[Correlation between beta-glucuronidase activity and serotype in the genus "Salmonella" (author's transl)]. Out of 4,114 cultures of Salmonella belonging to various serotypes, 1,241 (30,1%) hydrolyze p-nitrophenyl-beta-D-glucuronide, the chromogenic substrate of beta-glucuronidase. Serotypes are found to be homogenous with respect to beta-glucuronidase activity. The qualitative test, as simple as the ONPG test, may be of diagnostic value to assign a strain to a serotype in case of a monophasic or not motile variant of a normally diphasic serotype. Phage conversion does not influence beta-glucuronidase activity.


[Chlamydosporulation in "Candida albicans": influence of sowing density and glucose, nitrogen, biotin and mineral salts concentration in rice-cream decoction (author's transl)]. Chlamydospore production by Candida albicans was investigated in rice cream decoction inoculated with a blastospore suspension. Effects of glucose, KH2PO4, (NH4)2SO4 and biotin addition were estimated using a "chlamydospore index" i.e. the mean-number of mature chlamydospores produced from a single blastospore development. If no other component was added, it appeared that better sporulation was obtained with both glucose concentration and inoculum size optimum (0.5 g/l and 7.8 X 10(3) blastospores/ml respectively). In glucose-KH2PO4 enriched medium, chlamydosporulation was the best with 0.5 g/l glucose and 0.5 g/l KH2PO4. The addition of biotin reduced the chlamydospore index at the concentrations tested (0.6 to 20 microgram/l). When glucose and (NH4)2SO4 were added together, it seemed that one of the C or N component had to be in low concentration for a good sporulation. The action of salts was also investigated; and finally a synthetic medium is proposed. The improvement of this medium is now in progress.


Acute laryngeal obstruction in children. A fifty-year review. When dealing with acute laryngeal obstruction, the first important consideration is the differential diagnosis of the cause. The author considers the following six types: acute laryngotracheobronchitis, acute epiglottitis, diphtheria, supraglottic allergic edema (angioedema), subglottic allergic edema (spasmodic croup) and foreign body in the larynx or trachea. He traces the development of the treatments that have in 50 years lowered the mortality rate from 70% to practically zero. High humidity, best supplied by an ultrasonic nebulizer, antibiotics, and corticosteroids in very high dose have been the prime effective measures. There still is controversy about the choice between tracheostomy and nasotracheal intubation if medical therapy is delayed or ineffective. The primary physician must know when and where to send these children in order to prevent the fatalities so frequent in previous years.


Surgical treatment of acute blunt laryngeal injuries. The best management of acute laryngeal fractures is an approach which promotes healing by primary intention and does not require the use of a lumen keeper or core mold. This presenentation outlines the theoretical considerations and practical steps which make it possible to achieve these goals. The basic emphasis is on proper diagnosis, good surgical exposure and proper wound management. Also included are some ideas about the special anatomical and physiological characteristics of the larynx which make it difficult to obtain primary healing. The material presented is a synthesis of pertinent basic laryngologic research, wound healing theory and clinical experience.


Urinary excretion of lysozyme and N-acetyl-beta-D-glucosaminidase in the diagnosis of renal allograft rejection. The urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG) has been claimed to be of high diagnostic value in indicating acute renal graft rejection. Similar claims have been made for the significance of urinary lysozyme excretion. Serial measurements of urinary lysozyme and NAG have been made in 35 patients after renal transplantation, and during 52 suspected rejection episodes. A close correlation has been found between these two parameters although they did not consistently change in parallel. The use of both assays may give a better indication of impending rejection than either assay alone.


Extrahypophyseal effects of somatostatin. The tetradecapeptide somatostatin is widely distributed in the body in endocrine-like calls and in nervous tissues. When produced in endocrine cells, it seems to inhibit the secretion of hormones at the site of production, for example, it inhibits the secretion of growth hormone in the pituitary gland, insulin and glucagon in the pancreas, and the local hormones in the gastrointestinal tract. Its role in the nervous system is not clear, but it may act as an inhibitory transmitter substance or modulator. As for its mode of action, the data suggest that interference with the levels of cyclic nucleotides, within the cells as well as with Ca++ fluxes may be important. The clinical uses of somatostatin have been explored, especially in diseases characterized by hypersecretion of hormones known to be inhibited by somatostatin. But the wide distribution and multipotent inhibitory action calls for attention. Analogues to somatostatin with specific actions have already been produced and await clinical evaluation.


[12 cases of polycystic ovary type 1 (Stein-Leventhal syndrome): complete hormonal study (author's transl)]. Twelve patients with clinic and anatomic features of polycystic ovary syndrome type 1 were investigated for gonadotropic and androgenic functions. Basal LH level was 3 times higher than in normal women in the beginning of follicular phase (16,2 +/- 3,0 mUI/ml). After LH-RH stimulation (100 microgram), peak LH level was excessively high (70 +/- 10 mUI/ml) whereas FSH was normal. Clomiphen citrate (100 mg X 5 days) was followed in all cases by temperature ascension and biologic evidence of luteinisation. Four pregnancies were obtained. Plasma androstenedione was elevated in most cases (320 +/- 30 ng/ml) with elevated urinary androstanediol (80 +/- 20 microgram/24 h.) These elevated levels were strikingly reduced in all cases after ethinyl-estradiol, 50 microgram X 20 days. Physiopathologic hypothesis consistent with these results are discussed.


Postganglionic cholinergic dysautonomia. A 9-year-old boy presented with symptoms and signs of marked postganglionic cholinergic autonomic dysfunction manifested by bilateral internal ophthalmoplegia, impaired secretion of tears and saliva, lack of gastrointestinal motility, atony of the bladder, generalized absence of sweating, and hypertension. Clinical and pharmacological studies confirmed that the abnormalities were restricted mainly to the postganglionic cholinergic autonomic system and showed evidence of postdenervation supersensitivity to parasympathomimetic drugs. The patient was treated in the early phase of his illness by the administration of carbachol, and eventually he made a slow and partial spontaneous recovery. Histoimmunofluorescent studies on a skin biopsy specimen suggested an autoimmune origin for his disease, with IgG antibodies being produced against postganglionic cholinergic autonomic fibers.


Ultrastructural lesions of muscle and immunofluorescent deposits in vessels in Reye's syndrome: a preliminary report of serial muscle biopsies. Thirteen sequential percutaneous skeletal muscle biopsy specimens from 4 patients with Reye's syndrome were studied ultrastructurally and by direct immunofluorescence. Prominent generalized intermyofibrillar edema, mitochondrial disruption, and swelling of vascular endothelium were demonstrated in 2 patients. In all 4 patients there were granular deposits of immunoglobulins G and M in intramuscular vessels, presumably representing antigen-antibody complexes. We postulate that these changes may be pathogenetically important in this syndrome and suggest that muscle biopsy tissue be utilized for further investigation of Reye's syndrome.


Importance of Holter monitoring in patients with periodic cerebral symptoms. Holter monitoring was used in 358 patients with symptoms of intermittent dizziness, blackout, or both. The neurological findings in all patients were normal. None manifested evidence of a pertinent arrhythmia on the one-minute resting ECG; 8.9% of the patients demonstrated arrhythmias known to correlate with cerebral symptoms; and 11.2% demonstrated "predisposing" arrhythmias, ie. rhythms that in and of themselves may not precipitate symptoms but predispose to arrhythmias that do. The latter category included short bursts of ectopic tachycardia and intermittent short-duration asystole. High-frequency ectopic beats were observed in 24.6%. In almost every patient in whom intermittent arrhythmias precipitated symptoms there were many "predisposing" arrhythmias of short duration that occurred during the asymptomatic period. A comparison of 12- and 24-hour recordings demonstrated an increase in pertinent arrhythmias from 13.8 to 22.7%.


Cysticercosis of the nervous system: diagnosis by means of specific radioimmunoscan. A new method for diagnosing cerebral cysticercosis by means of specific radioimmunoscan is described. The method has been evaluated in 24 patients with encouraging results. The technique involves a single intravenous injection followed by brain scanning and thus far has been entirely nontoxic. The accuracy of the scan was confirmed in 6 patients by surgical removal of lesions that coincided in location and number with the diagnostic scan of the parasites.


[Pharmacokinetic characteristics and the effectiveness of using carbenicillin on newborn premature infants with infectious and inflammatory diseases]. The results of the study on the pharmacokinetics peculiar properties of carbenicillin premature infants treated with the drug administered intravenously or intramuscularly are presented. The maximum antibiotic blood levels after intravenous administration exceeded the MIC for most of the causative agents isolated from the children. The schemes for the antibiotic use in treatment of purulent septic processes of children are recommended.


[Effect of lipid preparations isolated from protozoa on experimental leishmaniasis in golden hamsters]. The effect of lipid preparations Nos. 59 and 60 isolated from Protozoa on experimental leishmaniosis of golden hamsters was studied. Preparations Nos. 59 and 60 are water-soluble derivatives of fatty acids isolated from Trypanosoma lewisi and Astasia longa respectively. It was found that both preparations administered to the animals simultaneously with the causative agent or after formation of the leishmaniosis infiltrates inhibited the development of the disease clinical signs. Such an effect of the preparations was observed only during the period of the animals exposure to them. After discontinuation of the treatment the development of the clinical process proceeded the same as in the control animals. The lipid preparation No. 60 differing from preparation No. 59 by high content of C20-poly-unsaturated fatty acids was more active and induced a significant decrease in the size of the leishmaniosis infiltrates formed. Preparation No. 60 was more effective in the treatment of the animals infected with L. tropica major than in the animals infected with L. tropica minor. Preparation No. 59 had no significant effect on multiplication and morphology of the parazites.


[Experimental study of the antimicrobial effect of lysozyme in combination with antibiotics]. A synergistic effect of the antibacterial action of the lysozyme combination with various antibiotics against gram-positive and gram-negative microorganisms was found. The intensifying effect of lysozyme on its combination with antibiotics was more evident with respect to gram-positive microbes and less evident with respect to gram-negative organisms. The combinations os lysozyme with methicillin, lincomycin, oleandomycin or erythromycin proved to be most active against staphylococci. A high synergistic effect against streptococci was observed with the use of the lysozyme combination with benzylpenicillin, morphocycline, oleandomycin, ceporin or monomycin. The combination of lysozyme with macrolides had a maximum inhibitory effect on Coli bacteria.


