[The usefulness of extended radical mastectomy in patients with breast carcinoma]. Extended radical mastectomy (EXT) in 1, 137 patients with breast carcinoma was devised to eradicate all possible sites of local nodal metastases including internal mammary chain during the years 1962 through 1990. The tumors had to be mainly any T located in medial-half, central (excluding less 1.0 cm or histologic type with good prognosis) and T size measuring 3.0 cm or more in lateral-half. Prognostic outcomes were analysed by the presence or absence of nodes involved. The 10-year relapse free survival in patients with internal mammary node (IMN) only invaded and both axillary (AX) & IMN invaded were 47.8% and 40.1%, respectively. The incidence of relapse free survival at 10 years in relation to the number of positive AX and IMN indicated virtually favorable--1 positive: 71.4%, 2 or 3 positive: 61.3% and 4-9 positive: 53.7%. In our study, the frequency of IMN involvement calculated 24.1% from all quadrants of the breast. In comparison of nonrandomized series with medial-half and lateral-half tumors which involved positive IMN, comprising 95 and 28 patients of the total (n = 495), relapse free survivals at 15 years were 36.5% and 17.5%, respectively. It was presumed that the benefit of EXT would be found in medial-central tumors.


[Attempt for modified radical mastectomy in the treatment of early breast cancer]. Until five years ago the Aichi Cancer Center had adopted the standard radical mastectomy where breast, major and minor pectoral muscles, and axillary adipose tissues were resected en bloc. In case tumor was found at the inner half or the center of the breast, parasternal lymph node dissection was concomitantly carried out. Recently, however, we begin to perform Auchincloss by which major and minor pectoral muscles can be conserved, as an attempt for modified radical mastectomy. In some cases, furthermore, breast conserving operation is made. The reasons why we changed the operation methods are: 1) Due to the advancement of diagnostic imaging and widespread self examination, percentage of early breast cancer has considerably increased. 2) Postoperative adjuvant therapy, especially adjuvant endocrine therapy, has remarkably progressed.


[The spread of breast cancer and indications of breast preserving operation]. Pathological investigations for intramammary spread of breast cancer of 205 partially resected specimens were performed by making continuous section every 5 mm width of the whole specimen. The materials were 167 quadrantectomized and 38 lumpectomized specimens. The results showed that the margin of 15.6% of quadrantectomized and 28.9% of lumpectomized specimens were positive for cancer, main causes of which were intraductal spread of cancer occupying 65% of positive margin in quadrantectomy and 91% in lumpectomy, multiple cancer, interstitial spread of cancer and so on. Multiple cancers were found in eleven (6.6%) quadrantectomized and in one (2.6%) lumpectomized specimens. Second cancers were 11 noninvasive ductal and 3 invasive cancers, including two triple cancers. Fifteen cases of quadrantectomy and 4 of lumpectomy were changed to be mastectomized because of positive margin or nodal involvements. Radiotherapy was performed for 33 cases. The median 31 months follow-up results of 186 partially mastectomized breast in 184 patients were as follows; one local recurrence in lumpectomy, two new cancers in residual breast, two distant metastases and one death for other cause of death. Quadrantectomy plus axillary dissection without radiotherapy assured of pathological complete resection was safe enough at the present.


[Breast preserving operation for early breast cancer--clinical implications and biological aspects of residual tumor]. Among the 93 surgical specimens of stage I breast cancer treated by breast preserving operation, surgical margins were less than 10mm in 23 (25%) cases because of intraductal spread. Such tumors with extensive intraductal component, however, were frequently positive for estrogen receptor (ER). Possible risk factors for local failure have been studied in an attempt to exclude high-risk patients not suitable for breast preserving operation. The results so far obtained suggest that biological markers such as c-erb B-2 or PS2 proteins may provide useful informations to select patients who are most likely to recur. In addition, adjuvant endocrine therapy may be recommended for the patients treated by breast preserving operation.


[Surgery in the treatment of stage I, II breast cancer]. Stage I and II breast cancer is thought to be operable cancer. Possible surgical methods for such breast cancer could be grossly divided total mastectomy and breast preserving surgery (BPS) with axillary node dissection. In is necessary to obtain clear surgical margin after performing BPS. However it is difficult to know preoperatively the exact resected margin which is either clear or not. In order to select the cases performing BPS, we intended to compare the degree of coexisting intraductal component with histologic types and some factors such as DNA ploidy. ER and expression of cerb B-2 which is concerned in the malignant potential of breast cancer. Intraductal component is more frequently seen in papillotubular carcinoma. Diploid tumor is increased with increasing intraductal component in breast cancer. Precise postoperative microscopic study of resected specimen and tight observation of the patients received PBS for long period should be emphasized. We used methylsalicylate packed method on 2mm slice in thick by Wellings for postoperative histological study and investigated the intraductal architectural spreading under the dissecting microscope. This method is useful to define the three-dimensional architecture of spreading of intraductal carcinoma. After proving clear surgical margin by this analysis, we usually do not recommend the radiation therapy.


[Breast conservation treatment of early breast cancer--eight-year experience]. In the period between 1983 and 1991, we experienced 80 cases of early breast cancer treated by conservative surgery and postoperative radiotherapy. Our indications for breast conservation treatment are the size of tumor (less than 2 cm), the location (more than 3 cm away from the nipple) and no palpable axillary nodes. Our surgical procedures of quadrantectomy and level I and II axillary dissection are presented. The shape of the skin incision should be elliptical with the major axis radial from the nipple for good cosmetic results. The entire quadrant of the breast containing the primary carcinoma is radically removed with the superficial pectoralis fascia. At present, one breast recurrence and two distant metastases have developed. A further follow up study is needed to prove the safety of breast conservation treatment.


[Application and technique of breast conservative treatment of breast cancer]. In patients with breast cancer who had undergone breast conservative treatment involving lumpectomy, axillary lymph node dissection and irradiation, the surgical results and risk factors for remaining cancer foci in the conserved breast were investigated. This form of surgery was performed in 25 patients with T1,N0,-Ia,M0 breast cancer. The cut end of the resected specimen was histopathologically positive for cancer cells in 48% of the patients, but there has been no local or distant recurrence in any patient to date with a mean follow-up period of 17.2 months after surgery. In most breasts resected after lumpectomy, slight cancer foci remained in mammary glands at the sites within 1.0cm distant from the cut end. Extensive remaining cancer lesions were found among cases with advanced intraductal spread or central necrosis in the intraductal foci. The above findings indicate that breast conservative treatment by lumpectomy may result in histopathologically remaining cancer foci, although these can be controlled by irradiation therapy. However, application for this treatment requires much caution when severe intraductal spread or central necrosis is present in the intraductal foci, because extensive cancer foci may remain postoperatively in such cases.


[Conservative surgery and radiotherapy of early breast cancer]. Radiation dose-effect relationship is reviewed. In conservative treatment of early breast cancer, necessity of radiation is discussed and surgery with radiotherapy technique in our hospital is presented. Our procedures consist of lumpectomy which is removal of 1 cm of normal tissue containing primary breast cancer and complete axillary dissection and postoperative radiotherapy of 46 Gy with 9 Gy of electron boost. Since November 1987, 51 patients treated by this procedure have shown a good local control and a satisfactory cosmetic results. We can conclude that the positive aspects of radiotherapy in the management of conservative therapy for early breast cancer are overwhelming.


[Postgraduate education in surgery and surgical specialties]. Accreditation system of postgraduate training in surgery was started by the Japan Surgical Society (JSS) in 1979, and since then more than 6,800 surgeons has been certified by examination performed by JSS in general surgery. This accreditation system has greatly contributed to the improvement of postgraduate training in general surgery in Japan; since the start of this system operative experiences of surgical residents have increased by 40% and reached to the level of 580 during the 4-year period. In order to further improve the level and quality of surgical care of the patients in Japan, it may be necessary to reform and adjust the accreditation system of postgraduate training in clinical specialties which was started quite separately by each medical specialty society. Probably, to establish a board for each medical specialties should become necessary in the near future to be officially recognized in the medical system of Japan.


[Recent progress in endocrine surgery: more refined and rational interventions for varied endocrine diseases]. Better understanding of pathogenesis and natural history of various endocrine diseases along with development of new and sensitive assays for accurate measurement of hormones and of modern image diagnostic procedures for clear demonstration of pathological tissues has enabled us to detect many kinds of endocrine diseases much more frequently than before and to perform a more refined and rational surgical intervention. Endocrine surgery is now a challenging and fascinating field of general surgery. This fact is best exemplified as follows: (1) different therapeutic approaches for intrathyroidal and extrathyroidal papillary carcinomas of the thyroid, (2) early detection and 131I treatment of clinically occult hematogenous metastasis of follicular carcinoma of the thyroid, (3) clinical usefulness of the serum calcitonin and CEA concentrations as sensitive tumor markers in the treatment of medullary thyroid carcinoma, (4) rational use of preoperative localization test and unilateral neck exploration for parathyroid adenoma, and specific surgical intervention for metastatic parathyroid carcinoma, (5) how to treat adrenal incidentalomas and a trial of subclassification of pheochromocytomas into epinephrine- and norepinephrine-secreting tumors, and (6) intraoperative localization of insulinoma by ultrasound and newly developed surgical approaches for gestrinoma.


[Infection of methicillin resistant Staphylococcus aureus (MRSA) in patients with cardiovascular operations: four years' clinical analysis of the effect of strategy of prophylactic and treatment]. To evaluate the effect of strategy to reduce the rate and outcome of MRSA infection, 940 patients with cardiovascular operations from 1988 to 1991 were analyzed in aspect of the infection rate and outcome per year. Prophylactics and treatments which have been done during recent four years were) to use the inhalation of vancomycin for MRSA carriers, to quit the intravenous administration of prophylactic antibiotics on third postoperative day, to limit medical staffs to treat the patient with MRSA infection, to choose the most susceptible combination of two antibiotics for MRSA in each patients by periodical checking of bacterial cultures. The percentages of patients with infection to all patients detected with MRSA per year were decreased from 80% in 1988, to 66% in 1989, 31% in 1990, and 30% in 1991. The numbers of patients who died from MRSA infection were 4 in 1988 and only one in 1989 and 1991. MRSA infection in cardiovascular surgery could be reduced by sophisticated methods of prophylactics and treatments.


