Transient high grade heart block following autologous bone marrow infusion. We performed a retrospective analysis of 42 consecutive patients undergoing autologous BMT to determine the incidence of second and third degree heart block following the infusion of cryopreserved autologous bone marrow and to identify any predisposing characteristics. A decrease in heart rate > or = 10 beats/min was observed in 80.5% of patients, with a mean decrement of 27 +/- 7 beats/min. 48.8% of patients developed absolute bradycardia (< or = 60 beats/min). Four of 41 patients (9.7%) experienced high-grade heart block: 9.7% second degree and 4.8% third degree. Heart block patients did not differ from the non-heart block group with respect to age, interval from diagnosis or bone marrow harvest to transplant, cardiac risk factors, pretransplant electrocardiograms or radionuclide angiograms, transplant chemotherapy regimens or serum chemistry values. There was an increased incidence of heart block in patients with prior exposure to cyclophosphamide (p < 0.05) and vinca alkaloids (p < 0.05). There appears to be a high incidence of transient second and third degree heart block following autologous marrow infusion. This may be related to prior chemotherapy, but more likely is an effect of the infusate itself. Predisposing factors were not identified.


A comparison of clinical criteria for the diagnosis of veno-occlusive disease of the liver after bone marrow transplantation. Two major studies have established clinical criteria for the diagnosis of veno-occlusive disease of the liver (VOD) after bone marrow transplantation (BMT). McDonald and co-workers defined VOD as the onset of two of the following occurring before day 30 post-BMT: (a) jaundice (bilirubin > 27 mmol/l), (b) tender hepatomegaly, and (c) ascites or weight gain. In contrast, Jones and co-workers defined VOD as the onset, before day 21 post-BMT, of hyperbilirubinemia (bilirubin > 34 mmol/l) as well as two of the following: (a) hepatomegaly, (b) ascites, and (c) weight gain. We retrospectively reviewed the occurrence of VOD in 101 patients transplanted primarily for hematologic malignancies between 1979 and 1990, applying both sets of criteria. Of the 101 patients, eight (7.9%) fulfilled the Jones criteria whereas 32 (31.7%) had VOD according to the McDonald criteria (p < 0.001). Early mortality (prior to 50 days post-BMT) was 75% (6/8) in patients who fulfilled the Jones criteria but only 28.1% (9/32) in the McDonald group (p < 0.005). Overall, mortality in each group was 75% (6/8) and 65.6% (21/32), respectively. All of the six patients with VOD according to the Jones criteria who died had evidence of hepatic failure. Of the 32 patients who fulfilled the McDonald criteria, eight have also fulfilled the Jones criteria and are described above. Of the remaining 24 patients, 22 had complete resolution of VOD as defined by these criteria within 50 days of BMT, none developed hepatic failure, and 15 died.(ABSTRACT TRUNCATED AT 250 WORDS)


Treatment of myelodysplastic syndrome with busulfan-cyclophosphamide conditioning followed by allogeneic BMT. Twenty-three consecutive patients undergoing related-donor BMT for myelodysplastic syndrome (MDS) were conditioned with a combination of busulfan (BU) and cyclophosphamide (CY). GVHD prophylaxis was with cyclosporine (CSP)/methotrexate (MTX) in 15 patients, CSP/methylprednisolone (MP) in six patients, and CSP/MP/MTX in two patients. The most frequent regimen-related toxicities were oral mucosal (87% of patients, 61% > or = grade II) and hepatic (82% of patients, 43% > or = grade II). The overall incidence of grade II-IV acute GVHD was 48% with eight patients dying of acute or chronic GVHD. There have been five relapses, with the cumulative risk of relapse being 35% (95% confidence interval [CI], 16%-66%). Eight patients remain alive and well (median follow-up 27 months, range 15-70 months), with an estimated 3-year event-free survival (EFS) of 35% (95% CI, 17%-54%). Univariate analysis of EFS by pretransplant variables indicated that only age < or = 35 years correlated with a favorable outcome (p = 0.04). BUCY is an effective, well-tolerated alternative conditioning regimen for MDS patients undergoing allogeneic BMT.


Anti-interleukin-2 receptor monoclonal antibody (BT 563) in the treatment of severe acute GVHD refractory to systemic corticosteroid therapy. Fourteen patients with corticosteroid-resistant acute GVHD were treated with a murine monoclonal antibody to the pp55 interleukin-2 (IL-2) receptor (MoAb BT 563). Nine of the 14 patients had also failed Xoma-Zyme-H65 as GVHD prophylaxis and/or treatment. Seven patients had received HLA-matched sibling donor bone marrow transplants, five had received HLA-matched transplants from unrelated volunteer donors, and two had received one-antigen mismatched transplants from unrelated volunteer donors. At the time of MoAb BT 563 therapy, the overall clinical grading of acute GVHD (Seattle grading system) was as follows: grade II--one patient, grade III--four patients, and grade IV--nine patients. MoAb BT 563 was administered as a short iv infusion of 5 mg daily for 10 doses, followed by 5 mg on alternate days for a further five doses. A complete response (CR) was observed in four patients (28%), and a partial response (PR) in four patients (28%). All four complete responders were treated within 28 days of first onset of grade > or = II acute GVHD. Four patients (three CR, one PR) remain alive. One complete responder subsequently died from chronic GVHD. MoAb BT 563 administration was well tolerated in all 14 patients; no significant toxicity was observed. We conclude that MoAb BT 563 directed against the IL-2 receptor on activated T lymphocytes may be useful in treating corticosteroid-resistant acute GVHD if given early, but that it is of limited value in attempting to rescue patients with far-advanced refractory acute GVHD.(ABSTRACT TRUNCATED AT 250 WORDS)


Escalating sequential high-dose carboplatin and etoposide with autologous marrow support in children with relapsed solid tumors. Thirty patients with relapsed pediatric solid tumors received high-dose carboplatin and etoposide with autologous marrow support in a dose-escalation trial. These patients had received extensive prior treatment, which included both cisplatin and etoposide in 25 cases. Six patient cohorts received carboplatin in doses of 1200-2100 mg/m2 and etoposide in doses of 960-1500 mg/m2. All courses were associated with severe neutropenia and thrombocytopenia. The median times from bone marrow infusion to granulocyte recovery (> 0.5 x 10(9)/l) and platelet recovery (> 50 x 10(9)/l) were 33 and 28 days, respectively, with similar findings for all dosage levels. The frequency of non-hematologic toxicities was generally low, although hyponatremia (Na+ < 129 mEq/l) was seen in one-third of the courses. Hepatoxicity was dose-limiting and was significantly associated with the cumulative prior cisplatin dose (p = 0.006). There were four toxic deaths (CNS hemorrhage, alfa-streptococcal sepsis, Candida sepsis, and enterocolitis). Eleven patients received a second course of therapy; toxicity profiles and times to hematologic recovery were similar for the two courses. Clinical responses were observed at all dosage levels. Eleven of 26 evaluable patients achieved a clinical response (one complete, 10 partial). The majority of responses were in patients with neuroblastoma (six of 16) or Hodgkin's disease (two of three). For phase II clinical trials, we recommend dosages of 2100 mg/m2 of carboplatin and 1500 mg/m2 of etoposide for children with prior cumulative cisplatin exposure < 960 mg/m2. This carboplatin dose represents a three- to four-fold increase over pediatric doses tolerated without bone marrow support.


Orthotopic liver transplantation for hepatic GVHD following allogeneic BMT for chronic myeloid leukaemia. A 21-year-old man who had an HLA-identical sibling donor BMT for chronic myeloid leukaemia developed grade IV acute GVHD of the liver that was unresponsive to corticosteroids and anti-IL2 receptor monoclonal antibody. He was treated with an orthotopic liver transplant and is currently well 6 months later with normal liver function and no evidence of GVHD in the transplanted liver.


Intensive radio-chemotherapy with peripheral blood stem cell transplantation in young patients with chronic lymphocytic leukemia. Two patients with previously treated CLL received autologous PBSC transplantation after total body irradiation and high-dose chemotherapy. Before intensification, a partial marrow response had been obtained in both patients, with disappearance of peripheral blood involvement as shown by immunophenotypic assessment using fludarabine. PBSC collection was performed after a single course of high-dose cyclophosphamide followed by GM-CSF administration. One patient failed to reconstitute normal hematopoiesis and died 3 months post-transplant. The other is in continuous complete remission 12 months after intensification. This strategy in young patients with poor prognosis CLL warrants further investigation.


Selective automatic neuropathy as a novel complication of BMT. Neurological complications have been reported in approximately 30% of patients following bone marrow transplantation. We report a case of selective sympathetic autonomic neuropathy following allogeneic BMT, a phenomenon not previously described.


Antilymphocyte globulin for treatment of pure red cell aplasia after major ABO incompatible marrow transplant. A patient developed pure red cell aplasia after ABO incompatible BMT for leukemia. He did not respond to plasma exchange. Antilymphocyte globulin therapy was followed by complete and permanent erythroid recovery with disappearance of recipient-derived isoagglutinins.


Successful removal of a curled central venous catheter using interventional radiologic techniques. We describe an unusual complication encountered in a patient with a refractory, bulky lymphoma whose 'subclinical' superior vena caval obstruction caused the central venous catheter to curl upon placement. We believe the invasive radiologic techniques described in this report represent a viable alternative to the surgical resolution of such a problem.


[Effectiveness of cisplatin with continuous infusion of 5-fluorouracil in neoadjuvant chemotherapy of carcinoma of the oral cavity]. The effect of neoadjuvant chemotherapy regimen utilizing cisdiaminodichloroplatinum (CDDP) with continuous 5-day infusion of 5-fluorouracil (5-FU) was analyzed during the treatment of 23 patients with histologically established epidermoid carcinoma of the oral cavity. All but four patients were in clinical stage III or IV of the disease and seven of them had an inoperable tumor. No one had been treated previously. Complete response after three courses of the chemotherapy was obtained in 13 patients (56.5%) and partial response in 8 (34.8%). The overall response amounted to 91.3%. Only two tumors showed no response to the therapy after one and two courses respectively. The former patient did not continue the chemotherapy due to toxicity and the latter because of its minimal effect. During therapy the disease did not progress in any of the patients studied. Neoadjuvant chemotherapy with CDDP and continuous 5-FU infusion lasting 120 hours was found to be both tolerable and an effective part of the complex management of oral cavity carcinoma without the risk of delaying subsequent definitive treatment. (Tab. 9, Ref. 18).