Pharmacokinetics of amoxicillin and ampicillin: crossover study of the effect of food. The effect of food on the absorption of amoxicillin and ampicillin was studied in 16 normal subjects in a double-blind crossover study after each subject was given a single oral 500-mg dose. Serum drug levels were analyzed, assuming a one-compartment linear model with first-order absorption and absorption delay, area under the curve, and urinary recovery. Variations in kinetic parameters were examined by using analysis of variance. The results showed little or no effect of fasting versus nonfasting on amoxicillin absorption, as evidenced by peak serum levels (8.9 mug/ml, fasting, 8.8 mug/ml, nonfasting), area under the curve (26.9 mug/ml per 70 kg, fasting, 22.2 mug/ml per 70 kg, nonfasting), and urinary recovery (47%, fasting; 44%, nonfasting). Ampicillin absorption was significantly decreased in the nonfasted group by the same parameters (peak level: 5.4 mug/ml, fasting, 4.0 mug/ml, nonfasting; area under the curve, 17.4 h.mug/ml, fasting, 12.0 h.mug/ml, nonfasting; urinary recovery, 37%, fasting, 29%, nonfasting). These results confirm the reliable absorption of orally administered amoxicillin in the fed as well as the fasted state.


Comparison of netilmicin and amikacin in treatment of complicated urinary tract infections. Netilmicin and amikacin, two recently developed aminoglycosides, were compared in a prospective, randomized study of 57 male patients with complicated urinary tract infections. Both drugs were administered intramuscularly every 12 h for 7 to 10 days, netilmicin at 2 mg/kg and amikacin at 7.5 mg/kg. The two groups were comparable as to infecting bacteria and underlying pathology of the urinary tract. No patients had indwelling catheters. All microorganisms isolated were sensitive to both antibiotics. A total of 69% of the patients treated with netilmicin and 57% of the patients treated with amikacin were cured of the infection, as defined by a negative culture at 7 days after discontinuation of treatment. No major side effects were recorded, and no significant changes were noted in parameters of renal function. Of the patients treated with amikacin, 21% experienced temporary local pain at the injection site; no such effect was noted in the netilmicin-treated group. Therefore, netilmicin appeared to be as effective and better tolerated than did amikacin in the treatment of complicated urinary tract infection.


Susceptibility of Eikenella corrodens to ten cephalosporins. The susceptibility of 24 strains of Eikenella corrodens was determined, by the agar dilution technique, to 10 cephalosporins, as well as to clindamycin, penicillin and dicloxacillin. All strains were uniformly very susceptible to penicillin G and cefoxitin and resistant to clindamycin and dicloxacillin. Cefazolin showed good activity. Cephalexin, cephradine, and cefadroxil showed poor activity, and cefamandole's activity was relatively poor. Cephalothin, cephapirin, cefaclor, and cephaloridine showed moderate activity, with some strains relatively resistant.


Photosensitivity due to a fluorouracil derivative. A photosensitive lichen planus-like eruption developed in a 63-year-old man during postoperative chemotherapy with 1-(2-tetrahydrofuryl)-5-fluorouracil. The action spectrum for the photosensitivity was in the long-wave ultraviolet light (UV-A). The reaction was reproducible on readministration of the drug and exposure to UV-A. At the time of this test, a flare-up phenomenon was observed at previously involved sites. The photopatch test was negative with the drug. Patients taking the drug who had no dermatitis were not sensitive to UV-A. These observations suggest that the photosensitivity that occurred in the patient was possibly photoallergic.


Bullous pemphigoid:therapy in patients with and without diabetes mellitus. Retrospective analysis of 34 patients with bullous pemphigoid (BP) showed 24 (70.6%) with widespread disease and 10 (29.4%) with localized disease. Initial prednisone therapy was significantly lower for patients with localized BP, mean dose of 38 mg vs 58 mg, compared with widespread disease. Fifteen of 24 (62%) patients responded; however, 9 (38%) failed to respond to moderate-dose prednisone therapy and ultimately required daily doses of 100 mg or more and/or an immunosuppressive agent. Adult-onset diabetes mellitus was present in 14 of 34 (41%) patients. Initial prednisone dose for diabetic patients with widespread BP was significantly greater, 70 mg vs 54 mg, compared with nondiabetic patients. Addition of methotrexate, 2.5 to 15 mg, every other day or twice-weekly oral administration, was effective when moderate-dose prednisone therapy failed. It is possible that insulin-dependent diabetic patients with BP should be preferentially started on immunosuppressive drugs.


Coexistent psoriasis and bullous pemphigoid: a report of six cases. Vesicles and bullae developed in six patients with psoriasis while they were undergoing therapy for their psoriasis. Skin biopsy specimens were taken from their lesions and studied for regular histopathologic findings and direct immunofluorescence. These studies confirmed that they had coexistent psoriasis and bullous pemphigoid. A review of the therapy that the patients were given incriminates anthralin, crude coal tar, ultraviolet light, and salicylic acid in the possible cause of the development of their bullous pemphigoid lesions.


Neonatal tetanus treated with high dosage diazepam. The combination of continuous intravenous infusion of diazepam (20--40 mg/kg per day) and intragastric phenobarbitone (10--15 mg/kg per day in 4 divided doses) was used to treat 19 cases of neonatal tetanus. Mortality was 2/19 (11%). This regimen was considered to have reduced the mortality and the need for artificial ventilation. The main side effects encountered were severe drowsiness, coma, and apnoeic episodes which were reversible when the level of diazepam was reduced.


Impairment of chemotactic activity of microphages in chronic mucocutaneous candidosis. The chemotactic microphage function in combating and eliminating microorganisms is one of the most important features of the cellular immune system. Using a modified Boyden method we studied in vitro the chemotactic activity of the granulocytes in 5 patients suffering from chronic muco-cutaneous candidosis (CMCC), 3 of the familial and 2 of the non-familial type. To obtain quantitative results comparable to those of a control group, we investigated the granulocytes of 55 clinically and immunologically healthy persons by the same way. So we found, in comparison with the results of the controls, a striking reduction of the chemotactic granulocyte activity in all CMCC patients. This impairment of chemotaxis is apparently due to an inborn defect of the microphages. It is pointed out, however, that a non-specific stimulation of the immune system might be a promising way in the treatment of patients affected by CMCC.


Allotransplantation of cultured human skin. Human skin allografts cultured for approximately 6 weeks on a solid medium were transplanted to non-related human recipients with skin defects of various etiology. Although based on this study no answer could be given to the question whether these grafts survived or were replaced by cells of the recipient, it was observed that in most cases the grafts were not rejected and remained in situ several months up to over a year.


Are Langhans giant cells precursors of foreign-body giant cells? Granulomas were induced in rats by subcutaneous implantation of pieces of Melinex plastic into the dorsum. The pieces of Melinex were removed at intervals varying from 16h to 14 days, and the adherent cells were studied morphologically and quantitatively. Giant cell formation started about 32h after implantation. The first giant cells to appear were of the Langerhans type. Two days after implantation, most of the giant cells are still Langhans-type cells. A few giant cells of the foreign-body type and transition forms between the Langhans and foreign-body types are also present. From the third day on, the foreign-body type gradually becomes predominant. Independent of the duration of implantation, giant cells with 3, 4, or 5 nuclei are virtually without exception of the Langhans type. The higher the number of nuclei between 6 and 30, the more cells are of the foreign-body type. Giant cells with 30 or more nuclei are all foreign-body type. The findings are discussed in the light of current knowledge concerning giant cell formation. It is concluded that under the present experimental conditions, Langhans-type giant cells are the precursors of foreign-body-type giant cells.


Effects of 8-methoxypsoralen plus 365 nm UVA light on Candida albicans cells. An electron microscopic study. Candida (C) albicans cells were exposed to 8-Methoxypsoralen (8-MOP) concentrations of 1.0 microgram/ml and 10.0 microgram/ml medium and irradiated with 365 nm light. The amount of energy emitted was 4.8 J/cm2. Two divergent types of cell damage occured concerning yeast cell cytoplasm and cell wall. Two hours after exposure cytoplasmic changes involving mitochondria, which showed irregularities in shape, blurred appearance or loss of mitochondrial cristae and outer membrane were seen. The number of vacuoles was increased. The cytoplasm showed large electron transparent areas, the cytoplasmic membrane disappeared in some areas completely. Nucleus and nuclear envelope usually remained intact in early stages. 24h after exposure conspicuous cell wall alterations were observed in addition to cytoplasmic changes. Newly produced cell wall material formed ball-like protrusions or was adherent sickle-shaped to the cell wall. The investigations strongly suggest that the results found after 8-MOP-UVA treatment of C. albicans cells can not be interpreted in the sense of a general cytotoxic effect. Apparently it takes the form of a combination of events involving regressive and progressive alterations.


[Physical forces in blister formation. II. Examination of total osmolality in blister fluid of suction blisters, "naturally" developed blisters and in serum (author's transl)]. Total osmotic pressure does not differ significantly in suction blister fluid or in serum of healthy persons from blister fluid, suction blister fluid or serum of patients with dermatitis herpetiformis, bullous contact dermatitis and pemphigus vulgaris. The average values reach about 293 mOsmol/kg. This results from direct measurement of total osmolality with an electronic semimicroosmometer based on the principle of freezing point reduction. In opposite to colloid osmotic pressure the total osmotic pressure does not take part in blister formation of human skin in spite of the different concentrations of the single electrolytes in the fluids of various blisters.


In vitro studies on microphage functions in chronic pyoderma vegetans. The function of microphages has been studied in two patients with chronic pyoderma vegetans by in vitro determination of phagocytosis as well as chemotaxis. The results showed a striking decrease in the chemotactic activity of the neutrophil granulocytes, a reduced phagocytosis of Candida albicans and Staphylococcus aureus, and a weakness of the intracellular killing of these microorganisms. However, the NADH-dependent oxidase activity appeared to be intact. No defect was found in the specific cellular or humoral immune system in either patient.


A comparative study of Butacote and Naprosyn in ankylosing spondylitis. A double-blind, cross-over comparison of Naprosyn (naproxen) 750 mg daily and Butacote (enteric-coated phenylbutazone) 300 mg daily was carried out in a multi-centre trial. Twenty-five patients, mostly male and under 40 years of age, were entered. After a 2-week period in which any existing anti-inflammatory drugs were tailed off, patients were entered into the trial and treated for 1 month with each drug. Patients were assessed at 4-weekly intervals. Both drugs significantly reduced morning stiffness. Morning pain and discomfort and wall-tragus distance were also significantly reduced by both drugs during the trial. Results of the Schober test showed improvement during both treatment periods. There were no overall statistically significant differences between the effects of the 2 drugs on objective parameters. However, overall subjective assessment of symptoms showed a greater improvement on Butacote. Treatment preferences by physician and subjective assessment by the patient both favoured Butacote but the difference between the 2 drugs was not statistically significant. Side effects were mostly of a minor nature. One patient had to discontinue the trial, due to indigestion while taking Butacote.


