Bacillus Calmette-Guérin immunotherapy in combination with DTIC (NSC-45388) for the treatment of malignant melanoma.

Combination studies of immunotherapy and DTIC chemotherapy for patients with malignant melanoma are described. At our institute the combination of bacillus Calmette-Guérin (BCG) with DTIC, or DTIC plus methyl-CCNU, has resulted in augmentation of remissions in areas regional to BCG immunization, prolongation of chemotherapy-induced remissions, and significant prolongation of survival compared to the use of DTIC chemotherapy without immunotherapy. Data from other centers have shown that augmentation of remissions for melanoma patients occurred when BCG was combined with DTIC, vincristine, and allogeneic tumor cells. Finally, another study has shown that DTIC combined with BCG has prolonged the disease-free interval and survival in melanoma patients with regional lymph node metastases. The potential inhibition of DTIC metabolism by BCG or Corynebacterium parvum is discussed.


Phase I evaluation of DTIC (NSC-45388) and other studies in malignant melanoma in the Central Oncology Group.

The major toxic effects of DTIC are nausea and vomiting, leukopenia, and thrombocytopenia. It is an effective drug in the treatment of malignant melanoma with better response rates in women than in men. The addition of other drugs to DTIC therapy does not produce a higher response rate. Preliminary results indicate that as an adjuvant to standard surgical therapy DTIC does not appear beneficial to patients with a high risk for recurrence.


DTIC (NSC-45388) studies in the southwest oncology group.

In a phase I study, the best antitumor/toxicity ratio for DTIC was reported to be at a dose of 250 mg/m2/day X 5 repeated at 28-day intervals. Nausea, vomiting, leukopenia, and thrombocytopenia were the major toxic effects noted. The best responses were seen in disseminated melanoma (19%), various sarcomas (22%), and Hodgkin's disease. A subsequent phase II study in refractory lymphomas showed a response rate in Hodgkin's disease of 56%. In disseminated melanomas, DTIC was then combined with vincristine and BCNU and demonstrated a response rate of 23% which did not improve with the addition of chlorpromazine (23%). A response rate of 31% was seen with the combination of DTIC, BCNU, and hydroxyurea which did not improve with the addition of vincristine (30%). Responders had a more significant survival rate as compared to nonresponders.


Eastern Cooperative Oncology Group studies with DTIC (NSC-45388).

In four randomized controlled clinical trials with DTIC involving patiens with malignant melanoma, the Eastern Cooperative Oncology Group has seen a consistent objective response rate of 16%. A study of combinations of DTIC with two nitrosoureas indicated no additive effect. Survival and response were strongly correlated with sex, ambulatory status, and site of disease involvement. As secondary treatment DTIC was less effective, indicating only a 3% response rate. Survival benefit could be related to response. Among the ten largest contributors of patients, response rates varied from 3% to 40%. Similarly, while the overall response rate was not different for the combinations as compared to DTIC alone, the variation noted from study to study reflects the need for controlled randomized trials of chemotherapy for patients with malignant melanoma.


Role of DTIC (NSC-45388) in the chemotherapy of sarcomas.

DTIC was one of the first of several new agents evaluated by the Southwest Oncology Group (SWOG) and the M.D. Anderson Hospital and Tumor Institute (MDAH) in the therapy of adult patients with metastatic sarcomas. It yielded an overall response rate of 17%. This is similar to that seen in a review of 138 patients who represent the total number reported in either published or unpublished data. The subsequent addition of DTIC to adriamycin in several studies carried out at the MDAH and in the SWOG has increased the complete and partial remission rate over that seen with adriamycin alone, and more importantly has increased remission duration and survival. DTIC is currently used in combination with adriamycin in the first-line therapy of patients with metastatic sarcomas and should be considered for patients who have relapsed on primary therapy not including DTIC.


Combination chemotherapy with DTIC (NSC-45388) in advanced malignant melanoma, soft tissue sarcomas, and Hodgkin's disease.

The results of three controlled studies with multiple-drug regimens which included DTIC are reported for advanced melanoma, soft tissue sarcomas, and Hodgkin's disease. In malignant melanoma, no statistical difference was observed between (a) DTIC, BCNU, and vincristine, and (b) DTIC, BCNU, and actinomycin D in terms of response rate, median duration of response, and survival. In sarcomas, the combination of adriamycin and DTIC produced a higher incidence of complete plus partial responders when compared to cyclophosphamide, vincristine, and methotrexate. However, no significant difference was seen in the median duration of response or survival. In Hodgkin's disease, a new four-drug combination including adriamycin, bleomycin, vinblastine, and DTIC (ABVD) produced an incidence of complete remissions and a median duration of response comparable to that achieved with mechlorethamine, vincristine (Oncovin), predinisone, and procarbazine (MOPP). Preliminary data indicate that ABVD is not cross resistant to MOPP. It is concluded that DTIC can be successfully and safely employed in combination with conventional agents in susceptible neoplastic diseases.


Methyl-CCNU (NSC-95441) in advanced colorectal carcinoma after failure of 5-fluorouracil (NSC-19893) therapy.

Twenty-seven patients with advanced colorectal carcinoma who had earlier failed to respond to 5-fluorouracil (5-FU) were given methyl-CCNU. Of the 21 patients who received methyl-CCNU alone, two (9.5%) responded partially with a mean duration of 28 weeks. Of the six patients who received methyl-CCNU and beta-TGdR, three had a partial response. There were no complete responses in either group. Response was not related to performance status, site of metastasis, or previous response to 5-FU treatment. It seems that the addition of beta-TGdR to methyl-CCNU might give better results after failure to previous 5-FU treatment. Hematologic toxicity was noted in 33% of the patients and gastrointestinal toxicity in 25%.


The ultrastructure of the cell types in the endocrine pancreas of the horse.

The islets of Langerhans of the equine pancreas were examined with the electron microscope after immersion or perfusion fixation. Five cell types could be distinguished after fixation by either technique: 1. A-cells, situated at the center of the islets, 2. B-cells, containing mostly "pale granules" and constituting the principal cell type of the periphery of the islets, 3. D-cells, also located mainly at the periphery of the islets, 4. G-cells, found at the edge of the islets and in the exocrine pancreas, and 5. S-cells, (small granule cells), which are relatively few in number and occur only in the islets. The function and age-dependent modifications of these cells are discussed. The formation of light and dark cells and of "mixed cells" are regarded as artifact, since cells of this type occur only under the condition of immersion fixation.


The gonadotropic cells in the pituitary of the black molly, Mollienisia latipinna, and other teleosts, identified by the immunofluorescence technique in normal and androgen-treated animals.

Using anti-carp-gonadotropic-gamma-globulin, the indirect immunofluorescence technique was applied to sections of the pituitary of 12 teleost species. From the investigated species, the Poeciliinae, and especially the black molly, showed a distinct localization of fluorescent cells in the mesoadenohypophysis. Strong fluorescence was observed in the ventralmost region, containing the presumed gonadotropic cells; weak fluorescence was observed in the dorsal region, in the presumed thyrotropic cells. The possibility of an unspecific reaction with TSH-cells in the latter region is discussed. Treatment of male black mollies for 38 days with methyltestosterone resulted in a loss of fluorescence in the ventral region of the meso-adenohypophysis, whereas there was no decrease of the fluorescence in the dorsal region. The results support the hypothesis that the ventral basophils in the mesoadenohypophysis produce gonadotropic hormone.


Transmission microscopy of freeze dried, unstained epiphyseal cartilage of the guinea pig.

The question of the initial mineralization in the epiphyseal plate has been investigated to date in specimens prepared by conventional electron microscopical techniques. As conventional techniques can produce artifacts, either a loss of mineral substance or a secondary nucleation, the mineralization process was investigated using freeze dried, vacuum embedded growth cartilage which was neither contrasted not stained and which had a very short contact with water. The prevailing theory that the first mineralization begins within extracellular matrix vesicles and that the mineralization outside these vesicles is a secondary process was confirmed. Mineralized matrix vesicles were found in the fully mineralized long septa down to the opening zone. In several cases a mineralization could be observed in those transverse septa in which organic substance was present between the cells. The typical radial arrangement of the apatitic needles and platelets in the matrix vesicles could be explained by the formation of a mineralization in an ionotropic gel, the orientation of the matrix macromolecules to be produced by a vectorial influx of calcium ions and phosphate groups coming from different directions. Thin strands of mineral substance with low contrast, which follow the direction of the longitudinal septum, were assumed to be the mineralized collagen fibrils. In several needles dot-like formations were seen and the distance between the middle of neighbouring dots was found to lie mainly in the range 30-56 A, while the lateral separation distance between the middle of closely packed parallel chains and needles was found to lie mainly in the range 30-42 A. Parallel periodic structures which could be visualized in apatitic chains and needles 20-40 A in diameter were assumed to be the 8.2 A-(100)-lattice planes of apatite, being an indication that these formations already possess criteria of the apatite lattice.


Some mechanisms operative in carcinogenesis a review.

Multiple factors contribute to the development of neoplasia. Sometimes a single agency can bring about a tumour if it has many different effects, but at other times a tumour arises more insidiously due to a succession of events [240,241] which by themselves may be innocent. Alterations in the genome of the cell are at the fore-front of our interest  because they can be brought about by most of the carcinogenic agents we know. The cell can repair some such alterations but both forward and destructive mutations do appear. The roles of cell proliferation, cell differentiation, the immune mechanism and carcinogen-activating enzymes are beginning to be understood. The effects of dose, route of administration, and of other agents given at the same time [242-245] must not be lost sight of. Other factors no doubt will be added as we begin to look at the structure and function of cell-surface membranes [246-248], at host susceptibility genetics [26, 249], and at the generation of carcinogens inside the body [250,251]. We are only beginning to understand carcinogenesis. In no single instance do we as yet know how a tumour comes about in full details of molecular biology. It is possible that fully rational treatment of cancer will not be possible until we have such an understanding. Once a tumour becomes independent of carcinogenic factors, it continues to develop in a bizarre fashion which makes its study and treatment by all means other than surgery difficult.


H2O2 generation during the redox cycle of mitomycin C and dna-bound mitomycin C.

Reduction of mitomycin C by NaBH4 or by NADPH in the presence of a cell extract followed by exposure to air results in the generation of H2O2. This phenomenon occurs not only with free mitomycin but also with mitomycin irreversibly bound to DNA. In view of these findings, the antibiotic activity of mitomycin was tested in two bacterial systems: a facultative aerobic bacterium grown in the presence or absence of oxygen and obligate anaerobic bacterium. No oxygen effect could be demonstrated in either case in the growth-inhibitory and bactericidal activity of the drug. Nevertheless, the H202 generating capacity of mitomycin-DNA complexes inside the nucleus may play a role in the drug-induced biological damage to the genetic material of cells.