[Postoperative infection with methicillin-resistant Staphylococcus aureus and its control measures in patients with esophageal cancer--prediction of high risk patients]. Whether or not compromised host defense prior to surgical treatment is related to development of infection with methicillin-resistant staphylococcus aureus (MRSA) following major surgery was examined. Of 22 patients undergoing esophagectomy for cancer between 1989 and 1990, 5 were free from MRSA colonization and infection (group I) while 8 had MRSA colonization without infection (group II) and 9 had MRSA infection (group III). Levels of neutrophil cytocidal functions, complements, immunoglobulins and cell-mediated immunity were not significantly different among these three groups. The mean of anti pneumococcal polysaccharide (PPS)-IgG was significantly lower in group III than in group I (p < 0.05) and tended to be lower than in group II (P = 0.08). All infected patients showed a titer < 600 EU while all but one non-infected patient did so > 600 EU. Serum IgG2 levels positively correlated with anti-PSS IgG levels. Thus, we concluded that a preoperative evaluation of antibody response against polysaccharides and serum IgG2 levels can serve to predict development of MRSA-related infection following esophageal surgery.


[Managements of hospital MRSA infections in digestive tract surgery]. 1444 cases if digestive tract surgery in our department were studied for the prevention of postoperative MRSA infections, during the period from September 1987 to February 1992. They were divided into nine stages each six months. There were no significant differences in incidence of postoperative MRSA infections, after some managements for hospital infection among each stage. We used the antibiotics as follows; the first generation of cefem families was used for the upper digestive tract operations and the second generation of cefem families was used for the lower digestive tract operation, and the incidence of postoperative MRSA infections were diminished significantly. This study showed that managements for hospital infections and appropriate selection of antibiotics for the purpose of postoperative prophylactic use are effective for the prevention of MRSA infections.


[Prevention and treatment of MRSA infection in emergency surgical ward]. During two years of 1990 and 1991, methicillin-resistant Staphylococcus aureus (MRSA) strains were isolated from 121 patients who were admitted to our emergency ward with infection rate of 7% (121/1843). The end results were miserable and the mortality rate was 28%. The infection rate was significantly higher than that of general surgical ward (2%). Specimens of respiratory tract such as coughed-up sputum and tracheal secretion showed high rate of MRSA isolation, especially in patients who underwent tracheal intubation or tracheostomy. All of these MRSA strains were strongly resistant to antibiotics and were typed as coagulase II. MRSA strains were isolated from nostril of medical staff and in-ward patients while admission with rate of 9% and 3%, respectively. Meanwhile, the MRSA strains were also isolated from the materials surrounding the infected patients such as bed, door, gowns and hands of in-charge nurses. To prevent inter-patient transmission, the infected patients were placed in isolation room and cared with isolation technique. Handwashing with benzalkonium chloride-alcohol is strongly recommended. Intravenous injection of IPM and CEZ or VCM is indicated. Local use of MINO was reported to be useful. In conclusion, MRSA infection often occurs in the immunodepressive patients and is hard to cure.


[Hospital acquired infection in surgical field and its countermeasure present situation of anaerobes, P. aeruginosa and MRSA infection]. The important organisms of hospital acquired infection in surgical field are anaerobes, P. aeruginosa and MRSA. From 1977, isolation frequency of anaerobes from pus has been increasing remarkably and it became 60% in 1978. The isolation rates of P. aeruginosa increased 20 to 30%. Since 1988, the incidence of isolation of MRSA has increased markedly. Almost of all the coagulase typing of MRSA were type II, and it suggested the possibility of hospital infection. In 1988, high antimicrobial activity against MRSA was observed with vancomycin, imipenem, minocycline and new quinolones, but in 1990 the multiresistant strains against imipenem, minocycline and new quinolones were increased. Among the various antimicrobial agents examined, vancomycin demonstrated the strongest activity to MRSA. The symptomatic infections were postoperative enterocolitis, respiratory infection, biliary tract infection and wound infection. The most severe symptom was observed in MRSA enterocolitis. We conducted a nation wide questionnaire of postoperative enterocolitis from January 1988 to June 1990. 169 institutes replied, and 126 cases were involved in postoperative MRSA enterocolitis in 53 institutes. Eighteen cases died, and mortality rate was 14.3%. In the treatment of MRSA infection vancomycin was the most effective antibiotics. It is the most important to prevent nosocomial infection.


[Investigation on central venous catheter related sepsis]. We made an investigation on central venous catheter related sepsis (CRS) in recent 5 years (1987-1991). The incidence of CRS was high; 16.0% (125 out of 782 cases) or 13.1% (135 out of 1029 catheters). CRS occurred frequently during 2-3 weeks after catheter insertion. The incidence of CRS was not affected by the kind of disease (malignant or benign), complication (diabetes, liver cirrhosis, collagen disease) operation or administration of antibiotics. Eight percent out of 91 organisms isolated from culture of catheter tips were so-called resistant strains; multi-drug resistant Staphylococci (16), Pseudomonas aeruginosa (5), fungi (49), etc. Complications (shock, acute renal failure, secondary pneumonia, fungal endophthalmitis) broken out in 18 patients (14.4% out of 125 CRS). Fungi were isolated from 14 out of 18 complicated cases, furthermore fungi were isolated alone in 11 cases. No complication were seen among cases from which gram positive cocci were isolated alone. Body temperature and white blood cell count of complicated cases were significantly higher than those of uncomplicated cases. The duration until removal of catheter from outbreak of fever in complicated cases was significantly longer than that in uncomplicated cases.


[Rapid detection of mecA gene by nested PCR for diagnosis of methicillin resistance in Staphylococcus aureus]. In recent years, the most common causative organism of hospital infections has been methicillin resistant Staphylococcus aureus (MRSA). The major mechanism of beta-lactam resistance in MRSA is attributed to the production of a specific penicillin binding protein (PBP2'), which is a product of mecA gene, with extremely low binding affinities to beta-lactams. In the present study, we have established a rapid identification method of MRSA by sensitive detection of mecA gene using nested PCR. Nested PCR method amplifying the target DNA in two steps enhanced the efficiency of the second round amplification. By means of this method, mecA gene was successfully detected in clinical samples, such as blood, pus, sputum and feces within 3-4 hrs. Rapid diagnosis of MRSA-bacteremia is particularly important for prevention of sever systemic infection. There are some strains of S. aureus which possess mecA gene in spite of low minimal inhibitory concentration of DMPPC. In these strains expression of mecA gene is induced by contact of beta-lactams and they obtain methicillin resistance. Using nested PCR method, these latent MRSA are rapidly and certainly detectable. This method should be useful for early and effective detection of MRSA hospital infections.


[Early diagnosis and management against perioperative fungus infection]. Candida spp. is frequently detected from perioperative patients, especially from those requiring central venous catheter (CVC) management. CAND-TEC could detect the blood fungus infection easily and quickly. We analyzed 438 specimens from 57 patients requiring perioperative CVC management for more than three weeks, and investigated usefulness of CAND-TEC comparing with other serological methods. The dilution rate of CAND-TEC showed a positive correlation with amount of beta-D-glucan which was measured by Toxicolor test and Endotoxin-specific assay. It also showed a positive correlation with amount of D-arabinitol. When blood samples diluted more than four times were positive with CAND-TEC, the patients were considered to be infected, and accumulative infection rate was up to 84.2% three weeks after introduction of CVC management. CAND-TEC would be useful in detecting Candida infection compared with conventional methods, and the fungus infection was much more frequent incidence than ever expected. It is important to detect fungus infection early and respond immediately to avoid nosocomial infection.


[Hospital infection and our policy to control]. Since we organized the committee to control hospital infection in April 1980, we have adopted various investigations and strategies. When wide spread of infection caused by Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa was recognized in 1981 our strategies were set up of water faucets with footpedal, frequent to wash of hands and use of disposable paper towels. Investigation of HBV markers revealed that the positive rate of HBV antibody was 21% in scrub and ward nurse group. Vaccination was performed to our HVB antibody negative members to protect them from the infection of HBV antigen. After HBV antigen positive patients were operated, we sterilized the operating room using our ultraviolet ray irradiation apparatus. In 1990, 50 cases infected with MRSA were detected bacteriologically, which occupied 68.5% of all those infected with Staphylococcus aureus. Therefore, we cultured MRSA from nose swabs and investigated numbers of bacteria in the air which were gathered with air sampler in the operating rooms and wards.


[A study of progression in hepatocarcinogenesis using cell transplantation system]. It is important to distinguish a precancerous lesion and hepatocellular carcinoma (HCC) with diploidy or aneuploidy nuclear DNA pattern, not only in clinical cases but also in experimental carcinogenesis models. Using liver perfusion technique, we detected early HCC from persistent hyperplastic nodules (HN) which were induced in Wistar rats by intermittent 5-6 months administration of 2-acetylaminofluorene. This investigation was undertaken to assess both promotive and progressive effects of liver regeneration following partial hepatectomy (PH). Results are as follows: 1) Isolated hepatocytes of precancerous HN, which were transplanted into the spleen, didn't develop to HCC by 2 months after 70% PH of host liver. 2) Diced tissues of HCC, which were transplanted into the liver via portal vein, grew many metastasis in 10/10 by 7 weeks after PH, while 5/19 in control. 3) Nuclear DNA patterns of early HN-late HCC in rat liver were diploidy at the rate of more than 90% each. But it changed to aneuploidy, when inoculation of HCC for one month was repeated 7 times in the spleen.


[Mode of origin of esophageal squamous cell carcinoma--serial histopathologic and immunohistochemical studies]. I. Serial histopathologic study of esophageal squamous cell carcinoma. A review of 335 cases of squamous cell carcinoma disclosed 55 cases (16.4%) with glandular components in addition to the ordinary component of squamous cell carcinoma, suggesting that this type of esophageal tumor had originated not only from the covering squamous epithelium but from esophageal gland or ductal epithelium. Intra-epithelial carcinoma concomitant with squamous cell carcinoma was seen in 95 cases (28.4%). The incidences of coexistence in such lesion were higher in the groups of early stage esophageal cancer. These observations support the concept of field carcinogenesis of esophageal cancer. II. Histopathologic study of squamous epithelial dysplasia. Among 91 cases without preoperative treatment, there were 40 dysplastic lesions in 23 cases (25.3%). The continuity of dysplasia to the carcinoma was 48.3% and it was often encountered in severe dysplasia rather than in moderate or mild dysplasia, suggesting some relationship between the severity of dysplasia and carcinoma. III. Immunohistochemical study of EGF and c-myc. Among 27 cases, EGF was positive in 10 (37.0%). c-myc was positive in 18 (66.7%) not only cancer but normal epithelium suggesting that some change of products of oncogene occurred also in the normal epithelium of the patients of esophageal cancer.


[Significance of p53 gene abnormalities in carcinogenesis of human gastrointestinal tract]. Allelic loss and mutation of the p53 gene are common events of esophageal, gastric and colorectal cancers occurring from an early stage. Moreover, p53 mutation takes place in intestinal metaplasia and adenoma of the stomach. p53 mutation spectra differ among esophageal, gastric and colorectal cancers suggesting exposure to different endogenous and exogenous mutagens. The in vivo and in vitro results indicate that the clonal expansion of p53 mutant cell may be associated with tumor progression. Although wild type p53 transfection technique may provide for cancer therapy, there is a rather serious problem about it.