Fungal infections in renal transplant recipients. Infection continues to be a major source of morbidity and the major source of mortality in renal transplant recipients who are susceptible to opportunistic infections. We recently reviewed all renal transplant recipients who had fungi cultured during a three year period. C. albicans and T. glabrata were cultured most frequently. Deep fungal infections occurred in many patients and were frequently observed late in the course of bacterial and viral infections. Ten patients had fungemia, and primary fungal pneumonia occurred in eight patients. Three patients had fungal infection of the central nervous system. Three of eight patients with fungal pneumonia and eight of ten patients with fungemia died as a result of their fungus infections. These patients frequently had poor renal function and were receiving high steroid doses or had recently been treated for kidney rejection. One patient with fungal pneumonia and six patients with fungemia had the fungus cultured from a superficial site. Several patients developed fungal infections late in the course of viral or bacterial infections. Amphotericin-B and 5-fluorocytosine remain the mainstays of antifungal therapy.


William Halsted and Theodor Kocher: "an exquisite friendship". William Halsted and Theodor Kocher were friends on both a professional and personal level. They greatly admired each other's surgical skills and judgment, and Halsted was a frequent visitor to Kocher in Switzerland. This is of interest in that both men have been described as introverted personalities with very few intimate acquaintances. An analysis of their personal correspondence, which has not been previously published, clearly demonstrates this unique relationship between the two reknowned surgeons.


Combination chemotherapy followed by skin grafts in the management of locally advanced breast cancer. A combined modality approach to the treatment of locally advanced, ulcerating carcinoma of the breast is discussed. The beneficial effects of combination chemotherapy have been amplified by split thickness skin grafts at the site of a large skin deficit, resulting in successful wound palliation. Criteria for assessing likelihood of graft acceptance by an ulcerating wound in the presence of local neoplastic disease are emphasized.


A model for planning health care in patients with end-stage renal disease. Treatment modalities of end-stage renal disease (ESRD) patients include in-center dialysis, home dialysis, and kidney transplant. We present a model to account for all aspects of modality use to aid in planning regional facilities. Five years of data for 979 patients on dialysis in Wisconsin between 1970 and 1975 are used. The model shows movement (transit probabilities) from one modality to another, eg, transition from in-center dialysis to transplantation, and data were used to derive all transitional probabilities characterizing patient movement from one modality to another. Model and probabilities were used to predict number of patients in each modality in 1976, and the model was used to predict number of patients in each treatment modality through 1990. These figures may be used for planning regional facilities. Extrapolation of this model and derived probabilities for nationwide projections may be possible.


Home hemodialysis review in Iowa: 1970--1977. One hundred fifty-two patients were accepted for home hemodialysis training from 1970 to 1976. Duration of training varied from 4 to 22 weeks. The primary reason for discontinuing home dialysis was patient and assistant psychological adjustment problems. Patient medical and technical problems were not related to the distance from the home dialysis training unit. Cumulative probability of survival was .948 at one year and .637 at five years. Cardiovascular disease accounted for 80% of the patients who died while receiving long-term hemodialysis. Fifty-three percent of the deaths in patients after renal transplantation were secondary to infectious causes.


Pharyngeal flora in ambulatory alcoholic patients: prevalence of gram-negative bacilli. The pharyngeal flora of a group of ambulatory alcoholic patients was studied and compared with the pharyngeal flora of a control group. Sixty-eight patients were studied, 34 alcoholics and 28 controls. Of the alcoholic patients, 59% had Gram-negative bacilli in their pharyngeal flora, while 14% of the control group had the same organisms. There were no differences in Gram-positive cocci colonization between the groups. Klebsiella pneumoniae was the most frequent isolate (40%) and the Klebsiella-Enterobacter group accounted for 76% of the isolates. Colonization rates of greater than 10 colony forming units/ml were found in 43% of the alcoholic patients. The high prevalence and higher colonization rates of Gram-negative bacilli in alcoholic patients might explain the higher incidence of Gram-negative bacillary pneumonia among alcoholics.


Minoxidil-induced bullous eruption.

Minoxidil-induced bullous eruption. A previously unreported side effect--serosanguineous bullae--was found in one patient receiving minoxidil. The eruption disappeared with cessation of the drug and reappeared on rechallenge. The question arises whether continuation of minoxidil treatment could have led to vasculitis.


Detection of small cryptic plasmids in Salmonella typhimurium strain LT2. Small cryptic plasmids of molecular weights ranging from 1 to 3 Mdal were detected by electron microscopy in Salmonella typhimurium strain LT2 (ColIb). They were divided into different size classes. Two of the cryptic plasmids were transferred simultaneously with ColIb to Escherichia coli.


Structure and function of small airways in health and disease. During the last two decades, great strides have been made in our understanding of the functional aspects of airflow in the periphery of the lung. It seems that the small airways are the important site of obstruction in a variety of chronic respiratory disorders associated with airflow obstruction. This review deals with the anatomic and functional aspects of small airways in normal and diseased lungs. In particular, the basis of obstruction or narrowing that is not dependent on intrinsic airway lesion is reviewed. The variety of pathologic changes in small airways observed in these diseases are outlined along with physiologic tests that are currently used to detect dysfunction at a stage long before they produce symptoms or alter standard tests of lung function.


ICLH cemented douple cup hip replacement. The ICLH (Imperial College-London Hospital) double cup hip replacement as it exists today consists of a metal femoral component which caps the femur and a polyethylene acetabular component which resurfaces the acetabulum. Both components are cemented in place. The first prosthesis was inserted in 1972 and during the following two years complications emerged which necessitated changes in the prosthesis and the operative technique. Since November 1974 there have been no changes in the prosthetic design or operative procedure and complications have been infrequent. An approach has been developed whereby trochanteric osteotomy is not necessary. Up to December 1977 159 hips were replaced in 154 patients. Clinically, the results of a successful double cup arthroplasty are similar to those obtained by conventional hip replacement. No deterioration in the quality of the results is suggested when early results are compared with those of patients who are now two years and longer since surgery. Technical errors to be avoided are varus placement of the femoral component and excessive vertical placement of the acetabular component. Conclusions drawn are tentative due to the fact that to date the follow-up period is inadequate.


Temperatures in human thighs after hot pack treatment followed by ultrasound. Temperature distributions were measured in the anterior thigh of human volunteers after preheating with a hot pack followed by ultrasound, a frequently used therapeutic procedure. The skin surface temperature is elevated by this procedure but the highest temperature is still produced at the bone-muscle interface at the end of the treatment session.


Metrifonate. Summary of toxicological and pharmacological information available. The organophosphorus compound 0,0-dimethyl-(1-hydroxy-2,2,2-trichloroethyl)-phosphonate was introduced as an insecticide, trichlorfon, in 1952 (Lorenz et al., 1955) and as a drug, metrifonate, in the treatment of schistosomiasis in 1960 (Lebrun and Cerf, 1960). This organophosphorus compound is unique in that it has been claimed not to be a direct acting cholinesterase inhibitor but being transformed nonenzymatically into an active component dichlorvos, 2, 2-dichlorovinyl dimethyl phosphate (DDVP). The evidence for this transformation has mostly been indirect. Recently it has been proved chemically and quantitatively that this transformation occurs in the animal body (Nordgren et al., 1978). Metrifonate is the sole organophosphorus compound currently studied clinically in schistosomiasis. A substantial therapeutic effect is obtained only in Schistosoma haematobium infections. In this review on available data of metrifonate it is suggested that further more detailed studies of both S. haematobium and S. mansoni are necessary. This should include studies of the enzymic properties of the worms and the reaction of their esterases towards both metrifonate and DDVP as well as the pharmacokinetics of these compounds in man. In addition there are still unsolved discrepancies reported regarding organ toxicity of the compound which may, however, be due to different grades of parity of the test material.


[Mechanisms of development of histological and cytochemical changes in mammals (rats) during orbital flights]. The review surveys the publications on morphological and cytological changes in different functional systems of mammals (rats) flown on board the biological satellites of the Cosmos series. The paper gives an evidence that major changes in rats during orbital flights are induced by motor deprivation resulting in distinct metabolic disorders which histologically are represented as osteoporosis of the spongy segments of the long tubular skeletal bones, their periosteocytic osteolysis and atrophic developments in skeletal muscles. The paper discusses neurotrophic nature of these changes and emphasizes the contribution of stress reactions at different flight stages, especially on return to 1 g.


Precautions in familial transmissible dementia: including familial Alzheimer's disease. Recent studies suggest that some cases of familial Alzheimer's disease may be associated with a transmissible dementia. Animal experiments show that presymptomatic carriers of "slow virus" agents can transmit disease. Because of these findings, we have extended the precautions previously delineated to include those at risk of acquiring transmissible dementia, specifically, to the descendants of those affected with familial Alzheimer's disease or familial Creutzfeldt-Jakob's disease. Blood donation from such persons may pose a danger, because transmissible spongioform encephalopathy has been passed from animal to animal by blood serum and by the WBC layer of frozen whole blood.


Treatment of parkinsonism with N-n-propyl norapomorphine and levodopa (with or without carbidopa). The effects of the concomitant administration of N-n-propyl norapomorphine (NPA) and levodopa, with and without carbidopa, were studied in 12 patients with unsatisfactory symptom control. Double-blind evaluation of the effects of NPA with suboptimal doses of levodopa or levodopa plus carbidopa (Sinemet) showed a mean overall improvement of 44% (20% to 74%) in nine patients and improvement of the "on-off" effect in five. Dyskinesia diminished in some patients after diminution of basal medication. In three patients, plasma dopa and growth hormone patterns did not differ substantially with and without NPA. The magnitude and timing of the therapeutic and side effects did not correlate with the pattern of growth hormone secretion, which suggests that this hormone might not be instrumental in the induction of these effects. N-n-propyl norapomorphine is a useful adjunct in the long-term management of patients with unsatisfactory response to levodopa.


The mechanism of timolol in lowering intraocular pressure. In the normal eye. A single-drop, double-masked, randomized, placebo-controlled study of the mechanism of the ocular-pressure lowering property of timolol maleate, a beta-adrenergic blocker, was carried out in 23 normal subjects, using fluorophotometry. Timolol suppressed aqueous formation in all subjects. The range of suppression was 13% to 48%, with a mean +/- SD of 34% +/- 9%. The drug had no effect on anterior chamber volume or endothelial permeability to fluorescein and, apparently, had no effect on outflow resistance. No differences were observed between its effect on men and women or between eyes with light and dark irides.