Early complications and late sequelae of induced abortion: a review of the literature.

An attempt to evaluate the recent literature on somatic complications of induced abortion shows that a great amount of data of widely varying quality has been collected. Many areas of research are still not covered in a way that permits valid conclusions to be drawn. Two important points emerge from the review: first, there is a need for uniform definitions of complications and some uniformity in the analysis of data collected; secondly, carefully selected control groups are needed in this kind of research. The areas most urgently requiring further study differ from country to country. From a global point of view, the existence or inexistence of significant late sequelae and the influence of the patient's health status on the complication rate seem to be of the highest priority.


The relative risks of sterilization alone and in combination with abortion.

In recent years many authors have claimed that the combination of tubal sterilization and induced abortion carries too high a morbidity and that the two operations should preferrably be performed separately. As this standpoint has serious practical consequences for many women undergoing abortion, a review of the literature was undertaken to see whether there are any data supporting it. No direct comparison of the morbidity accompanying the combined procedure and the total morbidity from the two procedures performed with an interval could be found. Indirect evidence suggests that the effect of an interval between the operations-if any-is negligible compared with the influence upon morbidity of factors such as methods of sterilization and abortion, health status, and age. To gain direct information on the problem, a multicentre study has been designed by the Task Force on Sequelae and Complications of Induced Abortion. The outlines of this study are briefly described.


[Was the regression of leprosy also a function of the economic development in the XIV century? (author's transl)].

During century of expansion, the bread was regularly more costly and the labour cheapness for surpopulation. During the XIVth century, the weather was too rainy and more cold, the corn harvest was too much deficient, the mortality increased; ground no fertile was deserted: dearth and famine consecutive. The rats followed peasantry and giving the pest, from which depopulation increased. The wages growed to twice as much while the bread was reduced. Life come better and the leprosy was coming to reduce.


[Bacteriological and epidemiological considerations on shigelloses in South Vietnam (1969-1973)].

During the period from 1969 to 1973, a rectal swabs from 13.943 diarrhea patients of all ages were examined by bacteriological surveys. From these specimens, 981 strains of Shigella were isolated. The most commonly encounted were Shigella flexneri 2. All of strains of Shigella isolated were highly resistant to Aureomycin, Tetracyclin, Terramycin, Novobiocin and Chloramphenicol. There has been an increase in the development of multiresistance of Shigella. Colimycin and Streptomycin were the most effective antibiotics. The greatest number of Shigella were isolated among children under 2 years and during the driest months (March, April, May, June).


Octopamine plasma levels and hepatic encephalopathy: a re-appraisal of the problem.

An investigation on the blood levels of octopamine was carried out on 70 adult individuals. There was a statistically significant correlation between the levels of octopamine and hepatic encephalopathy. Normal subjects had values below 1 ng/ml, while patients with grade 3 or grade 4 encephalopathy constantly showed values above 3.2 ng/ml. In these two groups the distribution was fairly homogeneous. Through the differences between cirrhotics without neurologic involvement and those with grade 1 or 2 hepatic encephalopathy displayed statistical significance, distribution of values in these groups was rather non-homogeneous. Octopamine levels paralleled variations in mental state in 3 out 4 cases. No difference was found between venous and arterial values. The reaction of transmethylation used in the assay of octopamine was constantly found to be inhibited by the presence of plasma. This inhibition is probably due to the presence of one or more beta-hydroxyphenylethanolamines other than octopamine.


[Value of indirect immunofluorescence in the diagnosis of "Trypanosoma gambiense" trypanosomiasis. 46 cases (author's translation)].

Several indirect fluorescents reactions were performed on 46 trypanosomepositive specimens which had been confirmed by parasitologic diagnosis. 38 sera analysed before the commencement of treatment were found to have a G.M.R.T. equal to I:I, 350 and the titer of antibody was, in all cases, greater than I:320. In all cases in the first phase, the  titer was I:640 or higher. Four cases were followed beyond the second month and their titers decreased slowly, becoming negative in the fifth month. In 12 cases in the second phase, the cerebrospinal fluid demonstrated fluorescent antibodies although 10 cases in the first phase were negative for these antibodies. Three cerebrospinal fluid were faintly positive and these corresponded to the second phase biologically. In the treatment of individuals (using the analysis of cerebrospinal fluid for antibody) the I.F.A.T. permits the classification of the patient into one of the two phases and enables the clinician to follow the course of therapeutic effectiveness. In mass screening, apparently healthy individuals possessing a serum titer of I:640 or higher are shown to need definitive treatment for trypanosomiasis, while those with titers of I:320 are those who may either be treated with Lomidine or tested regularly for change in serum titer.


Autoantibodies to cardiac conducting tissue and their characterization by immunofluorescence.

(1) A new antibody has been found by immunofluorescence which reacts with cardiac conducting tissue using ox heart false tendons. It was detected in eight out of ninety-three cases of idiopathic heart block (8-6%), in one out of twenty-two cases of secondary heart block (4-5%) and in seven of 165 normal controls (4.2%), in titres varying from 1:10 to 1:40. Previous authors had indicated that this tissue might contain unique antigens. (2) Sera reacting with type I fibres in skeletal muscle (red zebra) were found to be of two varieties, one of which stained conducting fibres diffusely while it gave minimal staining of cardiac muscle; the other reacting with myofibrils in Purkinje cells and heart. (3) Some sera with high titre smooth muscle antibodies (SMA) reacted with conducting tissue together with skeletal and cardiac muscle, suggesting that the four tissues have at least one antigen in common. (4) Known non-organ specific antibodies behaved as expected on beef conducting fibres: striational fluorescence of myasthenia gravis sera reacted with the same patterns on Purkinje myofibrils; AMA and ANA produced IFL in expected locations; ribosomal antibodies reacted strongly, while LKM and reticulin antibodies showed no reactivity. (5) Although the incidence of specific Purkinje fibre antibodies was not significantly raised in idiopathic heart block, the clinical associations suggest that some cases might be related to autoimmunity possibly involving cell-mediated mechanisms as in polymyositis.


The role of macrophages in the adjuvant effect on antibody production of Corynebacterium parvum.

Spleen cells from mice pre-treated with C. parvum gave an enhanced in vitro antibody response to SRBC, but not to DNP-POL. This enhancing activity was associated with the adherent, but not the non-adherent spleen cell population and was found to be radioresistant. It is concluded that macrophages are directly involved in the adjuvant effect of C. parvum and the possible mechanisms of action are discussed.


Prevalence of hypertension in a renal transplant population on alternate-day steroid therapy.

A study of the prevalence of hypertension in a group of renal transplant patients on alternate-day maintenance steroid therapy was conducted. Twenty-four percent of the transplant clinic population was hypertensive. The factors that were associated with a lower prevalence of hypertension were good graft function, bilateral nephrectomy of the patients' own diseased kidneys (although the majority of our patients without bilateral nephrectomy are normotensive), and use of a living related donor. We conclude that the prevalance of hypertension in transplant patients on alternate-day steroid therapy is low. In the presence of all these favorable factors, only 6% of allograft recipients were hypertensive.


Use of allogenous articular bone implants as substitutes for autotransplants in adult patients.

The series of 212 patients with transplanted articular ends of bones of different localization, and 16 patients with transplanted bone cartilaginous fragments are evaluated in terms of technical errors, complications, and 11-year follow-up results of organ allotransplantation. Special attention is focused on the indications and contraindications in the individual patient. Various kinds of osteotomy and schemes of transplant attachment and fixation are possible.


Antibiotic prophylaxis in orthopedic surgery.

Antimicrobial prophylaxis, to reduce the incidence of postoperative infections, has been a controversial subject. The administration of antibiotics just prior to and up to 3 hours after operation has been demonstrated to be effective in preventing experimental staphylococcal wound infection in animals. However, the use of prophylactic antibiotics in orthopedic surgery has been tested in appropriate clinical trials only in the treatment of fractured hip. In that setting, prophylactic antibiotics were found to be efficacious. Although used extensively in other procedures such as total hip replacement, statistically valid evidence of the efficacy of prophylactic antibiotics has not been produced. Suggestions are offered for the conduct of clinical trials to study this important question.


Surgical treatment of chronic osteomyelitis complicating fractures; 6 year interim follow-up.

The loss to continued follow-up of 25 of the 50 patients with chronic osteomyelitis complicating fractures who were treated surgically during the period January 1969 to December 1971 as previously reported, compromises the value of even a 6-year follow-up study. The group of patients lost to follow-up had dry asymptomatic wounds when last examined. However, late recurrence of drainage in 4 patients with dry healed wounds for as long as 15 to 36 months (Procedure 1 in 3, Procedure 2 in 1) demonstrates the uncertainty of the end results of even the most exacting techniques and appropriate antibiotics for the treatment of chronic osteomyelitis.


Hand injuries from sea urchin spines.

Sea urchin spine injury is usually a benign process that rarely comes to the attention of a physician. Aside from the transient episode of excruciating pain which responds dramatically to hot water soaks, there is usually no residual disability. As in any puncture wound, tetanus prophylaxis and observation for latent infection is advised. Complications arise, however, when spines are embedded over bony prominences, within joints, or in contact with nerves. Cases are reported herein of the latter two problems occurring in the hand. A case of a neuropathy associated with sea urchin injuries has not been previously reported in the literature. When such injuries necessitate exploration, aseptic surgical technique is required.


The pathogenesis and long-term end results of pigmented villonodular synovitis.

Pigmented villonodular synovitis, bursitis and tenosynovitis may present a variable picture depended upon the site and whether they are localized or diffuse. In the localized form of the disease occurring in association with tendon sheaths, complete surgical excision should result in a cure. Local recurrences probably result from incomplete excision. The localized disease within the knee joint may represent a picture of internal derangement. Simple excision should result in complete cure. Diffuse pigmented villonodular synovitis (DPVS) poses a diganostic and therapeutic problem. Where possible, complete synovectomy is indicated but the recurrence rate after such surgery is unfortunately quite high. Radiation therapy should probably be considered only in patients in whom lesions recur after surgical synovectomy. Despite three decades of study, the etiology and pathogenesis of the lesion remains obscure. It is apparent, however, that the lesion is not a neoplasm and probably represents an inflammatory process.


