Intrapleural and intravenous Corynebacterium parvum in patients with resected stage I and II non-small cell carcinoma of the lung

Summary A prospective randomized trial compared the administration of intrapleural plus intravenous Corynebacterium parvum (C. parvum) versus placebo in patients with resected Stage I and Stage II non-small cell bronchogenic carcinoma. Treatment consisted of 7 mg C. parvum injected into the pleural space and 7 mg C. parvum intravenously once between days 6 and 12 postoperatively and 7 mg intravenously every 3rd month during the 1st year. Intrapleural administration of 35 cc of saline served as the placebo and the flush after intrapleural C. parvum.Of the 303 patients entered into this study, 286 were evaluable, with an average follow-up time of 3.5 years. More complications, especially fever, were observed in patients receiving C. parvum. A fever greater than 38 °C was observed in 9% of the patients assigned to placebo and 76% of the patients assigned to C. parvum. There was no significant difference between the treatments with respect to disease-free interval or survival.M. Kaufmann, J. Stjernswärd**, A. Zimmermann (Ludwig Institute for Cancer Research, Bern Branch); K. Stanley**, M. Isley, M. Zelen (Frontier Science & Tech. Research Foundation, Brookline, MA, USA); C. Mouritzen, P. Paulsen, U. Henriques (Dept. of Thoracic and Cardiovascular Surgery and Institute of Pathology, Kommunehospital, Aarhus, Denmark); N. Konietzko, W. Maassen, W. Hartung, W. Wierich (Ruhrland Clinic, Essen-Heidhausen, and Pathology Institute, Ruhr-University, Bochum, FRG); P. Oehl (Innere Klinik und Poliklinik Tumorforschung, Essen, FRG); J. Vogt-Moykopf, H. Toomes, W. Hofmann (Rohrbach Hospital, Clinic for Thoracic Medicine and Pathology Institute, Heidelberg, FRG); F. Krause, R. Rios, R. Spanel (Klinik Löwenstein, Löwenstein, and Pathology Institute, Ulm, FRG); J. Orel, B. Hrabar, D. Ferluga, T. Rott (University Medical Center, Thoracic Surgery and Pathology, Ljubljana, Yugoslavia); H. A. Rostad, J. R. Vale, P. Lexow (Rikshospital, Oslo, Norway); S. Hagen, S. Birkeland (Ulleval Hospital, Oslo, Norway); T. Harbitz, R. Nissen-Meyer (Aker Hospital, Oslo, Norway); E. Aspevik, H. Engedal, A. Mykin (Haukeland Hospital, Bergen, Norway); V. O. Björk, L. Rodriguez, K. Böök, J. Willems (Karolinska Sjukhuset, Thoracic Surgical Clinic and Pathology Department, Stockholm, Sweden); E. Grädel, J. Hasse, P. Dalquen (Kantonsspital, Dept of Surgery, Div. of Cardiac & Thoracic Surgery & Pathology Institute, Basel, Switzerland); L. Eckmann, K. Hänni, K. Zimmermann (Tiefenauspital Surg. Clinic, Univ. of Bern, Switzerland); B. Nachbur, H. U. Würsten, H. Cottier, A. Zimmermann (Inselspital Dept. of Thoracic and Cardiovascular Surg. and Pathology Institute, Bern, Switzerland); W. Maurer, M. Kaufmann (Bürgerspital, Surgical Department, Solothurn, Switzerland); H. Denck, E. Zwintz, St. Wuketich (Krankenhaus der Stadt Wien-Lainz, I. Chir. Dept., and Path. Inst., Vienna, Austria); N. Pridun, H. Hackl (Pulmonologisches Zentrum der Stadt Wien, and Path. Inst., Vienna, Austria); E. Moritz, W. Schlick, H. Holzner (II. Chir. University Clinic and Path. Inst., Vienna, Austria); K. Karrer (Institute for Cancer Research, Vienna, Austria); R. G. Crispen (ITR-Biomedical Research, University of Illinois, Chicago, USA); D. S. Freestone, R. Bomford, M. T. Scott, T. Priestman, L. Toy (The Wellcome Research Laboratories, Beckenham, England)** Present address: Cancer Unit, World Health Organization, Geneva, Switzerland Offprint requests to: K. Stanley, Ludwig Institute for Cancer Research, Inselspital, CH-3010 Bern, SwitzerlandLudwig Lung Cancer Study Group:     

Search for the possible role of ‘immunotherapy’ in operable bronchial non-small cell carcinoma (stage I and II): A phase I study with Corynebacterium parvum intrapleurally