The removal of scleral buckles.

The removal of scleral buckles. Six hundred consecutive retinal detachments were operated on with silicone scleral buckles. Twenty-three implants, or 3.8%, were removed. There was one solid silicone implant and 22 silicone sponges. Removal was required because of infection (seven eyes), foreign-body sensation without infection (seven eyes), recurrent subconjunctival hemorrhages (four eyes), impingement on the optic nerve (one eye), and distortion of the macula (four eyes). The postoperative infection rate was 1.8% for silicone sponges, and there were no infections in eyes buckled with solid silicone. Only one retina redetached after buckle removal, yielding a recurrence rate of 4%. The one recurrence was successfully managed by a second scleral buckle.


Transplant size and elevated intraocular pressure. Postkeratoplasty. Elevated intraocular pressure after keratoplasty is a well-recognized phenomenon both in aphakia and in combined lens extraction and penetrating keratoplasty. Ninety-two consecutive cases of penetrating keratoplasty procedures were studied. These were randomly assigned to group A or B. Group A received a donor transplant 0.5 mm larger than the recipient bed. Group B received donor buttons equal in size to the recipient bed. Intraocular pressure was measured preoperatively and daily until the patients were discharged. Group A, which had aphakic penetrating keratoplasty or the combined procedure (0.5-mm larger button), also had significantly lower intraocular pressures (P less than .001) than group B (same size button). There was no difference in postoperative intraocular pressure between groups A and B for those who had phakic penetrating keratoplasties. A larger donor size can alleviate induced "aphakic keratoplasty glaucoma."


Subepithelial infiltrates: a probable sign of corneal transplant rejection. A previously undescribed slit-lamp manifestation of a probable corneal transplant rejection reaction was found in 22 patients among 145 who underwent penetrating keratoplasty during a two-year period. The reaction consisted of subepithelial infiltrates that were located only in the donor tissue; were without associated conjunctivitis; and that occurred six weeks to 21 months postoperatively, either alone or in association with epithelial and/or endothelial rejection; and that responded well to topical corticosteroid treatment. In one case, the subepithelial infiltrates preceded a severe endothelial rejection by only a few days. The lesions are a warning that all is not well and that corticosteroid therapy should be instituted or increased.


Orbital mass as a complication of contact lens wear. A 26-year-old woman, who was a hard contact lens wearer, had a cystic mass in the right orbit that was increasing in size. Although initial ultrasound evaluation was negative for a foreign body, a history of trauma to the right orbit, two years prior to examination, and clinical findings suggested a retained migrated contact lens with a secondary orbital cyst. Surgical exploration revealed the contact lens embedded in the supratarsal space with an associated sterile, inflammatory intraorbital cyst.


A comparison of 4 serological tests in the detection of humoral antibodies to anaplasmosis in cattle. A capillary agglutination (CA), a complement fixation (CF), a plate agglutination (PT) and an indirect fluorescent antibody (IFA) test to detect humoral antibodies to Anaplasma marginale are described. Serums from 3, 4 or 5 groups of cattle were used to examine the efficiency of the tests. Agreement between all 4 tests was 86.6%. Agreement between pairs of tests was greater. The CF test was the most sensitive while the PT test was the least sensitive. However the PT could be carried out very rapidly and was suggested as the best screening test, providing improved antigen preparation techniques could increase sensitivity. The CA, CF and IFA tests all showed a stronger homologous antibody reaction when A. marginale antigen was tested against serums obtained from cattle infected with either A. marginale or A. centrale. Antibodies in the A. marginale serums were first detected by day 7 post-inoculation, rose to peak around day 29 and were still present on day 200. Antibodies in the A. centrale serums were first detected by day 29 rose to a peak around day 50 and had disappeared by day 150.


A morphological study of the endocrine pancreas in human pregnancy. During human pregnancy an enlargement of the islets of Langerhans and hyperplasia of the beta cells is present. These morphological changes indicate that the endocrine pancreas is able to adapt to the metabolic changes of pregnancy.


[Institutional epidemic gastroenteritis caused by Salmonella enteritis Worthington type in 1976]. An outbreak of gastroenteritis that occurred in a boarding school in Mexico is described. The attack rate was 100% since all of 151 pupils were involved. Salmonella enteritidis serotype Worthington was identified as the only etiologic agent. This microorganism was isolated from 60% of the samples studied. Due to the delay in notification the food ingested during the outbreak could not be analyzed.


Menopausal hot flushes: a double blind comparison of placebo, ethinyl oestradiol and norgestrel. A double blind crossover study was planned in order to compare the effects of oestrogen, progestogen and placebo on hot flushes. The 49 women studied had previously undergone hysterectomy and bilateral oophorectomy. The drug regimen consisted of three months each of ethinyl oestradiol 50 microgram/day, d norgestrel 250 microgram/day, a combination of these two substance ("Nordiol") and a placebo. The predominant reason for requesting a change of medication was intolerable hot flushes associated with placebo use. All hormonal preparations were found to be significantly more effective than placebo in reducing hot flush frequency and intensity. Oestrogen containing medications were more effective than the progestogen (norgestrel).


[Recurrent bacteriuria in children].

[Recurrent bacteriuria in children]. Fifty-four children admitted to the Children's Hospital of Cordoba City with recurrent bacteriuria were followed for 12 to 97 months. Most of them were females and Escherichia coli was the most frequent microorganism isolated in recurrent attacks of bacteriuria. Ninety percent of the patients showed urological abnormalities. Vesicoureteral reflux was the most frequent. Residual urine was considered the main cause of recurrent bacteriuria. After antibioticotherapy and surgical treatment, over one half of the patients persisted with relapses. Recurrent bacteriuria appeared more frequently in male infants with severe urologic abnormalities. Finally, three groups of patients in relation to uropathy and functional renal impairment are described and a different prognosis for each one is proposed. For this reason, each patient must have a complete urologic examination done at the onset and a carefull follow-up, carried out.


Binding of recrystallized and chromatographically purified 8-anilino-1-naphthalenesulfonate to Escherichia coli lac repressor. 8-Anilion-1-naphthalenesulfonate (Ans), recrystallized from water as the magnesium salt, contains a fluorescent impurity representing 0.3% of the absorbance at 351 nm. This impurity can be removed by Sephadex LH-20 chromatography. The chromatographic and spectral properties of this impurity suggest that it is bis(Ans), a dimer of Ans. This bis(Ans) impurity makes a significant contribution to the fluorescence increment observed when lac repressor is added to recrystallized Ans. This occurs because bis(Ans) binds much more tightly to this protein than does Ans. The dissociation constant divided by the number of binding sites per subunit is 3.1 X 10(-6) M for bis(Ans); the corresponding value for Ans is greater than 1 X 10(-4) M. Because of their differing absorption spectra, the impact of this bis(Ans) impurity is especially large with excitation wavelengths above 400 nm. Users of recrystallized Ans should consider the potential consequences of this impurity whenever working with a protein to which Ans binds weakly.


Association of Escherichia coli lac repressor with poly[d(A-T)] monitored with 8-anilino-1-napthalenesulfonate. The association of lac repressor with poly[d(A-T)] was monitored with the fluorescent prob 8-anilino-1-naphthalenesulfonate (Ans). Excess poly[d(A-T)] decreased the emission intensity of the repressor--Ans complex by 30%. Fluorescence titrations indicated that 33 +/- 4 base pairs were required to bind all of the repressor. Sedimentation studies indicated, however, that all of the repressor sedimented as a protein--DNA complex with as few as 10 to 15 base pairs per tetramer, even in the presence of Ans. These data are interpreted with two models: one where repressors bind to both sides of the DNA (Butler, A. P., et al. (1977) Biochemistry 16, 4757: Zingsheim, H.P., et al. (1977) J. Mol. Biol. 115, 565), the other where a double layer of repressors bind to a single side of the DNA. Removal of the amino-terminal regions from the repressor decreased the fluorescence from bound Ans by 77%. The amino-terminal fragments alone did not enhance Ans fluorescence.


Joining of 3'-modified oligonucleotides by T4 RNA ligase. Synthesis of a heptadecanucleotide corresponding to the bases 61--77 from Escherichia coli tRNAfMet. Chemically synthesized fragments corresponding to the 3' end of tRNAfMet from Escherichia coli were joined by T4-induced RNA ligase to yield a heptadecanucleotide (bases 61--77). The 3' terminus of C-C-A was modified by introduction of the ethoxymethylidene group to prevent intra- and intermolecular self-joining reactions at the 3' end. The terminal trimer was phosphorylated using polynucleotide kinase and joined to C-A-A with RNA ligase. The hexamer [C-A-A-C-C-A(ethoxymethylidene)] corresponding to bases 72--77 was obtained in a yield of 60%. An undecanucleotide (bases 61--71) which had been synthesized in a yield of 34% by similar enzymatic joining of U-C-C-G-G to pC-C-C-C-C-G was allowed to react with the 5'-phosphorylated hexamer (bases 72--77) using an excess of RNA ligase to yield the heptadecanucleotide U-C-C-G-G-C-C-C-C-C-G-C-A-A-C-C-A (bases 61--77). The product was identified by homochromatography and nearest neighbor analysis.


Isolation and identification of yeast messenger ribonucleic acids coding for enolase, glyceraldehyde-3-phosphate dehydrogenase, and phosphoglycerate kinase. Yeast poly(adenylic acid)-containing messenger ribonucleic acid isolated from two strains of Saccharomyces cerevisiae was fractionated by preparative polyacrylamide gel electrophoresis in the presence of formamide. Three messenger ribonucleic acids, present at high intracellular concentration, were electrophoretically eluted from the polyacrylamide gels and translated in a wheat germ cell-free extract. The in vitro synthesized polypeptides were identified by tryptic peptide analysis. Messenger ribonucleic acids coding for enolase and glyceraldehyde-3-phosphate dehydrogenase were isolated from commercially grown baker's yeast (strain F1), and messenger ribonucleic acid coding for phosphoglycerate kinase was isolated from Saccharomyces cerevisiae (ATCC 24657). Significant differences in the spectrum of abundant messenger ribonucleic acids isolated from commercially grown baker's yeast (strain F1) and strain 24657 were observed. When both strains were grown under identical conditions, however, the spectrum of messenger ribonucleic acid isolated from the cells is indistinguishable.


