Clinical trial of six-month and four-month regimens of chemotherapy in the treatment of pulmonary tuberculosis. In a study in Singapore, Chinese, Malay, and Indian patients with pulmonary tuberculosis received 2 months of daily treatment with streptomycin, isoniazid, rifampin, and pyrazinamide followed either by daily treatment with isoniazid, rifampin, and pyrazinamide (SHRZ/HRZ regimen) or by daily administration of isoniazid and rifampin (SHRZ/HR regimen) allocated at random. Both regimens were given for either 6 or 4 months by random allocation. All 330 patients with drug-sensitive tubercle bacilli before treatment had a favorable bacteriologic response during chemotherapy. During the first 6 months after the end of chemotherapy, there was only a single bacteriologic relapse among 84 SHRZ/HRZ and 80 SHRZ/HR patients treated for 6 months, but 8 (10 per cent) of 80 SHRZ/HRZ and 4 (5 per cent) of 74 SHRZ/HR patients treated for 4 months relapsed. Of a total of 33 patients with bacilli resistant to isoniazid, streptomycin, or both drugs before treatment, only one had an unfavorable response during chemotherapy, and none of 31 patients relapsed during the first 6 months after stopping chemotherapy. The incidence of adverse reactions was low; 11 (3 per cent) of 397 patients had hepatitis, but not all episodes were attributable to drug toxicity, and one patient had thrombocytopenic purpura.


The use of antiepileptic drugs.

The use of antiepileptic drugs. The use of antiepileptic drugs has become increasingly effective through several factors: new techniques that allow better diagnosis of the seizure disorder and its underlying cause; the development of new medications and increased knowledge of old ones; and the widespread use of antiepileptic drug-level determinations. The choice of a drug depends heavily on an accurate diagnosis of seizure type, which may determine the response to the medication. Because of better diagnostic criteria and intensive monitoring procedures, the correct seizure disorder can be more easily diagnosed and, therefore, the proper medication selected. Minimal efficacious and toxic blood concentrations have now been identified for most antiepileptic drugs. Several, including primidone, carbamazepine, methsuximide, and mephenytoin, have pharmacologically active metabolites that affect both the toxicity and efficacy of the prescribed drug and can now be measured in the plasma. The most effective use of the antiepileptic drugs depends on a combination of reliable blood level measurements, clinical observation, and knowledge of their pharmacokinetics and biotransformation.


Physician manpower expansionism: a policy review. A lack of national health goals has allowed physician manpower policy to be dominated by an expansionist philosophy. Scarce resources have been channeled into the production of specialist physicians trained to provide complex and expensive care for uncommon diseases, using other scare and expensive resources and adding to the steep rise in medical care costs. Society seems to want access to primary care--a lack it views with dismay--and simultaneously fears increasing costs of care. Lack of access plus high cost might lead to rash implementation of other inappropriate policies. Success of policy decisions is pure serendipity if made without reliable and relevant information or based on inappropriate data, such as opinions alone. If information is unavailable, then physician manpower decisions should be delayed or, if made, implemented cautiously.


Neoantigen response in patients successfully treated for lymphoma. A Southwest Oncology Group study. To ascertain the cellular immune function of patients successfully treated for lymphoma, we measured skin-test reactivity to a battery of recall antigens, phytohemmagglutinin (PHA), and the neoantigens keyhole limpet hemocyanin (KLH) and dinitrochlorobenzene (DNCB). Seventy-four patients with Hodgkin's disease and 31 patients with non-Hodgkin's lymphoma were studied from 3 to 186 months after cessation of therapy for lymphoma. Although reactivity to recall antigens and PHA was normal, the number of patients responding to the neoantigens was significantly (P less than 0.01) lower than normal (KLH, 35%; and DNCB, 34%). This impairment in reactivity to neoantigens could not be correlated with specific diagnosis, stage of disease, or type of treatment. Reactivity to DNCB was significantly (P less than 0.01) improved in those patients studied more than 3 years after treatment, but the number who reacted was still markedly abnormal (17 of 33). Thus, successfully treated patients with lymphoma seem to have difficulty in responding to new foreign antigens.


Effect of hypertonic glucose on the muscular cramps of hemodialysis. The effect of hypertonic (50%) glucose injected for relief of hemodialysis-induced muscular cramps was studied in 15 chronically uremic, nondiabetic patients who experienced a total of 44 cramp episodes. In a double-blind trial either 50 mL (or less) of hypertonic glucose or physiologic (0.9%) saline solution was injected, and the therapeutic response was evaluated. Of a total of 44 episodes of cramps, 26 were treated with hypertonic glucose and 18 with normal saline. Treatment with hypertonic glucose relieved 17 of 26 episodes, in contrast to only five of 18 episodes relieved with 50 mL of normal saline (P less than 0.016). No complications related to hypertonic glucose administration were observed. Hypertonic glucose seems to be safe and effective for the relief of dialysis-induced cramps. It also avoids undesirable loading with sodium and mannitol, which have been suggested for treatment of dialysis-induced cramps.


Hypomagnesemia and renal magnesium wasting in patients receiving cisplatin. We studied renal function and serum electrolytes in 51 patients receiving cisplatin chemotherapy by retrospectively reviewing the charts of 44 patients and prospectively following seven patients. Hypomagnesemia developed in 23 of 44 evaluable patients who were receiving cisplatin. We documented inappropriate renal magnesium wasting in four patients. Two patients required hospitalization for symptomatic hypomagnesemia. We conclude that cisplatin can induce a renal tubular defect in magnesium conservation and serious clinical syndromes of magnesium deficiency.


The decreasing incidence of primary intracerebral hemorrhage: a population study. This population study describes the experience with primary intracerebral hemorrhage (PIH) in residents of Rochester, MN, for the 32-year period from 1945 through 1976. The average annual age-adjusted incidence rate for PIH was 12.1 per 100,000 population, and the incidence for all cases of spontaneous intracerebral hemorrhage was 15.2 per 100,000. The PIH rates were higher for males than for females, and they increased steadily with age. When patients on long-term anticoagulant therapy were excluded, there was a steady decrease in the average annual age-adjusted incidence rate for PIH in each succeeding 8-year interval since 1945. Prehemorrhage hypertension, present overall in 89% of patients, was much more frequent and severe in the earlier years of the study. The frequency and severity of prehemorrhage hypertension also varied inversely with age in the population with PIH. The median age at the onset of PIH increased from 65 years for the period 1945 through 1952 to 71 years for 1969 through 1976.