Thyroid hormone-like effects without thyrotoxicosis during one year's therapy with NA-DT3 for hypercholesterolemia.

A thyroid hormone analogue, sodium dextro-triiodothyronine (NaDT3), at a dosage of 1 mg/day for 1 or 2 yr, decreased serum cholesterol levels about 30% in 26 hyperlipidemic adults. There were less sustained decreases in the serum phospholipids, and occasional lowering of the serum triglycerides, but no effects on body weight, blood pressure, or pulse rate. Changes recognized as variable concomitants of spontaneous or induced thyrotoxicosis, such as transient increases in fasting blood glucose, calcium, and globulin, persistent rises in alkaline phosphatase, and nonsustained decreases in hematocrit are consonant with the fact that Na-DT3 exerts about one tenth of the thyroid hormone activity of LT3. These changes, however, appear to represent actions of iodinated amino acids apart from those effects that result in clinical thyrotoxicosis.


Influence of magnesium and aluminum hydroxide mixture on chlordiazepoxide absorption.

Ten healthy male subjects ingested 25 mg of chlordiazepoxide hydrochloride (Librium) with 100 ml of water or with 100 ml of magnesium and aluminum hydroxide (Maalox) in a single-dose crossover study. Multiple venous blood samples drawn during the first 24 hr after each dose were assayed for concentrations of chlordiazepoxide and its major metabolite, desmethylchlordiazepoxide. The antacid prolonged the mean chlordiazepoxide absorption half-time from 11 to 24 min, and in 6 of 10 subjects delayed achievement of the peak blood concentration by from 0.5 to 3.0 hrs. The formation of desmethylclordiazepoxide was also slowed. The areas under the 24 hr blood concentration curve for chlordiazepoxide and for its metabolite were not influenced by the antacid. The apparent elimination half-life of chlordiazepoxide (8.4 and 8.2 hr) was not significantly affected. Administration of chlordiazepoxide with antacid reduces the rate of its absorption but does not alter the completeness of absorption or the apparent rate of elimination.


Combination chemotherapy of adult acute lymphocytic leukemia.

Seventeen adults with previously untreated acute lymphocytic leukemia (ALL) received thioguanine, vincristine, dexamethasone, and pyrimethamine for remission induction. Nine patients (53%) achieved complete and 5 (30%) partial remissions. Once in complete remission patients were given two closely separated consolidation courses followed by monthly maintenance courses of the same regimen with the addition of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) every other month during maintenance therapy. The median duration of complete remission was 176 days (range 38 to 605). The median survival for all patients was 405 days (range 30 to 1,058+). Oral pyrimethamine and CCNU were used for their potential activity as prophylactics against meningeal leukemia. Dexamethasone was used instead of prednisone because of the potential enhancement of granulocyte mobilization by the former. Six patients (35%) developed meningeal leukemia while on initial induction or maintenance therapy, 5 within 6 months of diagnosis. Life-threatening infections occured in 10 patients (59%) during induction. Whereas the regimen effectively induced complete remission in about half of previously untreated adults with ALL, it was not effective in maintenance. The incidence of meningeal leukemia and infection during induction was high. Oral pyrimethamine was inadequate prophylaxis for meningeal leukemia; dexamethasone did not reduce the incidence of serious infection.


Influence of cannabidiol on delta-9-tetrahydrocannabinol effects.

Experiments investigating the possible interaction of tetrahydrocannabinol (THC) and cannabidiol (CBD), two major components of marihuana, were conducted under controlled laboratory conditions in a double-blind manner. In one study, 15 male volunteers were given placebo or 25 mug/kg of THC together with either placebo or 150 mug/kg of CBD by inhalation of the smoke of a single cigarette. All four treatments were assigned to each subject according to a series of Latin-square designs. CBD significantly attenuated the subjective euphoria of THC. Psychomotor impairment due to THC was not significantly altered by the simultaneous administration of CBD, but a trend indicating a decrease in THC-like effects was observed after the combination. When administered alone CBD was inactive for all the parameters measured. In a second study, 8 male subjects were given CBD (0 or 150 mug/kg) by smoke inhalation 30 min before THC (0 or 25 mug/kg) in a second cigarette. In contrast to the simultaneous administration of both drugs, CBD pretreatment did not alter the effects of THC on the parameters observed.


Plasma levels and antidepressive effect of imipramine.

The relationship between the antidepressive effect of imipramine and the plasma concentrations of imipramine and the active metabolite desipramine was studied in 24 patients suffering from endogenous depression. After a placebo period of 7 days, the patients received imipramine, 75 mg 3 times a day. The dose was reduced in patients with pronounced side effects. Blood samples for drug assay were drawn in the morning, 15 hr after the last drug intake. Imipramine and desipramine in plasma were assayed by quantitative in situ thin-layer chromatography. Individual variations in plasma concentration were 20- to 30-fold in both imipramine and desipramine. Severity of depression was assessed on the Hamilton Rating Scale (HRS). Eleven of 12 patients who responded satisfactorily to the treatment (HRS post-treatment score less than 8) had plasma concentration of imipramine greater than or equal to 45 mug/L, and desipramine greater than 75 mug/L, whereas the 12 patients not responding satisfactorily (post-treatment score on HRS greater than or equal to 8) all had concentrations of imipramine or desipramine or both below these limits.


Antihypertensive effectiveness of oxprenolol administered twice daily.

Oxprenolol, a beta-blocker, is an effective antihypertensive when administered 3 or 4 times daily. We evaluated the antihypertensive effect of oxprenololgiven twice daily (bid). The subjects were 15 ambulatory men whose standing diastolic blood pressure (BP) was at least 100 mm Hg after 3 wk of treatment with hydrochlorothiazide and oxprenolol placebo. Oxprenolol 40 mg twice daily was then substituted for the placebo. On subsequent weekly vists oxprenolol was titrated to 80 and 160 mg bid if the standing diastolic BP was greater than 90 mm Hg. BLood pressures on the last visit on placebo were compared to those on the last visit on oxprenolol. Standing BP declined from 145 +/- 4/108 +/- 1 to 130 +/- 4/98 +/- 4 on a mean dose of 256 mg of oxprenolol (p less than 0.001 syst.; p less than 0.01 diast.). Recumbent BP fell from 146 +/- 4/107 +/- 1 to 138 +/- 5/93 +/- 2 (p less than 0.06 syst.; p less than 0.01 diast.). During the final week, 13 of the 15 patients were admitted to the hospital for 24-hr monitoring of BP. The 24-hr BP readings showed a mean coefficient of variation of 6.6% recumbent and 7.2% standing. we conclude that bid oxprenolol will maintain 24 hr BP control in most patients.


Oral and intramuscular phenytoin.

Twelve epileptic patients, after 1 wk on their previously established oral dose of phenytoin, were given a 50% larger dose intramuscularly for 1 wk after which they were put on an oral phenytoin regimen for 1 wk on a dose one-half the original oral dose. Plasma phenytoin levels did not show a sharp decrease in the intramuscular (im) nor a dangerous increase in the second oral phase. Those receiving drug once a day maintained remarkably stable levels. By the end of the im phase, patients receiving drug in multiple daily doses had some increase in plasma levels proportional to the dose increase. Seizure control was maintained equally well throughout all phases of the study. Except for local irritation at the site of injection, there were no adverse experiences. The data indicate that the dosage regimen described is suitable for patients on oral phenytoin who have to be transferred to im drug for short periods.


Mammary augmentation--surgical techniques, evaluation of results, and complications.

Mammary augmentation is the procedure of choice in all forms of breast underdevelopment, atrophy or absence due to disease (subcutaneous or radical mastectomy), or trauma (burns of the thoracic region). Old reconstructive methods employing autogenous tissues have no substitute in special cases, but they are used less and less in favor of the new alloplastic materials (silicones) introduced as implants under the mammary gland. These implants have been shown to be permanently accepted by body tissues, the aesthetic results are good, and the operation is easier, faster, and less dangerous. Complications, however, do exist and the most frequent is the constriction of the fibrous envelope surrounding the implant. The etiology of this complication is still unknown. The patient should be advised of such a possibility. Circular incision of the fibrous envelope seems to be the best treatment for this complication. Silicone implants do not modify the physiology of the mammary gland and there is no reason to believe they have any influence on the incidence of breast cancer.


Ankylosing spondylitis and related diseases: significance of HL-A W27 histocompatibility antigen.

The HL-A W27 tissue antigen provides a laboratory tool that has already been of great use in explaining the clinical relationships and overlaps among the true seronegative arthritides. In addition, the W27 antigen is useful to the clinician in differential diagnosis and is becoming more readily available through various laboratories. Tissue typing can be done with a tube of blood sent to the laboratory. As and its variants are becoming better understood and thus will be better managed as a result of the recent developments in transplantation antigen typing.


Malignant hyperpyrexia.

Malignant hyperpyrexia is a dangerous complication of general anesthesia occurring in individuals with an underlying disease of muscle. The essential clinical features of the syndrome are a drastic and sustained rise in body temperature, metabolic acidosis, and widespread muscular rigidity. The results of experiments on susceptible pigs and in vitro studies of human muscle have shown that all the clinical features of the syndrome can be explained by a raised level of calcium ions in the myoplasm. This is caused by a massive and sudden release of calcium into the myoplasm from the calcium-storing membranes in the muscle cell when exposed to general anesthetic agents. Two myopathies predisposing to malignant hyperpyrexia have been identified. One is usually subclinical, dominantly inherited, and manifested only by raised serum CPK levels. The other occurs in young boys with a number of physical abnormalities, whose relatives are unaffected. The serum CPK is a useful screening test in families in which malignant hyperpyrexia has occurred. Unfortunately, though, the serum CPK is not a specific test, and in doubtful cases the only unequivocal method of establishing susceptibility to malignant hyperpyrexia is to carry out an in vitro muscle test in which the muscle is exposed to caffeine, halothane, succinylcholine, and potassium chloride. Susceptible individuals should be given local, regional, or spinal anesthesia if an operation is needed. If this form of anesthesia is unsuitable, barbiturates such as thiopentone, tranquilizers such as diazepam, narcotics such as Pantopon, and neuroanaleptics such as fentanyl, nitrous oxide, d-tubocurarine, and althesin appear to be safe. By far the most important aspect of treatment is prophylaxis. Early diagnosis and immediate cessation of the offending anesthetic agents are the most important factors in trying to reduce the very high mortality of the syndrome.