[Initial clinical experience with cardiopulmonary bypass and auto-oxygenation]. Cardiopulmonary bypass has been successfully used in cardiac surgery for more than 39 years. In spite of many technical improvements oxygenators are still damaging to blood. We therefore decided to change the conventional CPB into biventricular bypass without an oxygenator, using the patient's lungs for autooxygenation. A double-pump system without an oxygenator was used in 11 patients undergoing aortocoronary bypass grafting. The cardiopulmonary pulmonary setup consisted of two separate systems with two roller pumps, plastic reservoirs and filters, and one cardiotomy reservoir and heat exchanger. Cannulations were performed through the right atrium and pulmonary artery for right-sided bypass, and through the left atrium and aorta for left-sided bypass. The results confirmed the full reliability of autooxygenation during CPB. All patients were well oxygenated with and inspired oxygen fraction of 30%. Values of arterial and venous O2 saturation as well as pO2 and pCO2 varied within the normal range. Platelet count decreased by 23% between interval I and IV, but only by 12% during biventricular bypass. Fibrinogen concentration decreased by 7% between interval I and IV, but only by 4% during biventricular bypass. No patient developed postperfusion syndrome. In our small series of patients, biventricular bypass was found to be simple to handle and to provide good conditions for the surgical procedure. It appears to be more physiological and less traumatic than conventional CPB. The described approach may further improve results of aortocoronary bypass grafting. (Tab. 2, Fig. 1, Ref. 12.).


[Bispecific antibodies: present status and possibile applications]. The review article information on the current status in the research regarding bispecific antibodies. The theoretical and methodical aspects of preparation of heterohybridomas secreting bifunctional monoclonal antibodies are discussed in details. The question of purification of the antibody fraction with dual specificity is also addressed. The possible application of bispecific antibodies in human medicine for immunodiagnostics and aimed immunotherapy is described. (Fig. 7, Ref. 22.).


[Expression of the blood group antigens A and B in carcinoma of the urinary bladder]. In 21 patients with transitional cell carcinoma of the urinary bladder expression of A and B blood group antigens was studied by means of monoclonal antibodies on using indirect immunoperoxide reaction. Expression of the investigated antigens was found to be dependent on the degree of histological differentiation of the tumor tissue. In well-defined carcinomas mostly pronounced expression of A and B blood group antigens was observed. Middle and low-differentiated carcinomas were characterized by reduction or even total deletion of expression. The expected favorable prognosis in transitional cell carcinomas expressing A and B blood group antigens failed to be confirmed. The expression of incompatible A-like antigens in 0 and B blood group patients with carcinoma is discussed. (Tab. 3, Fig. 3, Ref. 22.).


[Effects of postural changes on blood pressure and pulse rate in neonates]. Information about the effect of body position on blood pressure (BP) in neonates is rare and controversial. The aim of the present study was to investigate the type of orthostatic reaction and it s incidence in a group of infants in relation to basal resting values of BP and heart rate (HR). BP and HR were measured in 125 full-term neonates in the supine position and in the 2nd minute after changing the position by lifting on arms to 90 degrees. The reactive values of BP and HR were significantly higher, p < 0.001 (71 +/- 12 compared to 66 +/- 9 Torr in systolic BP, 41 +/- 8 to 37 +/- 6 in diastolic BP, and 127 +/- 19 to 120 +/- 8.min-1 in HR). In the head-up position an increase in BP and HR was observed in about 70% of neonates, whereas their basal supine BP values of 66/36 Torr were significantly lower compared to the values of 69/40 Torr recorded in neonates whose BP decreased in the head-up position. These differences were significant (p < 0.01 syst. and p < 0.001 diast.). (Fig. 3, Ref. 15.).


[Detection of mitochondrial contact sites and creatine kinase activity in Ca(2+)-stimulated rat hearts]. Contact sites are created by fusion of the inner and outer mitochondrial membrane. They represent a dynamic microcompartment for creatine kinase activity. In this microenvironment the active sites of creatine kinase, oxidative phosphorylation, and ADP/ATP transport interact during basal and stimulated metabolism. Electron microscopic studies showed that the occurrence rate of contact sites was changing according to the energy state of the mitochondrion (Biermans et al., 1989). The aim of this study was to stimulate the metabolism of the rat heart by extracellular calcium and to investigate the formation of contact sites and creatine kinase activity. Isolated rat hearts were perfused with Krebs-Henseleit solution containing 1.6 mmol Ca. After 15 min stabilization the hearts were subjected to 0.6, 0.9, 1.8, 2.2, 2.6, or 3.6 mmol Ca for a period of 15 min. Stimulation of the isolated rat hearts with increasing calcium concentrations was physiologically manifested by enhanced dp/dt and LVP values. A higher frequency rate of mitochondrial contact sites was observed and these were associated with higher creatine kinase activity. We suppose that induction of contact site formation is related to the functional activity of the Ca stimulated rat heart and thus efficient production and transport of energy is provided. (Tab. 1, Fig. 6, Ref. 18.).


[The physician and personal computers--applications of the personal computer]. Nowadays the personal computer is being used in a broad range of activities. Its user is also its operator, directly controlling the process. The paper describes both hardware and software, the structure of the PC, its input and output equipment, basic principles of work with a PC, programs and programming, as well as computer languages. A classification of personal computers is presented and the most frequently used types are assessed. (Fig. 6, Ref. 15.).


[Primary axillary subclavian venous thrombosis]. The thoracic outlet syndrome includes primary axillary-subclavian vein thrombosis. The subclavian vein is compressed in a narrowed costo-clavicular space during certain movements of the arm. The intermittent compression of the vein results in position-dependent symptoms often misinterpreted. When occlusion of the vein occurs, the diagnosis of deep venous thrombosis suffers no difficulty. Non invasive vascular diagnostic techniques are nowadays paramount. Treatment, medical and/or surgical (removal of the first rib), depends on the venous state of the upper extremity.


[Mechanism of action and catabolism of atrial natriuretic factor in cultured human and animal kidney cells]. Cultures of renal cells from human or animal origins have allowed the modes of action and the degradation pathways of atrial natriuretic factor (ANF) to be characterized. Human glomerular mesangial and epithelial cells possess ANF receptors of both types, only clearance receptors (C) in mesangial cells, receptors with guanylate cyclase activity (A) and C receptors in epithelial cells which are, in addition, equipped with ectoenzymes rapidly degrading extracellular ANF. Epithelial cells which have been stimulated by ANF secrete cyclic guanosine monophosphate (cGMP) at their apical side. Vascular smooth muscle cells prepared from the rabbit renal cortex also possess A receptors of high affinity and C receptors. Neutral endopeptidase (NEP), an enzyme of which ANF is a specific substrate in the kidney, is expressed at the cell surface. Its expression is inhibited by factors present in the serum and is increased by glucocorticoids. Principal cells of the collecting duct are also a target for ANF via A and C receptors. Taken together, these studies demonstrate that the kidneys are sites both for the physiological effects and the degradation of ANF. Production of cGMP results in vasodilation in the renal cortex, increase of glomerular filtration rate and decrease of sodium reabsorption in the collecting duct. Degradation of ANF occurs via two different ways, its conversion into inactive peptides by NEP and its internalization after binding to C receptors.


[Extracapillary glomerulonephritis with fibrin deposits. Role of thrombin]. Fibrin deposits are frequent in experimental and human glomerulonephritides. In rapidly progressive glomerulonephritis (RPGN), fibrin deposits are considered as a factor generating glomerular sclerosis leading to renal failure. In RPGN, proliferation of epithelial cells associated with macrophages and fibrin deposits is observed in Bowman's space. It is known that 1) thrombin may be locally generated by tissue factor from glomerular cells and macrophages, 2) active thrombin may be adsorbed into the plug. Using immortalized human glomerular epithelial cells, we have tested the hypothesis that active thrombin may be a potential mediator of cellular proliferation, fibrin deposits persistency and collagen IV synthesis. The results clearly demonstrate that epithelial cells bind specifically iodinated alpha-thrombin. Thereafter, these cells synthesize specific inhibitors of plasminogen activators, converting their fibrinolytic activity into antifibrinolytic activity. Moreover, they synthesize soluble form of type IV collagen. A new therapeutical approach of RPGN may be taken into account based on the ability of new drugs inhibiting the binding of thrombin on its specific receptors, and/or inhibiting the extrinsic pathway of the fibrin formation.


[Non-atherosclerotic aorta in Cynomolgus after a hypercholesterolemic regimen]. The authors present a study on atherosclerotic and non atherosclerotic lesions of aortas of Cynomolgus (Macaca Fascicularis) on high fat diet (HFD) (6-12-24 months), after regression and after resumption. At 6 months 2 aortic responses are seen: an edema, deep and superficial clumps of foam cells, few synthetic smooth muscle cells (SSMC) few collagenic fibers--an edema, few superficial foam cells, many SSMC, secreting collagen and elastin. At 12-24 months, after regression and resumption, two aortic lesions are observed: a pronounced atheroma (47 animals) and no atheroma (14 animals). In this case, in the inner part of the aortic wall there are a fibrosis and an elastogenesis, SSMC and just some superficial lipids deposits. These aortic responses of SSMC are certainly the consequence of the environment (the diet) but also the possible consequence of the genetic determinism of SSMC since some animals only present this early, constant fibro-elastic response during this experimental follow up even though all the animals have been subjected to the same lipidic stress.


[Endo-brachy-esophagus, Jean-Louis Lortat-Jacob and Barrett's esophagus. The history of a modern disease]. The term-endo-brachy-oesophagus (EBO) became part of the specialised vocabulary of all those interested in the pathology of the esophagus at the beginning of the 1950s, when J. L. Lortat-Jacob identified the existence, in certain patients suffering from reflux, of the endo-esophageal ascent of a mucosa of gastric type, without any change in the gross topography of the gastro-esophageal junction. A similar lesion had been described at this same time in Great Britain by N. R. Barrett. Since then, numerous clinical and experimental arguments have been advanced in support of the acquired origin of this disorder, in the context of progressive gastro-esophageal reflux disease. However, the possibility of its regression after the treatment of reflux and its histologic presentation, though of major importance, remain unsettled. It is now generally acknowledged that EBO is one of the digestive lesions most prone to malignant degeneration, to the extent that it is now generally classified as precancerous, and all the most sophisticated procedures have been currently suggested and exploited in the attempt to detect EBO of potential malignancy. Within this field, ultramicroscopic study, enzyme assays and the techniques of flow cytometry are being used to better define this problem and to specify the point after which it is legitimate to abandon the medical treatment of reflux in favor of more aggressive methods, even surgical resection of the esophagus. The OESO Group, created in 1978, is currently conducting an international multidisciplinary and multicenter study to deal with the epidemiologic and therapeutic problems of this disorder.


[The role of immunotherapy in the treatment of cancer]. The first attempts to use immunotherapy in patients were made in the late sixties. At this time great hopes were raised by the theory of immunosurveillance developed by Burnet and the promising results obtained against transplantable tumors in mice. However, these expectations were not fulfilled and the results of most randomized clinical trials were disappointing. Since 1985, a better understanding of the fundamental bases of tumor immunology and the results of new human studies has led to a reappraisal of immunotherapy. The data which have accumulated since demonstrate that in certain types of tumors immunotherapy can, in a small proportion of patients, control even disseminated cancers. These new developments give a renewed interest to the results which had been obtained with immunomodulators such as poly A-poly U or levamisole in the adjuvant setting.