Cefazolin prophylaxis in head and neck cancer surgery. This study evaluated the prophylactic use of cefazolin in reducing the incidence of infection in patients undetgoing cancer surgery where the upper aerodigestive tract was entered from the neck. A prospective, randomized, double-blind design was conducted in a single hospital. The patient was given placebo or cefazolin, 1 gm intramuscularly with  the preoperative medications, then 0.5 gm every six hours for four doses. Of 55 determinate patients, 32 received antibiotics and 23 placebo. Infection rate was 38% (12/32) and 87% (20/23) respectively, representing a statistically significant reduction in infection (P less than 0.001/. There were 30 wound and two nonwound (sinusitis and pneumonia) infections. In conclusion, the perioperative use of cefazolin in patients undergoing cancer surgery where the oral cavity or pharynx has been entered from the neck is useful in reducing the incidence of wound infection.


Loss of secretory activity in the glands of nasal polyps. Secretory activity in the glands of the nasal polyps from 15 patients with or without allergic histories was examined by using a direct (anti-IgA) or indirect (anti-SC) immunofluorescent technique. Glands were found in all the examined nasal polyps, but in many, only a few were present. Active secretory process of IgA was observed in the intraepithelial glands of the ciliated, pseudostratified epithelium. Most of the dilated glandular ductules, on the contrary, appeared to be lacking in secretory activity of IgA. Such were commonly filled with mucus materials and their epithelial glandular cells showed no fluorescence for anti-SC, whereas the active IgA secreting cells showed a heavy fluorescence for secretory component (SC) in indirect immunofluorescent studies. Activity of the glandular ductules, therefore, was determined by examining the SC producing activity. In some extracted polyps, SC containing dilated glands were present. These active glands, however, lost the SC producing activity followed by the stasis of mucus flow in the lumen. Thus, when the glands were occupied by mucus materials, stretched and cystic degeneration ensued.


Thrombocytopenia associated with sodium valproate treatment. The administration of sodium valproate to 30 patients in daily doses of 1,200 to 3,000 mg was associated with a significant reduction of the platelet count. Thrombocytopenia without any concomitant bleeding abnormalities occurred in 10 of the patients. Platelet counts returned to baseline levels after withdrawal of the drug.


Prostaglandin effect on lymphokine production in multiple sclerosis. Production of the lymphokine LIF (leukocyte migration inhibitory factor) by normal lymphocytes on stimulation with mitogens in inhibited by E type prostaglandins (PGE). It has been reported that the leukocytes of patients with multiple sclerosis (MS) are resistant to this inhibitory effect. We have studied 10 MS patients using an indirect technique for measuring LIF activity. This allows separation of the effects of PGE on lymphocyte LIF production from the effects on the action of LIF on the target granulocytes. There was no difference between normal subjects and MS patients in the effects of prostaglandins E1 and E2 in concentrations of 2.5 and 0.25 microgram per milliliter, on LIF production, or on granulocyte response to LIF. These results do not support the hypothesis that the chronic inflammatory process in MS is the result of resistance to the normal inhibitory effects of PGE on the cellular immune response.


[Idiopathic hemochromatosis and gonadotropin secretion (author's transl)]. 18 patients suffering from idiopathic hemochromatosis were studied. Plasma testosterone was stimulated by HCG. Basal plasma LH and FSH were significantly lower for the male group compared to healthy men. Lack of reponse of plasma LH and FSH after IV LHRH (100 microgram) was observed in 10 cases; among the patients with a normal response there were a postmenopausal woman, a 47 XXY syndrome, and the two youngest patients. Repeated infusions of LHRH (200 microgram/day x 5 d) did not increase LH in 8 out of 9 cases. Repeated plasma LH determination for 4 hours showed no pulsatile pattern in 6/7 patients. Plasma LH-RH measured in 7 cases was not elevated. The conclusion is that hypogonadism in hemochromatosis, when present, is related to a gonadotropin secretion defect, presumably of pituitary origin.


[Streptococcus group A resistance to tetracycline: its spread and transduction]. The study on antibiotics resistance of group A streptococci isolated in 1977 showed that the number of the antibiotic resistant strains had significantly increased as compared to the data of 1960. High percentage (53%) of the cultures with multiple resistance was noted. It was observed that the number of the streptococcal cultures resistant to erythromycin and chloramphenicol decreased while the number of the strains resistant to tetracyclines increased. The level of resistance to tetracycline increased more than 2 times from 1960 and in some cases reached 125 and 250 gamma/ml. The wide spread of tetracycline resistance was evident of the presence of the mechanism of the marker transduction. Possible transduction of this feature was studied. Microbe-free phagolysates obtained by induction with UV-light from the strains with multiple antibiotic resistance were used as the donor material in the experiment on transduction. Principal possibility of transducing resistance to tetracycline from 2 donors to 4 recipients at a frequency of 10(-6) was shown.


[Plasmid-determined streptococcal resistance to antibiotics]. The analysis of the genetic organization of the determinant ERLI by means of obtaining and studying the antibiotic sensitive mutants from the strain resistant to erythromycin and lincomycin provided experiment data in favour of the fact that inducable resistance to erythromycin and lincomycin determined by the plasmid might be defined by the same or closely linked genes.


[Clinical study of oxacillin for intravenous administration in heart surgery after operations with artificial circulation]. Intravenous sodium oxacillin of Soviet production was subjected to clinical trials in 50 cardiosurgical patients. The patients were operated under conditions of artificial circulation. The antibiotic was used for prophylaxis and therapy of the postoperative pyo-inflammatory complications. In the group of the patients observed no lethal cases due to the post-operative infection were registered.