Salmonellosis in Equidae: a study of 23 cases.

Salmonellosis in Equidae is a serious global problem. The prevalence may range from 0.36% to 27%. Probably 5% to 10% of the equine population in the U.S. is or has been infected. Over 40 serotypes of Salmonella have been cultured from Equidae. S. typhimurium (66.31%), S. enteritidis (9.6%), S. newport (5.16%) and S. heidelberg (4.89%) have been the most common equine isolates. The clinical and bacteriological studies of 23 naturally occurring infections in a large veterinary hospital were studied. Nine patients were infected with S. typhimurium, 8 with S. anatum, 1 with S. newport, 4 with dual serotype infections (3-S. typhimurium/S. anatum and 1 S. anatum/S. newington) and 1 untyped Salmonella. The roles of physical, surgical and environmental stress factors were analyzed. The fecal shedder state persisted for over 3 1/2 months. Feces of a foal contained 10(5) salmonella per gram. Salmonella were isolated from the feces of horses during and following antimicrobial therapy regimes. Supportive and symptomatic treatment as well as good nursing care are imperative in handling cases of equine salmonellosis.


Hair follicle mites (Demodex spp.) of medical and veterinary concern.

A review is presented, including some new data, of the biology of ten species of hair follicle mites (Demodex spp.) of man and domestic animals. Major topics covered are taxonomy, host-parasite interactions, and problems in need of resolution. To this last, some information is provided on the question of species validity, the degree of host specificity and synhospitaly, population dynamics and incidence, transference and host behavior, pathogenesis and disease transmission, and control. A key is provided for species discrimination.


The distribution and characterization of group B streptococci in New York State.

Group B streptococcal isolates from bovine mastitis were classified according to the Lancefield typing scheme. Three types were found in New York State dairy herds; Ia, II and III. The distribution within these types varies from area to area, however, on a state-wide basis Ia is the most prevalent (69.6%) followed by type II (16.7%) and type III (10.1%). Within a single herd one type predominants with only slight incidence of other types. Attempts to determine distinguishing characteristics between isolates of human and bovine origin resulted in no entirely consistent differences.


The pharmacology and pharmacokinetics of azapropazone - a review.

The pharmacology and pharmacokinetic properties of azapropazone, a new anti-inflammatory analgesic, are reviewed. Anti-inflammatory activity was assessed by standard in vivo and in vitro methods. The drug was shown to have definite activity with a potency approximately half that of phenylbutazone. Azapropazone was absorbed from the gastro-intestinal tract in animals and man, although wide inter-species variation in pharmacokinetic properties was noted. In man, the biological half life was approximately 20 hours. Azapropazone is not extensively metabolised and most metabolites have been identified and assayed.


Clinical assessment of azapropazone in rheumatoid arthritis.

Azapropazone was investigated in a 2-week double-blind clinical out-patient trial against placebo in 23 patients with definite rheumatoid arthritis. The drug was given at a dose of 1200 mg. per day before food. At this dose level the drug was shown to have an antirheumatic effect in terms of pain relief, articular tenderness and duration of morning stiffness.


An assessment of the therapeutic potential of azapropazone in rheumatoid arthritis.

A trial to compare the therapeutic potential of 1200 mg. azapropazone daily with 3.9 g. aspirin daily was carried out in 108 out-patients with rheumatoid arthritis over a 14-day period. Analysis of results from the 85 patients with completed assessment data (49 on azapropazone; 36 on aspirin) showed that azapropazone was better than aspirin from the point of view of pain relief, number of days patients were withdrawn prematurely from the trial, and patient satisfaction with treatment. The differences, however, were not statistically significant. The authors compare the results obtained in this trial with those obtained previously from trials with 13 other antirheumatic drugs using the same method.


A comparative study of azapropazone and ibuprofen in the treatment of osteoarthrosis of the knee.

A double-blind crossover study in 41 patients with knee joint osteoarthrosis was carried out to compare the efficacy of 1200 mg. azapropazone per day with 1600 mg. ibuprofen per day. After an initial week on placebo, patients received one or other of the active medications for 2 weeks, then a 1-week placebo wash-out period before being crossed over to the alternative drug for a further 2 weeks. Objective assessments were made of knee joint movement, and of knee joint and thigh circumference. Patients made daily assessments of pain and a final overall assessment of preference for one or other treatment period. The results showed that azapropazone produced a significant improvement (p less than 0.05) in knee joint mobility over placebo, both active drugs were more effective than placebo (p less than 0.05) in providing pain relief, but that there was no significant difference between the two in these parameters. There was a highly significant patient preference, however, for azapropazone (p less than 0.01) compared to ibuprofen treatment periods. The most commonly reported side-effect with both drugs was dyspepsia, but in no case was it sufficiently severe to necessitate stopping treatment.


A comparative pilot study of azapropazone and indomethacin in the treatment of psoriatic arthritis and Reiter's disease.

A double-blind crossover study to compare 1200 mg. azapropazone per day with 100 mg. indomethacin per day was carried out on 34 patients with psoriatic arthritis and 16 patients with Reiter's disease. The same comparison was initiated in 8 additional patients but withdrawn in 3 cases because of side-effects to indomethacin, in 2 cases because of side-effects to azapropazone, and in the 3 remaining cases for other stated reasons. The results of the study showed no overall difference between the treatment results of azapropazone and indomethacin, but azapropazone seemed to be more effective in cases with psoriatic arthritis and indomethacin in cases with Reiter's disease. Indomethacin caused more side-effects than azapropazone. Neither drug seemed to influence the skin manifestations of either disease.


A comparative study of azapropazone and indomethacin in the treatment of rheumatoid arthritis.

A double-blind crossover trial in 50 patients with rheumatoid arthritis was carried out to compare the clinical effectiveness of 800 mg. azapropazone per day with 100 mg. indomethacin per day. Blood plasma concentrations of azapropazone were also studied in 10 patients. Patients received both drugs for a 3-week period, the order of treatment being determined at random. Although there were no marked differences in any of the objective measurements, patients assessments of response to and preference for each drug treatment period suggested that azapropazone produced significant improvement compared to that with indomethacin. Plasma levels of azapropazone remained remarkably constant for each patient. Side-effects with both drugs were mild and transient, and there were no marked variations from normal in any of the laboratory parameters measured.


Prolonged treatment with azapropazone.

In an open study of 40 patients with chronic rheumatoid polyarthritis, azapropazone was given alone in daily doses between 1200 mg. and 1800 mg. over a period of 6 months. Six patients were withdrawn, 2 because of side-effects (1 with an allergic-type rash; 1 with ankle oedema) and 4 because azapropazone alone did not provide adequate control of their pain and discomfort.  Objective assessments showed that 18 patients had a 'good' or 'very good' response to treatment, and a further 16 showed moderate improvement. These findings were supported by the patients' subjective assessment of the change in their condition from the start of the trial. In addition, 22 patients showed a significant decrease in erythrocyte sedimentation rate. There was no evidence of adverse effects on any of the haematological and biochemical parameters measured, and the incidence of side-effects was low.


Azapropazone - its place in the management of rheumatoid conditions.

The authors review the trials carried out on azapropazone in rheumatoid arthritis and other rheumatoid conditions. They comment that in terms of efficacy azapropazone would appear to be a useful non-steroidal anti-inflammatory analgesic which compares favourably with other established antirheumatic agents against which it has been tested. Its main advantages are its low incidence of side-effects and the fact that in the majority of the trials reported azapropazone treatment was preferred by patients to that with other agents.


A short-term double-blind trial of aerosol triamcinolone acetonide in steroid-dependent patients with severe asthma.

Twenty-five steroid-dependent severely asthmatic patients, ranging in age from 20 to 67 years, were hospitalized. Baseline laboratory and pulmonary function testing was followed by reduction of prednisone therapy to 5 mg daily and by entry into a randomized double-blind study of placebo vs active aerosol triamcinolone acetonide (300mug four times daily). In this four-week trial, aerosol triamcinolone acetonide further reversed airway obstruction and proved to be an effective substitute for large oral doses of steroids in steroid-dependent patients with severe asthma. No significant improvement occurred in the maximum midexpiratory flow or the maximum velocity of air flow after 50 percent or 75 percent of the vital capacity had been expelled. There was no significant difference in the frequency of untoward effects between the groups taking aerosol triamcinolone acetonide and its vehicle. No patient demonstrated any definite return of adrenal function.


Phagocytosis and cutaneous delayed hypersensitivity in patients with chronic obstructive pulmonary disease.

The objective of this study was to determine if the phagocytic and intracellular killing capacity of peripheral granulocytes or an expression of cellular-mediated immunity, delayed cutaneous reactivity, as measurements of native and acquired immunity, might be risk factors associated with chronic obstructive pulmonary disease (COPD). Over 100 patients with a value for their forced expiratory volume in one second (FEV1) less than or equal to 70 percent of normal were carefully matched with healthy participants having an FEV1 greater than or equal to 86 percent of normal, and together they served as the study group. Phagocytosis and intracellular killing were normal in patients with COPD; however, these patients demonstrated a significant impairment in the ability of their peripheral leukocytes to reduce nitroblue tetrazolium. The delayed-hypertensitivity response rate and the degree of reactivity were similar in the two groups, except for the patients with COPD having a significantly greater degree of reactivity to Monilia albicans extract ("canadin.") This finding is thought to be a consequence of reduced mucociliary clearance rather than a risk factor. The significance of decreased resting and stimulated cells' reduction of nitroblue tetrazolium in patients with COPD is not clear.


The psychosomatic approach to heart disease. A study in premodern medicine.

Prior to the discovery of the circulation of the blood and the advent of modern mind-body dualism, heart disease was conceived as invariably psychosomatic. Familiar vestiges of premodern theory are found in such expressions as "bighearted," "warmhearted," "brokenhearted," and so on. The original medical meanings of such expressions are explained. The historical disappearance of the psychosomatic approach to cardiovascular disease is explained as a product of dualistic thinking and the mechanistic physiopathology it created. It is urged that contemporaries could profit from a return to a holistic view of the disease process.


[Identification of Group A streptococci from throats of school children in Northern Taiwan].

A total of 2,750 strains of beta-hemolytic streptococci was isolated from 10, 510 throat swabs collected from school children in the Taipei area. Most of them were classified as group A (61%) and group G (34%), and about 5% belonged to other groups or unclassified strains. 1, 772 strains of beta-hemolytic streptococci isolated from healthy school children, and 69 strains from sick school children were grouped by the Lancefield precipitin test, fluorescent antibody technique and/or bacitracin paper disk method. A satisfactory correlation (over 90%) was obtained by the latter 2 methods, when compared with the results of the Lancefield technique. The occurrance of false negative or false positive was only 1 or 5%, respectively.