[Cancer immunotherapy: from fundamental concepts to clinical applications]. Cancer immunotherapy is based on the idea that anti-tumor immunity in its various aspects can usefully complement the effect of conventional treatments (surgery, chemotherapy, radiotherapy). Immunotherapy can be active when one stimulates the function of cells capable to contribute to tumor regression, notably by using interleukin-2 or immunomodulators. It can be adoptive when patients' lymphocytes are collected and incubated in vitro in the presence of cytokines to enhance their function and reinjected in the patients. It can be passive when one administers anti-tumor monoclonal antibodies.


[Immunotherapy study in the treatment of metastatic renal cancer in the adult]. Metastatic renal cancer is one of the models used for immunotherapy study. Interleukin-2 is active in this chemo and radioresistant tumour and induces the regression of diffuse metastatic lesions sometimes completely through the activation of the immune system. Interleukin-2 has also led to the development of new therapeutic approaches in oncology such as TIL and gene transfer.


[Radiolabeled monoclonal antibodies as anti-tumor missiles, their diagnostic success and therapeutic potential]. While it is now well accepted that radiolabeled antibodies can be useful for tumour detection by immunoscintigraphy, the use of larger doses of more aggressive radioisotopes coupled to antibodies for radioimmunotherapy is still in its infancy. At the experimental level, our group has shown that the intravenous injection of large doses of 131I labeled F(ab')2 fragments from monoclonal anti-carcinoembryonic antigen (CEA) antibodies can eradicate well established human colon carcinoma xenografts in nude mice. At the clinical level, in a dosimetry study performed at the Institut Gustave Roussy, the same anti-CEA monoclonal antibodies and fragments, labeled with subtherapeutic doses of 131I, were injected in patients with liver metastases from colorectal carcinomas. Direct measurement of radioactivity in surgically resected liver metastases and normal liver confirmed the specificity of tumour localization of the antibodies, but also showed that the calculated radiation doses which could be delivered by injections of 200 to 300 mCi of 131I labeled antibodies or fragments, remained fairly low, in the range of 1,500 to 3,000 rads. This is obviously insufficient for a single modality treatment. An alternative approach is to inject radiolabeled antibodies intra peritoneally to treat peritoneal carcinomatosis. Several clinical studies using this strategy are presently under evaluation and suggest that positive results can be obtained when the tumour diameters are very small. In systemic radioimmunotherapy, positive results have been obtained in more radiosensitive types of malignancies such as B cell lymphomas by intravenous injection of antibodies directed against B cell differentiation markers or against idiotypic antigens from each lymphoma, and labeled with 131I or 90Y. The major directions of research for improvement of radioimmunotherapy include the design of genetically engineered new forms of humanized antibodies, the synthesis of original chelates for coupling new radioisotopes to antibodies and the development of two step strategies for immunolocalization of radioisotopes.


[Immunotherapy: current problems and outlook]. Cytokines are soluble mediators involved in cell-cell regulations in the immunological and the hematopoietic systems-genetic engineering now allows the characterization and the availability of recombinant molecules, some of them being recently introduced in clinical trials. Interleukin-2 (IL-2) is the first factor having demonstrated a reproducible therapeutic effect on human metastatic solid tumors, mainly chemo-resistant. High-dose IL-2 alone could achieve about 25% objective responses in metastatic renal cancers, with a low but significant number of complete, relatively long-lasting complete responses. Similar data are obtained in metastatic melanoma. IL-2 is probably acting through the activation of cytotoxic lymphocytes (LAK: lymphokine activated killers). The interest for simultaneous administration of autologous LAK cells generated through in vitro culture with IL-2 is discussed. Cytokine therapy is currently limited by important systemic toxicities, including mainly general symptoms with fever, and a capillary leak syndrome with weight gain, edema and functional renal failure-cardiac are respiratory troubles can be life-threatening. Future trends for cytokine therapy will be: The discovery of new cytokines, and a better knowledge of their synergies, allowing the design of rational associations. A letter understanding of the mechanisms of toxicities, allowing specific prevention of adverse effects not needed for efficient anti-tumor action. Gene therapy, which will lead to important and prolonged release of cytokines by the tumor cells themselves, or by infiltrating peri-tumoral cells, in order to achieve local efficacy and to circumvent systemic toxicities.


[Evolution of the application of nutrition in French nursing infants. Comparison of two national investigations performed in 1981 and 1989]. A national dietary survey was carried out in 1981 among a representative sample of 499 French infants 1 to 18 months of age. When compared with the French Recommended Dietary Allowances, the diets were found to be well supplied in energy, protein and calcium, and inadequately supplied in iron and linoleic acid, mainly for infants aged 4-10 months. A comparative dietary survey conducted by using the same methodology (food consumption was measured by means of a 3-day food record) in 1989 showed that linoleic acid intake was higher in the 6-month-old group. Similarly, iron intakes were higher in 1989 for infants aged 1-7 months. The mean daily iron intake increased between 1981 and 1989 from 5.0 +/- 1.9 mg to 7.3 +/- 2.6 mg in the 6-month-old group and was similar for the older infants. The data suggest that educational programs were partially efficient but it is clear that more effective efforts are needed.


[Brief overview of Franco-Iranian medical relations during the 19th and 20th centuries]. The continuation and the development of medical cultural relations between France and Iran are highly desirable in the interest of both of our countries. After twelve year interruption these relations should become fruitful again. We have attempted to outline several proposals which should enable us to reach this proposed aim.


[Experimental vaccination against HIV-1 protects the chimpanzee against challenge with injections of infected lymphocytes]. Two chimpanzees were immunized against HIV-1 : C-339, using whole inactivated HIV-1 followed by purified recombinant gp160 and a KLH-V3 peptide conjugate; and, C-499, using purified recombinant gp160 and p18gag followed by a mixture of uncoupled V3 peptides. The antigens were emulsified prior to use with one volume Syntex adjuvant SAF-1 containing 1 mg/ml Threonyl-MDP. The animals were challenged twice one year apart by the intravenous route, the first time with cell-free virus, using 40 chimpanzee infectious units (100 TCID50) of the titrated HTLV-IIIB virus stock from the National Cancer Institute, Frederick, MD; and the second time with cell-associated virus, using 6 x 10(5) viable peripheral blood mononuclear cells (PBMC) from a naive chimpanzee which had been injected 3 months earlier with cell-free virus and had become virus-positive within a few weeks. From end-point titration of C-087 PBMC on indicator human PBMC, using a reverse transcriptase assay, this represented the equivalent of at least 15 infected cells. A 3rd chimpanzee, C-435, a näive animal, was also injected with 6 x 10(5) PBMC from C-087 to serve as a positive control. The PMBC of the animals were co-cultivated with fresh human PBMC and assayed for reverse transcriptase on a regular basis. In parallel, ELISA and Western blot analyses were carried out. Virus was detected in the PBMC from C-435 beginning at 4 weeks after challenge. This was followed by seroconversion of the animal to the Env and Gag antigens. By contrast, no virus could be detected in the PBMC from chimpanzees C-499 and C-339 during 7 and 12 months, respectively. Lymph node biopsies and bone marrow aspirates from these animals remained virus-negative upon co-cultivation with human PBMC. PBMC, bone marrow aspirates and lymph node biopsies also scored HIV-negative by polymerase chain reaction. Finally, no anamnestic antibody response of the animals could be detected by ELISA, and no modification of their western blot profiles that could have signed HIV-infection were observed during the follow-up period. C-499 accidentally developed an infectious endocarditis with congestive liver and kidney failure and had to be euthanized at 7 months post-challenge. Specimens from its brain, kidneys, liver, mesenteric nodes, pancreas, salivary glands, and spleen were processed for co-cultivation with human PBMC. No evidence for the presence of virus could be detected by reverse transcriptase assays in any of these co-cultures.(ABSTRACT TRUNCATED AT 400 WORDS)


Investigation of the tendency to wheeze in pollen sensitive patients. We have undertaken a double blind placebo controlled study of the effect of nasal beclomethasone on the tendency to wheeze in 20 unselected hay fever sufferers, half with a history of previous seasonal wheezing. We found no difference between either bronchial hyperresponsiveness, as measured by methacholine challenge, home-monitored PEFR, nor recorded wheeze nor cough between treated and placebo groups although the numbers were small. All were allowed the antihistamine cetirizine hydrochloride 10 mg daily. Eighteen out of the 19 patients had either bronchial hyperresponsiveness (PD20 methacholine < 8 mumol or a > 2 doubling dose change in their PD20 during the pollen season). We have shown a significant positive correlation between a hay fever score (HFS) (created by taking the sum of the home scored; nasal discharge, nasal blockage, eye irritation, sneeze and antihistamine use) and peak seasonal specific IgE to mixed grass pollen (Spearman correlation coefficient 0.5 P < 0.02). There was also a positive correlation between the rise in specific IgE from pre to peak season and the HFS, correlation coefficient 0.6 P = 0.03).


In vitro evaluation of acaricidal and fungicidal activity of the house dust mite acaricide, Allerbiocid. The acaricidal and fungicidal activities of a new acaricide Allerbiocid, formulated for the control of house dust mites, were investigated. The components of the Allerbiocid formula are: 3% benzyl benzoate (acaricidal and fungicidal), 1% tannic acid (protein denaturant) and 0.5% Tween (surface active agent) in 70% isopropyl alcohol (antiseptic). At application rates of both 150 and 250 ml/m2, the acaricidal activity appeared to work by both ingestion and contact and remained active upon contact with mites for up to 10 weeks. When Sabouraud agar was treated with Allerbiocid at a concentration of 5 ml/20 ml medium, the preparation was also found to have fungicidal and fungistatic activities on various species of fungi commonly found in house dust. Allerbiocid contains tannic acid which is a protein denaturant. Levels of Der p I in dust were found to be reduced by over 90% after treatment with Allerbiocid. The importance of denaturation of mite faecal allergens and allergens associated with dead fungal spores is discussed.


