Intraoperative enteroscopy: ten years' experience at a single tertiary center.

BACKGROUND AND METHODS: Intraoperative enteroscopy is an invasive technique for small bowel investigation. It enables us to investigate the entire small intestine and to treat pathological findings by endoscopic or surgical means at the same time. The investigation is invasive and that is why the proper indication is mandatory. RESULTS: Forty-one intraoperative enteroscopies were performed at our center within a 10-year period. The procedure was diagnostic in 37/41 patients (90.2%); in 3 patients no pathology was found, and in 1 patient we found only previously diagnosed celiac disease. The investigation was therapeutic in 35/41 (85.4%) patients; 2 patients with small bowel ulcers did not require any intraoperative therapy. The pathological findings were arteriovenous malformations (found in 12 patients), small bowel NSAID-induced or Crohn's ulcers (8 patients)--ulcerations and arteriovenous malformations were simultaneously found in three patients; carcinoid of the small intestine (5 patients); Peutz-Jeghers syndrome (5 patients); bleeding polyps (2 gastrointestinal stromal tumors, 1 paraganglioma, and 1 lipoma--in 4 patients); Rendu-Osler-Weber disease (2 patients); multiple cavernous hemangiomas in blue rubber bleb nevus syndrome (1 patient); Henoch-Schönlein purpura (1 patient); aortoenteral fistula (1 patient); and retrograde intussusception of Meckel's diverticulum (1 patient). In five patients with Peutz-Jeghers syndrome, 6-22 hamartomas (median of 18 per session) were removed by means of endoscopic polypectomy during intraoperative enteroscopy. There were no major procedure-related complications in our series. CONCLUSIONS: Intraoperative enteroscopy is accepted as the ultimate diagnostic procedure for complete investigation of the small bowel. Despite the introduction of double-balloon enteroscopy into clinical practice, intraoperative enteroscopy will be reserved for those cases where double-balloon enteroscopy cannot be performed or fails to investigate the entire small intestine, especially to prevent excessive bowel resection.

Gastric banding: intraoperative and early postoperative complications and their prevention.

BACKGROUND: Intraoperative and early postoperative complications in patients operated for morbid obesity were assessed. METHODS: 114 morbidly obese patients underwnent gastric banding (non-adjustable or adjustable). The influence of body mass index (BMI), age and preoperative morbidity on the occurrence of postoperative ventilatory disturbances was evaluated. RESULTS: Risk of postoperative ventilatory disorders increased with preoperative cardiovascular disease (p < 0.01), diabetes (p < 0.05), and increasing BMI (p < 0.01). Age, hypertension and pulmonary disease did not influence significantly the risk of complication in this series. There have been no deaths. CONCLUSION: In patients undergoing banding for morbid obesity, the potential for ventilatory complications increases with higher BMI, diabetes and cardiovascular disease in the preoperative comorbidities. Principles for postoperative care were developed.

Calf serum- and cell surface proteins released from cultured avian sarcoma virus-transformed and untransformed rat fibroblasts.

Cell surface and calf serum proteins were released in vitro from cultured virus-transformed, chemically transformed and normal rat embryo fibroblasts in 0.2 M urea containing serum-free culture medium. Fibronectin (LETS protein) was found in considerably higher amount in medium from normal rat embryo fibroblasts than in that from transformed cells. Major calf serum protein released from cultured normal and transformed rat cells corresponded by its electrophoretic mobility to calf serum albumin. No substantial differences in electrophoretic patterns of calf serum proteins released from normal and transformed cells into the medium were found. Time course of release of serum proteins from cell surface was measured quantitatively with the aid of radioiodinated Staphylococcus aureus protein A radioimmune assay. More than 50% of cell bound calf serum proteins remained on cell surface after 24 hours of incubation in serum-free medium.

Growth of mammalian cell lines in a spinner culture.

Several mammalian cell lines, derived from embryonic, normal adult and malignant tissues were adapted for long-term growth in agitated fluid medium. A simple method of spinner culture is described, employing a portable magnetic stirrer. Adaptation of cells for growth in spinner culture took a relatively short time, usually a few weeks, depending on the type of cell line used. Characteristics of cell lines maintained in spinner culture are described in details.

Complement-fixing soluble antigen from rat lymphoma (C58NT)D. Preliminary characterization of the antigen and some positive human sera.

