procedures with a higher morbidity and mortality than the immediate operative repair. To reduce the invasiveness of major surgery and avoid the risk of failure, we introduced laparoscopic techniques to deal with iatrogenic colonic perforations. METHODS: Each colonic perforation was identified by diagnostic laparoscopy. The perforation was then characterized by size and extent of thermal damage into one of three types, followed by type-dependent treatment (suture, tangential resection, segmental resection, or open procedure). Operative time, complications, clinical outcome, and patient satisfaction were recorded. RESULTS: Seven patients underwent diagnostic laparoscopy for colonic perforations. Laparoscopic treatment was performed on five patients (one simple closure by suture, three tangential resections, and one segmental resection). Two cases required open procedures. There was one intraoperative complication that necessitated conversion. There were no postoperative complications. All laparoscopically treated patients were satisfied with their clinical outcome and cosmetic results. CONCLUSIONS: Laparoscopic treatment seems to reduce the invasiveness and morbidity of major surgery. At the same time, it is more definitive than conservative treatment, so that we now prefer to use laparoscopic techniques to treat colonic perforations related to colonoscopy.

Adult↗
PubMed1999

Effects of pH on kinetic parameters of the Na-HCO3 cotransporter in renal proximal tubule.

The effects of pH on cotransporter kinetics were studied in renal proximal tubule cells. Cells were grown to confluence on permeable support, mounted in an Ussing-type chamber, and permeabilized apically to small monovalent ions with amphotericin B. The steady-state, dinitrostilbene-disulfonate-sensitive current (DeltaI) was Na+ and HCO3- dependent and therefore was taken as flux through the cotransporter. When the pH of the perfusing solution was changed between 6.0 and 8.0, the conductance attributable to the cotransporter showed a maximum between pH 7.25 and pH 7.50. A similar profile was observed in the presence of a pH gradient when the pH of the apical solutions was varied between 7.0 and 8.0 (basal pH lower by 1), but not when the pH of the basal solution was varied between 7.0 and 8.0 (apical pH lower by 1 unit). To delineate the kinetic basis for these observations, DeltaI-voltage curves were obtained as a function of Na+ and HCO3- concentrations and analyzed on the basis of a kinetic cotransporter model. Increases in pH from 7.0 to 8.0 decreased the binding constants for the intracellular and extracellular substrates by a factor of 2. Furthermore, the electrical parameters that describe the interaction strength between the electric field and substrate binding or charge on the unloaded transporter increased by four- to fivefold. These data can be explained by a channel-like structure of the cotransporter, whose configuration is modified by intracellular pH such that, with increasing pH, binding of substrate to the carrier is sterically hindered but electrically facilitated.

Animals↗
PubMed1999

Duodenal duplication cyst manifested by duodeno-jejunal intussusception and hyperbilirubinemia.

A rare case of duodenal duplication cyst containing stones in a 17-year-old patient is presented. The cyst, acting as a leading point for duodeno-jejunal intussusception caused proximal small bowel obstruction and hyperbilirubinemia. Preoperative diagnosis was based on abdominal computerized tomography. At operation, the cyst wall was unroofed creating free drainage into the duodenal lumen without damaging the biliary and pancreatic ducts with resolution of symptoms.

Adolescent↗
PubMed1999

Analysis of the costs of surgery for Hirschsprung's disease: one-stage laparoscopic pull-through versus two-stage Duhamel procedure.

The purpose of this report is to evaluate the cost-effectiveness of a single-stage laparoscopic pull-through for Hirschsprung's disease compared to the traditional two-stage Duhamel procedure. In this series of 33 children, the length of hospitalization (2.5 +/- 3.5 vs 10.6 +/- 3.9, p < 0.01), cost (19,088 +/- 13,075 vs 34,110 +/- 19,443, p < 0.05), and complications were all significantly less with the laparoscopic assisted pull-through compared to the more traditional open approach.

Child↗
PubMed1998

Single blood donor exposure programme for preterm infants: a large open study and an analysis of the risk factors to multiple donor exposure.

UNLABELLED: As the need for blood transfusions of very preterm infants remains considerable, various strategies are considered to minimize exposure to multiple blood donors along with blood wastage. In a large population of very preterm infants born between 24 and 31 weeks' gestation, we undertook an open study to assess the efficacy of a single blood donor exposure programme and to determine, among the population enrolled in this programme, the risk factors for exposure to multiple donors. One hundred and forty-two neonates were included in a single donor exposure programme with a 35-day expiry date blood unit. Though no inflation in the total number of transfusions was noticed, there was a 55% overall reduction in the total number of required donors. To determine the risk factors for exposure to multiple donors in this population, 114 neonates alive after the expiry date of the first unit of packed red blood cells were selected. The greatest and the most extending transfusion requirements were observed in very preterm infants born before 28 weeks' gestation and in those born after but with an intra-uterine growth retardation below the 10th percentile. Indeed, 70% of those high-risk infants were exposed to a second blood donor and more than 85% of the group exposed to a second donor belonged to this high-risk population. CONCLUSION: Neonates with a very high risk of a more-than-one donor exposure were born before 28 weeks' gestation or between 28 and 31 weeks but with an intra-uterine growth retardation below the 10th percentile.

Blood Donors↗
PubMed1998

Voltage and cosubstrate dependence of the Na-HCO3 cotransporter kinetics in renal proximal tubule cells.

The voltage dependence of the kinetics of the sodium bicarbonate cotransporter was studied in proximal tubule cells. This electrogenic cotransporter transports one Na+, three HCO3-, and two negative charges. Cells were grown to confluence on a permeable support, mounted on a Ussing-type chamber, and permeabilized apically to small monovalent ions with amphotericin B. The steady-state, di-nitro-stilbene-di-sulfonate-sensitive current was shown to be sodium and bicarbonate dependent and therefore was taken as flux through the cotransporter. Voltage-current relations were measured as a function of Na+ and HCO3- concentrations between -160 and +160 mV under zero-trans and symmetrical conditions. The kinetics could be described by a Michaelis-Menten behavior with a Hill coefficient of 3 for HCO3- and 1 for Na+. The data were fitted to six-state ordered binding models without restrictions with respect to the rate-limiting step. All ordered models could quantitatively account for the observed current-voltage relationships and the transinhibition by high bicarbonate concentration. The models indicate that 1) the unloaded transporter carries a positive charge; 2) the binding of cytosolic bicarbonate to the transporter "senses" 12% of the electric field in the membrane, whereas its translocation across the membrane "senses" 88% of the field; 3) the binding of Na+ to the cotransporter is voltage independent.

Amphotericin B↗
PubMed1998

Dissatisfaction encodes a tailless-like nuclear receptor expressed in a subset of CNS neurons controlling Drosophila sexual behavior.

The dissatisfaction (dsf) gene is necessary for appropriate sexual behavior and sex-specific neural development in both sexes. dsf males are bisexual and mate poorly, while mutant females resist male courtship and fail to lay eggs. Males and females have sex-specific neural abnormalities. We have cloned dsf and rescued both behavioral and neural phenotypes. dsf encodes a nuclear receptor closely related to the vertebrate Tailless proteins and is expressed in both sexes in an extremely limited set of neurons in regions of the brain potentially involved in sexual behavior. Expression of a female transformer cDNA under the control of a dsf enhancer in males leads to dsf-like bisexual behavior.

Amino Acid Sequence↗