Summary The possibility of giving C. parvum intrapleurally (i.p.) was investigated. C. parvum was given post-operatively either i.p. only or i.p. and intravenously (i.v.) simultaneously. The dose varied from 0.1–10 mg i.p. All patients had been operated for a bronchial carcinoma. Results: (1) Subjective complaints of either dyspnoea, thoracic pain, chills or nausea occured in 31 of 63 patients. No clear dose relation was found. A feeling of discomfort and fever could occur for another 3–4 days after the above more acute symptoms had disappeared. (2) Increased fever (0.5° C) occurred in 71% of the patients injected i.p. only. (3) No anaphylactic reaction was observed. (4) Increased total white blood cell counts (<20%) occurred in 38 patients. The WBC increase was mainly due to higher number of neutrocytes and granulocytes. Total lymphocyte, monocyte, eosinophilic, and basophilic granulocytes values per mm³ circulating blood remained unchanged, except at the dose of 7 mg C. parvum i.p. when monocyte values were increased significantly from 576±247 to 1100±578/mm³. (5) Moderate to severe effusions were observed radiologically in three patients after C. parvum intrapleurally.The study group is: M. Kaufmann, J. Stjernswärd (Ludwig Institut for Cancer Research, Lausanne Branch, Switzerland), M. Zelen, K. Stanley (Frontier Science and Technology Research Foundation, Inc. Amherst, New York, USA), D. S. Freestone, R. Bomford, M. T. Scott, T. Priestman (The Wellcome Research Laboratory, Beckenham, England), C. Mouritzen, G. Ahlbom (Dept. of Thoracic and Cardiovascular Surgery, Aarhus Kommunehospital, Aarhus, Denmark), N. Konietzko, D. Greschuchna (Ruhrland Klinic, Essen-Haidhausen, Germany), P. Hilgard (Innere Klinik und Poliklinik [Tumorforschung] Essen, Germany), J. Vogt-Moykopf, D. Zeidler, H. Toomes (Thoraxchirurgische Spezial-Klinik, Heidelberg-Rohrbach, Germany), F. Krause, R. Rios (Thoraxchirurgische Abt., Fachkrankenhaus für Lungen- und Bronchialerkrankungen, Löwenstein, Germany), J. Orel, M. Benedik, B. Hrabar (Clinical Center, Dept. of Thoracic Surgery, Ljubljana, Yugoslavia), S. Plesnicar (The Institute of Oncology, Ljubljana, Yugoslavia), H. A. Rostad, J. R. Vale (Rikshospital, Oslo, Norway), S. Hagen, S. Birkeland, (Ulleval Hospital, Oslo, Norway), T. Harbitz, R. Nissen-Meyer (Aker Hospital, Oslo, Norway), L. Rodriguez, V. O. Björk, K. Böök (Karolinska Sjukhuset, Thoracic Clinic, Stockholm, Sweden), E. Gradel, J. Hasse, P. Holbro (Kantonsspital, Thoraxchirurgische Klinik, Basel, Switzerland), L. Eckmann (Tiefenauspital, Chir. Univ.-Klinik, Bern, Switzerland), B. Nachbur, T. Liechti (Inselspital, Dept. of Thoracic and Cardiovascular Surgery, Bern, Switzerland), H. Cottier (Inst. of Pathology, Inselspital, Bern, Switzerland), W. Maurer, M. Kaufmann, P. Froelicher (Bürgerspital, Surgical Dept., Solothurn, Switzerland), H. Denck, N. Pridun (Krankenhaus der Stadt Wien-Lainz, Chir. Abt., Vienna, Austria), K. Karrer (Institute for Cancer Research, University of Vienna, Austria) Reprint requests should be addressed to any of the members listed above, or to the Ludwig Lung Cancer Trial, Operation Office, LICR, CH-1066 Epalinges, Switzerland. (For Current Contents, etc., please use above address)     

Transgenic animals as models for human diseasereport of an EC Study Group

ContributorsThis report results from the discussion of an Expert Group convened in Edinburgh on 29–30 October 1992 for a workshop on that subject sponsored and organized by the Commission of the European Communities, Directorate General XII (CEC-DG XII). The experts taking part in the workshop were: R. Lathe and J.J. Mullins, Coordinators (AFRC Centre for Genome Research, University of Edinburgh); G.N. Fracchia, Secretary (Medical Research-Pharmaceuticals, CEC-DG XII, Brussels); and the participants; C. Babinet (Dept d'Immunologie, Institut Pasteur, Paris); P. Eliard (EFPIA, Brussels); C. Benoist (LGME du CNRS/INSERM, Strasbourg); G. Bianchi (Ospedale San Raffaele, Universita di Milano, Milan); E. Boncinelli (DIBIT, Ospedale San Raffaele, Milan); G. Brem (Universitat München); G. Cossu (Institute of Histology, School of Medicine, University of Rome); N. Dillon (MRC National Institute for Medical Research, London); V. Episkopou (Dept of Biochemistry & Molecular Genetics, St Mary's Hospital Medical School, London); M. Evans (Wellcome/CRC Institute, Cambridge); R. Forster (Italfarmaco Research Centre, Cinisello Balsamo, Milan); D. Ganten (Max-Delbrück-Zentrum für Molekulare Medizin, Berlin); A. Gossler (Max-Delbrück-Laboratorium in der Max-Planck-Gesellschaft, Köln); J. Gray (Dept Psychology, Institute of Psychiatry, London); R. Hammer (Howard Hughes Medical Institute, University of Texas, Dallas, USA); A. Hobden (Genetics Unit, Glaxo Group Research Ltd, Middlesex); G. Kollias (Laboratory of Molecular Genetics, Hellenic Pasteur Institute, Athens); D. Lamy (Transgène SA, Strasbourg); D. Lincoln (MRC Reproductive Biology Unit, Edinburgh); J. Mallet (CNRS/LNCM, Gif-sur-Yvette); D. Melton (ICMB, University of Edinburgh, Edinburgh); J.M. Moalic (U127 INSERM, Hôpital Laraboisire, Paris); S. Mockrin (Dept Health & Human Services, National Institutes of Health, Bethesda, MA, USA); J. Ottesen (Biopharmaceuticals Division, Dept of Gene Technology and Virology, Novo Industrie, Denmark); D. Porteous (MRC Human Genetics Unit, Western General Hospital, Edinburgh); P. Rae (Pharmaceutical Division, Miles, West Haven, USA); F. Theuring (Schering AG, Pharmaceutical Research, Berlin); G. Tremp (Rhone-Poulenc Rorer SA, Centre de Recherche de Vitry-Alfortville, Vitry-sur-Seine); H. Van der Putten (Dept Biotechnology, Ciba-Geigy AG, Basel); R. Wolf (ICRF Molecular Pharmacology Group, Biochemistry Dept, Edinburgh). Further supporting documentation and commentary were received from P. Dement (Amsterdam), U. Habenicht (Berlin), P. Grüss (Gottingen), M. Lyon (Oxford), C.C.J. Miller (London), W.-D. Schleuning (Berlin) and R. Williamson (London) and their contributions are gratefully acknowledged.     