[Trial study of ampiox sodium used intravenously in the clinical picture of internal diseases]. Clinical efficiency of intravenous ampiox was shown. The drug was used mainly in the treatment of patients with bronchopulmonary diseases and other infections. The results of the treatment proved to be good and satisfactory in 12 out of 15 patients. In 3 severe patients the antibiotic therapy produced a temporary effect. The side effects were of allergic genesis and rapidly disappeared after discontinuation of the treatment and administration of tavegi or dimedrol. No phlebitis cases were observed.


[Expression of the gene for tetracycline resistance of plasmids R6 and RP4 in bacteria of the family Enterobacteriaceae]. It was found that manifestation of the tetracycline resistance gene depended on the type of the plasmid containing the gene. The tetracycline resistance gene was subject to less repression in plasmid R6 than in plasmid RP4. Sensitivity of the initial plasmid-free bacteria varied within lower dose ranges than that of the plasmid-carrying strains. Regulation of the tetracycline resistance gene manifestation in the given plasmid may change in different bacterial hosts, i.e. in different cytoplasmic environment at different gene background.


[Experimental characteristics of the antileishmaniasis action of amphotericin B]. The efficacy of amphotericin B powder of the Soviet production was studied as a therapeutic agent in experimental zoonotic leishmaniosis of the skin form of mice and hamsters. Amphotericin B was administered per os and its action was compared with that of monomycin injected subcutaneously (50 mg/kg). Amphotericin B in a dose of 150 mg/kg administered for 30 days protected 96+/-0.07 and 76.4+/-0.9 per cent of the mice and hamsters respectively from development of the leishmaniosis clinical signs, 68.8+/-0.04 per cent of the animals being cured (in the treatment experiments). The effect of the antibiotic was analogous to that of monomycin administered parenterally. The study of the kinetics of amphotericin B showed that the antibiotic was well absorbed from the digestive tract of the animals into the blood which provided the chemotherapeutic effect.


[Effect of the functional state of the internal organs on the pharmacokinetics of benzylpenicillin in angina]. The levels of benzylpenicillin on its intramuscular administration in doses of 100000 or 200000 units, as well as the renal and hepatic functions and the antibiotic binding by the host protein components were studied in 44 angina patients and 29 healthy persons. The benzylpenicillin levels were always markedly higher in the angina cases than those in the healthy persons. The ratio of the free and protein bound fractions of the antibiotic in the above groups was practically the same. At the same time the amount of benzylpenicillin excreted with the urine was higher in the angina patients than in the healthy persons approximately by 2 times. The study of the function of the internal organs revealed a marked decrease in the daily diuresis in all the angina patients and microhematuria in the 1/4 of the cases. The impairment of the renal function was determined by the laboratory methods practically in all the patients. On the basis of the data obtained it was concluded that the increased benzylpenicillin levels in the angina cases were connected with a decreased capacity for inactivating this substance by the liver enzymatic systems due to the changed function of this organ. The fact of the increased antibiotics levels in the host is considered as a positive phenomenon providing inhibition of the disease causative agents resistant to usual concentrations of the drug.


[Structure of the microflora of suppurative wounds and its sensitivity to antibiotics]. Clinico-bacteriological examination of patients with purulent infections showed that Staphylococcus was the predominating microflora in the wounds. Simultaneously an increasing role of gram-negative conditionally pathogenic bacteria was shown. Multiple drug resistance was found in the organisms tested. The highest sensitivity levels were observed to gentamicin, kanamycin, tetracycline, levomycetin. It was shown by means of special typing methods that staphylococci of phage group III and Ps. Aeruginosa of serotype II predominated in the infected wounds. When the pathological material contained the antibiotic resistant cultures of Ps. aeruginosa, Proteus, Klebsiella and toxigenic strains of Staphylococcus, a tendency for prolongation of the suppurative process was observed.


[Comparative experimental study of cyclacillin and ampicillin]. Cyclacillin was compared with ampicillin by its bacteriostatic efficiency in vitro, chemotherapeutic efficiency in experimental infections of mice and rats and pharmacokinetic characteristics. It was found that cyclacillin was not superior to ampicillin by its antibacterial action. By a number of characteristics it was even significantly inferior. The pharmacokinetic advantages of cyclacillin were not considered significant.


[Mechanisms of Proteus resistance to chloramphenicol]. Data on chloramphenicol sensitivity of clinical Proteus strains isolated within 1970--1975 and some mechanisms of their resistance to this antibiotic are presented. It was found that most of the Proteus strains (62.82 +/- 2.15 per cent) were resistant to chloramphenicol. 75 per cent of the isolates had resistance of transmissive character. Resistance of the Proteus cultures to chloramphenicol was not a stable feature and was lost during storage under laboratory conditions. Direct correlation between stability of the antibiotic resistance in the Proteus, the resistance level and the period of the culture storage was found. It was shown that the transmissive resistance to chloramphenicol in the Proteus cultures was due to synthesis of a highly active constituitive chloramphenicol-inactivating enzyme. Direct relation between the Proteus resistance level to chloramphenicol and the rate of the enzyme synthesis was noted. A number of the Proteus strains phenotypically sensitive to this antibiotic was capable of its inactivation. Still, the activity of the enzyme was low. The rate of the enzyme synthesis and the level of the acquired resistance in the chloramphenicol resistant mutants depended on the presence or absence of the enzyme in the cells of the initial sensitive strain. The capacity for chloramphenicol accumulation in a number of the chloramphenicol resistant mutants of the Proteus was decreased.


[Interrelationship between the serum lysozyme level and the leukocyte count in Sonne and Flexner dysenteries]. Interrelation between the level of the serum lysozyme and the count of the leucocytes, as well as the severity of the disease was found in testing of the peripheral blood of 133 patients with acute Sonnei and Flexner dysentery. It is recommended to use the lysozyme index in the clinical practice as one of the additional criteria for estimation of the severity of acute dysentery.