Release of a special fraction of the outer membrane from both growing and phage T4-infected Escherichia coli B. Growing Escherichia coli release envelope material into the medium. Upon infection with T4 phage increased amounts of this material are released and at a greater rate. In order to determine whether both inner and outer membranes are present in this material, and whether the material released by growing cells differs from that released by infected cells, we have examined the protein composition of envelope released by growing and T4-infected E. coli B. Our results show: (a) the protein composition of envelope released from growing or infected cells is similar, (b) the proteins present are representative of the outer membrane, (c) the major outer membrane protein of E. coli B, protein II, is deficient in the released material. We therefore conclude that the envelope material released from growing or infected E. coli represents a special fraction of the outer membrane. This finding is discussed in relation to outer membrane structure and function. In addition, data are presented on the differing outer membrane protein composition of substrains of E. coli B obtained from different laboratories.


Effect of membrane cholesterol on potassium transport in Mycoplasma mycoides var. Capri (PG3). Relationships between membrane lipid composition and physiological properties, particularly intracellular potassium levels, have been studied at 37 degrees C in Mycoplasma mycoides var. Capri (PG3). Native organisms grown on medium supplemented with either oleic acid plus palmitic acid or elaidic acid have identical growth characteristics, acidification properties and intracellular K content. On the other hand, when the cholesterol normally present in the membrane (20--25% of total lipids) is reduced to less than 2%, we observe: (1) the intracellular K content decreases (20 microgram K/mg cell protein instead of 40) and is independent of the phase of growth; (2) K passive permeability is drastically increased but K distribution remains in equilibrium with the transmembrane potential (delta psi); (3) organisms stop growing at pH 6.5 (instead of 5.2) and acidification is reduced by 40%, suggesting a large increase in H+ permeability, and (4) intracellular Na contents rise from 3 to 9 microgram Na/mg cell protein. Replenishing cholesterol in membranes of depleted cells results in a recovery of the high intracellular K level (35--40 microgram K/mg cell protein) and acidification properties. It is suggested that cholesterol affects the cation content via the increase in proton permeability which in turn controls the value of the delta psi responsible for the value of intracellular K equilibrium. Changes in K passive permeability, although related to the amount of cholesterol present in the plasma membrane, are probably not involved in the control of the intracellular K level.


Interactions between lipopolysaccharide and phosphatidylethanolamine in molecular monolayers. Lipopolysaccharide and phosphatidylethanolamine are the two major lipid constituents of the membrane of Salmonella typhimurium. Interactions between the purified lipopolysaccharide and phosphatidylethanolamine were studied in molecular monolayers at air-water interfaces. The equilibrium surface pressures of mixed films of lipopolysaccharide and phosphatidylethanolamine were determined as a function of the film composition. The plot of the equilibrium surface pressrue vs. the area occupied by phosphatidylethanolamine molecules exhibited two distinct regions. Below a phosphatidylethanolamine surface concentration at which 55% of the surface was occupied by phosphatidylethanolamine molecules, the equilibrium pressure was invariant and had the value of a pure lipopolysaccharide monolayer at maximum compression. At phosphatidylethanolamine surface concentrations in excess of 55% surface area occupation (phosphatidylethanolamine/lipopolysaccharide (mol/mol) greater than 16), the equilibrium surface pressure was a function of the surface concentration of phosphatidylethanolamine. The results suggest a simple model in which lipopolysaccharide and phosphatidylethanolamine form a complex in which each lipopolysaccharide molecule is surrounded ("lipidated") by a shell of approx. 16 phosphatidylethanolamine molecules.


Replication of bacteriophage G13 DNA in dna mutants of Escherichia coli. Host functions required for replication of microvirid phage G13 DNA were investigated in vivo, using thermosensitive dna mutants of Escherichia coli. In dna+ bacteria, conversion of viral single-stranded DNA into double-stranded replicative form (stage I synthesis) was resistant to 150 microgram/ml of chloramphenicol or 200 microgram/ml of rifampicin. Although multiplication of G13 phage was severely inhibited at 42--43 degrees C even in dna+ host, considerable amount of parental replicative form was synthesized at 43 degrees C in dna+, dnaA or dnaE bacteria. In dnaB and dnaG mutants, however, synthesis of parental replicative form was severely inhibited at the restrictive temperature. Interestingly enough, stage I replication of G13 DNA was, unlike that of phiX174, dependent on host dnaC(D) function. Moreover, the stage I synthesis of G13 DNA in dnaZ was thermosensitive in nutrient broth but not in Tris/casamino acids/glucose medium. In contrast with the stage I replication, synthesis of G13 progeny replicative form was remarkably thermosensitive even in dna+ or dnA cells.


Rifampicin and chloramphenicol effects on DNA replication in thymine-prestarved Escherichia coli B/r WP2 thy trp. When cultures of Escherichia coli B/r WP2 thy trp were prestarved for thymine for 30 min, DNA replication after readdition of thymine was limited to an increase of about 100% in the presence of rifampicin, an antibiotic which inhibits DNA-dependent RNA polymerase. However, chloramphenicol, an antibiotic which blocks protein but not RNA synthesis, did not limit replication. After prolonged thymine prestarvation (55 min) DNA increased only about 50% in the presence of rifampicin, but no such limitation occurred in the presence of chloramphenicol. The ability of a high concentration of rifampicin to limit DNA replication was eliminated by addition of either high or low concentrations of chloramphenicol, indicating that stoichiometric interaction of the antibiotics is not responsible for this effect.


Inhibition of DNA synthesis by griseolutein in Escherichia coli through a possible interaction at the cell surface. Griseolutein acts as a bactericidal agent, its toxicity decreasing with increase in cell density. There is no evidence that griseolutein acts at a specific stage of cell cycle. Inhibition of incorporation of radioactive thymidine into acid-insoluble fraction of cells is marked and rapid, while inhibition of incorporation of uridine also takes place. Incorporation of 32Pi into the acid-insoluble fraction of cells is inhibited while the incorporation into the nucleotide pool is not. Concentration of any of the four deoxyribonucleoside triphosphates in griseolutein-treated cells are similar to or higher than those in untreated cells. No extensive degradation of cellualr DNA is caused by griseolutein. DNA synthesis in plasmolyzed cells in not inhibited by griseolutein.


Specific interaction between mouse liver non-histone chromosomal proteins and mouse DNA demonstrated by a sequential DNA-protein binding procedure. The binding of mouse liver chromosomal proteins to DNA has been investigated using the nitrocellulose filter binding technique. Careful purification of the DNA involving nuclease S1 digestion and prefiltration through a nitrocellulose filter is used to reduce background binding in the absence of protein to less than 1%. Procedures involving direct binding of protein to labeled DNA, competition of binding of labeled DNA by unlabeled DNA, and dissociation of DNA . protein complexes with time do not indicate significant preference for binding to mouse DNA relative to Escherichia coli DNA. This specificity is demonstrated much more clearly by a novel type of procedure, which we call a sequential binding procedure. In this procedure non-specific binding proteins are sequestered by incubation with an excess of unlabeled E. coli DNA prior to addition of labeled DNA. Under these conditions, labeled mouse DNA is bound to filters to a 3- to 4-fold greater extent than labeled E. coli DNA.


A calorimetric investigation of melting of tRNAAsp from brewer's yeast. The thermodynamics of tRNAAsp unfolding was studied using a precision scanning microcalorimeter. The overall heat of melting was found to be about 55 J/g irrespective of the ionic strength and magnesium activity. The analysis of complex melting curves obtained in the absence of Mg2+ reveals four successive two-state transitions. The first was identified as the cooperative melting of the tertiary structure and the D region and the others as the melting of individual helical arms.


Structure of the yeast ribosomes. Proteins associated with the rRNA. Polyamines have been shown to bind to doubled stranded regions of rRNA [3]. Therefore, ribosomal proteins that can be cross linked to these molecules in the ribosomes structure must be bound to or located in the vicinity of the RNA. This technique is the first to yield results on the proteins associated with the rRNA in the eukaryotic ribosome where the lack of purified ribosomal proteins does not allow the use of direct binding studies as in bacterial systems. Proteins S7, S10, S13, S21, S22 and S27 in the small subunit and L2/3, L5, L10/12, L19/20, L22, L23, L36/37, L42 and L43' in the large subunit are labelled when cross linked to [14C]spermidine using 1,5-difluoro 2,4-dinitrobenzene and are good candidates to be RNA-binding proteins in ribosomes from Saccharomyces cerevisiae.


Expression of bacteriophage M13 dna in vivo. I. Synthesis of phage-specific RNA and protein in minicells. It is demonstrated that after infection of the appropriate minicell-producing strain of Escherichia coli with the filamentous bacteriophage M13, its replicative form DNA is segregated into minicells. Consequently these minicells have acquired the capability to direct the synthesis of phage-specific RNA and protein. Comparision of the electrophoretic mobilities of phage-specific RNA species made in vitro with those made in M13 replicative form DNA harbouring minicells, have indicated that almost all in vitro synthesized G-start RNAs have an equivalent among the in vivo synthesized RNA products. Furthermore it could be demonstrated that in M13 replicative form DNA harbouring minicells the phage-specific proteins encoded by genes III, IV, V and VIII are made. In addition the synthesis of a phage-specific polypeptide (molecular weight approx. 3000) co-migrating with the recently discovered capsid protein (designated C-protein) could be demonstrated. The meaning of these results for the resolution of the regulatory mechanisms operative during the life cycle of this phage will be discussed.


An enhanced thermostability in thermophilic 5-S ribonucleic acids under physiological salt conditions. The secondary structure of 5-S rRNAs of Thermus aquaticus (an extreme thermophile), Bacillus stearothermophilus (a moderate thermophile) and Escherichia coli (a mesophile) was compared using thermal denaturation techniques under varying ionic conditions. At a low ionic strength (10 mM K+), the Tm of T. aquaticus 5-S RNA differed by only 1 degrees C from that of E. coli RNA and the molecule was fully denatured well below the optimum growth temperature of the thermophile. The internal Na+, K+ and Mg2+ concentrations of T. aquaticus cells were determined to be 91 mM, 130 mM and 59 mM, respectively. Under these salt conditions, T. aquaticus 5-S RNA was significantly more stable than E. coli RNA and the 5-S RNA from B. stearothermophilus was intermediate as is its optimum growth temperature. The results suggest that the thermostability of macromolecules from thermophilic organisms may be specially dependent on the internal salt concentration. Furthermore, under these salt conditions, most of the secondary structure of the RNA remained stable at the optimum growth temperatures suggesting that ribosomal RNAs of thermophilic organisms contribute more to the thermostability of the ribosome than previously thought.