A soluble complement-fixing antigen prepared from the Gross virus-induced rat lymphoma (C58NT)D reacted in complement-fixation with the sera of some patients with different malignancies (particularly acute leukemia) and with the sera of some healthy individuals. Results of gel filtration of crude (C58NT)D antigen on Sephadex G 200 indicate that the antigenic activity is eluted with the void volume of the column, i. e. in fractions corresponding to proteins of low molecular weight. Gel filtration of some positive patient sera indicate that complement-fixing activity of tested sera is associated with fractions corresponding to IgM and/or IgG.

Trends in morbidity on the basis of newly-reported cases of malignant skin melanoma (172 ICD) and other skin neoplasms (173 ICD) in Czechoslovakia during the period 1961--1972.

According to newly-reported cases of all malignant neoplasms of the skin (172 + 173 ICD) morbidity rate for the period 1961--1972 showed a rising trend of statistical significance only in Bohemia; while in Slovakia it persisted at a practically steady level throughout the period followed. Malignant skin melanomas (172 ICD) were on the increase in both regions, the rate being of statistical significance only in men in Bohemia. Morbidity from other malignant skin neoplasms (173 ICD) during the same period showed a stable pattern in Slovakia while it rose steadily in a significant degree in both, males and females in Bohemia. The causes for these varying trends must be further analyzed.

Trends in mortality rate from malignant skin melanoma and other malignant skin neoplasms in Czechoslovakia during the period 1921--1970.

Mortality from malignant skin melanoma (172 ICD) has an ascending trend in Czechoslovakia (CSSR) the rate being higher after the year 1960 in Bohemia (CSR) than the Slovakia (SSR). Mortality from other malignant neoplasms of the skin (173 ICD) declines; this decline for the whole of Czechoslovakia has been statistically influenced mainly by the lower death rate from this cause in Bohemia, for in Slovakia this mortality rate has risen, although significantly only in women. The proportion of deaths from malignant skin melanoma (172 ICD) out of the total number of deaths from malignant skin neoplasms (172 + + 173 ICD) is relatively low in Czechoslovakia--lower in Slovakia than in Bohemia.

Cell surface alterations of avian sarcoma virus B77- and 3,4-benzo(a)pyrene-transformed rat fibroblasts. I. Cell surface proteins characterized by two-dimensional electrophoresis.

Plasma membrane proteins of avian sarcoma virus B77-, 3,4-benzo(a)pyrene- and spontaneously transformed rat fibroblasts were metabolically- and cell surface radiolabeled and analyzed by electrophoresis under denaturing conditions (SDS-PAGE) or by two-dimensional electrophoresis. Most extensive series of protein alterations on transformed cells was detected by two-dimensional electrophoresis of cell surface proteins radiolabeled by lactoperoxidase catalyzed radioiodination: a fibronectin-like 220k protein with two isoelectric forms (pI 5.9 and 6.2-6.6), a 180k protein of pI 6.2-6.6 and two proteins of 48k and 50k with pI approximately 5.4-all decreased on examined transformed cells. In addition to these proteins, a series of proteins more markedly detectable on transformed cells was found: a 110-120k (pI 5.6) protein found particularly on 3,4-benzo(a)pyrene- and spontaneously transformed cells, a series of proteins in the region of molecular weights 46-54k with pI 5.5-6.0, two proteins of 35k (pI 6.2 and 6.4) and two further, basic proteins of 38k (pI approximately 7.2 and 7.8). Some of these changes were visualized also by further techniques for radiolabeling of proteins, i.e. reductive methylation of cell surface proteins, metaboling radiolabeling by 3H-leucine followed by plasma membrane isolation, as well as by protein staining after two-dimensional electrophoresis of isolated plasma membrane proteins. A 70k and a 120k radioiodinated proteins were immunoprecipitated from all transformed cells and, to a markedly lesser extent from untransformed fibroblasts by the immune serum against rat type-C endogenous virus. A similar 70k protein was immunoprecipitated from avian sarcoma virus B77-transformed cells also by the antiserum against Friend murine leukemia virus envelope glycoprotein gp70.

Cell surface alterations of avian sarcoma virus B77-and 3,4-benzo(a)pyrene-transformed rat fibroblasts. II. Cell surface glycoproteins characterized by two-dimensional electrophoresis.