[Cellular and molecular biological study of the laminin-binding protein and its clinical application]. Tumor invasion and metastasis involve the interaction between tumor cells and basement membrane, which is mediated in part by laminin receptors. To search for tumor-associated-genes which can be used as new markers in colon cancers with known poor prognosis, cDNA libraries from a colon cancer cell line and colonic tissues were constructed and screened. We selected a cDNA clone which encodes 32-kD laminin-binding protein (LBP-32), and showed increased mRNA expression of LBP-32 in colon carcinoma. This mRNA expression was also correlated with clinical tumor staging. Furthermore, to investigate the role of LBP-32 in cancer invasion and metastasis, cell adhesion assays and in vitro invasion assays were performed, using anti-sense RNA of LBP-32 to block the synthesis of LBP-32. Results showed that anti-sense RNA of LBP-32 inhibits tumor cell attachment and invasiveness in vitro in transfectants of a colon cancer cell line. These data suggest that LBP-32 may play an important role in colon cancer progression, and that LBP-32 may be used as a marker of biological aggressiveness. These findings also imply that laminin receptors may provide a target for novel therapeutic strategies: modulating LBP-32 expression by anti-sense RNA or monoclonal antibodies may have clinical application in colorectal cancer therapy.


[A new method of in vitro chemosensitivity test using multicellular spheroids of cholangiocarcinoma cell line cocultured with fibroblasts]. We applied the multicellular spheroids which consist of cholangiocarcinoma cell line (MEC) and human dermal fibroblasts (HDF) to in vitro chemosensitivity test. Five-day multicellular spheroids were incubated with 1.5 micrograms/ml of mitomycin C (MMC) for 24 hrs. Then, cell kinetics of MEC and HDF in a spheroid was determined by flow cytometric analysis. Twenty four hrs after treatment with MMC, both MEC and HDF were accumulated on S phase. Seven-day after treatment, DNA histogram in MEC returned to normal, but that of HDF was disappeared. These results showed that the multicellular assay could be more like on in vivo like chemosensitivity test.


[Insulin receptor Arg1131-->Gln: a novel mutation in the catalytic loop of insulin receptor observed in insulin resistant diabetes]. A novel mutation Arg1131-->Gln in the catalytic loop of insulin receptor (IR) associated with insulin resistant diabetes was detected. A 56-year-old male with hyperinsulinemia (fasting IRI 92 microU/ml) showed moderate impairment in glucose tolerance (HbAlc 7.0%, fructosamine 258 mumol/l, fasting glucose 119 mg/dl, maximum value of blood glucose during 75 g OGTT 220 mg/dl). While insulin binding to erythrocytes IR was normal, the insulin-induced autophosphorylation of the patient's erythrocytes IR in vivo showed marked decrease, suggesting this patient had some defect in the kinase domain (exon 17-21) of IR. PCR-SSCP analysis of kinase domain with a genomic DNA obtained from the patient's leucocytes indicated the presence of some mutations in exon 19. Sequencing analysis in M13 revealed a heterozygous mutation at a position 1131 (CGG-->CAG) substituting Gln for Arg. Four people of patient's family analyzed are revealed to have an identical missense mutation at the same position with the patient.


[Therapeutic inventions of perioperative metabolic care in diabetic patients]. Perioperative metabolism was evaluated in diabetic patients and animal models. Experimental model of diabetes mellitus (DM) model was obtained by streptozotocine injection. Each 14C-labeled substrates and combination of carbohydrates mixture was administered intravenously in a dose of 1 g/kg before and after laparotomy and cumulative curves of expired 14CO2 was calculated. Biochemical analysis in the liver, blood and urine were performed following infusion of two different solutions, GFX (glucose, fructose, xylitol) solution, 4:2:1 ratio by weight vs. glucose solution, with 12% amino acids solution. In a clinical study, incidence of DM in digestive surgery was examined in 3481 cases. Retrospective evaluation of GFX solution was performed in cases after total gastrectomy. Incidence of DM was 14% and it has been increased recently, Glucose metabolism in DM rats was disturbed after surgery. Amino acids (alanine and valine) and lipid metabolism was slightly suppressed after surgery in DM rats. The best combination of carbohydrate mixture was determined as GFX solution in animal and clinical experiments. GFX solution improved not only glucose metabolism but also lipid and amino acid metabolism in DM rats. Decreased insulin requirement and increased total caloric intake were observed in DM and borderline DM cases treated by GFX solution.


[Management of patients with impaired glucose tolerance following esophagectomy in carcinoma of the esophagus]. We demonstrated the effectiveness of the "sliding scale" insulin infusion in diabetic patients undergoing esophagectomy. Fifty-eight patients were followed after esophagectomy to clarify differences in energy expenditure and caloric contributions of substrates. Energy expenditure was measured by indirect calorimetry on the 1st, 3rd, 5th, and 7th days after esophagectomy. Out of 58 patients, 7 were divided into diabetic (D), 30 into borderline (B), 21 into normal (N), according to a 75 g oral glucose tolerance test, preoperatively. Forty-four patients underwent esophagectomy by means of right thoracotomy, and blunt esophagectomy was done on 14 patients. The results were as follows: 1) No relation was found between postoperative morbidity and severity of glucose intolerance. 2) There was no significant difference in changes in energy expenditure following operation among groups D, B, and N. No difference was found in ratio of caloric intake to energy expenditure among groups D, B, and N. Caloric contributions of substrates seemed to be comparable among groups D, B, and N. These results suggest that "sliding scale" insulin infusion with total parenteral nutrition enable us to control not only blood glucose level but energy metabolism following esophagectomy in diabetic patients.


[Diabetes mellitus and coronary artery bypass surgery]. To determine the influence of diabetes mellitus on the results of coronary artery bypass surgery, a review of 163 diabetic patients operated on during 8 years, of whom 146 were receiving no drugs or receiving oral hypoglycemic agents, and 17 were receiving insulin. They were compared with 337 nondiabetic patients operated on over the same period. Higher incidences of hypertension and cerebrovascular disease for the diabetic group were found. The extent of coronary artery disease as judged angiographically was significantly greater in the diabetic group than in the nondiabetic group. Perioperative mortality was similar in the two groups. The incidence of perioperative myocardial infarction, sternotomy complication, neurological complication, and renal insufficiency was equal in the two groups. Early graft patency was comparable in the two groups. Overall 8-year actuarial angina free ratios were 88.5% for the diabetic group, 93.2% for the nondiabetic group. Overall 8-year survival rates were 95.6% for the diabetic group, 98.6% for the nondiabetic group. Results indicate that diabetic patients have quantitatively more coronary artery disease than the non-diabetic patients but have no higher perioperative morbidity and mortality than nondiabetic patients. Long-term results revealed a lower angina free rate in diabetic patients than in nondiabetic patients.


[Problems and management in kidney transplant recipients with impaired glucose metabolism]. Fourty two (12%: Group-I) out of 358 kidney transplant (Tx) recipients showed impaired glucose tolerance in pre-Tx O-GTT and the other 49 (14%: Group-II) showed that only after Tx. Insulin therapy was more frequently required and earlier started after Tx in Group-I than in Group-II, and total dosages of steroids before the initiation of insulin therapy were also smaller in Group-I. These trends were more markedly observed in the patients treated with ciclosporin (CsA) than in those with the conventional therapy. Poor graft survival rate of Group-I in long-term follow-up (> 5 years) was demonstrated. Thus, pre-Tx impaired glucose tolerance and the subsequent immunosuppressants, especially steroids and CsA, were the key factors for deteriorating glucose metabolism after Tx, and meticulous controls of drug doses and glucose levels are needed in diabetic patients, since diabetes could change the fate of kidney grafts.


[Combined pancreas and kidney transplantation for IDDM patients with diabetic renal failure]. We performed 7 cases of pancreas transplantation (PTX), simultaneous pancreas and kidney transplantation in 4 cases, and PTX after kidney transplantation in 3 cases. The pancreas and kidney were extirpated after in situ perfusion using UW solution and stored in UW solution. The pancreas was transplanted in the left iliac fossa with bladder drainage, and the kidney was placed in the contralateral iliac fossa. The immunosuppressive regimen consisted of cyclosporine, methylprednisolone, azathioprine and antilymphocyte globulin. Gabexate mesilate (30-40 mg/kg/day) and PGE1 (5 ng/kg/min) was administered intravenously to prevent the vascular thrombosis. The original diseases of 7 patients were insulin-dependent diabetes mellitus (IDDM) with chronic renal failure, retinopathy and neuropathy. Six out of 7 patients became insulin-free after PTX, while one patient developed the vascular thrombosis in the pancreatic graft which was removed after 12 hours after the transplantation. All patients became dialysis-free and serum creatinine was ranging from 1.5 to 2.0 mg/dl. HbAlc remained within normal range in 6 out of 7 patients, who showed normal to borderline glucose tolerance in 75g oral glucose tolerance test. Although further investigation will be required, PTX from cardiac-arrest donor will be promising as one of the therapeutic modalities for IDDM patients.


[Site-specific mutagenesis of residue Lys-172 of phage T7 RNA polymerase: characterization of transcription properties of mutant proteins]. Lys-172 residue of bacteriophage T7 RNA polymerase (T7RP) was substituted for Leu and Gly and Lys-172, Arg-173 were deleted by the site-directed mutagenesis using synthetic oligonucleotides. The specific activity of all mutant enzymes did not differ significantly from that of the wild-type (w.t.) T7RP while for Gly-172 mutant (G172) it was somewhat lower. Leu-172 (L172) and deletion (DEL172-3) mutants were able to direct RNA synthesis on the templates lacking the T7 promoter. DEL172-3 was not able to synthesize extraneous RNA sequences in addition to the expected run-off transcripts. L172 and DEL172-3 mutants revealed altered template specificity toward various DNA templates and showed the lower stability of enzyme-promoter complexes. The possible role of Lys-172 likely belonging to an interdomain "stretch" is discussed.


[Stably bent and low-melting DNA sequences at the origin of replication of chicken alpha-globulin gene domains]. The distribution of specific DNA sequence elements in a 2.9 kb HindIII fragment of chicken DNA containing the replication origin and the upstream matrix attachment site of the alpha-globin gene domain has been studied. The fragment was shown to contain the CR1-type repetitive element and two stable bent DNA sequences.