Biosynthesis of Saccharomyces cerevisiae glycoproteins: nature of some participating glycolipids. A particulate membrane preparation from Saccharomyces cerevisiae catalyzed the incorporation of mannose from GDP-mannose into lipids that were extractable in chloroform-methanol. One lipid has been previously characterized as dolichyl phosphomannose. Another one was purified by chromatography on silicic acid, DEAE-cellulose and Sephadex LH-20 was found to be alkali unstable. The lipid moiety was shown to be dolichol and the glycosydic part contained mannose, glucose and glucosamine. Radioactive mannose was also incorporated at a slower rate into more polar compounds. They were soluble in chloroform-methanol-water and were seen to liberate neutral oligosaccharides after alkaline hydrolysis. Radioactive mannose was also incorporated into substances which behave chemically as glycoproteins since they were insoluble in organic solvents, water and trichloroactic acid. Pronase treatment of the trichloroacetic acid-insoluble material released water-soluble oligosaccharides. When the particulate preparation which had been extracted with chloroform-methanol at-20 C, was incubated with GDP-(U-14C)mannose, radioactivity was incorporated into glycolipids that were soluble in chloroform-methanol-water and into glycoproteins. This result suggests that at least part of the mannose was transferred to endogenous acceptors independent of dolichyl phosphomannose.


L-Alanine as an end product of glycolysis in Saccharomyces cerevisiae growing under different hypoxic conditions. Saccharomyces cerevisiae growing under anaerobic or other hypoxic conditions releases L-alanine into the culture medium as an end product of glycolysis. Although the production of alanine is not as high as that of other fermentation products (ethanol, glycerol, succinic acid), consideration of the pathways leading to alanine in fermenting yeasts indicates that the release of alanine is advantageous to the cellular economy and may be considered as a safety device for excreting reducing equivalents derived from NADPH. No significant changes in the activity of alanine aminotransferase are found in the yeast when grown under different conditions.


Compression as a treatment of cancer, a historical survey. Compression in medicine goes far back in history. In ancient Rome and during the Renaissance compression by means of leaden plates was a well-known treatment of cancer. Compression flourished particularly in the first half of the 19th century, in Great Britain encouraged by the publications of Young and Arnott and Walshe, in France especially by the work of Récamier. Around 1850 this treatment fell into discredit: on the one hand because of improved surgical possibilities, on the other because not all the alleged cures concerned true malignancies. The results were, on the whole, disappointing. Quite recently, compression re-appeared as a treatment of carcinoma of the urinary bladder.


Low-dose heparin prophylaxis in herniorrhaphy? A prospective trial in bleeding complications. Low-dose heparin (L.D.H.) prophylaxis gives good protection against deep venous thrombosis (D.V.T.). In the case of subjects presenting for herniorrhaphy the literature is less unanimous regarding the chance of wound hematoma. In this prospective randomised, matched trial in 86 patients, a wound hematoma incidence of 36 percent was noted in the treatment group as against 7% in the control group. This is statistically significant at the P less than 0.001 level.


Photochemotherapy for psoriasis. A clinical cooperative study of PUVA-48 and PUVA-64. A clinical cooperative study involving 14 centers evaluated photochemotherapy (psoralen and high-intensity long-wave ultraviolet light [PUVA]) for psoriasis. Results from 465 patients treated with a PUVA-48 unit (equipped with 48 high-intensity UVA bulbs) and 110 patients treated with a PUVA-64 unit (equipped with 64 high-intensity UVA bulbs) confirmed the effectiveness of photochemotherapy for psoriasis. Clearing of psoriasis occurred in 85% of patients on PUVA-48 therapy. Mean number of treatments, joules per square centimeter, to clear, and total joules at clearing were similar to other reported trials. The plateau method of clearing resulted in lower joules per square centimeter at clearing, total joules per square centimeter, and number of treatments than the nonplateau method. Maintenance therapy groups were mainly M1 (once weekly) or M4 (no treatment for more than 60 days). No meaningful laboratory abnormalities were detected and ophthalmologic examinations showed a few abnormal results following PUVA. Short-term side effects were mainly erythema, nausea, and pruritus. The effectiveness and short-term safety of PUVA for psoriasis has now been confirmed by a second large cooperative study.


Juvenile dermatitis herpetiformis.

Juvenile dermatitis herpetiformis. Juvenile dermatitis herpetiformis occurred in a 20-month-old girl. She had granular lgA, C3, and fibrin bound to the basement membrane zone of the skin by direct immunofluorescence and negative serum antibodies against the skin on indirect immunofluorescence. The HLA typing of peripheral lymphocytes was A1, Aw30, B8, Bw51 without clinical evidence of malabsorption syndrome. A rapid improvement was observed on dapsone therapy. These findings strongly suggest that juvenile dermatitis hepetiformis is a disease entity different from chronic bullous dermatosis of childhood.


Immunopathology of psoriasis.

Immunopathology of psoriasis. Immunofluorescence (IF) studies by the direct and indirect methods demonstrate immunoglobulins and complement bound in vivo in psoriatic scales. The IF pattern is comparable to that of stratum corneum antibodies (SCAb) bound in vitro on specific substrate, as visualized by the indirect IF method. Formation of immune complexes can be responsible for the "squirting papilla" phenomenon, and conversion of the stratum corneum - which is normally an inaccessible antigen - into its reactive form seems to be brought about by proteases of polymorphonuclear leukocytes. Stimulation of protease production by polymorphonuclears appears to be an important factor in the pathogenesis of psoriasis. The stratum corneum of the epidermis is probably the target, and becomes an antigen for SCAb present in the circulation.


Treatment of rheumatoid arthritis with levamisole: long-term results and immune changes. We treated 29 rheumatoid arthritis patients with levamisole. on the basis of a 25% improvement in any 3 of 6 measurements 95% of the patients had a favourable response within 20 weeks. However, 64% of the patients discontinued levamisole by 40 to 60 weeks because of rash or secondary treatment failures. Delayed skin reactivity to streptokinase-streptodornase increased significantly in the entire treatment group, but there was in inverse correlation between skin test enhancement and clinical response. There was no overall change in lymphocytes response to phytohaemagglutinin (PHA) after 4 and 16 weeks of treatment, but seven patients with enhanced lymphocyte responsiveness to PHA experienced an earlier clinical response to levamisole. Treatment with levamisole frequently results in clinical improvement in rheumatoid arthritis, but this is not clearly related to a stimulatory effect on cell-mediated immunity. Its long-term usefulness may be limited by a high incidence of relapse and rash.