Effect of topical application with capsaicin on skin responses to bradykinin and histamine in man. Pre-treatment with topical capsaicin is known to induce neuropeptide depletion from sensory nerve endings and it is a useful pharmacological tool to evaluate the contribution of these nerves to skin injury and inflammation. To investigate the relative contribution of sensory neural stimulation to the action of bradykinin and histamine, a randomized, double-blind study has been undertaken evaluating the effect of topical capsaicin pre-treatment on the responses to intradermal injections of both agonists in 12 healthy volunteers. Capsaicin pre-treatment caused significant inhibition of the immediate mean flare responses (95% CI) to both bradykinin (from 51.5 [39.7-63.3] mm2 to 16.2 [8.0-24.5] mm2) (P < 0.01) and histamine (from 108.4 [80.4-136.4] mm2 to 52.3 [37.1-67.1] mm2) (P < 0.01). Topical capsaicin elicited a significant inhibition of the weal response induced by histamine, the mean weal area being reduced from 14.8 (12.6-17.0) mm2 to 12.1 (10.1-14.1) mm2 (P < 0.05). In addition, the effect of topical capsaicin was to completely inhibit the bradykinin induced weal response compared to control, the mean weal area (95% CI) being reduced from 13.4 (11.4-15.4) mm2 to 8.2 (5.3-11.0) mm2 (P < 0.01). Our findings show that repeated topical application with capsaicin led to a significant reduction of the skin responses to intradermal injections with both agonists, and particularly with bradykinin. The weal responsiveness to bradykinin may entirely follow neuropeptide release from sensory nerves within the skin and the same applies to the flare response, although this is not completely inhibited by topical application with capsaicin.


Longitudinal changes in skin-prick test reactivity over 2 years in a population of schoolchildren with respiratory symptoms. As part of a larger epidemiological study, 114 children with respiratory symptoms, born between 1978 and 1980, were skin-prick tested to Dermatophagoides pteronyssinus (DP), mixed grass pollens (G) and cat dander (C), and to histamine and saline controls (Bencard, U.K.) using 1 mm prick-lancets (Dome/Hollister-Stier), between July and September 1987 and again in October 1989. A weal > or = 2 mm to one or more allergens was regarded as a positive result. Each child was tested by the same investigator on each occasion, using similar techniques. Three children were excluded from analysis as they had failed to respond to histamine testing on one of the two occasions. In 1987, of the 111 children analysed, 58 (52%) children were skin-test positive, and 53 (48%) skin-test negative, while in 1989 62 (56%) were positive and 49 (44%) negative. Twelve children (11%) changed status from negative to positive, while eight (7%) changed from positive to negative. For the group as a whole the percentage agreement between the results obtained 2 years apart was 82%. In comparison to previous studies a greater number of subjects in this population than expected changed atopic status. We therefore further examined the data from those who had changed status and classified as borderline those subjects with no difference in weal size of greater than 2 mm for any allergen between 1987 and 1989. Only five children then changed status from negative to positive, none from positive to negative and 15 demonstrated only borderline changes. The coefficients of repeatability for the 106 children who did not change status were 3.37 mm, 2.80 mm and 2.33 mm for D. pteronyssinus, mixed grass pollens and cat dander respectively. The good short-term repeatability of the testing method was demonstrated in a group of 29 similar children; the coefficients of repeatability were 0.38 mm for DP and G, and 0.72 mm for C. These data demonstrate that, in a population of children with respiratory symptoms, skin-prick testing within individuals is highly repeatable over the short term, but poorly repeatable over a 2 year period. However, the percentage agreement in skin-prick test status for the group as a whole was high (82%). While no child became unequivocally skin-test negative having been previously positive, a small number of children changed status from negative to unequivocally positive, suggesting a genuine but small (4%) increase in the prevalence of skin-test positivity in this population.


Respiratory allergies (bronchial asthma and rhinitis) due to sensitization of type I allergy to red spider mite (Panonychus ulmi KOCH). The inhabitants of a fruit growing area often report spontaneously of sensitization to the red spider mite (RSM) (Panonychus ulmi KOCH). These are for the most part sensitizations with low clinical symptoms (rhinitis, conjunctivitis and erythema). Severe clinical developments with bronchial asthma have been observed. We investigated six patients working in a fruit growing area sensitized by RSM. The sensitizations corresponded to a Type I allergy. Skin tests and provocation tests (nasal as well as bronchial) with RSM showed immediate reactions and RAST positive results were obtained using RSM allergen disks. RAST measurements of sera from nine house-dust mite Dermatophagoides pteronyssinus allergic patients using RSM allergen disks showed RAST-class > 1 for eight patients. RAST inhibition and immunoprint suggest a possible cross-reaction between RSM and D. pteronyssinus.


The antigenicity and allergenicity of microparticulated proteins: Simplesse. New technologies are allowing the food industry to develop products from standard foods which may not be recognized in its modified form by food allergic patients. One such product, Simplesse, has been formulated by microparticulation of egg white and/or cows' milk proteins and is used as a fat substitute in many fat-laden foods. The purpose of this study was to determine whether the process of microparticulation altered the allergenicity/antigenicity of egg white and cows' milk proteins compared to the starting materials. Soluble protein fractions of Simplesse and its respective starting materials were compared to egg white, cows' milk protein, an ultra-filtered egg white/condensed milk mixture, and/or a whey concentrate by SDS-polyacrylamide gel electrophoresis. In addition, sera from 16 patients with documented egg and/or cows' milk hypersensitivity and two controls who were not allergic to egg or milk were used to assess potential allergenicity/antigenicity of these products by immunoblot (Western blot) analysis. There were heterogeneous IgE and IgG binding patterns to the food fractions among these food allergic patients suggesting differing sensitivity patterns among the individuals tested. However, utilizing both SDS-PAGE and immunoblot analyses, the major allergens in the microparticulated products were the same as those found in the starting materials, egg and cows' milk. In addition, there was no evidence of 'novel' protein fractions in the Simplesse test materials compared to the starting materials.


[Emotional states of mothers and behavior of the child during minor surgery]. The objectives of this descriptive correlational study were to measure the degree of fear-anxiety in mothers and in their children who were having minor surgery, and then to verify if there was a relationship between these two variables. Other variables examined were: child's age, rooming-in, premedication and child's experience with surgery. Fifty (50) mothers described their emotional states both on the day before and during the surgery. Fifty (50) children between the ages of two (2) and ten (10) years old were observed at five different times (one hour preoperatively, leaving for the OR, pre-induction, coming back to the room and one hour post-operatively). Their behaviours were noted on a new scale, the ECEO: Echelle des comportement de l'enfant opéré. This scale was developed to describe the behaviour of children experiencing fear, anxiety and pain. Results show that the mother's fear-anxiety score was significantly higher during the surgery than before it (t df.49 = 3.77, p < 0.001). Rooming-in significantly affected mothers' emotional states (F = 4.07, p < 0.05). Younger children were more affected than older ones; they also showed more distress when the mother was present (t df.49 = 5.31, p < 0.001). Rooming-in contributed significantly to the child's stress score; children whose mother roomed-in at least one night showed more distress behaviours than children whose mothers did not room-in (t df.48 = 2.03, p < 0.05). A correlation was found between the degree of mothers' fear-anxiety and the children's fear-anxiety-pain score or distress behaviours (r = .28, p < 0.05). A strong relationship was established between the total distress score of the child and his or her score when the mother was present (r = .81, p < 0.01). These results support the theory of social referencing: the child is influenced greatly by the mother's emotional state and he refers to her for the interpretation of events provoking anxiety. Therefore, nurses should inform and support mothers, especially those who are rooming-in; they should also make more use of parents in the preparation and the management of the child for surgery. The development and validation of an instrument measuring fear-anxiety-pain in younger children will facilitate evaluation of the effect of surgery or other procedures on the child, and the planning adequate interventions.


Purification, properties, and partial amino acid sequence of chitinase from a marine Alteromonas sp. strain O-7. Chitinase (EC 3.2.1.14) was isolated from the culture supernatant of a marine bacterium, Alteromonas sp. strain O-7. The enzyme (Chi-A) was purified by anion-exchange chromatography (DEAE-Toyopearl 650 M) and gel filtration (Sephadex G-100). The purified enzyme showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The molecular size and pI of Chi-A were 70 kDa and 3.9, respectively. The optimum pH and temperature of Chi-A were 8.0 and 50 degrees C, respectively. Chi-A was stable in the range of pH 5-10 up to 40 degrees C. Among the main cations, such as Na+, K+, Mg2+, and Ca2+, contained in seawater, Mg2+ stimulated Chi-A activity. N-Bromosuccinimide and 2-hydroxy-5-nitrobenzyl bromide inhibited Chi-A activity. The amino-terminal 27 amino acid residues of Chi-A were sequenced. This enzyme showed sequence homology with chitinases from terrestrial bacteria such as Serratia marcescens QMB1466 and Bacillus circulans WL-12.


Cloning of genetic loci involved in endoprotease activity in Streptomyces lividans 66: a novel neutral protease gene with an adjacent divergent putative regulatory gene. A skimmed-milk clearing assay was used to identify, in a multicopy Streptomyces lividans 66 genomic library, DNA fragments that lead to increased expression of protease activity in S. lividans 66. Three independent loci were identified. The majority class (slpA, which represented 68 of 71 clones) produced large zones of clearing. Two other classes (designated slpB and slpC) showed smaller zones than slpA. Subcloning and deletion analysis of the slpA locus delineated the relevant DNA to within a 2.5 kilobase pair fragment. DNA sequence analysis revealed a structural gene associated with the appearance of an extracellular protein in the culture medium. The derived amino acid sequence indicated the presence of a zinc-binding motif, which was previously noted to be characteristic of metalloprotease enzymes. However, the relatively small size of the protein (apparent molecular weight 20,000-24,000) suggests that it represents a novel class of neutral proteases distinct from the thermolysin-type enzymes. An adjacent divergent open reading frame was identified and shown to cause a significant increase in protease activity when present together with the protease structural gene on a multicopy plasmid in S. lividans 66. The derived amino acid sequence of this open reading frame showed homology with previously characterized regulatory proteins of the LysR family of transcriptional regulator proteins.


Glyphosate degradation by immobilized bacteria: laboratory studies showing feasibility for glyphosate removal from waste water. To evaluate immobilized bacteria technology for the removal of low levels of glyphosate (N-phosphonomethylglycine) from aqueous industrial effluents, microorganisms with glyphosate-degrading activity obtained from a fill and draw enrichment reactor inoculated with activated sludge were first exposed to glyphosate production wastes containing 500-2000 mg glyphosate/L. The microorganisms were then immobilized by adsorption onto a diatomaceous earth biocarrier contained in upflow Plexiglas columns. The columns were aerated, maintained at pH 7.0-8.0, incubated at 25 degrees C, supplemented with NH4NO3 (50 mg/L), and exposed to glyphosate process wastes pumped upflow through the biocarrier. Glyphosate degradation to aminomethylphosphonic acid was initially > 96% for 21 days of operation at flows yielding hydraulic residence times (HRTs) as short as 42 min. Higher flow rate studies showed > 98% removal of 50 mg glyphosate/L from the waste stream could be achieved at a HRT of 23 min. Glyphosate removal of > 99% at a 37-min HRT was achieved under similar conditions with a column inoculated with a pure culture of Pseudomonas sp. strain LBr, a bacterium known to have high glyphosate-degrading activity. After acid shocking (pH 2.8 for 18 h) of a column of immobilized bacteria, glyphosate-degrading activity was regained within 4 days without reinoculation. Although microbial growth and glyphosate degradation were not maintained under low organic nutrient conditions in the laboratory, the low levels of degradable carbon (45-94 mg/L) in the industrial effluent were sufficient to support prolonged glyphosate-degrading activity. The results demonstrated that immobilized bacteria technology is effective in removing low levels of glyphosate in high-volume liquid waste streams.