The distribution and effects of intrapleural Corynebacterium parvum in mice

Summary After intrapleural (IPl) injection of ¹²⁵ I or fluorescein labelled C. parvum, most was confined to the pleural and mediastinal spaces. The pleural phagocytes and mediastinal lymph nodes were heavily labelled, but very little was found in the lung. The amounts of C. parvum taken up by the liver and spleen were less than after IV injection and splenomegaly was also less after IPl than IV injection. A large proportion (>90%) of cells in pleural washouts following IPl C. parvum was activated macrophages which inhibited, nonspecifically, the growth of tumor cells in vitro. No similar activity was detected after IV C. parvum. IPl injection of C. parvum mixed with irradiated tumor cells conferred strong, specific systemic immunity against tumor challenge, and this immunity was also demonstrable using mediastinal lymph node cells in a Winn assay. The immunity resulting from IV C. parvum and IPl irradiated tumor cells was significantly lower. IPl C. parvum has been compared with IV C. parvum for its effect against tumors growing either in the lung or pleural cavity. Tumors growing in the pleural cavity were inhibited more effectively by IPl than IV C. parvum. With tumors growing in the lung (caused by tumor cells injected IV), although IV C. parvum was more effective at reducing the number of lung nodules during the first two weeks, the mice consistently survived longer after IPl C. parvum.M.T.S. is a member of the Ludwig Lung Cancer Study Group. The present work arose out of discussions with other members of the group and is presented on their behalf. The study group is: M. Kaufmann, J. Stjernswärd (Ludwig Institut for Cancer Research, Lausanne Branch, Switzerland), M. Zelen, K. Stanley (Frontier Science and Technology Research Foundation, Inc. Amherst, New York, USA), D. S. Freestone, R. Bomford, M. T. Scott, T. Priestman (The Wellcome Research Laboratories, Beckenham, England), C. Mouritzen, G. Ahlbom (Dept. of Thoracic and Cardiovascular Surgery, Aarhus Kommunehospital, Aarhus, Denmark), N. Konietzko, D. Greschuchna (Ruhrland Klinik, Essen-Haidhausen, Germany), P. Hilgard (Innere Klinik und Poliklinik [Tumorforschung] Essen, Germany), J. Vogt-Moykopf, D. Zeidler, H. Toomes (Thoraxchirurgische Spezial-Klinik, Heidelberg-Rohrbach, Germany), F. Krause, R. Rios (Thoraxchirurgische Abt., Fachkrankenhaus für Lungen-und Bronchialerkrankungen, Löwenstein, Germany), J. Orel, M. Benedik, B. Hrabar (Clinical Center, Dept. of Thoracic Surgery, Ljubljana, Yugoslavia), S. Plesnicar (The Institute of Oncology, Ljubljana, Yugoslavia), H. A. Rostad, J. R. Vale (Rikshospital, Oslo, Norway), S. Hagen, S. Birkeland, (Ulleval Hospital, Oslo, Norway), T. Harbitz, R. Nissen-Meyer (Aker Hospital, Oslo, Norway), L. Rodriguez, V. O. Björk, K. Böök (Karolinska Sjukhuset, Thoracic Clinic, Stockholm, Sweden), E. Gradel, J. Hasse, P. Holbro (Kantonsspital, Thoraxchirurgische Klinik, Basel, Switzerland), L. Eckmann (Tiefenauspital, Chir. Univ.-Klinik, Bern, Switzerland), B. Nachbur, T. Liechti (Inselspital, Dept. of Thoracic and Cardiovascular Surgery, Bern, Switzerland), H. Cottier (Inst. of Pathology, Inselspital, Bern, Switzerland), W. Maurer, M. Kaufmann, P. Froelicher (Bürgerspital, Surgical Dept., Solothurn, Switzerland), H. Denck, N. Pridun (Krankenhaus der Stadt Wien-Lainz, Chir. Abt., Vienna, Austria), K. Karrer (Institute for Cancer Research, University of Vienna, Austria)Visiting Investigator. Recipient of an American Cancer Society Fellowship     