Diflunisal in osteoarthrosis of the hip and knee. Diflunisal (750 mg per day) has been compared with acetylsalicylic acid (ASA) (3000 mg per day) in the treatment of osteoarthrosis of the hip and knee in a double-blind, randomised, multicentre, outpatient study. Thirty-one patients entered the diflunisal group and 29 the ASA group. The response of the 2 groups was comparable, but the incidence of side effects was higher in the ASA group. At the end of the 12-week period more patients in the diflunisal group chose to remain in a further, open study.


Indomethacin in rheumatoid arthritis: clinical effects, pharmacokinetics, and platelet studies in responders and nonresponders. Twenty patients with definite or classical rheumatoid arthritis entered and completed a sequential study of placebo for 1 week, oral indomethacin 25 mg 3 times a day for 3 weeks, and oral indomethacin 25 mg 3 times a day plus 100 mg indomethacin suppository at night for 3 weeks. Twelve of the patients had previously been classified as responders and eight as nonresponders to indomethacin by an independent assessor. At the end of each period patients were assessed by a blind observer for duration of morning stiffness, pain score, digital joint size, grip strength, articular index, analgesic tablet usage, and the patient's own overall global assessment and comparative global assessment. In 8 of the 9 tests used responders improved on indomethacin in comparison with placebo, while nonresponders did not improve. There were no significant differences between responders and nonresponders in the plasma half-life, plasma clearance of indomethacin, protein binding of indomethacin, or urinary excretion of free or conjugated indomethacin. There were no significant differences between responders and nonresponders in the urinary excretion of 7HDPA or in the platelet aggregation or platelet malonyldialdehyde production tests. In responders there was a significant positive correlation between the plasma indomethacin concentration (r=0.44, P<0.05) and the percentage inhibition of malonyldialdehyde production by the platelets. However, in nonresponders this correlation, while significant (P<0.05), was negative (r=-0.498). Both for responders and nonresponders there was a significant correlation between plasma indomethacin concentration and the percentage reduction in 7HDPA. There was no correlation between the clinical response and the plasma concentration of indomethacin. There appears to be a biochemical difference between responders and nonresponders, which, while not necessarily causally linked with the clinical response to indomethacin, is worthy of further study.


Naproxen in juvenile chronic polyarthritis. Naproxen at 10 mg per kg body weight was compared with aspirin at 80 mg per kg body weight in children suffering from juvenile chronic polyarthritis. It was found to be as effective as aspirin, with certainly no more and possibly fewer gastrointestinal side effects. A long-term tolerance study up to 12 months confirmed that naproxed was a satisfactory nonsteroidal anti-inflammatory drug in the management of various types of juvenile chronic arthritis.


How long should long be? Long-term trials in rheumatic diseases. In the interests of the safety of patients it is necessary to collect long-term clinical data. It is often assumed that the longer the trial the better. However, the longer the trial the more scientific compromises are necessary if it is to be carried out in a practical way. Because the therapeutic efficacy of and tolerance to the nonsteroidal anti-inflammatory drug diclofenac was established after 6 months' trial the optimal duration for a comparative long-term trial of this preparation is 6 months. The small amount of information gained from trials of diclofenac longer than this did not justify the reduction in quality of trial design and performance that they required. Thus it would appear that at least for 1 antirheumatic drug the assumption that 'the longer the trial the better' is not true.


Experimental pulmonary edema. The effect of unilateral PEEP on the accumulation of lung water. The effect of PEEP in retarding the development of pulmonary edema produced by elevation of left atrial (LA) pressure to 25cm water was studied in dogs, Using two synchronized volume respirators connected to a double lumen endotracheal tube, 10 or 25cm H2O PEEP was applied to one lung while the contralateral lung was ventilated with an equal tidal volume without PEEP. LA pressure was then elevated by inflating a Foley catheter ballon in the LA until the desired LA pressure was reached. After three hours of pulmonary edema with ventilation of one lung with PEEP lung water was quantitated by wet-to-dry weights. There were no differences in wet to dry weights of PEEP and nonPEEP lungs at either 10 or 25cm H2O level. Additional blood flow studies showed that 25cm H2O of PEEP reduced blood flow to the PEEP lung by 25% and to the nonPEEP lung by 7%. This study shows that PEEP will not mechanically retard the accumulation of lung water due to increased pulmonary capillary hydrostatic pressure.


Factors determining the mortality and morbidity in hepatic injuries. Analysis of 108 cases. The experience with 108 cases of hepatic injuries was reviewed retrospectively and a classification based on the severity of the hepatic injury was used to group these patients accordingly. Using this criteria, the factors determining the observed morbidity and mortality were analyzed. Hepatic injuries amenable to relatively simple surgical treatment were found in 72 patients (67%). Nine deaths occurring in this group were attributed to the commonly encountered associated injuries inside and outside the abdomen. Moderately severe imjuries requiring suturing of the hepatic parenchyma and ligation of injured vessels and bile ducts were seen in 15 patients (14%). Two of these patients died. Severe hepatic injuries resulting in major blood loss requiring partial hepatectomy were seen in 21 patients (19%). Seven of the eight deaths (7.4%) attributable to the hepatic injury occurred in this group. As noted, ten other patients died from causes other than the hepatic injury for an overall mortality of 16.7%. A pre-established patient management program based on the classification of hepatic injuries is proposed as a method for improving the management of patients with hepatic injuries.


Presidential address: the woman in the case. Jane Todd Crawford, 1763--1842. The 1978 Presidential Address of the Southern Surgical Association is dedicated to the wives of the members, past and present, in acknowledgment and appreciation of the enduring contribution that they have made to the quality and character of this association.


The role of HLA tissue matching in cadaveric kidney transplantation. The role of HLA match in donor--recipient selection has been studied in 271 patients who received cadaver transplants during a 10-year period. This series included 36 four-antigen matches, 181 three-antigen matches and 54 two-antigen matches. Our results support the concept that better results can be expected when better matched kidneys are utilized for transplantation.