Studies of native plants in Taiwan for lectin content.

The lectin content of some native plants in Taiwan was studied. The majority of the plants possessing lectin belonged to Leguminosae. Five lectins which are readily available were further studied for their hemagglutination titers, specificity, susceptibility to heat and proteolytic enzymes. The lectins of Ricinus communis had hemagglutination titer of 1:512 which was the highest among the five tested. All lectins were heat labile, destroyed by heating at 100 degrees C for 30 min, but resistant to trypsin and pepsin. The resistance was not due to the presence of any inhibitor in the crude extract, but possibly due to the special configuration of the lectin molecule or other unknow reasons. Among the five lectins only the lectin of Phaseolus vulgaris could not be inhibited by any of the carbohydrates tested. It seems that the lectin of Phaseolus vulgaris is nonspecific, or not specific for ABO blood group, or specific for other blood groups. The inhibiting activity of carbohydrates seems to be specific. The configuration of carbohydrates at carbon atom 2 and/or 4, the type of linkage (alpha or beta), or the carbon number where the linkage is formed seem to be important to the potency of carbohydrates to inhibit the activity of lectin.


A congenital contribution to emotional response in early infancy and the preschool period.

Four follow-up studies carried out from the newborn period in three different laboratories have shown that a slow, low-magnitude reaction of the newborn to interruption of sucking is associated in later infancy and the preschool period with the expression of pleasurable emotional response. The basic test of the newborn's response varied slightly in the four studies, but essentially it consisted of establishing a baseline of infant behaviour during 30 to 120 seconds of sucking on a pacifier, then recording amount of (a) bodily activity, (b) crying, (c) the latency to either or both, in the 30- to 135-second period after removal of the nipple. The three aspects of the reaction are consistently intercorrelated, even when allowances are made for artifactual interdependence. Newborns who respond quickly to interruption of sucking also continue to cry and move a lot. Up to the first month a slow, low-magnitude reaction of the newborn is associated with a later low rate of smiling, but beyond the third month there is an inversion and the association is consistently with various manifestations of postive emotional expression.


Metabolism of radiolabeled insecticides in insects and related arthropods: a critical study of various techniques.

The metabolism of radiolabeled insecticides in insects and acarina is studied largely by coupling radiotracer techniques with analytical methods, such as TLC, paper and column chromatography, gel-permeation chromatography, and enzymatic assays. These techniques in various combinations yield both the identification and quantification of the metabolites. Other analytical methods such as gas chromatography or IR spectrometry may also be used to obtain additional support for identification of metabolites. In the absence of authentic chromatographic standards, however, NMR and mass spectrometry are necessary in the identification of the unknown compound. The quantity of the radiolabeled insecticide administered should be within the toxicological range of the insect. Therefore, the dosage-mortality response of the insect using unlabeled material should be determined. A dose should be selected that keeps insect mortality to a minimum in order to avoid complications in the computation of the balance data. The radiolabeled insecticide is usually applied topically to the insect. Alternately, the material may be administered by dipping in a solution containing the radiolabeled compound or by exposure to filter paper impregnated with radiolabeled material. Administration of the radiolabeled material by the oral route presents several problems. Sterile rearing conditions are mandatory to avoid contamination of treated diet with microorganisms. Some knowledge of the insect's feeding rhythm is desirable so that the labeled diet is given at peak feeding time. Synthetic diets should be adjusted to pH 7.0. These precautions minimize degradation of the insecticide in the diet prior to consumption by the insect. Precise doses of radiolabeled materials may be administered by injection. The technique is mainly useful in metabolism studies of intermediate materials resulting from the biotransformation of the parent compound.


Treatment of parkinsonism.

Although the mortality of Parkinsonism is negligible, it is one of the major causes of progressive and often pitiful disability in the elderly. Treatment of the condition, whether medical or surgical, is still only palliative. The successes and limitations of the various forms of treatment are discussed. Despite the spectacular gains which have been achieved during the past 25 years by sterotactic thalamotomy and treatment with levodopa, the patient's disabilities tend to progress slowly and the fundamental enigma of Parkinson's disease and of its cure still awaits solution.


Clinical use of immunosuppressive drugs: part I.

While immunosuppressive drugs are principally used in the treatment of malignant disease, their use in non-malignant disease and transplantation has become commonplace. The mechanisms of action of immunosuppressive agents differ. Alkylating agents react with nucleophilic centres of D, and rna. Folic acid antagonists prevnt the conversion of dihydrofolic acid to tetrahydrofolic acid. Antibiotics act in a variety of different ways; the        alkaloids cause metaphase arrest. Cytarabine (cytosine arabinoside), methotrexate, hydroxyurea and  thioguanine act during mitosis and the latter also acts during replication is not known...


[Methods for the macroscopic diagnosis of pulmonary emphysema].

The author examined 124 dead persons with clinicaly and anatomicaly diagnosed emphysema in order to prove and demonstrate the possibilities of the the existing methods for macroscopic diagnosis of the lung emphysema. The method of Gough and Wentworth, barium-sulfate method of Heard and simple original method for examination of the changes by a naked eye or with only a magnifying glass in fixed total lung sections were used and the results described. He constructed a special table and knife to cut the lung at equal sections. An inference was made that the method of Gough and Wentworth, although more complicated, revealed the best advantages. Both the impregnation method of Heard and the newly proposed method were very suitable for an examination of a very large number of lungs, since they assure enough information for the diagnosis, distribution and degree ofthe pathological process and allow current morfological classification of the disease.


[Morphological changes in the endocrine pancreas of the rabbit after the administration of a Morus alba extract].

The authors examined the effect of an extract, obtained from the leaves of the plant Morus alba on beta-cells of the endocrine pancreas of a rabbit after subcutaneous and parenteral administration. The most manifested morphologic changes were observed after subcutaneous administration under the conditions of a chronic experiment. There were degranulation of beta-cells of the Langerhans islets of these animals developed at various degree and discovered by means of a light and electrone microscope.


[Study of the effect of centrophenoxine on experimental animals poisoned with chlordiazepoxide].

The author administered per os chlordiazepoxide in a toxic dose of 600 mg/kg of body weight/LD50 of chloridazenoxide for rats was 680 mg/kg of body weight in 20 white male rats, weighing 150 gm. Two hours later, on the background of advanced intoxication, manifested by adynamia ataxia, lateral position, somnolence reaching stupor, the author administered in half of the animals (10) per os centrophenaxine in a dose of 50 mg/kg of body weight. The signs of intoxication disappeared for 24 hours after treatment with centrophenoxine. Three of the control animals died, but in the remaining rats the intoxication worsened, but then slowly disappeared till the end of the fifth day. The obtained experimental data supported the observations of the clinical practice on the favourable effect of centrophenoxine in the treatment of acute poisonings with chlordiazepoxide.


[Determination of antigens M and N in blood strains by an absorption-elution method].

The author examined blood spots with known M and N antigens lasting up to two years by means of method of absorption elution. Positive results were obtained in 99 samples. The method was very sensitive and filament with a length of 2 to 4 mm were sufficient for determination of antigents.


[Various current trends in the study and interpretation of asphasic disorders].

This review of the present tendencies in aphasiology intends to put in evidence some features of to-day's aphasiology, in relation both to the methods used to study the language disturbances and to the concepts advanced to explain the clinical and experimental findings. From the methodological point of view, two important novelties are to be stressed : 1) the clinical approach is gradually substituted by a more strict experimental method ; 2) the pathology of language is to-day generally studied by means of linguistic methods, models and techniques. In the interpretation and classification of the aphasic troubles many tendencies are to be noted :--some Authors tend to classify the aphasic disorders on the  ground of empirical criteria (such as the distinction between a "fluent" and a "non-fluent" form of aphasia) ;--other writers tend to classify the aphasic disturbances on the ground of linguistic, of neuro-anatomic, or of neuro-linguistic criteria. The results of a few experimental investigations on some dimensions of auditory language comprehension in aphasia are reported and the implication of these results on the concepts advanced by various Authors are discussed.


[The clomipramine-lithium combination: controlled trial].

A controlled study on a pragmatic type was performed on 30 patients of both sex, all having recurrent depressions. The efficacy of Clomipramine plus Lithium Carbonate was compared to that of Clomipramine plus placebo. The control consisted in a double-blind study with random sampling of patients. Results were recorded by an independant observer using a rating scale and were analysed statistically. In spite of the bias which was observed it seems possible to conclude. The association of Clomipramine plus Lithium Carbonate has no greater global antidepressive efficacy as compared with Clomipramine plus placebo which means that Lithium does not potentialize nor antagonize the anti depressant effect of Clomipramine. In respect of the number of patients : bipolar depressions (10 cases) unipolar (20 cases) it has not been possible to study the results in these 2 sub groups.


[Amitriptyline in psychiatric therapeutics. Apropos of 8 years of its use in hospital practice].

A hundred and thirty patients have been treated with Amitriptylin in the "Clinique de Psychiatrie" in Marseille during the last eight years. The review of the hundred cases for which the information are the most detailed show that Amitriptylin remains one of the most powerful, manageable and well-tolerated anti-depressant drugs available at the present time. Its therapeutic indications are numerous, but the best ones are proceeding from depressed states with high anxious participation in subjects of less that 60 years of age.


[Study of the clinical effects of sulpiride and perphenazine in 82 schizophrenic patients by the double blind method].

Our clinical and statistical evaluation of Sulpiride and  Perphenazine was carried out over a 10 week period, in 82 patients of both sexes, aged 20 to 56 years. All were chronic patients with a symptomatic profile of apathy, lack of initiative but with the personality relatively well preserved in 56 patients. The daily dosage varied from 300 to 1,200 mg Sulpiride orally and 12 to 48 mg Perphenazine. The effects of therapy have been received globally and individually at the beginning, 4th, 8th, and the end of the 10th week using the B.P.R.S. modified by the clinic of Neurology and Psychiatry of the University of Hiroshima. Using "Armitage's Restricted Sequential Design", there was no significant difference between the therapeutic effects of these 2 neuroleptics. However, Sulpiride appeared more beneficial following the review during the 8th week and the 4th week.


Pituitary responsiveness to gonadotrophin releasing factor (GnRH) in puerperium.