A large nucleoprotein fragment of the 50-S ribosomal subunit of Escherichia coli. A large nucleoprotein fragment was isolated from a nuclease digest of Escherichia coli 50-S ribosomes and purified to gel electrophoretic homogeneity. Conditions were employed under which the fragmentation pattern was reproducible and the various fragment fractions were stable and maintained their sedimentation and electrophoretic properties throughout the several preparative and analytical procedures used. Fractions that appeared homogeneous in sucrose gradient centrifugation were found to be heterogeneous by gel electrophoresis. The large fragment was purified to homogeneity by preparative gel electrophoresis. It contained 21 proteins, the 5-S RNA, and two large oligonucleotides which together total about two thirds the molecular weight of the 23-S RNA. Because it can be prepared reproducibly in large quantities and because of its size and stability, the fragment appears suitable for functional and structural studies and as the starting material for further fractionation. An important contributing factor to the observed stability and reproducibility was the maintenance of an unchanging ionic environment. A single buffer was employed throughout all the procedures, and the fragments produced by nuclease digestion were dissociated from each other by heat rather than by changing the medium.


Regulation of acid phosphatase synthesis in Saccharomyces cerevisiae. In Saccharomyces cerevisiae-136ts (Hutchison, H.T., Hartwell, L.H. and McLaughlin, C.S. (1969) J. Bacteriol. 99, 807--814) derepressed acid phosphatase was almost exclusively located outside the permeability barrier. Only a minor part of the activity was associated with the protoplasts; about half of it (48%) in the soluble fraction, the rest bound to the internal (45%) and plasma (7%) membranes. The activity found in the membranes of derepressed cells decreased by 30--40% after addition of inorganic phosphate or cycloheximide suggesting that this activity is the precursor of the external enzyme. The alkaline phosphatase activity level could not be modified by changes in the concentration of inorganic phosphate. Acid phosphatase was not synthesized if the cells were transferred to a low phosphate medium at the moment of incubation at 37 degrees C or in the presence of cycloheximide at 23 degrees C. The data suggested that enzyme formation is the result of the transcription and translation of a specific gene(s) and not the activation of a proenzyme. Inorganic phosphate did not inhibit the translation of mRNA though it may act at the level of the transcription.


The kinetic complexity of Acetabularia chloroplast DNA. The kinetic complexity of Acetabularia cliftonii chloroplast DNA is 1.52 +/- 0.26 . 10(9) daltons, compared to 0.2 .10(9) daltons for Chlamydomonas chloroplast DNA. There is an average of three genomes per chloroplast. The unusually large size of the Acetabularia genome may reflect the ancient evolutionary history of this organism.


[Kinetics of beta-lactamase inhibition by clavulanic acid]. The mechanisms of action of 3 R-factors on beta-lactamases (penicillin amido-beta-lactamhydrolase, EC 3.5.2.6) (TEM-1 pI = 5.4, TEM-2 pI = 5.6 and Pitton's type 2 pI = 7.7) have been kinetically analyzed for clavulanic acid inactivation. Clavulanic acid appears as a competitive and irreversible inhibitor (Kcat inhibitor) reacting in two steps: a, formation of a reversible enzyme . inhibitor complex (characterized by a Ki); b, evolution of the reversible complex into a new derivative (covalent, stable and inactive) by monomolecular kinetics characterized by a k6 (or Kcat) related to half-life. The kinetic constants are: TEM-1: Ki = 0.8 micrometer, k6 = 0.027 s-1; TEM-2: Ki = 0.7 micrometer, k6 = 0.03 s-1; type 2: Ki = 0.6 micrometer, k6 = 0.046 s-1. These results justify the 'progressive irreversible' character of the inhibition generally described.


The affinity chromatography of transketolase. A number of possible affinity adsorbents for transketolase (sedoheptulose-7-phosphate:D-glyceraldehyde-3-phosphateglycolaldehydetransferase, EC 2.2.1.1) were prepared. The behaviour of the enzyme from Candida utilis and from Baker's yeast on columns of these and of Blue Sepharose CL-6B was examined, together with the behaviour of the contaminating enzyme, ribulose 5-phosphate 3-epimerase (EC 5.1.3.1). A procedure for removing bound thiamine pyrophosphate by dialysis against EDTA was developed. The competitive inhibition of transketolase by oxythiamine and neopyrithiamine was measured and the Ki values obtained of 1.4 and 4.3 mM, respectively, were compared with the affinity of adsorbents prepared from these two inhibitors. Adsorbents containing bound thiamine pyrophosphate were relatively ineffective but those containing epoxy-linked neopyrithiamine and D-ribose 5-phosphate adsorbed the enzyme at pH 7.4 and it could be eluted in a specific manner.


Subcellular localization and levels of aminopeptidases and dipeptidase in Saccharomyces cerevisiae. Three aminopeptidases (L-aminoacyl L-peptide hydrolases, EC 3.4.11) and a single dipeptidase (L-aminoacyl L-amino acid hydrolase, EC 3.4.13) are present in homogenates of Saccharomyces cerevisiae. Bassed on differences in substrate specificity and the sensitivity to Zn2+ activation, methods were developed that allow the selective assay of these enzymes in crude cell extracts. Experiments with isolated vacuoles showed that aminopeptidase I is the only yeast peptidase located in the vacuolar compartment. Aminopeptidase II (the other major aminopeptidase of yeast) seems to be an external enzyme, located mainly outside the plasmalemma. The synthesis of aminopeptidase I is repressed in media containing more than 1% glucose. In the presence of ammonia as the sole nitrogen source its activity is enhanced 3--10-fold when compared to that in cells grown on peptone. In contrast, the levels of aminopeptidase II and dipeptidase are less markedly dependent on growth medium composition. It is concluded that aminopeptidase II facilitates amino acid uptake by degrading peptides extracellularly, whereas aminopeptidase I is involved in intracellular protein degradation.


Studies on the carboxypeptidase Y-inhibitor complex of yeast. We report in vitro studies on the interaction of several substrates with the carboxypeptidase Y-inhibitor complex of yeast. Inhibition of carboxypeptidase Y cleavage of two peptides by carboxypeptidase Y-inhibitor is shown to be competitive. The experiments show a wide variation in the degree of cleavage of a variety of peptide substrates by carboxypeptidase Y, despite the presence of the inhibitor protein. The most likely explanation for this behaviour is a different capacity for the peptides to dissociate the inhibitor protein from the substrate-binding site of carboxypeptidase Y. While the carboxypeptidase Y-inhibitor is insensitive to proteolytic inactivation when complexed with carboxypeptidase Y, it is sensitive when in the free state. Addition of the substrate, N-Cbz-Phe-Leu, to the carboxypeptidase Y-inhibitor complex, however, allows proteolytic inactivation of the inhibitor protein. We suggest that the proteinase-inhibitor may play a crucial role in the regulation of proteinase activity. The inhibitor protein generally protects proteins from unwanted proteinase action. However, it will allow cleavage of proteins which, by some signal triggered metabolically, become substrates due to the exposure of amino acid sequences normally buried, and exhibiting a high affinity for the proteinase.


Properties of the activation by pepsin of inactive renin in human amniotic fluid. 1. The renin present in human amniotic fluid was found to have an apparent Mr of 58 000 by gel filtration and is thus bigger than renin in untreated kidney extracts and plasma (Mr approximately 40 000). 2. Treatment with pepsin (40 microgram/ml pH 4.8, 2 h, 22 degrees C) caused a 6-fold increase in activity of this renin species, although Mr was not very different (57 000). 3. Unlike renal renin, renin in human amniotic fluid was not a glycoprotein and behaved similarly on concanavalin A-Sepharose before and after activation by pepsin. 4. Ion-exchange chromatography demonstrated a small change in the ionization properties of human amniotic fluid renin after activation by pepsin. 5. Pepsin-mediated activation resulted in a five-fold increase in V, but only a small decrease in the Km of renin to 39% of normal, so that the increase in activity observed was not due to an increase in the affinity of the enzyme for its substrate. The kinetic data were consistent with the theory of noncompetitive inhibition. 6. The activation of human amniotic fluid renin by pepsin may be caused by a change in the tertiary structure of the molecule subsequent to a proteolytic action that does not remove detectable polypeptide components.


Modulation of 32Pi incorporation into phospholipids of polymorphonuclear leukocytes by ionophore A23187. The effects of ionophore A23187 on the incorporation of 32Pi into phospholipids and on 45Ca2+ uptake and release by polymorphonuclear leukocytes were examined. A23187 increased 32Pi incorporation into phosphatidic acid, phosphatidylglycerol, phosphatidylserine, and the phosphoinositides. It also promoted a rapid burst uptake and release of 45Ca2+ by leukocytes. External Ca2+, but not Mg2+, was required for full stimulation of 32Pi incorporation into phosphatidic acid and the phosphoinositides. In the absence of external Ca2+, the increased radiophosphorus activity of phosphatidic acid, phosphatidylserine and the phosphoinositides was grossly reduced but not eliminated, and the decreased radiophosphorus activity of phosphatidylcholine became pronounced. In addition, the ionophore effect on 32Pi incorporation into leukocyte phospholipids was not abolished by ethyleneglycol bis(beta-amino-ethylether)-N,N'-tetraacetic acid. ATP radiophosphorus activity was also enhanced by the presence of A23187, but the enhancement was much less than that of the acidic phospholipids. Based on these findings, it is suggested that the increased 32Pi incorporation into the acidic phospholipids of leukocytes induced by A23187 was not solely derived from the higher radioactivity of ATP, increased Ca2+ fluxes and perturbation of cellular Ca2+ distribution of leukocytes exposed to A 23187 may trigger part of the altered 32Pi incorporation into phospholipids.


Some aspects of protein folding.

Some aspects of protein folding. The mechanisms by which a polypeptide chain reaches the three dimensional structure which generates its functional properties are not yet totally understood. The great amount of data now available in the field of protein structure and the data already obtained on protein folding by in vitro experiments allow some schematic representation as, at least, a working hypothesis. In this respect the emphasis is put on the hierarchy in protein structure and the possibility of a folding by stages, some elements or "building blocks" being able to reach independently a native structure and to refine this structure by interacting with each other in the whole molecule.


[Study of the lipopolysaccharide from a thermosensitive mutant CR-34 T 83 of Escherichia coli K 12]. The structure of the core of the lipopolysaccharide from T 83 mutant of Escherichia coli K 12 CR 34 was partially determined. Using dephosphorylation, enzymic hydrolysis, Smith degradation, methylations and analysis by gas chromatography/mass spectrometry an oligosaccharide sequence was determined with D-glucose, D-galactose and L-glycero-D-mannoheptose as sugar components. The structure which was demonstrated could be that of the characteristic core fragment of the K 12 type lipopolysaccharides from Escherichia coli.