Classification of acute non-lymphocytic leukemia according to the distribution picture of peroxidase activity and cell size: correlation between the classification and therapeutic response

A distribution picture was prepared on the basis of the correlation between peroxidase activity and cell size in leukemic cells using an automated leukocyte differential counter (Hemalog-D). From this, acute nonlymphocytic leukemia was classified into three groups in which the therapeutic response was examined. The leukemic cells of Group I were medium or large and were negative or weakly positive to peroxidase. These cells were characterized by their location in the upper part of the normal lymphocyte distribution. The leukocyte differential count, measured by a computer on the basis of the distribution picture, showed an increase in large unstained cells (LUC) and lymphocytes. The leukemic cells of Group II were large and positive to peroxidase and were characterized by their location in the right upper part, across the region of LUC, monocytes, basophil and neutrophil leukocytes as seen in the distribution picture. The findings of Hemalog-D showed an increase in LUC, remainder and neutrophil leukocytes. The leukemic cells of Group III were medium-sized and moderately or strongly positive to peroxidase. This group was characterized by their location in the lower part of normal neutrophil leukocytes and Hemalog-D showed an increase in neutrophil leukocytes. A total of 71 patients with acute nonlymphocytic leukemia were assessed according to this classification. Group I (14 patients): 11 with acute myelogenous leukemia (AML), 2 with acute monocytic leukemia (AMoL) and 1 with acute myelomonocytic leukemia ( AMMoL ); Group II (17 patients): 7 with AML and 10 with AMoL; Group III (40 patients): 28 with AML, 4 with AMoL, 1 with AMMoL and 7 with acute promyelocytic leukemia (APL). These groups were treated with the protocol (DCMP two step, BH-AC DMP, BH-AC AMP) established by the Yamada Leukemia Study Group of the Japan Welfare Ministry Cancer Research Project (chairman Yamada, K). The complete remission rate was 35.7% in Group I, 58.8% in Group II and 85.0% in Group III. The difference between Groups I and III was statistically significant (P less than 0.005), as was the difference between Groups II and III (P less than 0.1), while that between Groups I and II was not significant. The median survival was 12 months in Group I, 9 months in Group II and 15 months in the Group III and the difference between Groups I and III was statistically significant (P less than 0.05). Group III included a small number of AMoL and APL patients in addition to AML, while Groups I and II consisted mainly of patients with AMoL and AML.(ABSTRACT TRUNCATED AT 400 WORDS)     

Efficacy of BCG presensitization in combination with cyclophosphamide (CY) on murine tumor growth and immunity

Summary The effect of BCG in combination with cyclophosphamide (CY) was studied in a solid tumor system in syngeneic mice. There was a marked effect of intradermal (ID) presensitization with BCG (10⁷ organisms) followed by a single intraperitoneal (IP) injection of CY (200 mg/kg) one day after the tumor inoculation on tumor suppression and tumor immunity, while there was no effect by either BCG or CY treatment alone. The optimal interval between BCG sensitization and tumor inoculation seemed to be about 3 weeks.Research supported in part by Grant-in-Aid Cancer Research from the Ministry of Health and Welfare, Japan     

Effectiveness of Omeprazole-Amoxicillin-Clarithromycin (OAC) Therapy for Helicobacter pylori Infection in a Japanese Population

Background. Omeprazole or lansoprazole, amoxicillin, clarithromycin (PPI/AC) therapy has been reported to provide a high cure rate of H. pylori infection with few adverse effects. Effectiveness of H. pylori therapy may vary among different geographic regions and patient populations. However, there are few reports in Japan as to its effectiveness. We have, therefore, studied the effectiveness of H. pylori therapy in a large group of Japanese patients. Methods. For this study, 366 H. pylori-positive patients with peptic ulcer disease or non-ulcer dyspepsia (263 men and 103 women, mean age 48.5 years) were assigned to 6 groups, each receiving a different PPI/AC regimen. Group 1 received omeprazole (OPZ) 20 mg, amoxicillin (AMOX) 1500 mg, and clarithromycin (CAM) 400 mg; Group 2 OPZ 40 mg, AMOX 1500 mg, and CAM 400; and Group 3 OPZ 20 mg, AMOX 2000 mg, and CAM 600 mg daily for 14 days. The group treated with lansoprazole (LPZ) 30 mg, AMOX 1500 mg and CAM 400 mg was used for 14 days in Group 1L. OPZ 20 mg, AMOX 750 mg, and CAM 200 mg were given to Group 4 for 28 days and OPZ 20 mg, AMOX 1500 mg, CAM 400 mg was administered to Group 5 for 7 days. Cure of infection was assessed by the ¹³C urea breath test one month after completion of therapy. Results. Cure rates calculated by excluding the patients who showed borderline value of ¹³C urea breath test (Δ¹³C value between 5 and 10‰ in Groups 1, 1L, 2, 3, 4, and 5 were 82.7% (95% CI; 74–90), 88.9% (76–96), 84.9% (72–93), 81.3% (67–91), 84.6% (72–93), and 85.1% (72–94) on an intention-to-treat basis, and 88% (80–94), 95.2% (84–99), 95.6% (85– 99), 90.7% (78–97), 95.7% (85–99) and 88.9% (76–96) on a per-protocol basis, respectively. Adverse effects that affected compliance were observed in 10 of 237 patients on 14-day regiments, one of 47 on a 28-day regimen and one of 46 on a 7-day regimen. Conclusion. Two weeks PPI/AC therapy is highly effective for cure of H. pylori infection in the Japanese population. The low dose one month regimen and the one week OAC regimen were also effective in our patient population.     