[Rare codons and gene expression in Escherichia coli]. Influence of rare codons upon gene expression in E. coli was investigated. The chimeric gene was created combining CAT gene and a fragment of the gene, encoding for alpha-domain of beta-galactosidase. The synthetic oligonucleotides were inserted in different parts of the chimeric gene. The constructed synthetic oligonucleotides encoded the same amino acid sequences and contained arginine codons AGG, AGA and CGT in various combinations. It was shown that the presence of rare arginine codons AGG and AGA in the template and their mutual arrangement significantly influence the level of gene expression. At the same time the presence of leucine, isoleucine, glycine and proline rare codons does not cause such an effect. Translation of AGGAGG and AGAAGA sequences was found to lead to the formation of a considerable amount of polypeptides of incomplete length. It was shown that the presence of such a cluster of rare codons effects on the length of specific mRNA.


[Enzymatic cleavage of superhelical DNA in a liquid crystal state]. Superhelical pBR322 DNA molecules form liquid-crystalline dispersions in water-salt solutions containing poly(ethyleneglycol). The formation of the liquid-crystalline dispersions from superhelical DNA molecules results in the appearance of two sites inside the DNA molecules that are split by Micrococcal nuclease. The first site of digestion does not differ from the standard site split by this enzyme in water-salt solutions, whereas the second one represents a new site specific only for the DNA molecules forming liquid-crystalline dispersions. Splitting of the DNA molecule through the first site is accompanied by formation of its linear form; splitting of a new site results in the formation of two linear DNA fragments with molecular masses equal to half of the initial DNA molecules. Enzyme digestion of superhelical DNA molecules forming liquid-crystalline dispersions induces a reformation of the "nonspecific" space organization of dispersions to the cholesteric one. A hypothetic model for packing of the superhelical DNA molecules inside liquid-crystalline dispersions and its transformation under enzyme action is suggested.


[A comparative study of chromatin from lymphocyte nuclei upon activation of transcription by irradiation from an He-Ne-laser or phytohemagglutinin]. The influence of He-Ne laser radiation (632.8 nm, 56 J/m2, t = 10 s) and phytohaemagglutinin (PHA, 2 micrograms/ml) on chromatin structure in human lymphocytes was studied by electron microscopy using ultrathin cell sections. Morphometric analysis of extranuclear condensed chromatin masses was performed 1 h after the irradiation or after the beginning of PHA treatment. In the irradiated cells the following insignificant changes were revealed: decrease in the relative area of the nucleoplasmic chromatin, increase in the relative area of decondensation zones as well as increase in the number of clumps of nucleoplasmic chromatin and relative length at their boundary with nucleoplasma. The tendency of these morphological changes may be interpreted as functional activation of extranucleolar RNA synthesis in response to irradiation by red laser light. Action of PHA results in significant changes of the surfaces of chromatin clumps, namely increase in relative length of nucleoplasmic chromatin boundary and decrease in relative length of perimembranous chromatin boundary with nucleoplasma as well as some less expressed delamination of the chromatin masses from the nuclear membrane. These essential changes may reflect chromatin activation by proliferative stimulus. Peculiarities of the ultrastructural reorganisation in the condensed chromatin after irradiation and PHA-treatment probably reflect the differences in the processes of gene activation caused by the two agents.


[The effect of intracellular concentrations of tRNA, corresponding to the rare arginine codons AGG and AGA, on the gene expression in Escherichia coli]. Influence of increased arginine concentrations of tRNA's corresponding to rare codons AGG and AGA was studied in the model system constructed earlier. The model system is a chimeric gene consisting of CAT gene fragment, part of the gene encoding for alpha-domain of beta-galactosidase E. coli and a series of synthetic inserts enriched with codons AGG and AGA. In order to increase the intracellular tRNA concentration the natural gene of AGA-specific tRNA and the artificial gene of AGG-specific tRNA were cloned in plasmid under the control of p15A ori compatible with co1EI ori and used for maintaining the model gene. It was shown that the artificial AGG-specific tRNA gene produces a functionally active tRNA. A steep rise in the synthesis of polypeptide encoded by the model template containing rare codons was demonstrated when the genes of tRNAs recognizing these codons were propagated in the multicopy plasmid. It was shown that AGA-specific tRNA efficiently translates both AGA and AGG codons while AGG-specific tRNA - only AGG codons.


[Design and expression of a diphtheria toxin hybrid protein and human interleukin-2 gene in Escherichia coli]. Recombinant fusion proteins consist of the N-terminal 488 or 513 amino acids of diphtheria toxin joined to human interleukin 2. Initially those fusion proteins were expressed in E. coli under the control of the tox promotor. Western blot analyses showed that E. coli strains bearing the hybrid genes produce 68 kDa or 72 kDa fusion proteins that retain the immunological determinants of both the diphtheria toxin component and the interleukin 2 component. The fusion protein with mol. mass 72 kDa was partially purified by affinity chromatography. The expression of the fusion proteins under the control of the strong promotors was increased (100-fold for tac- promotor) compared to that under the control of the tox promotor. DT-IL2 might be a useful cytotoxic agent in the treatment of diseases involving IL2 receptor-positive cells, such as allograft rejection, graft-versus-host disease, multiple sclerosis et al.


[Adducts of 3'-azido-2,3'-dideoxythymidine 5'-phosphate or 5'-phosphonate as inhibitors of cytopathic effect and transformation of cells under the influence of retroviruses in cell culture]. Inhibition of HIV-1- or HIV-2-induced cytopathicity and (Moloney) murine sarcoma virus (MSV)-induced cell transformation by amino acid and amino alcohol adducts of either 3'-azido-2',3'-dideoxythymidine 5'-monophosphate (AZTMP) or 5'-hydrogenphosphonate (AZTHP) were investigated. Both types of nucleotide adducts inhibited replication of HIV-1 and HIV-2 in MT-4 cells at a 1.5- to 3-fold higher EC50 (50% effective concentration) than AZT; and, also, selectivity indexes of these adducts were approximately 1.5 to 3-fold lower than that of AZT. The activity of the AZTMP and AZTHP adducts against MSV-induced transformation of C3H/3T3 cells was equal to or only slightly inferior than that of AZT, but their toxicity was 10-fold lower, so that their selectivity indexes were 2- to 7-fold higher. The nature of the aminoacyl component of the adducts significantly influence the antiretroviral activity of the test compounds.


[The mechanism of action of pressure on intramolecular protein dynamics]. The physical mechanisms of pressure influence on the protein dynamics and parameters of Mossbauer spectra were investigated. The pressure effects measured for human serum albumin using Rayleigh Scattering of Mossbauer Radiation techniques were described using the model of local diffusion. Parameters of this model were determined by analysing the experimental data. As a result an estimation of approximate values for activation volumes was performed. It was shown that the obtained values are well consistent with the previous experimental results.


[Tandem DNA repeats in the vertebrate genome: structure, possible mechanisms of formation and evolution]. Possible models for the generation and the evolution of tandem repeats are discussed. The model of A.J. Jeffreys and co-workers as well as facts, supporting or contradicting this model are discussed. Facts supporting the hypothesis of the generation of the tandem repeats as the result of mitotic recombination are described. On the basis of an analysis of the structure of the tandem repeats containing loci, it is supposed that there exist space and time relations between the multimerization of the tandem repeats and tandem gene duplication. On the basis of this supposition, the generation of majority of the tandem repeated gene as a result of sister chromatids recombination in mitosis is proposed. Factors determining the existence of recombination hotspots of are discussed. Some specific features of the evolution of tandem repeats of the coding region are also described.


[Nuclear envelope of frog diplotene oocytes has telomere-binding activity]. A telomere-binding activity was found in the nuclear envelope of frog oocytes Rana temporaria by methods of binding on nitrocellulose filters and retardation. At present it is hardly possible to detect the proteins responsible for this activity. However, the results of this study allowed us to identify a specific mechanism of DNA-protein interaction, which provides the binding of the telomere to the envelope in meiotic cells.


[Renal cell cancer in childhood].

[Renal cell cancer in childhood]. Renal cell carcinoma is a rare disease in children and difficult to distinguish from Wilms-tumor before surgery. We present case histories of two children with renal cell carcinoma and discuss the problems of differential diagnosis versus nephroblastoma, therapy and prognosis. In contrast to Wilms-tumors, the most common kidney-tumor in children occurring mostly in young infants, renal cell carcinoma is rare in childhood and predominantly manifests in school-age. Only a few cases of renal cell carcinoma in younger children are described in the literature. Diagnostic imaging cannot reliably distinguish renal cell carcinoma from other neoplasm of the kidney. However, hematuria in patients with small tumors or no response to preoperative chemotherapy may indicate the presence of renal cell carcinoma rather than nephroblastoma. The determination of "tumor-associated trypsin inhibitor" (TATI) might give further contribution of differential diagnosis. It was measured only in one of our patients and was markedly elevated. Complete surgical resection (nephrectomy with lymphadenectomy) is a curative therapy in patients with tumors limited to the kidney. Chemotherapy and irradiation show no convincing effect. In metastatic tumors therapy with interleukin 2 may be successful.


[Neonatal diabetes mellitus and microcephaly. Indications for autosomal recessive inheritance]. Still, there are a lot of questions about the pathogenesis of neonatal diabetes mellitus. In the author's opinion neonatal diabetes mellitus is a distinct entity which differs from the well-known types of diabetes in children (type 1 diabetes, MODY-diabetes) and transient neonatal hyperglycemia regarding pathogenesis, pathophysiology and prognosis. Casuistics of three children two of whom were sibs are reported in detail to demonstrate the characteristics of neonatal diabetes mellitus. Regarding the reported sibs we suppose genetic origin of the disease. Autosomal-recessive mode of inheritance must be assumed.


[Formation of two types of enzyme-substrate complexes during the interaction of EcoRII restriction enzyme with synthetic DNA-duplexes]. Binding of EcoRII restriction endonuclease to synthetic oligodeoxyribonucleotide substrates of 11-30 base pairs long was investigated by polyacrylamide gel electrophoresis under nondenaturing conditions in the absence of Mg2+ ions. Irrespective of the length of a substrate, two types of specific DNA-protein complexes were shown to be formed. Their mobility in gel was close to that of the monomer (45 kDa) and dimer (90 kDa) of marker protein, ovalbumin. The ratio of these complexes in solution depended on that of the molar concentrations of EcoRII restriction endonuclease and DNA duplexes. The possible structure of the complexes is discussed.


[Extrapyramidal movement disorders after "Isostar Cocktail"]. Two children developed extrapyramidal symptoms after taking an electrolyte drink. There was a good response to Biperidon (Akineton). Only exact investigations showed that both children had accidentally been poisoned by a neuroleptic drug which was added to the electrolyte mixture.


[A method of isolation and properties of DNA-dependent DNA-polymerase epsilon from human placenta]. DNA polymerase epsilon was purified to near homogeneity from human placenta. The enzyme has one subunit (170 kDa, sedimentation coefficient 8.2S), intrinsic 3'-5'-exonuclease activity, it is independent on PCNA and high processivity on poly(dA)-oligo(dT) template-primer without PCNA. It was shown, that the enzyme incorporates 3'-amino-2',3'-dideoxythymidine 5'-triphosphate in DNA, after that synthesis is stopped. Simultaneously DNA polymerase alpha was purified.