Administration of a single dose of Gondotrophin Releasing Factor (GnRF) failed to induce an increase in serum FSH immediately postpartum suggesting pituitary refractoriness. However, multiple injection of GnRF resulted in significant FSH increments but these were less than the FSH responses to a single injection of GnRF in normal menstruating women.


The onset of puberty in the female rat: changes in plasma prolactin, gonadotropins, luteinizing hormone-releasing hormone (LHRH), and hypothalamic LHRH content.

In order to study the sequence of hormonal changes that accompany the onset of puberty in the female rat, immature animals were sacrificed by decapitation between days 32 and 38, and plasma titers of gonadotropins, prolactin, and LHRH, and the hypothalamic content of LHRH were determined by specific radoimmunoassays (RIA). Animals were decapitated at 10:00 and 16:00 h throughout the pubertal period, the uterine weight was recorded, and the ovaries were inspected for signs of ovulation. Animals with the vagina closed were grouped according to the condition of the uterus as anestrus (A), early proestrus (EP) and late proetrus (LP), the uterus being unstimulated, dilated with some fluid, or ballooned, respectively. Vaginal opening was usually associated with ovulation and in most cases occurred at the end of the late proestrous phase. Animals were studied up to 3 days after vaginal opening and were grouped according to vaginal cytology. Uterine weight, taken as an index of estrogen secretion, was low during A, increased during EP, and reached a peak at LP, declining thereafter. Plasma LH and FSH were low from A until the afternoon of LP. At this time, uterine weight was maximal and both gonadotropins increased dramatically. The following morning (estrus), LH but not FSH, had returned to basal values. FSH returned to basal levels on the afternoon of estrus. Plasma prolactin was low in the morning during the entire period, but showed peaks on the afternoon, which reached a maximum at LP and declined thereafter following the pattern of changes in uterine weight. Plasma LHRH was uniformly low throughout the entire pubertal period, whereas hypothalamic LHRH content declined on the morning of estrus (day of vaginal opening). We suggest that the onset of puberty in the female rat is brought about by a gradual increase in estrogen secretion which, acting at the CNS-pituitary level, triggers a preovulatory proestrus-like surge of gonadotropins and prolactin.


The existence of a possible short-loop negative feedback action of LH in proestrous rats.

The responsiveness of the hypothalamus to electrochemical stimulation (ECS) and of the pituitary to lutenizing hormone-releasing hormone (LHRH) was examined following either endogenous LH release or exogenous LH infusion on the afternoon of proestrus in rats that had received Nembutal to block the LH surge. In all animals, sequential jugular blood samples were taken and LH was determined by radioimmunoassay. Unilateral ECS of the medial preoptic area (MPOA) resulted in a peak plasma LH of 121.1 +/- 8.6 ng/ml 120 min after stimulation; ECS of the contralateral MPOA of the same animals at 200 min resulted in a peak plasma LH of 66.9 +/- 5.6 ng/ml or half that obtained following the first ECS. To eliminate a pituitary role in this phenomenon, 250 ng LHRH was given iv at 200 min following unilateral MPOA-ECS. The peak in plasma LH following LHRH (510.7 +/- 60 ng/ml) was not depressed and, in fact, was significantly greater than that obtained in response to LHRH without prior ECS-induced LH release (143.9 +/- 19.6 ng/ml). To compare the responses obtained following exogenous LH, purified rat LH (4 mug) was infused iv over 60 min. When plasma LH had returned to pre-infusion levels, an iv pulse of 250 ng LHRH was administered; the peak in plasma LH was similar to that obtained in response to LHRH in animals without prior LH infusion. Finally, in another group of LH-infused rats (4 mug/60 min), MPOA-ECS was performed, resulting in a plasma LH peak half that obtained without prior exogenous LH exposure. These studies suggest decreased hypothalamic sensitivity to stimulation on proestrus due to a negative feedback of either LH or LH-induced steroids.


The effect of surgical isolation of the hypothalamus on its luteinizing hormone-releasing hormone content.

The luteinizing hormone-releasing hormone (LHRH) content of the medical basal hypothalamus was determined by radioimmunoassay 10 days after the isolation of this region. Between 70 and 90% of the LHRH disappeared from the hypothalamus after the surgical procedure. Thus, it seems that most of the LHRH present in the medial basal hypothalamus arises from, or is controlled by, cells elsewhere in the brain.


Effect of hypophysectomy on immunocytochemically demonstrated gonadotropin-releasing hormone in the rat brain.

The objective was to determine the effect of hypophysectomy on the store of gonadotropin-releasing hormone (GnRH) in certain parts of the brain as revealed by immunocytochemistry. The antiserum used was prepared against synthetic GnRH conjugated with limpet hemocyanin. No change was observed in the store of GnRH in the organum vasculosum of the lamina terminalis or in the cephalic segment of the median eminence GnRH was depleted severely from the central and caudal (junction with the infundibular stem) segments of the median eminence. GnRH was not found in the axons of magnocellular neurons that regenerate during repair of the median eminence-pituitary stalk after hypophysectomy.


Reduced luteinizing hormone release by synthetic luteinizing hormone-releasing hormone (LHRH) in postpartum lactating rats.

The ability of the pituitary to release LH in response to synthetic LHRH was tested in lactating female rats on days 7 and 17 post partum, and compared with that of normal cycling female rats on diestrous day 2 (controls). Three consecutive injections of LHRH (100 ng/100 g BW, sc), each 50 min apart, were given to each rat and sequential blood samples were collected at 25-min intervals by cardiac puncture under light ether anesthesia. In all 3 groups, the 2nd and 3rd injections of LHRH produced much greater increases in serum concentrations of LH than the 1st injection. However, this self-priming effect of LHRH on LH response was markedly attenuated in the postpartum lactating rats (PPL), compared with the normal cycling female rats on diestrous day 2. Three consecutive injections of LHRH produced significantly less LH release in PPL rats than in normal cycling female rats on diestrous day 2. Both day 7 and day 17 PPL rats released equally small amounts of LH in response to LHRH administration. The total amount of LH released by anterior pituitaries (APs) during a 5 h incubation in medium-199, from day 7 or day 17 in PPL rats, was significantly less than that released by the APs from normal cycling female rats on diestrous day 2. APs from day 7 and day 17 PPL rats also released less LH in vitro in response to LHRH (50 ng) stimulation than APs from normal cycling female rats. When APs from normal cycling female rats on diestrous day 2 were incubated with LHRH, the increments in LH release were greater at the end of the 2nd and 3rd h than after the 1st h of incubation. However, such increments in LH release were relatively small when APs from day 7 or day 17 PPL rats were similarly incubated with LHRH.


The therapeutic activity, post-treatment fertility and safety of prostaglandin F2alpha-Tham salt in clinically anoestrous mares: a review.

Of 275 mares receiving prostaglandin F2alpha-Tham salt for its luteolytic effect upon the corpus luteum, 231 (84 per cent) exhibited signs of oestrus (range 73-95 per cent). Some mares not exhibiting overt oestrus may, nevertheless, ovulate and post-treatment rectal palpation of mares apparently failing to respond is suggested in these cases. Of 210 mares for which subsequent history was available. 104 (49.5 per cent) became pregnant from breeding at the post-treatment oestrus that immediately followed the induced regression of the corpus luteum (range 42.2-55.5 per cent). Fertility is comparable at this oestrus to that of naturally occurring oestrus in Thoroughbred mares. Side effects with PGF2alpha-Tham are physiological responses to the prostaglandin and include various degrees of sweating, some hypermotility of the gastrointestinal tract, increased pulse and respiratory rates and, occasionally, abdominal discomfort. These signs only occur in a proportion of mares, are transitory in nature and are resolved without treatment. A brief review of a comprehensive toxicity study using 20-160 times the recommended dose level, daily for 8 days, concludes that PGF2alpha-Tham has a 20 times safety margin for a single injection.


The similarity between the primary structures of two non-histone chromosomal proteins.

A comparison has been made of two non-histone chromosomal proteins, high-mobility-group proteins 1 and 2. Peptides produced from the two proteins by various chemical and enzymic cleavages have been compared by polyacrylamide gel electrophoresis and peptide mapping. In addition the cysteine content, C-terminal sequences and amide content have been determined for both proteins. The results obtained, together with previous results, show that high-mobility-group proteins 1 and 2 have closely related primary structures, although obviously being different molecules. When analysed by isoelectric focusing both proteins exhibit complex banding patterns. In the case of high-mobility-group protein 1, this could be caused by aggregation, but with high-mobility-group protein 2 there are four distinctive subfractions focusing between pH 7.8 and 8.9.


Replication of Escherichia coli DNA in vitro: inhibition by oxolinic acid.

Oxolinic acid, a quinolone antibacterial agent, inhibits reversibly the ATP-dependent replicative DNA synthesis in permeable cell systems as well as in cellophane disk lysates. It is about 10-fold more active than the structurally related nalidixic acid. Both drugs have no effect on the ATP-independent DNA repair, but interfere to some extent with RNA synthesis in permeable cells. They appear to interact with the same target since spontaneous nalidixic-acid-resistant mutants of nalA phenotype are also resistant to oxolinic acid. Full sensitivity to oxolinic acid can be conferred to lysates from resistant cells by addition of extracts from sensitive cells.


A simple method for measuring specific radioactivities of ribonucleoside triphosphates using RNA polymerase.

We describe a method for the rapid, one-step determination of the specific radioactivity and pool size of ATP, UTP, CTP or GTP. Escherichia coli RNA polymerase and poly[d(A-T)] or poly[d(G-C)] are used to synthesize an alternating copolymer from a [3H]nucleoside triphosphate of unknown specific activity and a [14C]nucleoside triphosphate of known specific activity. The fact that [3H]nucleotide and [14C]nucleotide are incorporated into poly[r(A-U)] or poly[r(G-C)] in equimolar amounts, coupled with a knowledge of the [14C]nucleotide specific activity, permits calculation of the [3H]nucleotide specific activity. The requirement for direct knowledge of the [14C]nucleotide specific activity may be bypassed by an isotope dilution procedure. The pool size of a nucleoside triphosphate can be estimated either from isotope dilution data or by determining the fraction of [3H]nucleotide polymerized, dividing the number of counts 3H/min in the polymer by this fraction and by the [3H]nucleotide specific activity. The method was successfully applied to acid extracts made from sea urchin embryos labeled with a [3H]RNA precursor.


Isolation and analysis of the lipid A backbone. Lipid A structure of lipopolysaccharides from  various bacterial groups.