Ovarian response to gonadotropin stimulation in juvenile rhesus monkeys

The objective of this study was to investigate juvenile rhesus monkeys responding to various gonadotropin regimen stimulations. Thirty-two prepubertal rhesus monkeys were randomly allocated into five groups for ovarian stimulation as follows: Groups I, II, and III were given 35, 18, and 9 IU recombinant human follicle-stimulating hormone (rhFSH), respectively, twice daily for 8 d; Group IV was given 18 IU rhFSH twice daily until the appearance of maximal increase in sex skin during the breeding season; and Group V was treated identically to Group II but during the nonbreeding season. In addition, nine menarchial monkeys (Group VI) were treated identically to Group II. Menarchial monkeys yielded two- to fivefold the numbers of MII oocytes (24.1) and almost twice the development potential of in vitro-fertilized oocytes (blastocyst rate: 50.0%) compared with those of the other groups. Moreover, prepubertal monkeys in Group V had approximately double the numbers of MII oocytes and in Groups IV and V twice the development potential compared with those of Groups I and II, whereas Group III did not respond to stimulation. The most prominent sex skin swelling was in association with peak serum estradiol concentrations, and good responses to stimulation were associated with reduced body temperatures. All stimulated monkeys had normal reproductive performance at adulthood, except those in Group I. In conclusion, gonadotropin stimulation of menarchial monkeys could be appropriate for addressing the high cost and limited availability of rhesus monkeys in studying reproductive biology in primates.     

Source of oestrogen in early pregnancy in the mare

Oestrogen secretion was determined by oestrogen conjugate (EC) analysis of urine in three groups of pregnant mares: Group I (N = 6), animals ovariectomized on Day 18-19 of gestation with pregnancy maintained by daily administration of an oral progestagen, altrenogest; Group II (N = 9), untreated, pregnant mares; Group III (N = 5) intact, pregnant mares treated daily with altrenogest. The mean EC concentrations in the ovariectomized mares in Group I increased in a constant linear manner from 17 ng/mg Cr on Day 20 to 291 ng/mg Cr on Day 70, with no apparent surge in oestrogen secretion around Day 39. Mean EC concentrations on Days 33, 39 and 44 were respectively 41, 48, and 73 ng/mg Cr. In the intact mares in Groups II and III (shown in parentheses), the mean urinary EC concentrations were 201 (171) ng/mg Cr between Days 20 and 33 of gestation, increased rapidly from 172 (77) ng/mg Cr on Day 33 to a peak of 1066 (895) ng/mg Cr on Day 39, followed by a decline to 637 (719) ng/mg Cr on Day 44. After Day 44, EC concentrations continued to increase in a linear manner to 1191 (842) ng/mg Cr on Day 70. The mean EC concentrations between Days 20 and 70 in Group I were significantly (P less than 0.05) lower than in mares in Groups II and III. EC concentrations in Group III mares were significantly lower (P less than 0.05) than in Group II mares between Days 28 and 34.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of estradiol supplementation on superovulation in Swamp buffalo

The effect of estradiol-17beta (E(2)) supplementation on superovulation with (PMSG) or (FSH) was investigated in Swamp buffalo. Sixty-eight buffalo were treated in seven groups. Group 1 served as control and was superovulated by standard PMSG or FSH treatment used in routine bovine embryo transfer protocols. Group 2 was superovulated by standard PMSG regimen plus two injections of E(2) at a 48 h interval beginning one day before the onset of gonadotropin treatment (short-term supplementation) for a total dosage of 2.5 mg E(2); Groups 3 and 4 received the same regimen as Group 2, but in doses of 5.0 and 7.5 mg E(2), respectively. Group 5 received the standard FSH regimen (40% LH). Group 6 received short-term E(2) (7.5 mg) supplementation of FSH-p. Group 7 was superovulated by standard FSH regimen (40% LH) plus three injections of E(2) at 48-72 h intervals beginning five days before the onset of gonadotropin treatment (long-term supplementation) for a total dosage of 7.5 mg E(2). The number of corpora lutea (CL) and follicles >/= 8 mm in diameter were recorded by palpation per rectum and after slaughter. The mean numbers of CL and follicles were 0.99, 5.8, 8.0, 10.6, 4.0, 3.9, 8.1 and 0.25, 6.8, 6.2, 6.2, 1.6, 0.0, 4.1 for Groups 1, 2, 3, 4, 5, 6, 7, respectively. In Group 7, the rates of nonsurgical and postmortem embryo recovery were 46 and 90.4%, respectively and 54.4% of the collected ova were fertilized. These results indicate the possibility of producing viable embryos in buffalo by using E(2) supplementation for the gonadotropin treatment.     