Effect of oxygen supply during growth on the production of cytochromes, enzymes, and acid end products by Haemophilus parasuis. Haemophilus parasuis, grown under conditions of high aeration, was found to lack a tricarboxylic acid cycle but to possess phosphoenolpyruvate carboxylase and a reductive pathway leading to the production of succinate. Such organisms contained approximately equal quantities of b-, c-, and d-type cytochromes and excreted acetate. When the oxygen supply for growth was either reduced or eliminated, the specific activities of phosphoenolpyruvate carboxylase, malate dehydrogenase, fumarase, fumarate reductase, and NADH: fumarate oxidoreductase were increased substantially, and the acid products were succinate, acetate, and formate. Organisms grown under the latter conditions also contained increased quantities of b- and c-type cytochromes, some of which were low-potential cytochromes. These low-potential cytochromes were reduced by NADH and oxidized by fumarate, and hence, appeared to be components of NADH: furmarate oxidoreductase. Our results indicate that in H. parasuis, growing aerobically in medium containing glucose, the sole function of the reductive pathway is to provide intermediates for biosynthetic processes, and oxygen is the preferred electron acceptor. As the supply of oxygen is reduced or eliminated, the reductive pathway becomes more involved in NAD+ recycling and fumarate becomes the acceptor. In effect, irrespective of the oxygen supply, the growth of H. parasuis is absolutely dependent upon the presence of an electron transport system.


Interaction between flocculent and nonflocculent cells of Saccharomyces cerevisiae. Interaction between nonflocculent and flocculent cells of Saccharomyces cerevisiae was studied. Adhesion experiments were done using three types of nonflocculent cells and a flocculent one. Two types of nonflocculent cells were obtained from the flocculent strain by changing environmental growth conditions. The integration of nonflocculent cells in the flocs was observed by two different methods: measurement of the sedimentation capacity of mixtures and microscopic observation of stained nonflocculent cells blended with flocculent cells. It was possible to verify that cell-cell interaction corresponds to a true stable binding and not to a simple entrapment inside the floc matrix.


Inhibition of microbial adherence by sphinganine. Sphingosines (precursors and degeneration products of complex sphingolipids) are mediators in membrane second-messenger cascades and in a wide variety of functions in eukaryotic cells. Sphingosines are also lethal for gram-positive microorganisms. In addition to its direct effect, sphinganine is here reported to affect the adherence of Streptococcus mitis to buccal epithelial cells and of Staphylococcus aureus to nasal mucosal cells after incubation for 90 min at 37 degrees C. When the bacteria were pretreated with 8.1, 16.2, 32.5, or (for Strep. mitis) 65 microM sphinganine for 60 min at 37 degrees C, adherence counts were reduced for Staph. aureus by 27, 37, and 60% and for Strep. mitis by 19, 44, 54, and 73%, respectively (p < 0.001). In contrast, pretreatment of buccal cells with 81.2 microM lipid increased adherence by 14% (p < 0.01), but no change occurred at either 16.2 or 325 microM lipid. These results further demonstrate the double-edged ability of sphingosines to regulate cellular activities and their potential as multifunctional therapeutic agents for infectious diseases.


[Antineutrophil cytoplasmic antibodies--ANCA]. Antibodies against cytoplasm of neutrophil leucocytes (ANCA) with a high sensitivity are found in idiopathic necrotizing vasculitis and idiopathic glomerulonephritis with formation of sickles (IGS). C-ANCA are directed against the myeloid lysosomal enzyme--proteinase 3 and are found above all in Wegener's granulomatosis and microscopic polyarteritis. P-ANCA are directed mainly against myeloperoxidase, less against leucocytic elastase or lactoferrin and are found above all in microscopic polyarteritis, IGS, polyarteritis nodosa or Churg-Strauss syndrome. The elevated ANCA level correlates with the activity of the disease or precedes a relapse. These autoantibodies activate probably neutrophil leucocytes for respiratory exacerbation and degranulation which may lead to extensive tissue damage.


[Systemic lupus erythematosus and pregnancy. Present status]. The authors summarize contemporary views on the relationship between pregnancy and systemic lupus erythematosus (SLE) in women of reproductive age. It is known that pregnancy influences in a marked way the basic disease and conversely the disease has an impact on the course of pregnancy. Risks ensuing from these in errelations for women--mothers with SLE and the foetus are exacerbation (flare-up) of the basic disease, loss of the foetus and damage of the foetus by so-called "lupus neonatorum" with skin symptoms and complete cardiac A-V block. The contemporary state of knowledge makes it possible under certain conditions to achieve by therapeutic interventions the development and delivery of a healthy foetus. Among these views it is most important for conception to occur in these patients with SLE during a period without signs of active SLE. To ensure a successful course of pregnancy in women with SLE is the task of a team of appropriate specialists.


[Pharmacotherapy of rheumatoid arthritis in 1992]. The author presents an overview of the state of pharmacotherapy of rheumatoid arthritis in 1992. In the area of non-steroidal antirheumatic drugs attention is drawn to some problems associated with their undesirable action on the digestive tract and articular cartilage. The problem of interindividual reactivity to this group of drugs has not been resolved so far. The mechanism of their action, in particular analgetic action, is also still open. It was found that they can exert not only a central but also a peripheral action. Drugs modifying the disease have been extended by sulphasalazine, methotrexate and cyclosporin. Interest in combined treatment with these drugs is increasing as well as attitudes to the strategy of their application. The corticoid group has been extended by the preparation deflazacort which does not have a catabolic effect on osseous tissue. In the posology of corticoids pulsed treatment is gaining support.


[Use of pulsed steroid therapy in active rheumatoid arthritis]. The authors implemented a prospective double blind, controlled study where they administered to a group of 34 patients with active rheumatoid arthritis either three pulses a 1000 mg methyl prednisolone in a small infusion or an infusion with placebo. They proved a very favourable effect--after five days a marked diminution of morning stiffness was recorded, as well as a diminution of pain during locomotion and rest and general improvement of the patient's condition. After two weeks the effect declines but even after 8 weeks the difference, as compared with the initial value, was still significant, while this was not the case after placebo. The sedimentation rate increased after the initial decline to the baseline level. A number of immunological indicators (decline of immunoglobulins, IgG rheumatoid factor, rise of C3 and C4 complement levels and rise of T lymphocytes) suggest an anti-inflammatory and immunosuppressive action persisting for several weeks although the mentioned results were not statistically significant (the majority at the very borderline of significance, probably significant in a larger number of patients). The authors confirm the reactive safety of this treatment, emphasize the necessity to investigate contraindications of treatment (diabetes mellitus) and the necessity to follow up patients in inpatient facilities.


[Replacement of hip joints with total endoprostheses in patients with ankylosing spondylitis]. In the Rheumatological Institute in Prague the authors made a retrospective investigation in patients with ankylosing spondylitis affecting the hip joints where one or two endoprostheses of the hip joints were made. In all these patients they evaluated the subjective complaints of the operated extremity, they made a clinical examination of the extent of mobility and evaluated X-ray pictures. The group of 20 patients was examined in this retrospective investigation approximately five years after operation. From the total of 32 endoprostheses 20 were cemented and 12 were without cement. From the total number of patients only four reported slight pain in the area of the operated hip joint without signs of loosening on the X-ray picture. The extent of mobility of all endoprostheses was very satisfactory, the mean abduction in men was 27.6 degrees, in women 30.7 degrees, flexion 84 degrees in men and 97 degrees in women. As revealed by X-ray, the hip joints were affected most frequently by coxitis, osteoplastic changes were detected in six patients. Enthesopathies of a small or medium extent were found in almost all patients. Heteroplastic bone was evaluated according to Brooker's criteria, in the investigated group only type I was found. It may be stated that the submitted group of results of endoprostheses in ankylosing spondylitis is a very satisfactory solution which improves the patient's mobility and thus restores the quality of life.


[The first isolation of Dermatophilus congolensis van Saceghem 1913 in Czechoslovakia]. The authors present a report on the first detection of the strain Dermatophilus congolensis in Czechoslovakia. The strain was isolated from ulcerations on the dorsal part of the foot of a 47-year-old patient who previously worked for several years in Libya. Based on examinations of smears from cultures of an isolated strain and four strains from a collection, from experimental dermatofilosis of a rabbit the authors describe the micromorphology of strains of D. congolensis under a light and electron microscope (transmission an and raster microscope). They describe the colonial morphology, haemolytic properties and in particular detail the haemolytic interaction of strains of D. congolensis with Streptococcus agalactiae, Rhodococcus equi and with strains of S. aureus (monoproducers of beta-haemolysin and delta-haemolysin resp.). They assessed the sensitivity of strains of D. congolensis to some staphylococcal bacteriocin-like substances.


[Legionella antibodies in domestic animals]. Serological examination of 420 domestic animals for the presence of antilegionella antibodies indicates their high exposure to legionellae. On examination by the microagglutination reaction with a serum dilution of 1:64 or more the highest positive values were recorded in horses which reacted with antigens of L. pneumophila 1-14 in 36.2% and with antigens of another 19 types of legionellae in 47.8%. In pigs positive values recorded in 16.2% and in 21.1%; in cattle in 3.8% and 29.5%, in sheep in 7.5% and 11.3% and laboratory rabbits were quite negative. The importance of these findings with regard to the possible role of animals in the ecology of legionellae is obscure.


[Personal experience with culture and isolation of urogenital mycoplasmas]. The authors describe the basic technique of cultivation and isolation of urogenital mycoplasmas, focused on their detection. They describe methods of collection of samples, principles of cultivation, dilution of samples and their sub-passage recommended to ensure a higher detection rate.


[Isolation of Nocardia asteroides using hemoculture (clinical report)]. The authors describe the finding of Nocardia asteroides from a haemoculture in a patient with right-sided pneumonia and several predisposing factors (TB, chronic bronchitis, anamnestic hepatitis, chronic hepatopathy, anaemia, ethylism and a neoplasm revealed in the final stage). Early bacteriological diagnosis and aimed antimicrobial therapy led to radical improvement of the very dramatic clinical condition. The authors discuss the possible higher incidence of the infectious agent, which previously was detected only very rarely.