[Trichobezoar. A rare cause of recurrent upper abdominal pain]. Recurrent abdominal pain in a two years old toddler and a sixteen years old girl were caused by huge trichobezoars, which in one case had a weight of 412 g and provoked a gastric ulcus. Removal was possible only by gastrotomy. Deprivation in infancy and childhood in combination with slight mental retardation had led the teenager to trichtillomania and -phagia for years and finally to a bezoar. A short historic survey of the variety and therapy of bezoars is added.


[Complex characteristics of the alterations of oncogenes HER-2/ERBB-2, HER-1/ERBB-1, HRAS-1, C-MYC and antioncogenes p53, RB1, as well as deletions of loci of chromosome 17 in colon carcinoma]. Abnormalities of some oncogenes, antioncogenes and losses of heterozygosity (LOH) of chromosome 11p, 17p, and 17q in colorectal carcinomas (CC) was studied. Amplification of ERBB-1/HER-1 oncogene was detected in 2 of 56 cases; ERBB-2/HER-2- in 4 of 62. There was a lack of evidence for C-MYC oncogene amplification (67 cases). LOH of chromosome 11p (HRAS-1 probe) was found in 2 of 37 informative (heterozygous) cases; such events were not accompanied by point mutations in "hot" codons (12th or 61st) in the remaining allele. Prevalence of A3 and A4 alleles of HRAS-1 oncogene (68 cases) as compared to healthy donors was noted. RB-1 (41 cases) and p53 (62 cases) suppressor genes did not show any alterations in Southern-blot analysis. LOH of chromosome 17p (YNZ-22 probe) was found in 15 of 26 heterozygous CC; 17q (THH-59 probe)--in 4 of 16. Analysis of 175th codon of p53 gene revealed only one case of mutation in 35 CC studied. Finally, we were able to detect genetic alterations in 23 of 40 (58%) CC, that were studied on each parameter using Southern-blot. We failed to find any correlation between various molecular abnormalities or clinical characteristics. The data obtained are in disagreement with the view concerning frequent involvement of p53 antioncogene in chromosome 17p deletions.


[Fructose malabsorption and dysfunctional gastrointestinal manifestations]. Individuals with impaired intestinal absorption of fructose may exhibit recurrent abdominal discomfort after the ingestion of fructose-containing foods. We report on patients with this disorder in whom the diagnosis was made by the fructose hydrogen breath test. We investigated 293 patients with recurrent abdominal pain, meteorism or diarrhea in connection with the ingestion of fruits, apple juice or soft drinks. Mixed expired air was collected before and at 30 minute intervals after a fructose load and analysed thereafter by a hydrogen sensitive electrochemical cell. Incomplete absorption of fructose was defined as a peak rise in breath hydrogen of > 20 ppm. 108 out of 293 patients showed an abnormal peak rise after fructose (mean 71.8 ppm, SD 31.4). This malabsorption of fructose was associated with clinical symptoms in 79 of them. Sensitivity and specificity of the fructose hydrogen breath test were 98 or 86 per cent respectively. 19 patients with an abnormal breath test and symptoms following fructose were reexamined after a load with equimolar concentrations of glucose and fructose. Hydrogen breath test was normal in all of them, none developed abdominal discomfort. A considerable number of individuals suffer from dysfunctional gastrointestinal problems due to fructose malabsorption. The fructose hydrogen breath test is a simple, sensitive and noninvasive method for the diagnosis for this disorder. Possible means of treatment are dietary fructose restriction or a modification of the diet in which fructose-containing foods are exchanged for those with equal concentrations of glucose and fructose.


[Hyaline membrane syndrome. Evaluation of sonographic criteria]. In preterm infants with hyaline membrane disease an echo increase above the diaphragm can be demonstrated by parasagittal sonography of the lower thorax using the liver or spleen as an acoustic window. In order to study the clinical practicability of this observation we made 145 investigations in 101 infants. The ultrasonographic evidence of an increased echo above the diaphragm could be found in all 85 infants with a clinically defined hyaline membrane disease. Ultrasonographic criteria have proved to be useful and reliable in assessing the severity of hyaline membrane disease. This method is also suitable for documenting the success of surfactant administration. Using sonography hyaline membrane disease can quickly be differentiated from other pulmonary diseases. Although sonography cannot serve as a substitute for chest X-ray it may be able to give valuable additional information for the differential diagnosis of respiratory disorder in the premature infant.


[Diagnosis and nosology of glycanosis CDG ("carbohydrate deficient glycoprotein syndrome")]. The "Carbohydrate-deficient glycoprotein syndrome" is a recently discovered inborn error of complex carbohydrate metabolism. The disease involves a number of organ systems and various deficient glycoproteins. An abnormal isoform of serum transferrin is of diagnostic value. We analysed the glycoprotein alpha-1-antitrypsin of two affected infants and their clinically healthy parents using high resolution isoelectric focusing technique. Besides normal isoforms of alpha-1-antitrypsin, we found an abnormal cathodic isoform ("CDG-alpha-1-antitrypsin") which represented almost half of the total amount of alpha-1-antitrypsin of the patients. This new marker-glycoprotein suggests a defect of the production of biantennary and of triantennary N-glycans during an early step of their synthesis, resulting in monoantennary N-glycans. Also this marker-glycoprotein seems to be a specific biochemical diagnostic tool for discovering glycanosis CDG (Carbohydrate-deficient glycoprotein syndrome). The mode of inheritance is probably incomplete autosomal dominant. The same genetic defect of N-glycan synthesis may be present in more than one type of the hybrid molecule glycoprotein, and was also found in transferrin, resembling a "genetic back-pack", that might explain the multitude of clinical symptoms. In view of these findings, we present novel systematics of those diseases that are due to inborn errors of N-glycan synthesis, and which we suggest to call "glycanoses".


[Bilateral pheochromocytoma. Is there stimulation of an extra-adrenal adrenaline liberation after bilateral adrenalectomy?]. In order to determine plasma and urine epinephrine levels after bilateral adrenalectomy, we examined a 16 year old young man with bilateral pheochromocytoma. Bilateral removal of the adrenals was performed because of a left sided pheochromocytoma relapse. An extra-adrenal pheochromocytoma could be excluded by scintigraphy. Macroscopically all adrenal tissue was removed during surgery. A modified radioenzymatic determination of the free catecholamines epinephrine, norepinephrine and dopamine was used according to Peuler and Johnson. Sulfoconjugated catecholamines were measured after addition of 60 microU arylsulfatase type VI. Urine catecholamine levels were determined fluorometrically. The elevated plasma andurine norepinephrine levels before surgery returned to normal after surgery. In contrast, plasma epinephrine levels returned to subnormal values thereafter but increases 5-fold at the end of anesthesia suggesting an extra-adrenal source of epinephrine. Urine epinephrine levels remained in the lower normal range. An insulin induced hypoglycemia was performed resulting in adrenergic symptoms of hypoglycemia and a subnormal increase of epinephrine. We conclude, that epinephrine mediated physiological regulations occur inspite of bilateral adrenalectomy probably by a regulated extra-adrenal source of epinephrine. The kidney could be the site of extra-adrenal epinephrine production.


[Minimally invasive surgery of sinugenic orbital complications in childhood]. Orbital complications of acute sinusitis are classified into inflammatory edema, orbital cellulitis, subperiostal abscess and orbital abscess. The diagnosis is based on endoscopy of the nose, computed tomography of sinuses and orbit and an ophthalmological examination. Endonasal sinus surgery improves drainage and ventilation of sinuses and is free of long-term complications as observed with previous surgical techniques. Thus, the early surgical treatment of orbital complications is indicated even in children. Inflammatory edema and orbital cellulitis will still be treated conservatively. Subperiostal abscess and orbital abscess are treated surgically.


Testifying in criminal court: emotional effects on child sexual assault victims. Child victims must cope not only with the emotional consequences of criminal acts but also with the potentially traumatizing effects of legal involvement. Dramatic increases in the reporting of child sexual abuse are bringing greater numbers of children into contact with the criminal justice system, raising fears that child victims of sex crimes will be further harmed by the courts. In the present study, the effects of criminal court testimony on child sexual assault victims were examined in a sample of 218 children. From this sample, the behavioral disturbance of a group of "testifiers" was compared to that of a matched control group of "nontestifiers" at three points following testimony: 3 months, 7 months, and after prosecution ended. At 7 months, testifiers evinced greater behavioral disturbance than nontestifiers, especially if the testifiers took the stand multiple times, were deprived of maternal support, and lacked corroboration of their claims. Once prosecution ended, adverse effects of testifying diminished. In courthouse interviews before and after testifying, the main fear expressed by children concerned having to face the defendant. Children who appeared more frightened of the defendant while testifying were less able to answer the prosecutors' questions; and later, after the cases were closed, they were more likely to say that testifying had affected them adversely. The two most pervasive predictors of children's experiences in the courtroom, however, were age and severity of abuse. Despite relevant laws, few innovative techniques were used to help the children testify. The results are discussed in relation to children's ability to cope with stressful situations, the interaction of the legal system with the child/family system, and debates about the need to protect child victims who testify in criminal court.


Turns-amplitude analysis of the electromyographic recruitment pattern disregarding force measurement. I. Method and reference values in healthy subjects. We used turns-amplitude analysis to characterize the EMG recruitment pattern disregarding force measurement. The electrical muscle pattern of the brachial biceps (BB), abductor pollicis brevis (APB), medial vastus (MV), and anterior tibial (AT) muscles was analyzed during progressive increase in force from rest to maximum using the mean amplitude as an indicator of the force of the muscle. The following parameters were obtained on-line: the maximal ratio of turns to mean amplitude (peak-ratio, PR), the mean amplitude, and the number of time intervals (TI) between turns at PR and at near maximum force (NMF). The highest PR values were obtained in BB, the lowest in MV. Analysis of the distribution of the TI between turns at different degrees of voluntary contraction showed fewer spikes with short duration and small amplitude at high force compared with low force.


Turns-amplitude analysis of the electromyographic recruitment pattern disregarding force measurement. II. Findings in patients with neuromuscular disorders. Turns-amplitude analysis of the electromyographic recruitment pattern was performed on-line in the brachial biceps muscle of 46 patients with neuromuscular disorders using the mean amplitude as an indicator of force. The parameters, peak-ratio (PR) and number of time intervals (TI) from 0 to 1.5 ms, were increased in patients with myopathy. In patients with neurogenic involvement, the characteristic pattern was a decreased PR and a decreased incidence of TI between 0 and 1.5 ms. The results indicate that the two parameters supplement each other as some of the patients were identified only by one or the other. In patients with myopathy, the method had a higher diagnostic yield than the individual motor unit action potential analysis. The method is objective, fast, and reliable.