A diagnostic approach to the adult with fever of unknown origin. The evaluation of the condition of a patient with fever of unknown origin requires a knowledge of those disorders that produce this syndrome, an awareness of the potential significance of subtle findings in the history and physical examination, and an appreciation of the value in this clinical setting of specific diagnostic procedures. In this report, we review these aspects of fever of unknown origin and outline a diagnostic approach to the persistently febrile patient.


Albuterol and isoproterenol in bronchial asthma. Efficacy and toxicity of drugs administered via intermittent positive pressure breathing. This study compared the efficacy and side effects of 1.25, 2.5, 5, 10, and 15 mg of albuterol and isoproterenol hydrochloride administered by intermittent positive pressure breathing (IPPB) to 12 patients with reversible airway obstruction. Equal doses of the two medications induced similar peak increases in pulmonary function, but the increase following albuterol persisted longer. The degree of bronchodilation was impressive; 15 mg of albuterol induced a mean increase over six hours of 82% in the forced expiratory volume in one second. Significant cardiovascular side effects were more common after isoproterenol than after albuterol. Albuterol is superior to isoproterenol as a bronchodilator when administered by IPPB because, for a given peak bronchodilation, cardiovascular side effects are fewer and bronchodilation persists longer with albuterol. The optimal dose of isoproterenol hydrochloride is 2.5 to 5.0 mg and the optimal dose of albuterol is 10 mg when these drugs are given by IPPB.


Oropharyngeal candidiasis treated with a troche form of clotrimazole. A randomized double-blind trial was conducted to assess the local effectiveness and safety of a troche form of clotrimazole in the treatment of oropharyngeal candidiasis in cancer patients. One half of the patients received one 10-mg troche and the other half received one 50-mg troche, five times a day for two weeks. Clinical cures were observed in 50 episodes, resulting in a cure rate of 96%. The median duration of oropharyngeal candidiasis after the start of therapy was three days in those treated with the 50-mg troche and four days in those who had received the 10-mg troche. Side effects were minimal, and only one patient experienced nausea and abdominal pain. Both the 10-mg and 50-mg troches appear to be efficacious and safe, but the 50-mg dose may be preferable because it is somewhat more effective without additional toxicity.


High blood pressure. A side effect of drugs, poisons, and food. Arterial hypertension, either transient or persistent, may be induced or aggravated by ingestion of various chemical agents, such as drugs, poisons, and food. Most of these agents either cause sodium retention and expand extracellular fluid volume or act as direct or indirect sympathomimetics. Others act directly on arteriolar smooth muscle. For a few agents, no precise mechanism has been ascertained. Hypertensive reactions may also occur as a result of drug interactions or food and drug interactions. In addition, paradoxical increases in pressure may be encountered during or after discontinuance of antihypertensive therapy. In general, these pressure increases are small and transient; however, a few have been associated with severe hypertension involving encephalopathy, strokes, and irreversible renal failure. Careful review of a patient's drug regimen, including over-the-counter preparations, may avoid chemically induced hypertension. Identification of any offending or incriminating agent will prevent the labeling of a chronic illness and obviate the need for lifelong antihypertensive therapy.


Pulvomycin, an inhibitor of prokaryotic protein biosynthesis. Antibiotic 1063-Z isolated from culture fluids of Streptoverticillium mobaraense was identified as pulvomycin. Pulvomycin was observed to inhibit protein biosynthesis in growing cells of Bacillus brevis. The poly(U)-directed poly(Phe) synthesis in cell-free systems of Bacillus brevis and Escherichia coli was highly susceptible to the antibiotic. Pulvomycin did not affect the transfer of Phe to tRNA. The results suggest that the target of pulvomycin action is the polypeptide chain elongation.


[Determination of fetal infectious risk by bacteriological examination of the amniotic fluid after premature rupture of the membranes]. Amniotic fluid was obtained per vagina from 228 mothers with premature rupture of the membranes and examined bacteriologically. The aim was to assess the importance of amniotic fluid contamination and the risk of foetal infection in the absence of systematic antibiotic therapy in the mothers. The incidence of amniotic fluid contamination was apparently greater in those mothers who had received antibiotics although the proportion of neonates with a true infection (3%) was almost identical. The absence of selection of resistant organisms is indicated by the marked preponderance of streptococci and the scarcity of Gram negative enterobacteriacae. Thus it is reasonable not to give systemic antibiotics but to culture the amniotic fluid. Infection can then be anticipated and in affected neonates the appropriate therapy started immediately.


Clozapine, chlorpromazine, and placebo in newly hospitalized, acutely schizophrenic patients: a controlled, double-blind comparison. Clozapine is a unique compound belonging to a relatively new group of antipsychotic agents, the dibenzazepines. To our knowledge, the present study represents the first double-blind, controlled comparison recorded in the United States. The data suggest that clozapine in the present population of newly admitted, acutely psychotic schizophrenic individuals, and in the doses employed, was more effective in overall improvement response, discharge rate, and ameliorating discrete symptoms across the different objective rating scales used than was chlorpromazine (Thorazine) hydrochloride. Placebo was ineffective. Unlike chlorpromazine, no extrapyramidal reactions occurred in those patients ingesting clozapine. Clozapine was also beneficial in reversing abnormal involuntary motor movements. It is an excellent anxiolytic and hypnotic agent. Sedation, hypotension, and hypersalivation are among the more common side effects observed.


Application of a combined rosette-fluorescent test in studies on leukemia cells. In 13 healthy blood donors and 42 patients with leukemia and lymphosarcoma, the combined rosette-fluorescent test allowed simultaneous evaluation of peripheral blood lymphocytes and leukemia cells for presence of membrane immunoglobulins and sheep erythrocyte receptors. In healthy subjects, the mean value for T cells was 68 +/- 9%, for B cells 16 +/- 5%, for nonreceptor cells 15 +/- 8%, and for cells possessing both markers 0.6 +/-0.8%. In cases of acute leukemia, nonreceptor cells predominated (59 +/-24%), and values for T cells were usually low (19 +/- 13%). In chronic lymphocytic leukemias and lymphosarcoma, B and "null" cells usually predominated more or less. In all patients values for double-receptor cells were low. The combined test proved to be a convenient method for simultaneous evaluation of both markers. It is also useful for recognizing blastic leukemia cells devoid of both receptors and for determing values for T and B lymphocytes.