A degradation procedure of lipopolysaccharides was worked out which allows the isolation of the reduced backbone of lipid A in a total yield of between 20 and 30%. This procedure was applied to lipopolysaccharides of S forms (Salmonella minnesota, Shigella flexneri 5b, Escherichia coli 086, E. coli 0111, Xanthomonas sinensis, Rhodopseudomonas gelatinosa) and R mutants (Salmonella minnesota, Shigella flexneri, 5b, E. coli BB9 and E. coli EH 100). Chemical analysis, reaction with beta-N-acetyl-glucosaminidase and application of methylation analysis revealed that the lipid A backbone of all strains contains beta 1', 6-linked glucosamine disaccharides carrying two phosphate groups, one in glycosidic and one in ester linkage, a structure, identified previously in the Salmonella minnesota Re mutant.


Specific irreversible inhibition of enzymes concomitant to the oxidation of carbanionic enzyme-substrate intermediates by hexacyanoferrate (III).

Four different enzymes, class I fructose-1,6-biphosphate aldolase from rabbit muscle, class II fructose-1,6-bisphosphate aldolase from yeast, transaldolase, and transketolase, are inactivated progressively in the presence of their specific substrates and hexacyanoferrate(III). The inactivation is strictly linked to the oxidation of the carbanionic enzyme-substrate intermediates of these enzymes reported previously [Healy, M. J. and Christen, P. (1973) Biochemistry, 12, 35]. However, the loss of activity is not due to the products of this oxidation, i.e. to hexacyanoferrate(II), or to the oxidation product of the substrate such as hydroxypyruvaldehyde phosphate in the case of aldolase [Healy, M. J. and Christen, P. (1972) J. Am. Chem. Soc. 94, 7911]. The inactivation is not reversed on removal of low-molecular-weight compounds by gel filtration or extensive dialysis indicating a covalent modification of the enzyme. The rate of inactivation obeys saturation kinetics with respect to substrate concentration. Hence, the modifying agent is a transiently reactive intermediate formed during the oxidation of the carbanionic enzyme-substrate intermediate by hexacyanoferrate(III).


[Study of the lipopolysaccharide from Escherichia coli K12 CR34].

The structure of the lipopolysaccharide from Escherichia coli K12, strain CR34 has been investigated. The lipopolysaccharide contains D-galactose, D-glucose, D-glucosamine, L-glycero D-mannoheptose, 2-keto-3-deoxyoctonate and lipid A. The core region does not contain D-glucosamine but contains galactose, glucose, and heptose in the molar ratios 1:3:6. Methylations were performed on the lipopolysaccharide and on the degraded polysaccharide obtained after acetic acid hydrolysis of the lipopolysaccharide. It was found that galactose is in terminal position partly in the form of galactopyranose, partly in the form of galactofuranose. A part of the heptose was found as unsubstituted heptofuranose. A mole of glucose was 1 leads to 2 linked and another mole of glucose was 1 leads to 6 linked. Periodate oxidation of the lipopolysaccharide followed by borohydride reduction eliminates galactose and only a part of glucose and of heptose. A mole of heptose gave mannose and thus it is unsubstituted in C6 and C7. One mole of glucose and one mole of heptose were not degraded by periodate oxidation.


The major proteins of the Escherichia coli outer cell-envelope membrane. Heterogeneity of protein I.

One of the major proteins of the Escherichia coli outer cell envelope membrane, protein I, can be separated electrophoretically into protein components Ia and Ib. Strain differences exist regarding presence or absence of component Ib and this component can selectively be lost by mutation to resistance against a phage. Both components Ia and Ib are further heterogeneous isoelectrically, and both together may contain at lease six separable isoelectric species. As judged by analysis of their cyanogen bromide fragments, Ia and Ib are almost identical concerning their primary structure; the difference (charge only or size and charge) was located in a part of the protein that does not correspond to the C-terminal or N-terminal regions. Components Ia and Ib thus represent essentially the same polypeptide and they may arise by a modification process in vitro or the existence of two almost identical genes.


The use of a cleavable crosslinking reagent to identify neighboring proteins in the 30-S ribosomal subunit of Escherichia coli.

A cleavable bifunctional reagent, dimethyl 3,3'-dithiobispropionimidate, has been used to crosslink proteins that occupy neighboring positions in the 30-S ribosomal subunit of Escherichia coli. The crosslinked proteins were identified, fully or partly, by their positions in two two-dimensional gel electrophoretic systems, one diagonal and the other quasi-diagonal, in which the complexes were cleaved after the first-dimensional run. It was found to be necessary to block the protein sulfhydryl groups in order to prevent artifactual disulfide crosslinking after extraction of the protein from ribosome. Eleven crosslinked complexes were detected. Four were fully identified: the triplet S4-S5-S8, and the pairs S2-S3, S4-S5, and S5-S8. In five others one component was identified unambiguously. No additional complexes were seen when the longer homologous butyro and capro reagents were used.


Functional hybrid enzymes reconstituted from Escherichia coli and Serratia marcescens RNA polymerase subunits.

RNA polymerase was isolated from Escherichia coli and Serratia marcescens. The subunits of both enzymes were separated by electrophoresis on cellulose acetate sheets in the presence of urea. Under conditions favouring reconstitution of the RNA polymerases, stoichiometric amounts of the subunits were allowed to interact. Active hybrid enzymes were formed if corresponding subunits of both enzymes were mutually exchanged. The analysis of the RNA products synthesized showed that the reconstituted enzymes are able to recognize the promoters for transcription and the termination signals on the DNA template. The transcription products can serve as messengers for cell-free protein synthesis.


Labelling of L-isoleucine tRNA ligase from Escherichia coli with L-isoleucyl-bromomethyl ketone.

L-Isoleucine: tRNA ligase from Escherichia coli could be irreversibly inactivated by L-isoleucyl-bromomethyl ketone but not by L-isoleucyl-chromethyl ketone. The inactivation rate exhibited a saturation concentration dependence typical for an affinity reagent. L-Isoleucine provided 100% protection against inactivation at saturating concentration, whereas ATP, AMP, and pyrophosphate offered partial protection and tRNAIle, no protection. The ligase was labelled in preparative scale with L-[14C]isoleucyl-bromethyl ketone. The molar ration of label incorporated to enzyme inactivated was close to unity. The protein was subsequently subjected to tryptic digestion and the radioactive peptide isolated and identified. The labelled amino acid proved to be the same cysteine previously reported as being labelled with N-[14C]ethylmaleimide [Kula, M.-R. (1974) FEBS Lett. 46, 130-133].


Interactions of guanosine triphosphate analogues with elongation factor G of Escherichia coli.

Previous studies from this laboratory have shown that the GTP analogue guanosine 3'-diphosphate 5'-triphosphate (pppGpp) was almost without activity in the translocation reaction catalyzed by elongation factor G (EF-G), while it was fully active with elongation factor T (EF-T) and initiation factor 2 (IF-2). To assess the importance of the 3'-ribose hydroxyl itself in the translocation reaction, we examined polyphenylalamine synthesis and EF-G-dependent formation of N-acetylphenylalanylphenylalanylpuromycin (Ac-Phe2-puromycin) supported by 3'-deoxyguanosine 5'-triphosphate (3'dGTP) and 3'-deoxy-3'-aminoguanosine 5'-triphosphate (3'dNH2GTP). Like pppGpp, these nucleotides were similar to GTP in EF-T and IF-2dependent reactions. We also examined the ability of the dialcohol derived from GTP by periodate oxidation and borohydride reduction (rroGTP) and of ITP to support translocation. These compounds had shown significant, although reduced, activity with EF-T and IF-2. A spectrum of activity was found with these compounds in both poly(Phe) synthesis and Ac-Phe2-puromycin formation. All had significantly reduced activity relative to GTP, but all were significantly more active than pppGpp. A surprising finding was that the activities of all analogues relative to GTP were dependent on the reaction temperature in both poly(Phe) synthesis and Ac-Phe2-puromycin formation; these relative activities were significantly lower at 8 degrees C after than 37 degrees C. Furthermore, the extent of poly(Phe) synthesis with the analogues relative to GTP could be significantly affected by the amounts of EF-G and EF-T in the reaction mixture. Differences were minimized in the presence of rate-limiting EF-T and saturating EF-G and maximized in the presence of rate-limiting EF-G and saturating EF-T. This effect of reducing the level of EF-G in the reaction mixture thus mimicked the effect of lowering the incubation temperature, and a similar observation was made for Ac-Phe2-puromycin formation. GTP-supported formation of Ac-Phe2-puromycin at 8 degress C could be inhibited by 3'dNH2GTP, 3'dGTP, pppGpp, and ITP: these compounds were better inhibitors than they were substrates. Inhibition by other nucleotides was also noted. The GTP analogues were compared to GTP as substrates in EF-G-dependent reactions uncoupled from protein synthesis. Fusidic-acid-dependent binding of nucleotides to ribosomes and ribosome-dependent catalytic nucleotide hydrolysis were both examined, and the activity of the nucleotides as substrates in these ractions showed little correlation with their ability to support translocation.


Membrane hybridization by centrifugation analysed by lipid phase transitions and reconstitution of NADH-oxidase-activity.

A procedure is described to hybridize cytoplasmic membranes of Escherichia coli. The method involves high-speed contrifugation of two different vesicle preparations at 37 degrees C in the presence of cations such as spermine or Mg2+. The occurrence of hybridization is shown by the following experiments. Firstly, formation of a mixed lipis phase starting from two membranes having a different hydrocarbon chain composition in their phospholipids. Secondly, formation of a hybrid membrane having an intermediate lipid to protein ratio from two membrane fractions having different lipid to protein ratios. Thirdly, reconstitution of NADH oxidase activity by hybridization of two membrane fractions, one lacking active cytochromes and the other being deficient in quinones. It is proposed that hybridization occurs by fusion of vesicles during the tight association of collapsed vesicles under high centrifugal forces. This interpretation is supported by electron microscopy of the membrane pellets after centrifugation. However, lipid transfer as the mechanism of hybridization cannot be excluded and attempts to reconstitute active galactoside transport by complementation of beta-galactoside transport-deficient membranes and cytochrome-deficient membranes have been unsuccessful.


Factors affecting the release of folded chromosomes from Escherichia coli.