Lambert-Eaton syndrome: antigen-antibody interaction and calcium current inhibition in chromaffin cells. Plasma and IgG obtained from 10 Lambert-Eaton myasthenic syndrome (LES) patients (5 with carcinoma, 5 without associated cancer), 6 healthy subjects, and 1 patient with small-cell lung cancer (SCLC) were examined in their ability to recognize chromaffin cell antigens on Western blots. The pattern of antigen recognition was compared with the magnitude of inhibition of voltage-dependent calcium and sodium currents recorded with the patch-clamp technique from chromaffin cells. Eight of the 11 patients with LES and/or SCLC recognized plasma membrane proteins and 9 of the patients' IgG interacted with cytoplasmic antigens with no apparent pattern of antigen recognition between patients. Also, there was no obvious band pattern distinguishing patients with LES from those with LES and concurrent SCLC. Eighty percent of the LES patients' antibodies were capable of reducing the calcium current (ICa) in chromaffin cells. One of the novel findings of this study is that 30% of the patients had produced antibodies which were able to inhibit both calcium and sodium currents (INa). The heterogeneous response of the IgG on the Western blots does not appear to correlate with the efficacy of reducing the inward currents.


Sensory testing versus nerve conduction velocity in diabetic polyneuropathy. We sought to evaluate the utility of quantitative sensory testing (QST) and nerve conduction velocity (NCV) studies as measures of distal symmetric polyneuropathy (DSP). We studied 36 diabetic patients divided into four clinical categories of increasing severity. QST included thermal testing and vibration thresholds. NCV studies included median, peroneal, and sural nerves. Results of QST and NCV were compared among clinical groups using survival methodology. The log-rank statistic showed significant differences among the groups; the direction of the differences were consonant with clinical severity. For each diabetic patient, the result of each measurement was classified as normal or abnormal; more diabetic patients had abnormal NCV than either vibration tests or thermal tests. In conclusion, findings of QST and NCV are in keeping with clinical categorization of patients, QST and NCV are complementary tests, and the sural sensory study is the best single predictor of DSP.


Differential effects of amitriptyline on sudomotor, cardiovagal, and adrenergic function in human subjects. Tricyclic antidepressants, especially amitriptyline (AMI), are widely used by patients who require tests of autonomic function. AMI effects autonomic function, but its quantitative and differential effects are not known. We prospectively evaluated the effect of AMI on sudomotor (M3 receptor), cardiovagal (M2) and adrenergic functions in 6 subjects, aged 20-40 years before, during, and 48 hours after AMI 75 mg/d. M3 receptor function was evaluated using QSART (quantitative sudomotor axon reflex test) recordings from four sites, M2 receptor function from cardiovagal studies [HR response to deep breathing (HRDB) and the Valsalva ratio (VR)], and adrenergic function from an analysis of beat-to-beat BP responses to tilt and the Valsalva maneuver. Plasma AMI was determined from blood samples obtained within 2 hours of autonomic testing. QSART volume was reduced by 47% by AMI (P = 0.01), and recovered to 81% of baseline following a 48-hour washout. HRDB was unaffected by AMI (P > 0.05). VR was increased by 10% (NS) with AMI and by 14% (P < 0.05) with washout. The percent change in hemodynamic parameters by AMI were: orthostatic reduction in SBP, 62% (NS); orthostatic HR increment, 66%, (P = 0.03); and phase IIe SBP decrement, 665%, (P = 0.02). We conclude that AMI in the moderate doses used, resulted in greater inhibition of M3 than M2 receptors. The BP, HR, and beat-to-beat BP alterations likely reflect adrenergic inhibition resulting in a reduction in effective plasma volume. A washout of 48 hours is adequate for muscarinic but not adrenoreceptors.


The silent period in pure sensory neuronopathy. "Normal" electromyographic silent periods (SPs), produced by electrical stimulation of digital nerves, were recorded in the voluntarily contracting abductor pollicis brevis (APB) muscle of a patient with pure sensory neuronopathy and absent sensory nerve action potentials (SNAPs). Such findings implicate the smaller, slower conducting fibers in the genesis of the cutaneous silent period. These same fibers may be activated during more proximal stimulation to contribute to the latter portions of the mixed nerve silent period.


Evidence for new isoform of fast myosin heavy chain in rat skeletal muscle. The purpose of this study was to investigate whether the previously demonstrated heterogeneity of myosin heavy chain (MHC) in type 2B fibers of the superficial portion of the lateral gastrocnemius (SLG) muscle of the rat may be due to presence of type 2D/X fibers. Immunohistochemical identification of MHC heterogeneity, histochemical identification of appendicular 2D/X muscle fibers, and immunoblotting of myosins were used. It was found that some, but not all, of the heterogeneity of rat SLG is correlated with the presence of type 2D/X fibers. Immunoblots with MHC from several muscles revealed the presence of at least two antigenically distinct isoforms of MHC with the same electrophoretic mobility as the "2B" band. These results show that a previously undetected type of MHC is expressed in rat skeletal muscle and raise the possibility that there may be as yet undetected MHC isoforms in human muscle which may be clinically important.


Recurrent potentials in human peripheral sensory nerve: possible evidence of primary afferent depolarization of the spinal cord. To study slowly conducted components of the orthodromic compound sensory action potential (CSAP), the response evoked at the lateral malleolus in the sural nerve was recorded through near-nerve needles at two to four sites along the nerve at midcalf. When 500 to 2000 responses were averaged at high gain, components with latencies of 30 to 80 ms were often recorded. In contrast to the main component and late components with latencies of less than 15 to 20 ms, the latencies of these extremely late components diminished the closer to the spinal cord that they were recorded. This suggested that the components were conducted antidromically from proximal to distal. This assumption was supported by abolishing the components by local anesthesia of the nerve proximal to the recording electrodes. These antidromic potentials therefore appear to be due to recurrent discharges in the sural nerve. Recurrent discharges were recorded from 65% of 60 subjects (18 normal subjects and 42 patients with peripheral or central nervous system disorders). The latencies of the recurrent discharges allowed conduction to and back from the spinal cord. Although the origin of these potentials remains unknown, we suggest that they are due to dorsal root reflexes within the spinal cord. In this case, the responses may be a direct expression of primary afferent depolarization (PAD) seen in presynaptic inhibition, and may be of value in further studies on the physiology and pathophysiology of presynaptic inhibition of cutaneous fibers in man.


Perineurial cell hypertrophic mononeuropathy manifesting as carpal tunnel syndrome. A 42-year-old man showed signs and symptoms suggestive of carpal-tunnel syndrome, but EMG showed an isolated motor axon-loss lesion affecting the right median nerve distally. After the MRI revealed a mass in the median nerve, surgical exploration showed a diffusely swollen median motor branch. Biopsy showed a lesion with marked onion-bulb formation composed of perineurial cells as identified by immunohistochemical analyses and electron microscopic examination. Although we previously coined the term "perineurioma" for this condition, re-reviews of our cases do not support the idea that the onion-bulb lesion is a benign tumor; instead, it appears to be reactive hyperplasia. Although rare, electromyographers and neurologists need to be aware of this problem because it is self-limited and does not require surgical resection.


Historical influences affecting the behavioral actions of abused drugs. The studies summarized briefly in this chapter provide several examples of experiments in which a behavioral or pharmacologic history produced profound changes in the effects of an abused drug. In many instances these changes were qualitative, that is, the direction of the effects that drug usually has on behavior was modified dramatically. AMPH and cocaine--both of which may normally decrease a particular behavior--will, after a suitable history, produce large increases in the behavior under study. If the behavioral effects of a drug are related to the abuse potential of that drug, then any condition that alters the behavioral effects of that drug could be an important component underlying vulnerability to drug abuse. A noteworthy aspect of the studies that have been described is the lasting influence that prior experience can have on behavior and the behavioral effects of drugs in the absence of overt changes in behavior itself. The traces of past experience may not be apparent in ongoing behavior until a drug is administered. This feature makes drugs an important tool for studying behavioral processes and raises several related questions pertaining to the mechanisms responsible for these effects (Barrett 1986). Although at present there is little information available to answer the several questions raised by these experiments, newer techniques available in the neurosciences promise to help reveal the neurobiological correlates of these changes that should be of general importance in understanding both behavioral and pharmacologic processes.


Cocaine self-administration: pharmacology and behavior. I would like to stress several major points. The first is that CNS pharmacology is important in drug self-administration. In the case of cocaine, the necessary effect seems to be blockade of reuptake of DA rather than blockade of reuptake of NE or 5-HT or local anesthetic effects. An action of that DA on the D2 subtype of DA receptors appears to play an important role. The second point is that increased DA in synapses in the CNS is not sufficient to explain drug-maintained behavior. The schedule of reinforcement critically determines rates and patterns of drug-maintained behavior. Barrett (this volume) has presented convincing evidence that behavioral history, i.e., the sequential exposure to alternative behavioral conditions, can dramatically alter drug effects. What I hope to have made clear is that the simultaneous opportunity to engage in other behaviors can dramatically alter the reinforcing effects of drugs. Indeed, under appropriate conditions, a drug can function as both a positive and a negative reinforcer simultaneously in the same animal. The drug-choice studies I have described emphasize the dynamic nature of the interaction between behavioral and pharmacologic variables. The frequency of choosing to self-administer cocaine varied with the magnitude of the dose of cocaine (a pharmacologic variable) and the magnitude of an alternative positive reinforcer that was available (a behavioral variable). Increasing the magnitude of the alternative reinforcer decreased cocaine choice, but increasing the magnitude of the cocaine dose reestablished drug preference. In a sense, this is not a surprising result. The rhetoric of the times, however, would have it that drug self-administration, particularly cocaine self-administration, is somehow a different class of behavior that is not subject to the usual laws governing the behavior of organisms. As our understanding of drug self-administration evolves, however, that position will become increasingly untenable. These experimental findings challenge simple neurobiological assumptions about the determinants of drug self-administration. Clearly, a reinforcing effect, like any other behavioral effect of a drug, is not simply an immutable effect of a pharmacologic property of the drug. Given the pivotal role of reinforcing effects in drug abuse, this realization is critical to a comprehensive understanding of drug abuse. Any complete neurobiological model of drug abuse will have to account for these effects. Apparently, increasing the concentration of DA in synapses in the brain is not sufficient to account for drug-maintained behavior.(ABSTRACT TRUNCATED AT 400 WORDS)


Brain-stimulation reward: a model for the study of the rewarding effects of abused drugs. The robustness of the findings of the BSR effects of abused substances indicates that almost any technique used to measure these effects will be useful. The simplest procedure, however, of selecting a single intensity of stimulation and determining the effects of drugs on the rate of response for that selected intensity is fraught with difficulty in interpretation. More than 25 years ago the interpretation of changes in response rate as a reflection of changes in the reward value of the stimulation was challenged (Hodos and Valenstein 1962). Today it is rare to see a single stimulation intensity used in a published manuscript. However, most studies still use procedures of which rate of response is an integral part. Because animals will press a lever more than 80 times per minute and--on a continuous reinforcement schedule--receive as many as 80 stimulations a minute, conclusions about the specificity of the effects of drugs are difficult. The results of BSR experiments on mechanisms of action of abused substances clearly indicate that the reinforcing effect of most, if not all, such substances is probably the result of activation of a reward system that originates in the cell bodies of the ventral tegmental area and courses rostrally to the limbic and frontal projection sites of the mesocortical system. Thus, we believe, the technique has clearly fulfilled its promise as a "window on the brain" (Olds 1977).