A one-stage test for simultaneous study of migration inhibition factor of peripheral leukocytes (LIF) and migration inhibition factor of mouse spleen cells (MSIF). A one-stage modification of the test of the migration inhibition is described, which permits stimultaneous study of the factor influencing in vitro migration of peripheral blood leukocytes and mouse spleen cells. The results of this correlate with the results of the two-stage test.


The influence of daunomycin on development of Yoshida sarcoma after nonspecific stimulation of the immunologic system. Injection of BCG in rats inoculated with Yoshida sarcoma had no influence on the development of the tumor or survival time of the rats in comparison with the control group. On the other hand, survival time was increased more than threefold in a group of rats injected with daunomycin in a single dose of 1 mg/kg i.p. Longest survival and lowest mortality were observed in a group of rats treated with daunomycin and BCG, in which survival time was six times longer than in the controls, and mortality after 50 days was 30% lower, and after 100 days even 50% lower than in the group of rats that were treated only with daunomycin.


Effect of freezing conditions in liquid nitrogen on biological properties of Treponema pallidum. The influence of various conditions of freezing in liquid nitrogen on the motility, virulence, antigens, and immunogenicity of Treponema pallidum was studied. The suspending medium, rate of freezing, kind and concentration of cryprotector, and duration of preincubation with cryoprotector were found to be critical. On the basis of the results obtained the optimal conditions of freezing of T. pallidum in liquid nitrogen were established. These are: the Nelson-Diesendruck medium with the addition of 10% DMSO; freezing rate 1 degree per min, and thawing rate between 12.6 degrees and 120 degrees per min.


Effect of immunosuppressive agents on the course of experimental syphilis in rabbits. Rabbits infected intradermally with T. pallidum at four sites on the clipped back were treated with cyclophosphamide, 1 mg/kg body weight, twice before and every two days after inoculation for five months, or 5-fluorouracil 2.5 mg/kg body weight, according to the same regimen. Humoral response, as measured repeatedly by VDRL, FTA, and hemagglutination tests, was not affected by the application of the immunosuppressors. In contrast, the cell-mediated response, as determined by the Macrophage Migration Inhibition test was completely abolished in the treated rabbits. In the group of rabbits which were given immunosuppressors, syphilitic lesions appeared not only at the challenge sites, as usual, but also multiple multiform lesions developed over the entire dorsum after healing of the primary lesions. Taking into account the fact that abolishment of the cell-mediated response led to an aggravated course of the syphilitic infection, the results of these experiments were interpreted as indicating the protective role of the cell mediated response in syphilis.


Isolation and characterization of low molecular weight antibodies for Treponema pallidum. A low molecular weight antibody which coated Treponema pallidum without producing any apparent reaction was isolated from human syphilitic serum. Its electrophoretic mobility was that of IgG immunoglobulins, and molecular weight was between 115,000 and 120,000.


Acute rejection episodes and long-term transplant function. From 1974 to 1976, 114 kidneys transplanted to 108 patients underwent acute rejection episodes. With repeated rejections there was an increased likelihood of recovery without subsequent rejections, eg, 59% of kidneys that had three rejections regained prolonged good function. Repetitive rejections did not increase mortality. Discriminate analysis of the best serum creatinine level or creatinine clearance within 30 days of rejection and the time interval between rejection episodes correctly predicted the outcome of second rejections in 20/25 cases and correctly predicted the outcome of third rejections in 23/29 cases. Treatment of a third rejection was successful in 70% of kidneys if the best serum creatinine level was less than 2 mg/dL after the second rejection and was successful in 77% of kidneys if the time interval between second and third rejections was longer than one month.


Hepatic dysfunction during isoniazid chemoprophylaxis in renal allograft recipients. Hepatitis is a frequent complication of dialysis and renal transplantation; therefore, the occurrence of drug hepatotoxicity is an additional important consideration in renal allograft recipients. Azathioprine, needed for immunosuppression, and isoniazid, used for antituberculous chemoprophylaxis, are both potentially hepatotoxic. A retrospective study of 119 patients who received 126 renal allografts was done to estimate the probable incidence of isoniazid-related hepatic dysfunction. All patients in this series were administered isoniazid chemoprophylaxis. Posttransplantation hepatitis developed in 13 patients. Circumstantial evidence supported a presumptive diagnosis of isoniazid hepatotoxicity in three recipients. We concluded that routine isoniazid chemoprophylaxis is not justified in renal allograft recipients based on the probability of hepatotoxicity as contrasted to the infrequent occurrence of tuberculosis.


[Morphology of the pancreas in peptic ulcer and the complications of stomach resection]. Morphological examinations of 115 biopsies of the pancreas showed the peptic ulcer and postgastroresection complications not to be the leading factors in the etiology of the pancreatic pathology. Gastric resection does not favour the affection of the pancreas but may aggravate the existing morphological lesions. Despite the lack of crude histological changes in the pancreatic gland in peptic ulcer, ultrastructural signs of a high functional intensity of acinary cells were found which may also be regarded as pathological manifestations. Most marked changes of the histostructure of the pancreatic gland were found in peptic ulcer of the gastroenteroanastomosis. A difference in the A-cell content of pancreatic insulae before and after gastric resection was found. As the pancreatic tissue is frequently intact in the afferent loop syndrome, the leading role of duodenostasis in chronic affection of the pancreatic gland may be disputed.


Tapia's syndrome. The erratic evolution of an eponym. The syndrome first described in 1904 by the Spanish otolaryngologist, Antonio Garcia Tapia, has been variously interpreted by subsequent authors such that there is little current agreement as to the site of the lesion responsible for the condition or the specific symptoms included in this disorder. The confusion arose in part because Tapia's original patient had associated neurologic findings. Careful review of Tapia's reports reveals (1) that he regarded the syndrome as consisting of ipsilateral hemiplegia of the larynx and tongue with normal function of the soft palate and (2) that he believed the lesion resulting in these signs was outside the CNS.