Cell surface proteins of avian sarcoma virus B77-, 3,4-benzo(a)pyrene- and spontaneously transformed rat fibroblasts were tritium-radiolabeled by mild periodate oxidation followed by tritiated sodium borohydride reduction and by galactoseoxidase/NaB3H4 technique. Radiolabeled cell surface sialoglycoproteins were analyzed by electrophoresis under denaturing conditions (SDS-PAGE), or by two-dimensional electrophoresis (isoelectric focusing, SDS-PAGE). Following major glycoproteins were quantitatively decreased on all examined transformed cells: a 220k glycoprotein with pI of approximately 6.2, a 180k glycoprotein with pI of approximately 6.0-6.2, a 110k glycoprotein (pI 6.2). A series of further sialoglycoproteins was quantitatively increased on all examined transformed cells, as follows: a markedly increased 70k glycoprotein (pI approximately 4.8) and a 120k glycoprotein (pI 5.0-5.2) increased more markedly on onco-virus-transformed and spontaneously transformed cells. Further sialogalactoprotein (28k, pI 5.8) was visualized as increased on all examined transformed cells only by galactoseoxidase/NaB3H4 technique. A 70k sialoglycoprotein was immunoprecipitated from transformed cells by the immune serum against Friend murine leukemia virus envelop glycoprotein gp70 and, to a markedly lesser extent, from untransformed cells. A similar glycoprotein, together with an another 120k glycoprotein were precipitated from transformed cells and to a minor extent from untransformed cells also by an antiserum against endogenous rat type-C virus.

Inhibition and promotion of growth of B77-virus-induced rat tumor with KCl-solubilized tumor cell components.

Rats were immunized with a single and repeated doses (1.0 mg of protein per dose per animal) of KCl-solubilized thymus and avian sarcoma virus B77-induced rat tumor cells LWF B77 prior to challenge with the same tumor cells. Results obtained showed that single administration of KCl-solubilized tumor cell material resulted in inhibition of growth of syngeneic tumor cells. Repeated inoculations of the same material, however, promoted tumor growth. Possible explanation of this dual effect is discussed.

Retrovirus like particles produced by human embryonal cells and cell lines derived from human malignancies. II. Protein structure.

Very small amounts of retrovirus-like particles were isolated from tissue culture media of various types of human cell lines derived from malignant as well as normal cells. The aim of the present study is the characterization of protein profiles of these isolates. The comparison of protein profiles of isolated human virus-like particles with the profiles of well characterized animal as well as human exogenous (LAV/HTLV-III) retroviruses revealed a 25k protein (p25) to be a major protein or at least one of the protein components of human retrovirus-like particles.

An attempt to induce formation of antibodies against endogenous retrovirus BP 6 in syngenic rats.

An attempt was made to induce production of antibodies against C-type endogenous rat retrovirus BP 6. Syngenic Lewis rats were immunized with viable tumor cells, induced with benzo(a)pyrene, continuously producing BP 6 virus. By use of immunoblotting technique, neither short- nor long-term immunization did cause formation of any detectable amounts of antibody against structural proteins of the virus. On the other hand, in immunoblotting, antibodies arised in rats immunized with purified mouse leukemia virus have been found to cross-react with endogenous rat retrovirus.

Concentration of retroviruses by low-speed centrifugation.

Concentration of retroviruses from volumes of up to 6 liters of medium by low-speed centrifugation is described. In contrast to pelleting, no damage or aggregation of particles was observed. Surface glycoproteins were also fully preserved. This method enables simple handling of relatively large volumes of medium. Highly purified mouse mammary tumor virus (MMTV) was obtained and its transframe protein p30 in SDS-PAGE was recognized as a double band.

Purification and protein composition of endogenous rat viruses.

Endogenous retroviruses are not in the majority of cases the cause of any neoplasia, except for the laboratory conditions. As far as they might serve for the evolution of pathogenic retroviruses more attention should have been paid to them. In this paper we introduce some approaches to the purification of rat endogenous retroviruses to such a degree of purity that enabled satisfactory SDS-PAGE analysis of its structural proteins. Purities of samples obtained by usual purification methods, long-term isopycnic centrifugation at a high gravity force and velocity centrifugation are compared. Protein profile of rat endogenous virus in SDS-PAGE is compared with the ones of other retroviruses. For the first time the evidence was obtained for the striking similarity between electrophoretic protein profile of rat endogenous virus WERC and feline leukemia virus. The major structural proteins of rat endogenous retrovirus and feline leukemia virus cannot be distinguished even when resolution long gradient PAGE had been employed. The accordance of electrophoretic mobilities of major structural proteins in SDS-PAGE can indicate the relatedness of retroviruses.

An in vitro study of the oncogenic effects of two variants of avian sarcoma virus B77 on rat embryonal fibroblasts.