G protein-mediated receptor-receptor interaction: studies with chemotactic receptors in membranes of human leukemia (HL 60) cells. Differentiated human leukemia (HL 60) cells contain high numbers of receptors for the chemotactic factors, N-formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe) and complement component 5a (C5a), both coupled to pertussis toxin-sensitive guanine nucleotide-binding regulatory proteins (G proteins). Agonist activation of either receptor stimulated binding of the GTP analog, guanosine 5'-[gamma-thio]triphosphate (GTP[S]), to membrane G proteins and by a similar extent in a non-additive manner. The possible interaction of the two receptors was studied by measuring agonist binding to one receptor in the presence of the other receptor agonist. fMet-Leu-Phe and C5a had no effects on [125I]C5a and fMet-Leu-[3H]Phe receptor binding, respectively, when studied in the absence of regulatory ligands. Similarly, the inhibitory effects of NaCl and GDP on agonist receptor binding were not altered in the presence of the other receptor agonist. In contrast, in the presence of the GTP analogs, GTP[S] and guanosine 5'-[beta,gamma-imino] triphosphate, fMet-Leu-Phe and C5a reduced the binding of [125I]C5a and fMet-Leu-[3H]Phe, respectively, in a concentration-dependent manner. The potencies of the GTP analogs to inhibit binding of [125I]C5a and fMet-Leu-[3H]Phe was increased about 3-fold by fMet-Leu-Phe and C5a, respectively. The data presented suggest that fMet-Leu-Phe and C5a receptors share the same G protein pool in membranes of HL 60 cells and that activation of these G proteins by one of the two receptors decreases the availability of G proteins for the other receptor.


5-Hydroxytryptamine increases contractile force in porcine right atrium but not in left ventricle. Positive chronotropic as well as inotropic effects of 5-hydroxytryptamine (5-HT) have been observed in pig atrial tissue, but no data are available about the direct effects of 5-HT on ventricular tissue. In the present study we investigated inotropic effects of 5-HT on atrial and ventricular trabeculae obtained from hearts of 3 months old pigs. The baseline isometric contractile force was significantly higher in ventricular (4.14 +/- 1.25 mN) than in atrial tissue (0.47 +/- 0.11 mN). A noradrenaline concentration-response curve (0.01 to 10 mumol/l) was used to check contractile responsiveness of the tissue and all responses were expressed as percentage of the response to 10 mumol/l noradrenaline. Noradrenaline caused a concentration-dependent increase in contractile force in both atrial and ventricular trabeculae. In contrast, though 5-HT (0.01 to 100 mumol/l) did increase force of contraction in atrial tissue (maximum: 72 +/- 20% of the response to noradrenaline 10 mumol/l), the contractility of ventricular trabeculae was not significantly affected (maximum: 12 +/- 6%). The present data show that, in contrast to atrial tissue, contractile force of ventricular tissue could not be significantly affected by 5-HT. To our knowledge, this is the first study to show that an agent which increased force of contraction in the atrium, did not have a corresponding effect on the ventricle. These findings may have important implications for a better understanding of the physiology and pharmacology of cardiac contractility.


Effects of serotonin and noradrenaline on superficial hand veins in patients with primary hypertension and in healthy volunteers. In vitro and animal experiments suggest that a constitutional increase in vascular responsiveness to serotonin (5-HT) may play a role in the development of essential hypertension. We have studied the potential alterations in vascular responsiveness to the direct and catecholamine-potentiating effects of exogenous 5-HT in patients with essential hypertension, by comparing in vivo responsiveness of superficial hand veins to local infusions of 5-HT or to coinfusions of 5-HT and noradrenaline in unmedicated hypertensive patients and in healthy control subjects. The dorsal hand vein compliance technique was employed. There was no significant difference between patients and control subjects in the maximal 5-HT-induced venoconstriction or in the doses required for half-maximal venoconstriction (ED50) for 5-HT. Coinfusions of a constant dose of 5-HT caused a significant leftward shift in the dose response curve for noradrenaline as compared with noradrenaline alone. This was indicated by a 9.7 +/- 13.7 fold and a 10.4 +/- 13.8 fold increase in the ED50 for noradrenaline in the patient and control groups, respectively (p = 0.89 between study groups). Our results argue against a generalized increase in responsiveness to the direct or catecholamine-potentiating effect of 5-HT in vivo in vascular smooth muscle, associated with hypertension.


A component of 5-HT-evoked depolarization of the rat isolated vagus nerve is mediated by a putative 5-HT4 receptor. This study describes a component of 5-HT-evoked depolarization of the rat isolated vagus nerve which was unaffected by the 5-HT3 receptor antagonist ondansetron. A grease-gap extracellular recording technique was used. Ondansetron (10-100 nmol/l) displaced the 5-HT concentration-response curve to the right yielding a pA2 value of 8.6 (8.5-8.8), consistent with 5-HT3 receptor antagonism, and revealing a component of the 5-HT response which was resistant to ondansetron blockade. In the presence of ondansetron (100 nmol/l) the maximum depolarization in the resistant phase was 15.5 (12.6-19.2)% of the initial maximum response to 5-HT and the pEC50 value was 7.0 (6.7-7.3). The mechanism of the ondansetron-resistant component of the 5-HT response resembled a 5-HT4-receptor-effect in being absent in preparations equilibrated with 5-methoxytryptamine (10 mumol/l) and antagonised by ICS 205930 (tropisetron, pA2 6.4). 5-Methoxytryptamine alone was an agonist in the vagus nerve with a maximum response similar to that of the ondansetron resistant phase of the 5-HT response. Similarly renzapride alone evoked small depolarizations of this preparation but antagonized the ondansetron resistant phase of the 5-HT response (pA2 7.3-7.4). These effects of 5-methoxytryptamine and renzapride are also consistent with a 5-HT4 receptor mechanism. Ketanserin (1 mumol/l) and methysergide (1 mumol/l) had little effect on responses to 5-HT. The depolarization evoked by this putative 5-HT4 receptor mechanism was small but prolonged and appears to mask and after-hyperpolarizing phase of the 5-HT response in this tissue.


Stimulation of the nigrostriatal dopamine system inhibits both heat production and heat loss mechanisms in rats. The effects of stimulating the pars compacta of the substantia nigra (SNC) on thermoregulation were assessed in normal rats, in rats with chemical lesion of the SNC dopamine (DA) pathways and in rats with striatal DA receptor blockade. Electrical stimulation of the SNC produced hypothermia, decreased metabolism and/or cutaneous vasoconstriction in rats at ambient temperatures (Ta) below 22 degrees C, as well as hyperthermia and cutaneous vasoconstriction in rats at Ta of 30 degrees C. Microinjection of an excitotoxic amino acid (kainic acid) at the same brain sites also produced the same thermal responses. In vivo voltammetric studies revealed that electrical or chemical stimulation of the SNC produced an increase in striatal DA release. The enhanced striatal DA release induced by SNC stimulation was attenuated in rats after selective destruction of the nigrostriatal DA pathway by administration of 6-hydroxydopamine into the medial forebrain bundle. In addition, the magnitude of the thermal responses produced by the SNC stimulation in the cold was attenuated by selective bilateral destruction of the nigrostriatal DA pathways or selective blockade of the striatal DA produced by intrastriatal infusion of haloperidol, a DA receptor antagonist. The results indicate that stimulation of the SNC inhibits both heat production and heat loss mechanisms in the rat.


Pituitary adenylate cyclase activating peptide, a novel VIP-like gut-brain peptide, relaxes the guinea-pig taenia caeci via apamin-sensitive potassium channels. Effects of pituitary adenylate cyclase activating peptide (PACAP-(1-27)) and vasoactive intestinal polypeptide (VIP) on the guinea-pig taenia caeci were studied in the presence of guanethidine and scopolamine. Both peptides (1 nmol/1-1 mumol/l) concentration-dependently relaxed the smooth muscle of the taenia. PACAP-(1-27) and VIP were nearly equipotent. Apamin (30 nmol/l), a selective blocker of calcium-activated potassium channels, abolished the relaxation induced by PACAP-(1-27) whereas the effect of VIP remained unaffected. PACAP-(1-27) may be a candidate for the noncholinergic, non-adrenergic inhibitory neurotransmitter which induces apamin-sensitive relaxation in the intestinal tract.


Inhibition of aconitase by alloxan and the differential modes of protection of glucose, 3-O-methylglucose, and mannoheptulose. Alloxan inhibited aconitase with a half maximal inhibitory concentration of 0.5 mM in sonically disrupted and 2.3 mM in intact isolated liver mitochondria. For dialuric acid the half maximal inhibitory concentrations were 1.1 mM and 2.5 mM, respectively. Ninhydrin and N-ethylmaleimide (NEM) also inhibited aconitase with half maximal inhibitory concentrations in the submillimolar range and t-butylhydroperoxide (BuOOH) in the millimolar range, which, however, were not different for disrupted and intact mitochondria. Only the aconitase substrate citrate, but not glucose provided protection of the enzyme against inhibition. In intact liver cells the half maximal inhibitory concentration for alloxan was 6.8 mM. Again, dialuric acid and BuOOH were less potent inhibitors while ninhydrin and NEM were more potent inhibitors of aconitase in intact liver cells. In intact liver cells, glucose and 3-O-methylglucose, but not mannoheptulose and citrate provided protection against alloxan inhibition. The results show that aconitase is not an enzyme particularly sensitive towards alloxan inhibition and thus apparently not a primary site for mediation of alloxan toxicity as it is the glucokinase. This makes a primary site of alloxan action in the mitochondria extremely unlikely. On the other hand the results demonstrate that both the intact mitochondrial and plasma membrane as uptake barriers provide protection against alloxan toxicity. In addition the results clearly show, that 3-O-methylglucose provides protection against alloxan action only at the level of the plasma membrane through inhibition of alloxan uptake into the cell, while the site of protection of mannoheptulose is only the sugar binding site of the glucokinase. In contrast, glucose is shown here to be the only sugar with a dual protective effect both through inhibition of alloxan uptake through the plasma membrane like 3-O-methylglucose and through protection of the glucokinase sugar binding site against alloxan inhibition of the enzyme like mannoheptulose. In the light of these results the unique protective potency of glucose as compared to that of other sugars is not surprising.