[Extraction of acid alpha-glucosidase from human spleen]. An efficient method for isolation of acid alpha-glucosidase from human spleen is developed. The method involves chromatography of the enzyme on rho-aminophenyl-alpha-D-glucopyranoside covalently bound to CH-Sepharose 4B, with subsequent gel filtration on Sephadex G-200. The enzyme was homogeneous by the polyacrylamide gel electrophoresis data; it was purified about 1500-fold, as compared with the crude extract (the total yield 12.5%). Besides acid alpha-glucosidase, the preparations of alpha-L-fucosidase, alpha-D-galactosidase and beta-N-acetylglucosaminidase were isolated and purified 200-, 130- and 280-fold, respectively. The nature of interaction between acid alpha-glucosidase and immobilized rho-aminophenyl-alpha-glucopyranoside is discussed.


[Interrelationship between metabolic and genetic regulation of alkaline and acid phosphatases in E. coli cells]. The effect of exogenous orthophosphate and mutations in regulatory genes of alkaline phosphatase on the level of nonspecific acid phosphatase was studied. The level of this enzyme as well as the level of alkaline phosphatase were shown to be regulated by exogenous orthophosphate being derepressed under phosphate starvation. The derepression of acid phosphatase is accompanied by more rapid secretion of enzyme from membranes to soluble fraction. Mutations in all the four regulatory genes decrease the level of enzyme in cells. Genes phoR and phoS, participating in regulation of alkaline phosphatase, are required for the derepression of acid phosphatase under the conditions of phosphate starvation.


[Induction of E. coli alkaline phosphatase synthesis in cells preincubated at low temperatures]. Cell preincubation at lowered t degrees was found to result in increased alcaline phosphatase synthesis. The ability of cells for increased alcaline phosphatase synthesis correlates with increased content of cis-vaccinic acid and higher liquidity of lipids. It has been ascertained that modifications caused by cell preincubation at lowered t degrees favour the greater stability of mRNA coding the alcaline phosphatase.


[Sensitivity of chromosomal and plasmid E. coli DNA to restriction endonuclease Eco RII]. It was shown that E. coli C, E. coli MRE 600 DNA, and also plasmid DNA of Col E1, RSF 2124 from E. coli K-12, and plasmid DNA from E. coli  MRE 600 were completely resistant against restriction endonuclease R. Eco RII. Plasmid DNAs of Col E1, RSF 2124 amplificated for 4 hours in the presence of chloramphenicol are sensitive to R. Eco RII but after 16-hour amplification in the presence of chloramphenicol these DNAs acquire complete resistance against R. Eco RII. These data point to the slower rate of modification of DNA in vivo by DC-methylases of Eco RII type in comparison with DNA methylase Eco RII.


A statistical procedure useful in evaluating time of onset and termination of response in clinical trials. A statistical model jointly characterizing the onset and termination of treatment response of a subject over a fixed observed time period is presented. The model requires that the observations for each subject are made at a set of pre-selected time points during the observed time period. A useful index characterizing the probability of being in response is developed along with maximum likelihood estimates and variances. A likelihood ratio test is developed to simultaneously compare two treatment groups with respect to this index for all times. The proposed procedure is applied to a set of data from a clinical trial of two bronchodilator drugs from which our procedure is motivated.


[Effect of sodium thiosulfate on the pancreas in experimental pancreatitis]. Histological and histochemical study of the pancreas of albino rats and experimental pancreatitis showed the use of sodium thiosulfate to considerably inhibit the progress of necrotic changes and circulatory disturbance. The preparation prevented recidivation of pancreatitis and inhibited sclerotic changes in the gland. Sodium thiosulfate stimulated the regenerative process including regenerative hypertrophy and expressed epimorphosis.


[Effect of serum fractions from cancer patients on leukocyte migration out of capillary tubes]. Blood serum of patients suffering from cancer of the stomach and urinary bladder inhibited in vitro migration of autologous leukocytes, leukocytes of donors and control patients, and also guinea pig macrophages in over half of cases. In chromatography of these sera on Sephadex G-100 the activity inhibiting the leukocyte migration was revealed in fraction I (Mol. wt. over 100000) and in fractions IV and V (Mol. wt under 30 000). The blood serum and its fractions from cancer patients failed to eliminate the leukocyte migration inhibition caused by the tumour antigens in comparison with the leukocyte migration in the medium with control serum without any antigens. As suggested, the activity of fraction I inhibiting the leukocyte migration was due to the antigen-antibody complex, and of fraction IV and V--to a factor similar by its properties to the factor produced in vitro by lymphocytes stimulated by the antigens or mitogens.


Successful use of multiagent immunosuppression in the bone marrow transplantation of sensitized patients. Of 23 patients with severe aplastic anemia, 17 were sensitized to histocompatibility antigens of HLA-A, -B, and -D loci-identical potential sibling donors as determined by cell-mediated lysis (CML) assays in vitro. Antibody-dependent sensitization was detected in 3 patients, antibody-independent cellular sensitization in 11, and both in 3. Fourteen sensitized patients were transplanted after initial multiagent immunosuppression consisting of rabbit anti-human thymocyte serum, procarbazine, and cyclophosphamide, eleven with a CML-positive donor and three with a CML-negative donor. Engraftment was achieved in each of 13 patients who were evaluable, and only 2 ultimately rejected their marrow grafts, 1 with subsequent return of his own marrow function. Five patients without evidence in vitro of sensitization were transplanted after immunosuppression with cyclophosphamide alone; none of these rejected their grafts. These studies show that sensitized bone marrow transplant recipients can be successfully transplanted after optimal donor selection and multiagent immunosuppression.


Alterations of the antibody response following in utero exposure to diethylstilbestrol. The antibody response to SRBC and E. coli 0127 lipopolysaccharide were determined in offspring from mice exposed in utero to diethylstilbestrol. The antibody response to SRBC, a T-cell dependent antigen, was similar in control and exposed animals. In contrast, the LPS antibody response was suppressed in treated females and enhanced in treated males. These studies indicated that in utero exposure to DES alters the humoral immune system to T-independent antigens.


Highly selective vagotomy or truncal vagotomy and pyloroplasty for chronic duodenal ulceration: a randomized, prospective clinical study. The results of a randomized, prospective clinical trial of highly selective vagotomy (HSV) versus truncal vagotomy and pyloroplasty (TVP) in 126 male patients undergoing elective surgery for chronic duodenal ulceration are presented. The operations were performed by surgeons of all grades of experience. At a mean follow-up time of just over 3 years a satisfactory result was obtained in 93 per cent of patients following HSV and 78 per cent of patients following TVP, the difference being probably statistically significant (P less than 0.05). The incidence of early and late dumping, bile vomiting, flatulence, post-prandial epigastric discomfort and wound infection was statistically significantly less after HSV than after TVP. Three patients have developed a recurrent duodenal ulcer after each type of operation (5.4 per cent). At this early stage HSV has advantages over TVP; it will be interesting to see if these are maintained with the passage of time.


Fasting and postprandial serum gastrin levels before and after highly selective gastric vagotomy, truncal vagotomy with pyloroplasty and truncal vagotomy with antrectomy: is there a cholinergic antral gastrin inhibitory and releasing mechanism? Fasting serum gastrin levels and postprandial gastrin response were measured before and 1 month after highly selective vagotomy, truncal vagotomy with pyloroplasty and truncal vagotomy with antrectomy. The three groups of patients, 12 in each group, were closely matched for age, sex, maximum acid output and completeness of vagotomy. After highly selective and truncal vagotomy an identical and significant increase in fasting gastrin was observed, whereas after truncal vagotomy with antrectomy the pre- and postoperative fasting gastrin levels were not different. The net postprandial gastrin output over basal value was significantly increased after highly selective vagotomy, unchanged after truncal vagotomy and significantly lowered after truncal vagotomy with antrectomy. These results suggest the presence in the intact subject of a cholinergic inhibitory mechanism in the gastric body and fundus for the release of antral gastrin in the fasting and postprandial states and a possible cholinergic facilitatory mechanism for the release of antral gastrin after meals.


Metronidazole prophylaxis for elective large bowel surgery in children: a prospective trial. In a controlled trial in children aged 2 days to 16 years undergoing large bowel surgery a group of 15 patients had preoperative mechanical bowel preparation and oral neomycin while a second group of 15 patients was similarly prepared but also had preoperative and postoperative metronidazole medication. Five of the 9 postoperative infections in the first group involved Bacteroides spp. but no anaerobe was found in the 3 wound infections in the metronidazole group.


Azathioprine plus prednisone in treatment of pemphigoid. Twenty-five patients taking part in a controlled trial to compare azathioprine plus prednisone with prednisone alone in the treatment of pemphigoid were followed up for three years. Results showed that the addition of azathioprine 2.5 mg/kg body weight daily reduced the total maintenance dose of prednisone needed by about 45%, with no increase in serious side effects or mortality. The suggestion that azathioprine might increase the risk of disseminated malignancy in elderly patients was not supported. We conclude that in future trials the combination of azathioprine with prednisone should be used as the standard treatment for comparison.


Cimetidine: an advance in gastric ulcer treatment? Sixty patients with endoscopically confirmed gastric ulceration took part in a single-blind trial of cimetidine 1 g daily compared with conventional treatment--namely, carbenoxolone in patients aged under 60 and Caved-(S) in those over 60. Twenty-nine patients received cimetidine: in 12 (41%) ulcer healing was complete after one month, in 26 (90%) healing was complete after two months, and all ulcers were healed after three months of treatment. In the under-60s, ulcers were healed in a greater proportion of patients given cimetidine than in those given carbenoxolone. The difference, however, was significant only at the 5% level, which owing to small numbers was of doubtful clinical validity. In the over-60s cimetidine and Caved-(S) were of similar efficacy. The 54 patients with healed ulcers are being followed up for two years; so far there have been 16 recurrences (30%). Further trials are needed, which should include maintenance treatment aimed at lowering the unacceptably high recurrence rate.


Trial of high-titre human rubella immunoglobulin. To test the efficacy of passive antibody for protecting susceptible pregnant women who have been exposed to rubella high-titre human rubella immunoglobulin (HRI) was given to 20 seronegative male adult volunteers simultaneously with rubella vaccine. After receiving the intramuscular injections of HRI (750 mg of IgG) and vaccine (10(3 . 92) median tissue culture dose, Wistar RA27/3 subcutaneously) eight of the 20 volunteers failed to show seroconversion. Antibody responses--and thus presumably viraemia--in the remaining 12 volunteers were delayed and reduced when compared with those in 19 volunteers given the vaccine alone. No significant responses occurred in volunteers given only HRI. We conclude that HRI may be of value for seronegative pregnant women who come into contact with clinical rubella, particularly when termination is likely to be refused.