A multi-stage model for border contrast

A multi-stage model has been constructed. In this model, the first level is expressed by a convolution of the physical luminance function with a line spread function whose parameters depend on luminance. The second level extracts the gradient of the response that is processed at the first level. The third level detects, as edges, the inflection points of the response that is processed at the first level. The fourth level averages the response gradients over each interval between the edges, while keeping the total magnitude of the final response within the interval the same as those of the initial response at the first level. The model was applied to border contrast phenomena as described by Bergström (1966)This work was presented at the XXIst International Congress of Psychology in Paris (July, 1976), and in Hamada (1976) and in Ph.D. Thesis of Hokkaido University, Sapporo, Japan. Prof. M. J. Morgan of University College London, England, provided many helpful comments and recommended revising the original paper (Hamada, 1976). Prof. T. S. Aiba of Hokkaido University and Prof. M. Akita of the Kyoto Institute of Technology showed a continued interest in this research. The author would like to express his grateful appreciations     

Effects of α-lipoic acid supplementation on maternal diabetes-induced growth retardation and congenital anomalies in rat foetuses

The mechanism of diabetic embryopathy is not known. Excessive reactive oxygen species (ROS) produced in diabetes may be causally related to foetal anomalies. The objective of this study was to determine whether supplementation with the antioxidant lipoic acid (LA) could prevent maternal diabetes-related foetal malformations and intrauterine growth retardation (IUGR) in rats. Pregnant rats were non-treated (Group I) or made diabetic on gestation day (GD) 2 by injecting streptozotocin (Group II). Group III was injected with 20 mg kg^–1 of LA daily starting on GD 6 and continued through GD 19. Group IV was administered only Tris buffer on the corresponding days. Group V was a set of STZ-treated animals, which were supplemented with a daily dose of 20 mg kg^–1 of LA from GD 6 through GD 19. All fetuses were collected on GD 20. Lipoic acid did not affect the blood sugar levels of diabetic animals significantly but improved their body weight gain and reduced food and water consumption. Diabetic group had a high incidence of embryonic resorption, IUGR, craniofacial malformations, supernumerary ribs and skeletal hypoplasia. Lipoic acid significantly reduced these abnormalities. These data support the hypothesis that ROS are causally related to fetal maldevelopment and IUGR associated with maternal diabetes in the rat. They also highlight the possible role of antioxidants in the normal processes of embryo survival, growth and development. (Mol Cell Biochem 261: 123–135, 2004)     

Biosystematic studies of theClosterium peracerosum-strigosum-littorale complex

Nine representative pairs of heterothallic (=self-sterile, cross-fertile) strains of theClosterium peracerosum-strigosum-littorale complex from the northern Kanto area in Japan have been studied under the defined standard culture conditions. Since a wall thickening at cell apices was observed in vegetative cells of all the strains, these strains turned out to belong to the morphological group II (Ichimura and Watanabe, 1976). The result of statistical analyses of their cell size variations corresponded well with the result of intercrossing experiments between them. It was shown that these strains are virtually composed of three biologically different groups which are morphologically distinct and reproductively isolated completely or at least partially from each other. For convenience, these three groups have been designated as II-A, II-B, and II-C in the order of from smaller to larger cell size. In intra-group crossings, a large number of zygospores were formed, and they germinated well to yield healthy populations of their progenies in all the three groups. In inter-group crossings, no sign of sexual reproduction was observed between Group II-A and Group II-C or Group II-B and Group II-C, and a marked decrease of zygospore formation was observed between Group II-A and Group II-B, especially between Group II-A minus and Group II-B plus. It was concluded that the distinctions between the three groups are biologically sound and that each represents an evolutionary unit. This paper represents a portion of a thesis submitted to the faculty of the Graduate School of Hokkaido University by the senior author in partial fulfillment of the requirements for the degree of Doctor of Science. This study was supported by the Grants in Aid, No. 174233 & No. 034036, from the Scientic Research Fund of the Ministry of Education, Japan.     

Effects on tissue and electrolytes of a mineralocorticoid blocker during DOCA-induced potassium depletion.