Three levels of dietary calcium-effects on blood pressure and electrolyte balance in spontaneously hypertensive rats. The effects of three levels of calcium intake on blood pressure (BP) and electrolyte balance were studied for 12 weeks in spontaneously hypertensive rats (SHR): the chow of the SHR-1 group contained 1.1% calcium, and that of the supplemented groups 2.1% (SHR-2) and 3.1% (SHR-3) calcium. Wistar-Kyoto rats on a 1.1% calcium diet (WKY-1) served as normotensive controls. After 10 and 12 weeks BP was significantly lower in both calcium-supplemented groups than in the SHR-1 group, the SHR-2 and SHR-3 groups not deviating from each other. Platelets and lymphocytes were used as experimental cell models to study the effects of the calcium diets on intracellular free calcium ([Ca2+]i) level, which was measured by the fluorescent indicator quin-2. At the end of the study [Ca2+]i was lower in both cell types in SHR-2 and SHR-3 than in SHR-1, the supplemented groups being comparable to each other. In platelets [Ca2+]i still remained higher in the calcium-treated than the WKY-1 group, while in lymphocytes the levels were similar between SHR-2, SHR-3 and WKY-1. Plasma sodium, calcium and magnesium levels did not differ in the SHR groups, but plasma potassium was higher in both supplemented groups than in SHR-1. Plasma renin activity was comparable in SHR-1, SHR-2 and WKY-1, but was suppressed in the SHR-3 group. Creatinine clearance in the SHR-3 group was higher than in SHR-1 and SHR-2, but still remained lower than in WKY-1.(ABSTRACT TRUNCATED AT 250 WORDS)


Photochemically induced thrombosis of the rat coronary artery and functional evaluation of thrombus formation by occurrence of ventricular arrhythmias. Effects of acetylsalicylic acid and a thromboxane A2 synthetase inhibitor of thrombus formation. During i.v. infusion of rose bengal (48 mg/kg/h), the proximal portion of the rat left coronary artery was illuminated from the outside of the myocardium by green light (540 nm) to produce a transluminal thrombus subsequent to endothelial damages. The primary endothelial damages within the illuminated vascular portion, which resulted from the photochemical reaction between the dye and green light, and the subsequent formation of transluminal platelet-rich thrombus, were easily revealed by both light and electron microscopy. The establishment of the thrombus was accompanied in all cases by the occurrence of ventricular arrhythmias due to myocardial ischaemia. The times required to initiate ventricular premature beats (VPBs) and ventricular tachycardia (VT) were 381 +/- 96 s and 444 +/- 114 s (mean +/- SEM, n = 10), respectively. Pretreatment of the rat with acetylsalicylic acid (3 and 10 mg/kg, i.v.) before the initiation of illumination had no effect on the times required to exhibit the first VPBs and VT, the incidences of both types of arrhythmias were not reduced, and the thrombus was finally formed. On the other hand, pretreatment with Y-20811, a novel thromboxane A2 synthetase inhibitor (0.3, 1 and 3 mg/kg, i.v.), delayed the onset of both VPB and VT in a dose-dependent manner. The incidences of VPBs and VT were significantly reduced at 1 and 3 mg/kg, and the thrombus formation was prevented. The formation of a transluminal thrombus in the left coronary artery by the present technique was highly reproducible and could be functionally evaluated by the occurrence of ventricular arrhythmias.


Cardiac effects of R 79595 and its isomers (R 80122 and R 80123) in an acute heart failure model. A new class of cardiotonic agents with highly selective phosphodiesterase III inhibitory properties. R 79595 (N-cyclohexyl-N-methyl-2-[[[phenyl (1,2,3,5-tetrahydro-2 oxoimidazo [2,1-b]-quinazolin-7-yl) methylene] amin] oxy] acetamide) and its isomers represent a novel class of compounds with phosphodiesterase (PDE) inhibitory and cardiotonic (positive inotropic) actions. The cardiac effects of this class of compounds were investigated in the hexobarbital-depressed heart-lung preparation of the guinea-pig. After induction of heart failure (reduction of cardiac output to 25% of the initial value) cumulative addition of R 79595 or its isomers R 80122 (E-isomer) and R 80123 (Z-isomer) concentration-dependently reversed the cardiac depressant effects of hexobarbitone-Na. With regard to reconstitution of contractility and cardiac function R 80122 (E-isomer) was 10 fold more potent than R 79595 (1:1 mixture of the isomers) and nearly 100 fold more potent than R 80123 (Z-isomer). Furthermore, the cardiotonic action of the most potent isomer (R 80122) was compared to the effects of several positive inotropic reference compounds. The order of cardiotonic potency was as follows: (-)-adrenaline > R 80122 = adibendan > digitoxin > milrinone = enoximone > theophylline. Adibendan (EC50 value: 6.7 +/- 1.8 x 10.-8 mol/l), which showed cardiotonic effects in the same concentration range as R 80122 (EC50 value: 6.1 +/- 1.3 x 10(-8) mol/l), was significantly (p < 0.01) less effective than R 80122 with respect to the maximally induceable increase in cardiac output (CO). The cardiotonic effects of R 80122 could be observed in the low concentration range of 3 x 10(-8) to 1 x 10(-6) mol/l, whereas enoximone (EC50 value: 1.2 +/- 0.1 x 10(-5) mol/l) and milrinone (EC50 value: 8.9 +/- 3.5 x 10(-6) mol/l) elicited positive inotropic effects at 100 fold higher concentrations. Digitoxin was 10 fold less and theophylline was 300 fold less potent than R 80122 with regard to reconstitution of heart function. The cardiotonic effects of R 80122 were not accompanied by an increase in heart rate as found with milrinone, theophylline or (-)-adrenaline in this model. Furthermore, the PDE inhibitory effect of R 79595 and its E-isomer R 80122 were investigated in partially purified isoenzymes from guinea-pig ventricles. The IC50 values of R 79595 and R 80122 on PDE I-IV were compared to the IC50 values of adibendan, milrinone, enoximone and theophylline. The selectivity of an inhibitor for PDE III was evaluated by division of its IC50 values on PDE I, II and IV by the IC50 value on PDE III. R 80122 was the most potent and selective PDE III inhibitor.(ABSTRACT TRUNCATED AT 400 WORDS)


Electrophysiologic and anticholinergic effects of pirmenol enantiomers in guinea-pig myocardium. Since it has been reported that several class I drugs stereoselectively block sodium channels, potassium channels and muscarinic receptors in cardiac tissues, electrophysiologic and anticholinergic effects of enantiomers of pirmenol, a class I antiarrhythmic drug, were examined. Both (+) and (-) pirmenol depressed the maximum upstroke velocity (Vmax) of the action potential in a concentration-dependent manner in guinea-pig papillary muscles driven at 1.0 Hz, and there was no significant difference in the potency of the class I effect between the enantiomers. The onset rates of use-dependent block (UDB) of Vmax at 2.0 Hz for 10 mumol/l (+) and (-) pirmenol were 0.30 +/- 0.03 and 0.29 +/- 0.01 per action potential, and the recovery time constants from UDB for (+) and (-) pirmenol were 27.0 +/- 2.7 and 27.7 +/- 1.9 s, respectively, indicating no difference in the binding and unbinding kinetics to the sodium channel between the enantiomers. Both (+) pirmenol and (-) pirmenol prolonged action potential duration (APD) at low concentrations (1-10 mumol/l) and shortened it at high concentrations (30-100 mumol/l). Again, there was little difference with respect to the effects on APD between the enantiomers. However, in the isolated guinea-pig left atria (-) pirmenol more potently antagonized the negative inotropic effect of carbachol than (+) pirmenol, and the pA2 values for (+) and (-) pirmenol were 6.41 and 6.71, respectively. The functional study was supported by the radioligand binding experiments using [3H]N-methylscopolamine ([3H]NMS) in guinea-pig left atrial membranes.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of the triazole derivative loreclezole (R72063) on stimulus induced ionic and field potential responses and on different patterns of epileptiform activity induced by low magnesium in rat entorhinal cortex-hippocampal slices. Effects of loreclezole (R72063), a triazole derivative with anticonvulsant properties, were studied on field potentials in rat hippocampal slices and on different patterns of low Mg(2+)-induced epileptiform activity in combined entorhinal cortex-hippocampal slices. Lowering extracellular Mg2+ induced recurrent (10-60/min), short (40-80 ms) discharges in hippocampal areas CA1 and CA3. In the entorhinal cortex (EC) up to 90 s long ictaform events associated with large negative field potential and changes in the neuronal microenvironment were generated. These seizure like events changed their characteristics after one to two hours to recurrent discharges of 0.8 to 10 s. 20 microM loreclezole blocked the seizure like events in the entorhinal cortex completely 30-80 min after onset of application. The recurrent short discharges in the hippocampus were reliably blocked by 40 muM loreclezole 60-90 min after bath application with incomplete recovery after washout of several hours. The recurrent discharges in the entorhinal cortex were reliably blocked by 80 microM loreclezole applied for 80-100 min. Decreases in [Ca2+]o and associated slow field potentials evoked by repetitive stimulation of the stratum radiatum were depressed in a dose dependent manner, while similar changes induced by alvear stimulation remained almost unaffected. A paired pulse stimulus paradigm used to test for effect of loreclezole on synaptically evoked transient field potentials in normal medium revealed interference with mechanisms involved in frequency potentiation. While responses to alvear stimulation were largely unaffected, the response to a paired pulse stimulus to stratum radiatum was depressed over the whole range of tested stimulus intervals (15 to 150 ms). The findings suggest that loreclezole has effects on different patterns of epileptiform activity induced by extracellular low Mg2+ possibly by interfering with processes leading to frequency potentiation.


Pentylenetetrazol-induced seizures in rats: an ontogenetic study. A quantitative description of motor seizures induced by pentylenetetrazol (PTZ, metrazol) was performed. Seizures were induced by PTZ in doses from 40 to 120 mg/kg s.c. in 477 male albino rats of the Wistar strain 7 to 90 days old. Two patterns of seizures were elicited: minimal, i.e. predominantly clonic seizures of facial and forelimb muscles with preserved righting ability, and major, i.e. generalized tonic-clonic seizures with a loss of righting reflex. Minimal seizures could be reliably elicited since the age of 18 days; the CD50 for these seizures did not significantly differ with age. Major seizures were elicited regularly at all developmental stages studied. Their CD50 did not significantly differ among 7-, 12- and 25-day-old rat pups but the value for 18-day-old rats was smaller and for adult animals larger than these three age groups.