[Virulence factors in strains of Escherichia coli isolated in urinary tract infections]. In 21 strains of E. coli isolated from patients with cystitis and asymptomatic bacteriuria the authors assessed the group, mannososensitive and mannoresistant agglutination of human, bovine, chick, guinea pig, sheep and pig erythrocytes, the production of haemolysis, colicins, aerobactin, the capacity of strains to induce oedema on mouse paws, the lethal effect in mice, the resistance to tetracycline, streptomycin, chloramphenicol, kanamycin, ampicillin and sulfamethoxidine, the transmission of resistance and sensitivity of strains to the action of fresh human serum. In ten strains the authors recorded also the production of haemolysin and the lethality for mice, in four strains the production of colicin V. production of aerobactin and serum resistance. In three strains the aerobactin production was recorded concurrently with the haemolysin production. In none of the samples P. fimbriae were found.


[Epidemiologic aspects of a new approach to monitoring hygienic food handling using the hazard analysis critical control points (HACCP) system]. The hitherto used traditional control of food hygiene focused on assessment whether the controlled sanitary and technological practice is consistent with requirements of regulations sometimes comprises also details of minor importance. To put it briefly, in the course of the production process are many check-up points, but only some or possibly only one is a critical control point. Moreover, by periodic supervision the hygienist is able to record the hygienic and technological state typical only for the time of control. Microbiological examination of final products can reveal only negative sequelae of microbial processes; it does not provide information on the conditions of contamination nor ensure protection against it. For these and other reasons the conclusion is reached that the hitherto used traditional approach of the hygiene supervision is not quite effective and must be replaced by a more active approach focused on the control of factors threatening the wholesomeness already during the production process. The new approach to supervision of food hygiene is the HACCP system (hazard analysis critical control points). The system works rationally as it is based on analysis of systematically assembled data on the causes and conditions which evoked the illness of the consumers by food products or meals. HACCP can be described as prompt, as health or quality problems are revealed immediately after their genesis during production or processing and are eliminated immediately. The system is also comprehensive as it comprises not only the basic technological process incl. processing or modification of ingredients but takes into account also the handling of the given food product after termination of production and in particular final culinar processing. The system can be applied to all pathogenic agents transmitted by foods to man from bacteria and their toxins, viruses, parasites, moulds and mycotoxins, biotoxins but also contaminants and radionuclides. The system requires reliable epidemiological statistics of alimentary infections and intoxications and in practice the participation of an epidemiologist, hygienist and food technologist.


[Relation of foci of tick-borne encephalitis to original plant associations in the Czech Republic]. Based on reports of the hygiene service on the morbidity from tick-borne encephalitis during the period of 1953-1987, data in the literature and results of the authors' field research on the incidence of ticks, the authors defined foci of the disease. In Bohemia there is an extensive important focus of tick-borne encephalitis in the Central Bohemian region in the Krivoklát area, along the Berounka river, along the mid Vltava river and lower Sázava river, in the area of the Brdy mountains and Czech Karst, linked in the South along the Vltava river to foci in the South Bohemian region in the districts of Písek and Ceské Budĕjovice and in the West linked along the Berounka river with the focus in the central part of the West Bohemian region. In the North Bohemian region and East Bohemian region, only minor separate relict foci of tick-borne encephalitis were found. In Moravia there are extensive foci of tick-borne encephalitis in the North Moravian region in the districts of Opava and Bruntál, in the South Moravian region in its central and southern parts. The foci in Bohemia are separated from foci in neighbouring countries, foci in Moravia are continuous with those in Poland and Austria. It was proved that ticks and the revealed foci of tick-borne encephalitis in the Czech Republic are in areas of original oak groves. Areas of beech woods, even those which are below areas where ticks are found, i.e. lower that 700 m above sea level, do not offer favourable conditions for ticks andare not foci of tick-borne encephalitis. Assessment of areas with regard to the prevailing flora and fauna proved a useful method of prediction of the occurrence of ticks and foci of disease they transmit.


[Serologic detection of antibodies to Helicobacter pylori in children and adolescents using ELISA]. Serum antibodies anti-Helicobacter pylori (H.p.) which are specific against antigenic determinants of the bacterial body, were determined in 183 non-ulcer dyspeptic children and adolescents, aged 7- to 20 years. 108 patients (59%) of the same group underwent fibroendoscopy for direct detection of H.p. in bioptic specimens of the stomach and duodenum microbiological methods. Discrimination analysis revealed a overall positive correlation between the results of the two methods: P < 0.01. For detection of specific IgG antibodies against H.p., evaluation by the P/N ratio was used, i.e. the absorbance of the examined serum (P) was compared with the mean absorbance of three negative serum controls (N). The specificity of the ELISA test in this examination was 96.1%, the sensitivity of 90.3%. The mentioned ELISA method for detection of specific IgG antibodies against H.p. is comparable with direct microbiological prove of H.p. presence in mucus specimens in our group examined. It is a very important non-invasive method useful for primary detection of H.p. infection.


[The effect of breast feeding on anthropometric indicators in very young children]. In a group of 4,217 children from Southern Moravia the authors evaluated growth from birth to the age of three years in relation to breastfeeding. The basic anthropometric data were evaluated in sub-groups made with regard to birth weight. Infants who were breastfed longer than four months were lighter during the first half year, as compared with those breastfed for a shorter period; at the age of one year this difference disappeared, in the second year they tended to have a higher weight, in the third year the differences were balanced. The same was manifested in weight increments. The influence on height was not so marked and was manifested mainly in the difference of increments. The head circumference and chest circumference was not much influenced. Evaluation of indices of proportionality revealed a more harmonious growth in children who were breastfed for a longer period.


[Long-term clinical experience with spherocytic hemolytic anemia in children]. The author investigated during a 15-year period 27 children with spherocytic haemolytic anaemia. In 20 children the disease was familial. The initial symptoms were jaundice and anaemia. In six children the disease was manifested by severe neonatal jaundice and in four an exsanguination transfusion was made. Of five older children three were at first treated for infectious hepatitis. The anaemic syndrome was in the foreground of the clinical picture in 16 children, incl. 10 where it was present already in infant age. In 24 children splenectomy was performed, usually after the age of 6 years. For prophylaxis of bacterial infection the splenectomized children were given penicillin preparations for a period of three years. The OPSI syndrome was not recorded.


[Sonographic screening of upper urinary tract obstruction and approach to its treatment]. Sonographic examination of the uropoietic tract (UPT) in neonates makes it possible to detect in time inborn disorders, in particular of the excretory pathways and to provide treatment before irreversible changes develop. The authors subject within the framework of screening of inborn developmental defects all neonates to sonographic examination of the UPT. In the submitted paper they evaluate a group of 2,103 children in the course of a one-year period; 89 neonates had changes as compared with normal findings. The largest group is formed (by as many as 57) by marginal dilatation of the hollow space. A dilated hollow system was recorded in 24 neonates and eight had different types of anomalies. By further comprehensive investigation conservative or surgical procedure is indicated to prevent further, possibly irreversible damage of the renal parenchyma.


[Analysis of the effect of 2 criteria in the creation of age groups on the value spread of selected somatic parameters]. In the paper we present the results from the statistic analysis of the effect of two age-group creation criterions (1-st criterion--f.e. the 7 years old--from 6,500 to 7,499 years, 2-nd criterion--f.e. the 7 years old--from 7,000 to 7,999) on value dispersion of somatic characteristics chosen. We analyzed the body weight and body height in the set consists of 846 Gypsy children from 7 to 13 years of age. The dispersions of individual values of parameters followed for both criterions were tested by F-test, average values by un-pair Student's t-test. We found that the both criterions have no effect on value dispersion of evaluated somatic characteristics, except of two cases. Further, average values of testing parameters by criterion No. 1 are somewhat lower in comparison with those by criterion No. 2. Statistically significant differences are however incidental and rare. We suggest that in the case of population studies concerning with body growth and development it is possible to compare the sets with different age-group creation criterions.


[Macroprolactinoma with galactorrhea in a 13-year-old girl]. The case-history demonstrates a 13-year-old girl with the unusual finding of a macroprolactinoma which was clinically manifested by galactorrhoea. By medicamentous treatment the prolactin (PRL) level was reduced in six weeks and the clinical condition improved.


[Pneumoperitoneum in a premature, non-ventilated neonate with respiratory distress syndrome leading to respiratory failure. What is the etiology?]. The authors describe the finding of extensive tensive pneumoperitoneum which developed during distension therapy (CNP) in an immature neonate. The absence of another extraalveolar cumulation of air made the authors use PO2 analysis in the peritoneal air by means of a transcutaneous electrode. Because the PO2 rose after connection of the electrode to the catheter draining the abdominal cavity, the authors assumed a respiratory aetiology of the pneumoperitoneum and did not consider laparatomy. They assume that the above test may be valuable in the differential diagnosis between respiratory and gastrointestinal pneumoperitoneum in those instances where it is not possible to assess the aetiology of pneumoperitoneum unequivocally.


[Juvenile granulosa cell tumor with subsequent occurrence of gastrointestinal polyposis, subcutaneous lipomatosis and nodular goiter]. A juvenile tumour from granulosa cells (M-8622/1), 13 x 8 x 6 cm, in the right ovary in a three-month-old girl produced some symptoms of pseudopubertas praecox isosexualis which disappeared after operation. Microscopic examination of the tumour revealed in addition to typical structures a less common differentiation to Sertoli cells. Despite actinotherapy and chemotherapy one and a half years after the onset of the disease X-ray examination revealed metastases in the lungs which were successfully cured by further doses of the above two types of treatment. Between the age of 6 and 15 years the girl developed successively polyposis of the stomach, small and large intestine (M-7564/0), subcutaneous lipomatosis of the trunk and left lower extremity (M-8881/0) and nodular goitre (M-7164/0), predominantly quiescent. In the literature a connection between gonadal stromal ovarian tumours and mesenchymal tumours, intestinal polyposis and disorders of the thyroid gland is described, but in different patients. The authors' observation is unique by the successive incidence of these changes in a single patient surviving 15 years after operation; and thus genetically conditioned associations could be involved.


[Adverse effects of Spophyllin Spofa Retard]. An account of reported undesirable effects of Spophyllin Spofa Retard based on the register of the Commission for monitoring undesirable effects of drugs of the Scientific Council of the Ministry of Health between December 1987 and November 1990. It is surprising that no undesirable effects of Syntophyllin and Spophyllin were reported from paediatric practice. The author submits questions associated with the discrepancy of notifications of undesirable effects of Spophyllin in adults and children.


[Tussigenic reaction after inhalation of Intal]. Sodium cromoglycate is the drug of choice in the preventive treatment of bronchial asthma. Its only undesirable effect is the tussigenic irritation of the respiratory pathways which may lead to termination of its administration. The author analyzes the causes and submits proposals for their solution.