Chronic experiments were carried out on three groups of rats to evaluate tissue and electrolyte effects of a mineralocorticoid blocker canrenoate potassium (SC-14266) during DOCA-induced hypokalemic metabolic alkalosis. Group I animals received DOCA alone, group II received DOCA plus canrenoate, while group III received canrenoate alone. The daily dose ratio (per kilogram of body weight) was 180 mg canrenoate-0.45 mg DOCA. All animals ate a synthetic diet and drank 0.15 N NaHCO3. Group II animals demonstrated a lesser degree of metabolic alkalosis and a higher muscle potassium content when compared with group I rats. The most conspicuous histological abnormality was myocardial necrosis, the degree and extent of which was impressively reduced by the blocking agent.     

Administration of Recombinant Rhesus Interleukin-12 during Acute Simian Immunodeficiency Virus (SIV) Infection Leads to Decreased Viral Loads Associated with Prolonged Survival in SIVmac251-Infected Rhesus Macaques

The ability of recombinant rhesus interleukin-12 (rMamu-IL-12) administration during acute simian immunodeficiency virus SIVmac251 infection to influence the quality of the antiviral immune responses was assessed in rhesus macaques. Group I (n = 4) was the virus-only control group. Group II and III received a conditioning regimen of rMamu-IL-12 (10 and 20 microg/kg, respectively, subcutaneously [s.c.]) on days -2 and 0. Thereafter, group II received 2 microg of IL-12 per kg and group III received 10 microg/kg s.c. twice a week for 8 weeks. On day 0 all animals were infected with SIVmac251 intravenously. While all four group I animals and three of four group II animals died by 8 and 10 months post infection (p.i.), all four group III animals remained alive for >20 months p.i. The higher IL-12 dose led to lower plasma viral loads and markedly lower peripheral blood mononuclear cell and lymph node proviral DNA loads. During the acute viremia phase, the high-IL-12-dose monkeys showed an increase in CD3(-) CD8 alpha/alpha(+) and CD3(+) CD8 alpha/alpha(+) cells and, unlike the control and low-IL-12-dose animals, did not demonstrate an increase in CD4(+) CD45RA(+) CD62L(+) naive cells. The high-IL-12-dose animals also demonstrated that both CD8 alpha/alpha(+) and CD8 alpha/beta(+) cells produced antiviral factors early p.i., whereas only CD8 alpha/beta(+) cells retained this function late p.i. Long-term survival correlated with sustained high levels of SIV gag/pol and SIV env cytotoxic T lymphocytes and retention of high memory responses against nominal antigens. This is the first study to demonstrate the capacity of IL-12 to significantly protect macaques from SIV-induced disease, and it provides a useful model to more precisely identify correlates of virus-specific disease-protective responses.     

Quantitative electron microscopic studies on the kinetics of secretory granules in G-cells

Summary Ultrastructural studies of secretory granules of rat antral G-cells and measurement of serum gastrin level were performed under the condition of fasting and administration of alkaline solution into the stomach. On electron micrographs, no qualitative difference was observed among those experimental groups. However, morphometrical analysis revealed significant quantitative differences. The population density of secretory granules of the rats treated once with alkali first increased and then decreased reaching that of the fasted group, while that of the repeatedly treated group remained nearly equal to the maximum value. The average sectioned surface area of secretory granules tended to decrease for 1.5h after the stimulation but the difference was not significant among those groups.From the results obtained at present, responding to chemical stimulation such as pH changes in the antrum, it seems probable that not only exocytosis but also migration of secretory granules from supra- and/or para-nuclear portion to the basal portion of the cell occurs rapidly in G-cells and that both these processes are inhibited immediately by antral acidification. Moreover, the present results apparently indicate that under the condition of no antral acidification G-cells have a capacity of secreting gastrin for a fairly long time, such as 4–8 h, responding to adequate stimulus. These findings are strongly suggestive of the existence of a capacious pool of granules in the supra- and/or para-nuclear cytoplasm or of fairly speedy production of secretory granules in the Golgi area.The author wishes to express thanks to Prof. R. Furihata, Department of Surgery, and Prof. T. Nagata, Department of Anatomy, Shinshu University School of Medicine, for their constant interest and guidance, and to Dr. F. Iida, Department of Surgery, who has followed the course of this work throughout     

Glycogen metabolism in human hormone-producing trophoblastic cells in continuous culture

Summary Glycogen metabolism was studied in human hormone-producing trophoblastic cells (BeWo line). Cells supplemented daily with high glucose (3 g per liter in medium) contained 5.5% glycogen and utilized glucose at an initial rate of 12.2 mμmoles per min per mg of protein. In cells supplemented daily with low glucose (1 g per liter), the initial rate of glucose consumption was 23 mμmoles per min per mg of protein and the glycogen content reached only 0.4% of wet weight 24 hr after medium replenishment. When glycogen-depleted cultures were refed glucose, an accumulation of glycogen was observed, with initial deposition occurring in areas near the cell surface. After exhaustion of extracellular glucose, cytoplasmic glycogen was utilized at a rate of 2.8 mμmoles per min per mg of protein. Addition of either low or high glucose to glycogen-depleted cells resulted in the same rate of glycogen synthesis (approximately 8 mμmoles per min per mg of protein). It was suggested that unique regulatory mechanisms function in the control of glycogen metabolism in glycoprotein hormone-producing cytotrophoblastic cells. This work was supported in part by Public Health Service Research Contract PH43-68-1010, Research Grant CA 05524 from the National Cancer Institute, and by grants from the Milwaukee Division of the American Cancer Society, Inc., and the Damon Runyon Memorial Fund for Cancer Research, Inc.     