Two variants of avian sarcoma virus B77 have been tested for their heteroinductive oncogenic in vitro effects on rat embryonal cells. Variant 22-B77V belonging to subgroup B, had been cultured for a long time exclusively on its original host, i.e. on chicks, before being used in the study. This variant of B77V yielded negative results in repeated experiments on rat cells. Variant 55-B77V belonging to subgroup C and which had been made to pass through rat cells, caused a malignant transformation in the latter. The resultant type of this interaction were virus-producing tumorous cells (designated LWF B55). This study present some of the basic characteristics of these cells.

Alterations in surface proteins of rat cells transformed by avian sarcoma virus B77.

Cell membrane proteins of avian sarcoma virus B77V--transformed cells LWF B55 and LWF B77 and uninfected rat embryo fibroblasts were analyzed by SDS acrylamide gel electrophoresis. Following alterations in cell surface proteins of LWF B55 and LWF B77 cells were found: one, a slight decrease of two high molecular weight proteins the larger of which corresponds presumably to "LETS" protein and a decrease of a protein with approximative molecular weight 50 000; two, increase in content of proteins with molecular weight of about 90--95 000 and 70--75 000 as well as a marked increase of a protein with molecular weight of 30--35 000.

Retrovirus-like particles produced by human embryonal cells and cell lines derived from human malignancies.

Very small amounts of retrovirus-like particles were obtained from tissue culture fluids of various human cell lines. The particles were found in almost all cultures of rapidly growing human cells (embryo fibroblasts, various types of leukemias, melanoma, urinary bladder, lung and mammary carcinomas). Morphology and some biochemical characteristics of purified samples of these particles are presented. The particles resemble C-type mammalian retroviruses.

High prevalence of circulating antibodies to MuLV p30 antigen in human sera. An autoimmune response?

To study the possible involvement of a murine leukemia virus (MuLV) related agent in human cancer, an extensive immunoblotting analysis of human sera (cancer, autoimmune as well as control normal ones) for the presence of antibodies to MuLV structural proteins was performed. Out of 350 sera, 89 reacted with gag precursor Pr65, 72 reacted with major viral core protein p30 and five with the matrix protein p15. Antibody reactivity to the env-encoded glycoprotein gp70 was detected in 7 cases out of 16 sera tested. There were no significant differences between pathological and normal sera concerning the patterns and the frequency of the reactivity. Sera from patients with various malignancies (mainly with breast cancer) generally displayed more intensive signals to MuLV p30 than normal sera. Epitope mapping revealed that MuLV p30-reactive antibodies recognize an antigenic determinant(s) located at the carboxyterminus of the protein.

Presence of complement-fixing antibodies against antigens of Gross virus-induced rat lymphoma and normal rat thymus in sera of patients with some forms of malignancies.

Presence of complement-fixing antibodies against soluble antigen(s) of Gross virus-induced rat lymphoma W/Fu(C58NT)D was determined in sera of 180 healthy donors, in sera from 100 healthy pregnant women and in sera obtained from 139 patients with acute myelosis, acute lymphadenosis, acute undifferentiated leukosis, chronic myelosis, chronic lymphadenosis and Hodgkin's disease. Antibodies against soluble complement-fixing antigen from W/Fu(C58NT)D rat lymphoma were found in 33 (5).5%) out of 64 sera of patients with acute leukemia, in 15 out of 60 sera of patients with Hodgkin's disease, in 17 (9.4%) out of 180 healthy donors and in 22 (22.0%) out of 100 sera from healthy pregnant women. None of 15 sera from patients with chronic leukemia were positive. Titers of complement-fixing antibodies in positive sera remained unchanged after absorption with pooled lymphocytes from healthy human donors. Some selected positive sera reacted also with a soluble antigen prepared from normal rat thymus. Nature of complement-fixing antibodies detected and of corresponding antigens is discussed.

Assessment of sphincterotomy results in patients treated for anal fissure.

OBJECTIVE: This paper summarizes results of a study of 90 patients operated on for chronic anal fissure with lateral sphincterotomy (LS) between 1985 and 1997. METHODS: Preoperative and postoperative complaints, anal examination and continence were evaluated. All patients were asked to rate the results of LS as very good, good, satisfactory or unsatisfactory. RESULTS: Sixty-six patients (73.3%) judged the results of LS to be very good. Only 4 (4.4%) judged the results unsatisfactory and 1 patient (1.1%) considered the results satisfactory. Recurrent anal fissure was found in 4 cases (4.4%). Two patients developed a fistula in connection with the procedure. CONCLUSIONS: Lateral sphincterotomy is a safe and simple operation that yields good results. However, LS should be diagnosed and performed by a surgeon skilled in proctologic surgery.