Amino acids and peptides. XXXV. Synthesis of mouse metallothionein I. (2). Synthesis of a nonacosapeptide corresponding to N-terminal sequence 1-29 (beta-fragment) of mouse metallothionein I and related peptides and examination of their heavy metal-binding properties. A nonacosapeptide corresponding to the N-terminal sequence 1-29 (beta-fragment) of mouse metallothionein I and related peptides were synthesized by the conventional solution method and their heavy metals (Cu2+, Cu+ and Cd2+)-binding properties were examined. The Cu(2+)- or Cu(+)-binding activities of various peptides were not greatly dependent on the peptide structure, so far as examined, as in the cases of N. crassa MT and A. bisporus MTs. On the contrary, the Cd(2+)-binding activities of these peptides were fairly structure-dependent.


Synthesis and biological activity of 5'-aminobenzoxazinorifamycin derivatives. Benzoxazinorifamycin reacted with various secondary amines to yield various 5'-substituted aminobenzoxazinorifamycin derivatives. The derivatives exhibited potent activities against gram-positive bacteria and mycobacteria. The antimicrobial activities of these compounds against Mycobacterium tuberculosis and Mycobacterium intracellulare were superior to those of rifampicin. Some of these compounds showed good plasma levels after oral administration in rats.


Syntheses of 2-(3,4-dimethoxyphenyl)ethylamine derivatives and their antiulcer activities. A series of acyl derivatives of 2-(3,4-dimethoxyphenyl)ethylamine (4) were synthesized and evaluated for their effectiveness to prevent water-immersion stress-induced gastric ulceration when given intraperitoneally to rats. Among them N-[2-(3,4-dimethoxyphenyl)ethyl]-2-phenylaminoacetamide hydrochloride (15) had significant antiulcer activity. Further modification of the four parts of 15 revealed that only the introduction of a carbamoyl group into 2- or 3-position of the phenylamino part gave compounds (49-51, 54 and 55) which retained antiulcer activity comparable to the lead compound. However, the compounds (49-51 and 54) did not exert a prophylactic effect when administered orally except for the 3-substituted bezamide derivative 55. Alkyl substitution on the nitrogen of benzamide gave 3-[[[2-(3,4-dimethoxyphenyl)ethyl]carbamoyl]methyl] amino-N-methylbenzamide (66, DQ-2511) and the related compounds (67, 70, 74 and 77) which all had potent antiulcer activities at oral doses of 50-400 mg/kg.


3 beta-Hydroxysialic acid glycosides. I. Calcium-binding ability and chemical and enzymatic stabilities. Methyl alpha- and beta-glycosides of N-acetylneuraminic acid (Neu5Ac) and N-acetyl-3 beta-hydroxyneuraminic acid (Neu5Ac beta 3OH) (1-4) were prepared to evaluate their calcium-binding ability. (Methyl alpha-glucopyranosidonyl) alpha- and beta-, and 4-methylumbelliferyl alpha-glycosides of Neu5Ac and Neu5Ac beta 3OH (5-10) were also synthesized for the comparison of chemical and enzymatic stabilities, respectively. Methyl beta-glycosides of Neu5Ac and Neu5Ac beta 3OH, 3 and 4, respectively, showed intense calcium-binding abilities, while no such ability was observed in the corresponding alpha-glycosides, 1 and 2. The Neu5Ac beta 3OH glycosides, 6, 8, and 10, showed much stronger resistance to acidic hydrolysis and sialidase digestion than the corresponding Neu5Ac glycosides, 5, 7, and 9.


Synthesis and cardiovascular activity of phenylalkylamine derivatives. I. Potential specific bradycardic agents. A series of acyclic amide derivatives of N-(omega-aminoalkyl)-N-methylhomoveratrylamine was synthesized and evaluated for their bradycardic activity in isolated guinea pig right atria. Among these compounds, (E)-N-[3-[N'-[2-(3,4-dimethoxyphenyl)ethyl]-N'-methylamino]propyl]- 4-[3,4-(methylenedioxy)phenyl]-3-butenamide (35) was the most potent in vitro and was also found to show dose-dependent bradycardia without remarkable reduction of left ventricular dp/dtmax or mean aortic pressure in anesthetized dogs.


Phenolic constituents in Erythrina x bidwilli and their activity against oral microbial organisms. Five flavonoid compounds, including two new isoflavanones, were isolated from the root bark of Erythrina x bidwilli. Their structures were determined to be 2,10-di(gamma,gamma-dimethylallyl)-3,9-dihydroxypterocarpan (erythrabyssin II), 6,8-di(gamma,gamma-dimethylallyl)-7,2',4'-trihydroxyisoflavanone (bidwillon A), 8-gamma,gamma-dimethylallyl-2',4'-dihydroxy-[6",6"-di-methylpyrano - (2",3":7,6)]isoflavanone (bidwillon B), 8-gamma,gamma-dimethylallyl-7,4'-dihydroxyisoflavone (8-gamma,gamma-dimethyl-allyldaidzein), and 8-gamma,gamma-dimethylallyl-5,2',4'-trihydroxy-[6",6"-dimethylpyrano+ ++- (2",3":7,6)]isoflavone (auriculatin), by means of spectroscopic analysis. Some potent activities against oral microbial organisms (Fusobacterium nucleatum and Prevotella intermedia) were shown in these flavonoid compounds.


Synthesis of deuterium-labeled 16 alpha,19-dihydroxy C19 steroids as internal standards for gas chromatography-mass spectrometry. [2 beta,7,7,16 beta-2H4]16 alpha,19-Dihydroxyandrost-4-ene-3,17-dione (14) and [7,7,16 beta-2H3]3 beta,16 alpha,19-trihydroxyandrost-5-en-17-one (16), with high isotopic purity, respectively, were synthesized from unlabeled 3 beta-(tert-butyldimethylsiloxy)-androst-5-ene-17 beta-yl acetate (1). The deuterium introduction at C-7 was carried out by reductive deoxygenation of the 7-keto compound 3 with dichloroaluminum deuteride and that at C-2 beta and/or C-16 beta by controlled alkaline hydrolysis of 16-bromo-17-ketone 11 or 12 with NaOD in D2O and pyridine. [7,7-2H2]3 beta-Hydroxyandrost-5-en-17-one (6), obtained from compound 1 by a five-step sequence, was converted to compound 14 or 16 by an eight-step or seven-step sequence, respectively. The labeled steroids 14 and 16 are useful as internal standards for gas chromatography-mass spectrometry analysis of the endogenous levels.


Inhibition of urease activity by dipeptidyl hydroxamic acids. A series of dipeptidyl hydroxamic acids (H-X-Gly-NHOH: X = amino acid residues) was synthesized, and the inhibitory activity against Jack bean and Proteus mirabilis ureases [EC 3.5.1.5] was examined. A number of H-X-Gly-NHOH inhibited Jack bean urease with an I50 of the order of 10(-6) M and inhibited Proteus mirabilis urease with an I50 of the order of 10(-5) M. The inhibition against Jack bean urease was more potent than that with the corresponding aminoacyl hydroxamic acids (H-X-NHOH).


The influence of chloro-substituent sites of hexachlorobiphenyl on the respiration of rat liver mitochondria. The actions of three hexachlorobiphenyls (HCBs) 2,3,4,2',3',4'-, 2,3,4,3',4',5'- and 3,4,5,3',4',5'-HCBs, on the respiration of rat liver mitochondria with succinate as the substrate were compared, and the effect of chloro-substitution sites in HCB on the respiration was examined. 2,3,4,2',3',4'-HCB strongly inhibited both state 3 and 2,4-dinitrophenol (DNP)-stimulated respiration with 50% inhibition dose of 52 and 54 microM for state 3 and DNP-stimulated respiration, respectively. The inhibitory action of 2,3,4,3',4',5'-HCB on both respiration was approximately half as potent as that of 2,3,4,2',3',4'-HCB. On the other hand, 3,4,5,3',4',5'-HCB did not inhibit any respiration at all. These results indicate that both inside (ortho) and outside (meta or para) positions in each phenyl ring of the biphenyl molecule should be replaced with chlorines for HCB to be an effective inhibitor. Either the actual position of chloro-substituent or steric conformation caused by its substitution or both can be considered as factors affecting the inhibition. On the basis of the conformational energy, calculated by AM1 (Austin model 1) method, with increases in chlorine number in ortho position, HCB molecule became angulated. Furthermore, calculated probability of the conformation distribution for HCB indicated that the probability of nonplanarity was higher for effective HCB than for less effective HCB. These structural features suggest the significance of steric conformation as well as chloro-substituent sites in determining the inhibitory ability of HCB.(ABSTRACT TRUNCATED AT 250 WORDS)


80 kDa mouse sperm protein as a substrate of protein kinase C. Calcium and phospholipid-dependent protein kinase (PKC) activity was detected mainly in the cytosol of the mouse sperm. The PKC in the cytosol fraction was partially purified by ion-exchange chromatography. Using the partially purified PKC, the phosphorylation of PKC substrates was examined in vitro. The phosphorylation of the 80 kDa protein was enhanced by phorbol ester treatment in vitro as well as in vivo. The partial amino acid sequence of this protein was homologous with that of guanosine 5'-cyclic monophosphate (cGMP)-dependent protein kinase and myosin light chain kinase, both of which are related to ligand-receptor-transduction. The present data suggest that the activation of PKC and subsequent specific protein phosphorylation might be involved in the regulation of the zona pellucida-induced acrosome reaction.


Three types of membranous ATPase on rat liver lysosomes. At least three types of vanadate-insensitive membranous ATPase were identified on rat liver lysosomes: bafilomycin A1-sensitive Mg(2+)-ATPase (H(+)-ATPase), N-ethylmaleimide (NEM)-sensitive but bafilomycin A1-insensitive Mg(2+)-ATPase (ATPase I), and NEM-insensitive Ca2+/Mg(2+)-ATPase (ATPase II). They showed different sensitivity to chemicals and ions with apparent molecular masses of 700-800, 500-650, and 360 kDa, respectively. Of these membranous ATPases, H(+)-ATPase seemed to constitute only one tenth of the ATPase activity on rat liver lysosomes and to be the only ATPase that exposed its active site to the cytoplasmic side of the lysosomal membranes.


Glycation and insolubility of human lens protein. To learn whether glycation plays a role in insolubilization or in senile cataractogenesis, the reactivity of lens protein from normal and senile cataractous lenses and individual crystallin prepared from human lens with various sugars [glucose, glucose-1-phosphate (G-1-P), glucose-6-phosphate (G-6-P) and fructose], and the insolubility of those proteins were determined. The reactivity of human lens protein to glucose was increased in a dose-dependent manner, and it was demonstrated that 17.9, 18.5 and 24 kDa proteins were susceptible to glycation with sugars. The study also showed that alpha-, beta-crystallins and high molecular weight (HMW) aggregate obtained from cataractous lens have some weak reactivity against sugars. It was demonstrated that the proteins obtained from normal lens of older age and from cataractous lenses have higher insolubilities to glucose than do normal younger ones. Measurement of glycosylated protein by affinity column chromatography revealed that cataractous lenses contained a larger amount of glycosylated protein than normal ones. These results suggest that there is an age-related increase of glycation in normal human lens protein, and that such glycation increases the amount of insolubilized protein with the effect of aging. The author also speculates that an abnormal acceleration of glycation in the human lens may induce senile cataract formation.