Polyunsaturated fatty acids in treatment of acute remitting multiple sclerosis. One hundred and sixteen patients with acute remitting multiple sclerosis (MS) took part in a double-blind controlled trial of treatment with polyunsaturated fatty acids and were randomly allocated to one of four groups. Two groups received linoleic acid, one alone as a spread and one with gamma-linolenic acid in capsules (Naudicelle); and two control groups received oleic acid, one as a spread and one in capsules. Rates of clinical deterioration and frequencies of attacks were not significantly different between treated and control groups. Exacerbations were shorter and less severe in patients receiving a high dose of linoleic acid than in controls, but those receiving a lower dose--that is, Naudicelle--showed no such difference. Thus supplementing the diet with 20 g linoleic acid marginally affected the duration and severity of relapses of MS but had no effect on overall disability. The dose of Naudicelle used provided insufficient supplementation.


The luteinizing hormone-releasing hormone (LH-RH) neuronal networks of the guinea pig brain. II. The regulation on gonadotropin secretion and the origin of terminals in the median eminence. Cell bodies synthesizing LH-RH are located throughout the central nervous system including the hypothalamic arcuate nucleus, the medial preoptic area and medial septal nucleus. The contribution of each of these cell groups to the LH-RH terminals in the median eminence was assessed by immunocytochemistry following placement of radiofrequency lesions in male guinea pigs. Lesions in the arcuate nucleus resulted in an almost complete absence of LH-RH fibers in the median eminence. Lesions in the medial preoptic area or suprachiasmatic nucleus produced a decrease in the amount of immunoreactivity throughout the median eminence; but there was only a small decrease in the numbers of labeled fibers, which was localized to the internal zone of the median eminence and the external zone on the ventral and lateral surfaces of the infundibular stalk. Lesions of the mammillary bodies, medial amygdaloid nucleus, septal nucleus and fornix had no effect. The effects of these lesions on pituitary gonadotropin secretion was also assessed. Only lesions in the arcuate nucleus diminished plasma concentrations of luteinizing hormone (LH) and testosterone in the male guinea pigs. Similary only arcuate lesions prevented the postcastration rise in plasma LH in ovariectomized female guinea pigs. These data strongly suggest that the arcuate nucleus is the major but not the sole source of LH-RH terminals in the median eminence, and these are responsible for the neural regulation of tonic gonadotropin secretion.


Multimodal immunotherapy of primary gastrointestinal tumors in rats. 1. Histologic correlation. The effect of multimodal immunotherapy was studied in rats bearing primary gastrointestinal tumors induced by 1,2-dimethylhydrazine dihydrochloride. Multimodal immune manipulation consisted of combinations of splenectomy, C. parvum, unblocking serum, unblocked lymphoid cells, and levamisole. Such immunologic intervention resulted in significant inhibition of tumor growth, and their metastasis. Ten of 10 untreated rats, 8 of 8 rats treated with splenectomy alone and 10 of 10 rats treated with normal rabbit serum died of progressive tumor growth. None of the rats treated with combinations of splenectomy, unblocking serum, unblocked lymphoid cells, C. parvum and levamisole succumbed to progressive tumor growth during the observation period. The histologic evidence of tumor destruction was obtained in 18 of 22 tumors in rats of groups receiving multimodal immunotherapy.


Variation of macrophage migration by a factor from regional lymph node cells of breast cancer patients. Using a standard heterologous assay in which lymphokines alter guinea pig peritoneal macrophage migration, it was found that regional lymph node cells (RLNCs) from patients with primary breast cancer elaborate soluble factors which variably affect such migration. Variation of migration resulted when soluble factors employed were obtained from different nodes in the same patient as well as from nodes from different patients. Some nodes from a patient elaborated migration inhibition factors (MIF) and other migration enhancing factors (MEF). The findings are in keeping with others obtained by us relative to the variation in lymphocyte transformation and thymidine uptake by RLNCs and further emphasize that all RLNs in patients with breast cancer are not biologically similar. They lend support to our previous hypothesis that the reason why some RLNs contain metastases and others do not is more likely due to biological differences than because of anatomical happenstance, i.e., transport of tumor cells to some nodes and not to others.


Childhood acute lymphocytic leukemia: study VIII. This controlled study of children with ALL was designed to test the efficacy and toxicity of one-, two-, three- and four-drug therapy during remission and whether more aggressive therapy in the first eight weeks prolongs remission in patients with features associated with a particularly poor prognosis. After inducing remission with prednisone, vincristine and asparaginase, patients received cranial irradiation and IT methotrexate and were randomized to receive: 1--methotrexate alone; 2--methotrexate plus mercaptopurine; 3--same as in group 2 plus cyclophosphamide; and 4--same as in group 3 plus arabinosyl cytosine. Patients with CNS leukemia at diagnosis received IT methotrexate weekly during the induction period and a higher dose of CNS irradiation. Patients with anterior mediastinal enlargement at diagnosis received radiotherapy to the mass during the induction period. Patients who failed to attain bone marrow remission after four weeks of therapy were given daunorubicin and prednisone for 2--4 additional weeks. Of the 282 patients entering this study between January 1972 and November 1975, 268 (95%) attained complete remission and 228 (85%) were randomized to receive continuation chemotherapy with 1, 2, 3 or 4 drugs. In Group 1 (methotrexate alone), 14 of 20 patients relapsed and 9 developed leukoencephalopathy without antecedent CNS leukemia apparently due to higher doses of intravenous methotrexate; in Groups 2, 3 and 4 the results were equivalent, but without leukoencephalopathy in initial CR. The addition of cyclophosphamide and arabinosyl cytosine increased toxicity and complications without demonstrably increasing the leukemocidal effect. In the 40 patients given additional early therapy, the modalties employed in this study did not prolong remission.


Adriamycin versus methotrexate in five-drug combination chemotherapy for advanced breast cancer: a randomized trial. Adriamycin is of noteworthy efficacy in the treatment of metastatic breast cancer. Its role in combination regimens is under investigation. One hundred seventy-five women with advanced breast cancer were entered into a prospectively randomized trial comparing two five-drug regimens. Regimen CMFVP consisted of cyclophosphamide (C), methotrexate (M), 5-fluorouracil (F), vincristine (V), and prednisone (P). Regimen CAFVP was identical but substituted Adriamycin (A) for methotrexate. Twenty-seven patients were disqualified; 148 were evaluable. With CMFVP the complete response rate (CR) was 11%, and the partial response rate (PR) was 46%; with CAFVP, CR was 13% and PR was 45%. Duration of response tended to be slightly longer for patients on the Adriamycin arm. The median survival for CR and PR patients with CMFVP was 20.2 months, which was shorter (p = .07) than the 33 month median survival with CAFVP. Although statistical significance was not reached at the 5% level, the increased survival of responders on the Adriamycin regimen supports the data of other studies which suggest that first line combination chemotherapy in advanced breast cancer should include Adriamycin.


Sequential chemoimmunotherapy of colorectal cancer: evaluation of methotrexate, Baker's Antifol and levamisole. Fifty-two untreated patients with colorectal cancer were randomized to receive 5-fluorouracil (5-FU) alternating either with methotrexate (MTX) or Baker's Antifol (BAF) with or without the immunostimulant, levamisole (Program I). Fifty-five patients who had received prior treatment were randomized to receive methyl-CCNU (Me) with MTX or BAF (Program II). Fifteen of these patients had failed to respond to initial therapy with 5-FU plus MTX or BAF and subsequently received Me plus the alternate antifol. Overall response rate for each of programs I and II was 10%. The responses were 1/11 with 5-FU-MTX plus levamisole, 2/12 with 5-FU-MTX, 1/8 with 5-FU-BAF plus levamisole, 0/8 with 5-FU-BAF, 2/20 with Me-MTX and 2/21 with Me-BAF. The median survival times (MST) for patients receiving Programs I and II were 10 and 5 months, respectively. The MST for all patients receiving MTX was significantly longer than that of patients receiving BAF Survival was not influenced by levamisole administration. Both chemotherapy programs were well tolerated. The sequential administration of 4 active agents failed to improve the results of treatment of colorectal cancer.


Adjuvant radiation therapy for regional nodal metastases from malignant melanoma: a randomized, prospective study. After nodal metastasis from malignant melanoma, approximately 80% of patients die from disseminated disease. To clarify the role of radiation therapy (XRT) following node dissection. 56 patients with biopsy-proven nodal metastasis participated in a randomized, prospective clinical trial which compares radiation therapy to the regional lymph node area following lymphadenectomy (27 patients) with lymphadenectomy alone (29 patients). Interesting differences in the survival curves (p = 0.09) and in the disease-free interval curves (p = 0.08) for the two treatment groups proved to be attributable to imbalances in the age and nodal distributions in the treatment groups. Covariate analysis identified age and sex as the factors having the most significant (p less than 0.04) effect on survival and identified the number of positive nodes as the covariate having the most significant (p less than 0.02) effect on disease-free interval. Treatment did not have a significant effect upon survival or disease-free interval.


Clinical trials of immunotherapy: present status. This brief review of the more promising clinical trials suggests that immunotherapy is indeed beneficial for selected cancer patients. Because of its limited potency, it should not be used as primary treatment for malignant disease except as local immunotherapy for certain accessible tumors. It is effective for eradication of primary neoplasms of the skin as well as cutaneous metastases of malignant melanoma and breast carcinoma. The most important role for immunotherapy is in combination with other modalities. It may help control occult micrometastases that cause recurrence and death following surgical procedures or irradiation. Results of adjuvant immunotherapy appear promising for malignant melanoma, for carcinoma of the lung, breast, and colon, and for soft-tissue sarcomas. In combination with chemotherapy, immunotherapy appears to prolong remission and survival in acute myelogenous leukemia and in disseminated tumors of the lung and breast. Clearly, immunotherapy is not a panacea for malignant disease, but it could become an important arm in a multimodality attack on cancer.


Immunotherapy with BCG administered by scarification: standardization of reactions and management of side effects. The method of administering BCG by scarification is described in detail. A system of classifying the intensity of local reactions is proposed to standardize administration. The experience of the M.D. Anderson Hospital involving over 2700 patients is reviewed. Administration of BCG by scarification has been accomplished with safety and has been well tolerated and accepted. The most commonly observed side effects are discussed as well as their management.