1. The envelope-bound folded chromosome prepared by gentle lysis of Escherichia coli K 12 DG 75 cells in the presence of 1.0 M sodium chloride was found to be a non-specific complex of DNA strands entangled in vesicle-like envelope structures. This envelope remnant is still more or less rod-shaped. 2. Both the envelope-bound and the envelope-free folded chromosomes arise directly from intact cells. Even under extreme conditions of lysis, envelope-bound chromosomes cannot be converted into envelope-free nucleoids.


The study of DNA-RNA-polymerase complexes by kinetic formaldehyde method.

A modification of the kinetic formaldehyde method has been proposed providing a possibility for locally denatured regions (defects) formed in DNA preincubated with RNA polymerase (in the absence of nucleoside triphosphates) to be detected. This modification consists in a previous fixation of DNA-enzyme complex with small concentrations of formaldehyde, which do not induce formation of defects in DNA alone. The method has been calibrated under the conditions favourable to RNA synthesis. Studies of the effect of the fixation conditions on the number of defects in DNA interacting with RNA polymerase have shown that the number of defects is constant with formaldehyde fixation concentration between 0.05% and 0.3-0.5% and with fixation time between 2 min and 100 min. The dependence of the number of defects in DNA on RNA polymerase concentration at low ionic strength (0.05 M KCl) is presented by a curve with a plateau. From the initial linear part of the curve it has been found that the enzyme bound to DNA as a monomer. At the excess of the enzyme the mean number of nucleotide pairs between defects is 400-500. Increase of ionic strength results in decrease of the number of defects in DNA. The number of defects depends on temperature of preincubation of the complex. There were no defects in DNA at temperatures below 20 degrees C. At temperatures above 30 degrees C the number of defects reaches saturation. A sharp transition occurs in the range of temperatures between 20 degrees C and 30 degrees C. Analysis of the experimental and literature data, concerning the interaction of formaldehyde and amino acid methylol derivatives with DNA bases, leads to the conclusion that the mechanism of the formation of defects in helical DNA most likely consists in its unwinding or sharp weakening upon binding of RNA polymerase, prior to addition of formaldehyde.


Dryander of Marburg's woodcut (the oldest transposition?)

A sixteenth century woodcut depicting atypical congenitally corrected transposition is described. The possibility that the artist saw an example of the condition is considered in the light of Galen's theory of the circulation.


The morphogenesis of transposition of the great arteries, 1875-1975.

A century of embryological theory with respect to the morphogenesis of transposition is reviewed. The differing opinions are discussed in the light of experimental study, and a suggestion for further experimentation is outlined.


Recovery from aplastic anemia following attempted marrow transplantation.

A 23-year-old man with severe idiopathic aplastic anemia was prepared for marrow transplantation by the administration of cyclophosphamide (CY) 50 mg/kg on each of 4 days. He then received an intravenous infusion of 9.5 x 10(9) marrow cells from an HL-A matched and mixed leukocyte culture non-reactive sister. The graft was successfully established as shown by cytogenetic studies but was rejected after approximately 4 weeks. In preparation for a second transplant he was given procarbazine 12.5 mg/kg and goat antihuman thymocyte globulin (ATG) 7 mg/kg administered on alternate days for a total of 4 doses of each agent. At the end of this therapy his white blood cell count was noted to be going up and the second transplant was not carried out. Complete hematologic recovery of host type marrow ensued and persists now 20 months later. The various pathophysiologic mechanisms that may be involved are discussed.


Animal models of diabetes and obesity, including the PBB/Ld mouse.

Diabetes mellitus occurs in many animals species. However, only a few have been utilized in systematic studies designed to answer unsolved problems associated with the disorder in man such as molecular basis, pathogenesis of the vascular and neural lesions, and the roles of diet, exercise and obesity. Among the animal models available, rodents have been studied most thoroughly for a number of reasons: a) short generation time (sexually mature at about 3 mo of age, gestation time 21 days) and life-span is approximately 3 yr; b) hyperglycemia and/or obesity is known to be inherited in several species; c) environmental factors can be controlled easily in the laboratory because of small size; and d) economic considerations. The better-known rodent diabetes/obesity syndromes may be categorized as follows: 1) hyperglycemic with ketoacidosis, nonobese (Chinese hamster, South African hamster); 2) hyperglycemic with insulin hypersecretion, moderate obesity and may develop ketoacidosis (diabetic mouse (db/db), spiny mouse, sand rat); and 3) less pronounced hyperglycemia with hyperinsulinemia, insulin "resistance" and marked obesity (obese (ob/ob), yellow (Ay) and New Zealand obese (NZO) mice, and the Zucker "fatty" rat). The PBB/Ld mouse, described here in detail for the first time, is a new strain of mouse that also fits into the latter category. Members of this strain following maturity develop an obesity that is characterized by increasing cellularity of adipose tissue, increased serum immunoreactive insulin, reduced glucose tolerance, fatty liver, and hyperlipidemia. Therefore, this strain of mouse represents another model for study of adult onset obesity.


Maintenance of a constant brain extracellular potassium.

The development of potassium specific ion exchanger microelectrodes has enabled investigators to measure directly brain extracellular potassium ion activity. Although serum potassium in various species ranges between 3.5 and 6 mEq/l, brain extracellular potassium is maintained at a level close to 3 mEq/l independent of fluctuations in serum values. Despite this buffering of the internal brain environment by extracerebral changes, local variations in extracellular potassium occur in response to evoked neuronal activity, seizures, and spreading depression. Mechanisms involved in the maintenance of this ionic homeostasis in the brain include mediated transport at the level of the cerebral capillary and the choroid plexus epithelium. In addition, there are ouabain-sensitive clearance mechanisms presumably involving Na,K-ATPase that participate in the removal of excess potassium. The relative roles of simple diffusion, high glial cell conductance of potassium, and active ionic pumps in restoring basal potassium levels after activity are still controversial.


Change of potassium activity associated with membrane current flow.

Potassium-sensitive liquid ion exchanger microelectrodes are capable of recording fast concentration changes. Under suitable conditions, changes in extracellular potassium can be quantitatively measured during spike activity or voltage clamp pulses of exposed neurons. Comparison between transferred charge during outward current and the amount of extra potassium outside the cell reveals a deficit in net outward current, which is brought about by a long-lasting inward current. This inward current is facilitated by preceding depolarization and its appearance adds to a true inactivation of the potassium system. This is an example of how ion exchanger microelectrodes can be used to differentiate between time ionic fluxes underlying membrane currents. They are also useful in examining relations between neuronal activity and extracellular changes in potassium ion concentration in the central nervous system. Due to restricted diffusion in its extracellular space transient changes in potassium activity remain quite localized for considerable time. This makes it possible to identify discrete fields of activated neurons and to compare changes of neuronal excitability with variations of extracellular potassium activity.


Events at the cardiac sarcolemma: localization and movement of contractile-dependent calcium.

At present there is a controversy about the appropriate model for excitation-contraction (E-C) coupling in mammalian heart muscle. There is an apparent conflict between the data derived from voltage clamp experiments and data derived from isotopic exchange studies. The maximum transsarcolemmal Ca influx as derived from voltage clamp studies is not more than 10 mumoles/kg wet weight heart per beat. This is barely enough to achieve 10% activation of contractile force. It has been proposed, therefore, that the remainder of the required Ca is derived from intracellular sites. However, ionic exchange studies, using specific E-C uncouplers such as lanthanum, indicate that the major source of coupling Ca is external to the sarcolemma. Its movement, during E-C coupling, would then have to be nonelectrogenic since influx of this quantity of Ca is not discerned with voltage clamp experiments. It is proposed that on excitation, Ca moves across the cardiac sarcolemma by two routes: 1) a small amount through a pore or channel and 2) a much larger amount via an electroneutral carrier. Movement through the pore would be electrogenic and therefore measurable with the voltage clamp whereas movement via the carrier would not be evident with the clamp. The model is consistent with the proposal that the immediate source of contractile-dependent Ca in mammalian heart is derived from sites external to the sarcolemmal (and T tubular) membrane.


Excitation-contraction coupling--cardiac muscle events in the myofilament.

The interaction of myosin and actin is by intracellular Ca2+ concentration, which in turn is controlled by the sarcoplasmic reticulum. In muscle--including cardiac muscle--of vertebrates, and some invertebrates, the site of Ca2+ control is in the thin, actin-containing filaments. These filaments contain tropomyosin and troponin; the latter is a complex of three subunits. When Ca2+ combines with troponin C, the Ca-binding subunit, a shift occurs in the position of tropomyosin that makes it possible for the myosin heads to bind to actin. This process is inhibited by a conformational change in troponin C, resulting in the release of the troponin complex from one of the binding sites on the thin filament. This process exhibits cooperative aspects which have been analyzed in terms of the Ca-binding process and the effect of Ca2+ on actomyosin ATPase activity.


Ultrastructural aspects of activation and contraction of vascular smooth muscle.

Ultrastructural studies of potential intracellular calcium storage sites and of the organization of contractile proteins in vascular smooth muscle are reviewed. The sarcoplasmic reticulum (SR) is a system of closed tubules present in every smooth muscle examined. The volume of the SR varies in different smooth muscles (from approximately 2.0 to 7.5% of cytoplasmic volume) and correlates with the ability of a given smooth muscle to contract in calcium-free media. The SR accumulates the divalent cation strontium and forms couplings with the surface membrane. Mitochondria are other potential sites of calcium accumulation in vascular smooth muscle, as indicated by the respiration supported accumulation of calcium (with a Km of approximately 17 muM) by isolated mitochondria and by the energy dependent accumulation of barium by mitochondria in situ. The presence of barium in mitochondrial granules in tissues contracted with barium and the presence of strontium and calcium in appropriately incubated preparations have been verified with electron probe microanalysis. Calcium has also been demonstrated in mitochondrial granules in frozen dried cardiac muscle sections and in cultured vascular smooth muscle cells. This technique appears suitable for eventual quantitation of mitochondrial calcium content in vascular and other smooth muscles. Thin (actin, 50-80 A), thick (myosin, approximately 155 A), and intermediate (approximately 100 A) filaments are present in suitably fixed vascular smooth muscle. In rabbit portal anterior mesenteric vein intermediate high voltage stereo electron microscopy shows the myosin filaments to be tapered and approximately 2.2 mum long: significantly longer than the myosin filaments in vertebrate striated muscle. Actin filaments insert on dense bodies. The ultrastructural findings are compatible with physiological evidence suggesting the contributions of intracellular organelles to the regulation of cytoplasmic free calcium levels and the operation of a sliding filament mechanism of contraction in vertebrate smooth muscle.