In vitro replication study of modified bases in ras sequences. DNA templates containing a modified base (O6-methylguanine, 8-hydroxyguanine, xanthine or hypoxanthine) which was located in nucleotide sequences corresponding to the 12th or 61st codon of a ras gene were synthesized and deoxynucleotide incorporation opposite the lesions was investigated. The templates were replicated by Taq DNA polymerase, recombinant rat DNA polymerase beta and mouse DNA polymerase alpha-primase complex. Sequence analysis of the replicated products indicated selective incorporation of nucleotide(s) opposite a modified base, depending on the kind of base and of DNA polymerase. This system is very useful to obtain results of in vitro replication of modified bases in ras sequences.


Effects of 32-oxygenated lanosterol derivatives on 3-hydroxy-3-methylglutaryl coenzyme A reductase activity and cholesterol biosynthesis from 24,25-dihydrolanosterol. The effects of 32-oxygenated lanosterol derivatives on 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity and cholesterol biosynthesis from [24,25-3H]24,25-dihydrolanosterol were studied. Among the derivatives, 3 beta-hydroxylanost-7-en-32-oic acid was the most active in depressing HMG-CoA reductase activity (IC50: 0.7 microM) and cholesterol biosynthesis (IC50: 0.4 microM) from 24,25-dihydrolanosterol.


Preparation and evaluation of eudragit gels. V. Rectal gel preparations for sustained release and avoidance of first-pass metabolism of lidocaine. The release of lidocaine from hydrogel and xerogel preparations was remarkably suppressed compared with polyethylene glycol (PEG) 2000 suppository. The release rate of lidocaine from hydrogel and xerogel increased with the increase in the amount of sodium hydroxide incorporated within the range of 3 to 7 milliequivalent (meq). After an oral administration of lidocaine HCl solution, the plasma concentration of lidocaine was considerably lower than that after intravenous administration for all time periods. The absolute bioavailability (F(oral)) was 5.63%. For the Witepsol S-55 and PEG 2000 suppositories, the plasma levels of lidocaine were higher than those for the oral preparation, and Cmax and area under the concentration-time curve (AUC) values significantly improved (p < 0.01). The absolute bioavailabilities were 21.3 and 29.6%, respectively. On the other hand, Eudispert hv-hydrogel and xerogel preparations showed the characteristics of a sustained-release preparation, especially the xerogel preparation with 5 meq NaOH. Absolute bioavailability for hydrogel and xerogel preparations increased significantly (p < 0.05) by approximately 1.7-3.4 folds compared with those of Witepsol S-55 and PEG 2000 suppositories.


A study of embolizing materials for chemo-embolization therapy of hepatocellular carcinoma: antitumor effect of cis-diamminedichloroplatinum(II) albumin microspheres, containing chitin and treated with chitosan on rabbits with VX2 hepatic tumors. As an effective therapy for hepatocellular carcinoma, hepatic arterial chemo-embolization therapy has been widely used, and many embolizing materials have been extensively investigated. In the present study, we prepared various types of cis-diamminedichloroplatinum(II) (CDDP) albumin microspheres using chitin and chitosan, both of which have attracted considerable attention as new non-toxic biological polymer materials having favorable characteristics such as immune adjuvant activity, biological compatibility, and biodegradation. Hepatic artery of rabbit hepatic cancer models, which had transplanted VX2 tumors, were embolized with various types of microspheres. The anti-tumor effects and tumor-targeting of the microspheres, and the effects of the microspheres administration on the hepatic tissue were investigated. As a result, anti-tumor activity of the microspheres was increased by the addition of chitin-containing or chitosan treated materials; tumor growth rates of chitin addition and chitosan treated groups were approximately 160% and 120%, respectively, and were significantly lower than that of the non-treatment groups with a rate of approximately 580%. However, complete inhibition of tumor growth might have been impossible. Anti-tumor activity was increased by the addition of chitin-containing or chitosan treated materials. Whereas the growth inhibitory effect was insufficient, in order to potentiate anti-tumor activity, higher CDDP contents and sustained release of CDDP at a high level from microsphere and so on should be essentially improved for the near future. The CDDP level in hepatic tissue following the administration of microspheres was increased by adding chitin to the microspheres or by treating the microspheres with chitosan.(ABSTRACT TRUNCATED AT 250 WORDS)


Synthesis and drug-release characteristics of the conjugates of mitomycin C with N-succinyl-chitosan and carboxymethyl-chitin. By condensation of mitomycin C (MMC) with N-succinyl-chitosan (Suc-chitosan) and carboxymethyl-chitin (CM-chitin) using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, Suc-chitosan-MMC conjugate (Suc-chitosan-MMC) and CM-chitin-MMC conjugate (CM-chitin-MMC) were prepared, respectively. The reaction conditions for 45 min at pH 5 and for 2 h at pH 5 were selected as the most appropriate for the preparations of Suc-chitosan-MMC and CM-chitin-MMC, respectively. Suc-chitosan-MMC was obtained as a water-insoluble product, while CM-chitin-MMC was partially water-soluble. When the ratio of MMC to the polymer supports changed in the conjugation reaction, the conjugates with 33% (w/w) and 23% (w/w) MMC contents were obtained as those most highly drug-loaded for Suc-chitosan-MMC and CM-chitin-MMC, respectively. At pH 7.4 at 37 degrees C, Suc-chitosan-MMC regenerated MMC very slowly, while the release of MMC from CM-chitin-MMC was relatively fast. Each drug release followed very nearly pseudo-first order kinetics, in which the apparent drug release rate constants (k(apps)) of Suc-chitosan-MMC and CM-chitin-MMC were 3.9 x 10(-3) and 1.1 x 10(-1) (h-1), respectively.


External control of drug release and penetration. VI. Enhancing effect of ultrasound on the transdermal absorption of indomethacin from an ointment in rats. The effect of an ultrasound (1 MHz) on transdermal absorption of indomethacin from an ointment was studied in rats. Ultrasound energy was supplied for between 5 and 20 min at a range of intensities (0.25, 0.5, 0.75, and 1 W cm-2), energy levels commonly used for therapeutic purposes. For evaluating skin penetration of indomethacin, the change of plasma concentration was measured. The pronounced effect of ultrasound on the transdermal absorption of indomethacin was observed at all ultrasound energy levels studied. The intensity and the time of application were found to play an important role in the transdermal phonophoretic delivery system of indomethacin; 0.75 W cm-2 appeared to be the most effective intensity in improving the transdermal absorption of indomethacin, while the 10 min ultrasound treatment was the most effective. Although the highest penetration was observed at an intensity of 0.75 W cm-2, 0.5 W cm-2 was preferred because intensities of less than 0.5 W cm-2 of ultrasound for 10 min did not result in any significant skin temperature rise nor did it have any destructive effect on rat skin. Progressively more skin damage was noted as the intensity and the time of application of ultrasound increased. When used at a proper intensity and time of application, ultrasound appears to be a safe technique for enhancing the passage of various drug molecules through human skin.


A fluorescent DNA probe prepared by the direct derivatization of the sugar moiety for hybridization assay. The preparation of a fluorescent DNA probe based on the derivatization of the terminal hydroxyl group of the sugar moiety of a DNA primer and its applicability to the DNA hybridization assay are described. M13mp8 plasmid primer reacts with 2-(5-chlorocarbonyl-2-oxazolyl)-5,6-methylenedioxybenzofu ran in the presence of sodium azide to form the corresponding fluorescent probe, which can be used for the hybridization assay to the target DNA, M13mp8 plasmid vector. The detection limit of the DNA with the naked eye is 10 ng (approximately 300 fmol)/spot on filters for the hybridization assay.


Purification and characterization of hamster hepatic microsomal N,O-acetyltransferase. A microsomal N,O-acetyltransferase which activates carcinogenic arylacetohydroxamic acids was purified 75-fold from hamster liver sequentially by anion exchange column chromatography, chromatofocusing, gel filtration, and hydroxyapatite column chromatography. The purified enzyme, AT-2, was a glycoprotein with a molecular weight of 60000 and a pI value of 5.4. The N-terminal amino acid sequence of AT-2 was: 60000 and a pI value of 5.4. The N-terminal amino acid sequence of AT-2 was: Asp-Ser-Pro-Ser-Pro-Ile-Arg-Asn-Thr-His-Thr-Gly-Gln-Val-Arg-Gly-Leu-Val- His- Lys-. This sequence was highly homologous to that of the form 2 carboxylesterase of rabbit liver, but not to that of major hepatic microsomal carboxylesterases of hamster and other species. AT-2 catalyzed the hydrolysis of 4-nitrophenyl acetate and the N,O-acetyltransfer of N-hydroxy-2-acetylaminofluorene. Both enzyme activities were strongly inhibited by paraoxon, but not by iodoacetamide. These results demonstrate that this N,O-acetyltransferase is a member of carboxylesterase (EC 3.1.1.1).


Contributions of various serum proteins to the binding of prasterone sulfate in humans. The binding of prasterone sulfate (PS) in human plasma was investigated. Binding percentages of PS to human plasma, human serum albumin (HSA), human alpha 1-acid glycoprotein (AGP) and human gamma-globulin (GGL) were independent of the PS concentration between 0.1 and 8.0 micrograms/ml. The mean binding percentages were 99.1% for human plasma, 98.3% for HSA, 12.6% for AGP and 8.1% for GGL. Though PS is an acidic drug, binding of PS to AGP was observed. From the binding index, it was found that PS mainly bound to HSA in human plasma and that the contributions of AGP and GGL to PS in plasma were negligible.


Distribution of pirarubicin in human blood. We investigated the distribution and stability of pirarubicin in human blood obtained from 12 healthy volunteers. The distribution of pirarubicin into blood cells showed marked temperature- and concentration-dependencies and the Arrhenius plot for pirarubicin uptake in blood was biphasic. Therefore, pirarubicin appears to be taken up into blood cells by a carrier-mediated system. Pirarubicin was mainly enzymatically metabolized to pirarubicinol in blood cells, but pirarubicin was not metabolized into doxorubicin in either blood or plasma. On the other hand, in plasma, pirarubicin was degraded to unknown inactive compounds instead of pirarubicinol. It is therefore suggested that blood cells serve to protect against the degradation of pirarubicin into inactive compounds in blood. Accordingly, when the monitoring of pirarubicin and its active metabolites is carried out in patients, both blood and plasma must be frozen immediately after blood collection.