Dissecting the forces that shape crops: an interview with Ryohei Terauchi

Ryohei Terauchi is a Professor at Kyoto University and a Group Leader at the Iwate Biotechnology Research Center, Japan, studying the evolution of crops and their pathogens. In this interview, Ryohei describes his research interests, how the revolution in sequencing technology helped improve our understanding of orphan crops, and who are the scientists that inspire him.     

Endocrine and superovulatory responses in heifers pretreated with FSH or bovine follicular fluid

This study examined the effects of altered serum FSH concentration on subsequent ovarian response to superovulation. Synchronized heifers were assigned randomly on Day 1 of the cycle (estrus = Day 0) to three pretreatment groups that consisted of 6-d of saline (7ml, s.c., b.i.d.; Group I), FSH-P (0.5 mg, i.m., b.i.d.; Group II) or charcoal-extracted bovine follicular fluid (BFF; 7 ml, s.c., b.i.d.; Group III) injections. Superovulation was initiated on Day 7 and consisted of FSH-P in decreasing dosages over 4 d (4,3,2,1 mg; i.m., b.i.d.), with cloprostenol (500 mug) on the morning of the third day. A second replicate with 14 heifers was conducted using the same protocol but twice the pretreatment dosage of FSH-P (1 mg) and BFF (14 ml). Endogenous plasma FSH decreased during BFF and FSH-P pretreatments compared to controls (P < 0.02). Endogenous FSH concentrations in both primed groups (II and III) were similar to control values (Group I) 12 h after the start of superovulation. Basal LH concentrations were not different between pretreatment groups. The interval from cloprostenol treatment to the preovulatory LH surge in Group III was 21.3 and 23.9 h longer (P < 0.0001) than it was in Groups I and II. The postovulation progesterone rise was delayed in Group III. The number of corpora lutea (CL) was lowest in the BFF-primed group (4.2 +/- 0.8) compared with the FSH-primed (7.4 +/- 1.3) and the control (12.0 +/- 1.8; P < 0.003) groups. In the FSH-primed group (0.68 +/- 0.06 cm(3)), CL volumes were larger than in the control group (0.45 +/- 0.03 cm(3)), whereas in the BFF-primed group (0.27 +/- 0.02 cm(3)) CL volumes were smaller compared with the control group (P < 0.0001). Mean FSH concentrations for 48 h preceding superovulation and the number of CL per cow were positively correlated (r = 0.55; P < 0.004; n = 26). We concluded that both FSH-P and BFF pretreatments decreased the superovulatory response of heifers to FSH-P. The mechanism for this would appear to be associated with reduced endogenous FSH prior to the start of superovulation.     

Prostaglandin E2 counteracts the effects of PGF2 alpha in indomethacin treated cycling gilts

Twenty crossbred gilts with at least 2 consecutive estrous cycles of 18 to 21 days in length were used to study the effects of prostaglandins E2 and F2 alpha (PGE2 and PGF2 alpha) on luteal function in indomethacin (INDO) treated cycling gilts. Intrauterine and jugular vein catheters were surgically placed before day 7 of the treatment estrous cycle and gilts were randomly assigned to 1 of 5 treatment groups (4/group). With exception of the controls (Group I) all gilts received 3.3 mg/kg INDO every 8 h, Groups III, IV and V received 2.5 mg PGF2; 2.5 mg PGF2 alpha + 400 micrograms PGE2 every 4 hr, or 400 micrograms PGE2 every 4 h, respectively. All treatments were initiated on day 7 and continued until estrus or day 23. Jugular blood for progesterone analysis was collected twice daily from day 7 to 30. Estradiol-17 beta (E2-17 beta) concentrations were determined in samples collected twice daily, from 2 d before until 2 d following the day of estrus onset. When compared to pretreatment values, estrous cycle length was unaffected (P greater than 0.05) in Group I, prolonged (P less than 0.05) in Groups II, IV and V; and shortened (P less than 0.05) in Group III. The decline in plasma progesterone concentration that normally occurs around day 15 was unaffected (P greater than .05) in Group I; delayed (P less than 0.05) in Groups II, IV and V; and occurred early (P less than 0.05) in Group III. Mean E2-17 beta remained high (31.2 +/- 4.9 to 49.3 +/- 3.1 pg/ml) in Groups III and IV, while the mean concentrations in Groups III and V varied considerably (17.0 +/- 2.0 to 52.2 +/- 3.5 pg/ml). The results of this study have shown that PGE2 will counteract the effects of PGF2 alpha in INDO treated cycling gilts. The inclusion of PGF2 alpha appeared to either stimulate E2-17 beta secretion or maintain it at a higher level than other treatments.