Treatment of Duchenne's muscular dystrophy with penicillamine. Results of a double-blind trial. Eleven boys with Duchenne's muscular dystrophy, randomly assigned to placebo (group A, n = 6) or penicillamine treatment (group B, n = 5), received three capsules per day containing lactose or 250 mg of penicillamine. All patients received pyridoxine, 50 mg daily. Mean age at entrance into study was similar for both groups (group A , 86.7 +/- 31.6 months; group B, 95.4 +/- 43.4 months). Clinical status was assessed with timed functional activities, manometric measurements of muscle force, and manual muscle testing. After 14 to 16 months of treatment, statistical analysis (analysis of variance) of data disclosed no significant differences in the overall performance of the two groups. A longer-duration trial, involving younger patients, is needed to determine whether there are palliative effects of penicillamine or other potentially therapeutic agents.


Effects of cerebellectomy on eye movements in man. A patient with a right hemicerebellectomy and recurrence of a tumor on the left cerebellar hemisphere was examined regarding oculomotor and vestibuloocular functions. The findings were as follows: inability to maintain eccentric gaze; defective smooth pursuit system; failure to suppress by visual fixation the nystagmus induced by sinusoidal horizontal rotations; moderate increase of the vestibulo-ocular reflex gain when rotated in the dark; relative preservation of the slow phases of the optokinetic nystagmus, as compared to the slow phases induced by tracking a pendulum; decreased gain of the vestibulo-ocular reflex when rotated with the lights on fixating the environment; square wave jerks; and no saccadic dysmetria.


Comparative trial of valproate sodium and clonazepam in chronic epilepsy. A crossover comparative study of valproate sodium and clonazepam in the treatment of 32 adult epileptic patients receiving multiple drug therapy is described. Serum concentrations of other anticonvulsant drugs were unchanged by the addition of clonazepam. However, patients receiving high doses of other anticonvulsant drugs had lower serum concentrations of clonazepam (p less than .01). With valproate sodium, phenobarbital concentrations increased (P less than .05) in patients receiving phenobarbital but not significantly in patients receiving primidone. Phenytoin concentrations were reduced (P less than .05) during treatment with valproate sodium. Both drugs significantly reduced the frequency of minor seizures, with valproate sodium having the greater effect. However, it is important to monitor serum concentrations of other anticonvulsant drugs during treatment with valproate sodium since changes in these may influence seizure control or cause side effects.


Limulus lysate assay for early detection of certain Gram-negative corneal infections. The limulus endotoxin assay has been previously demonstrated to be the most sensitive method available for detection of bacterial endotoxin. A commercially available form of limulus amoebocyte lysate was used in this study for detection of Gram-negative corneal infections in both experimental animals and in a group of nine patients. The limulus assay enabled rapid detection of Gram-negative infections in both the experimentally induced ulcers in rabbits and in the patients studied. False-positive reactions did not occur in corneal infections due to either Gram-positive bacteria, fungi, or herpes simplex keratitis. The limulus test proved to be more sensitive than examination of Gram-stained smears of corneal scrapings and became positive earlier than bacterial cultures. The limulus test was helpful in the diagnosis of partially antibiotic-treated corneal infections but could not be used to assess the response to antimicrobial therapy, since endotoxin persisted in the corneal scrapings for some time after initiation of therapy.


Corneal ulceration secondary to intestinal Shigella sonnei. A 34-month-old boy had a corneal ulcer due to Shigella sonnei. To my knowledge, this is the first documented case reported where the same organism was recovered from the stool. One other case of S sonnei and two cases of S flexneri corneal ulcers have appeared in the literature. All cases have been in young black boys between the ages of 23 months and 6 years. The exact pathogenesis remains unclear, but direct fecalocular inoculation into a preexisting epithelial defect, a probable herpetic dendrite in this case, is most likely.


Bone formation in a periosteal graft.

Bone formation in a periosteal graft. Bone formation occurred in periosteal graft that was used to anchor a Cardona keratoprosthesis.


Vertical mattress suture for strabismus resection operation. The suturing technique for strabismus resection has been simplified with the use of a single double-armed suture in the form of a vertical mattress. Synthetic ophthalmic suture material has made this technique still more desirable because of the tensile strength and tissue-gripping characteristics of these materials.


Slit-lamp attachment for examination of donor corneas in McCarey-Kaufman medium. An attachment permits slit-lamp examination of excised donor cornea in McCarey-Kaufman medium. The bottle that contains the excised cornea is placed in a holder and examined with the slit lamp by means of a mirror placed at a 45 degree angle. This mirror reflects the slit beam up through the bottom of the bottle. This allows accurate examination of the donor cornea in a stable manner that was not previously possible.


Determinants of graft clarity in penetrating kerotoplasty. A total of 240 consecutive penetrating keratoplasties with a minimum six months' follow-up were examined to determine the influence of preoperative prognosis, donor age, death-enucleation time, death-surgery time, and surgical experience on graft clarity. Graft results, classified as clear, nebulous, or edematous, were compared within two prognostic categories in aphakic, combined, and phakic keratoplasty groups. Preoperative prognostic groups and surgical experience appeared to be most important in affecting graft clarity, whereas the donor age, death-enucleation time, and death-surgery time seemed to have little effect.


Aponeurotic ptosis surgery.

Aponeurotic ptosis surgery. The recognition of defects in the levator aponeurosis associated with a renewed interest in eyelid anatomy has led to a revival of aponeurotic surgery. We describe our approach with emphasis on techniques that help obtain and maintain avascular planes throughout surgery. The advantages of this approach include preservation of (1) tear-producing structures, (2) Müller's muscle and Whitnall's ligament, and (3) normal anatomical planes and structures of the eyelid. Our results indicate that this technique is the procedure of choice for acquired ptosis cases. It also gives good results in congenital ptosis cases with at least 5 mm of function. Overcorrection at surgery is necessary in all cases. Tucking of the aponeurosis is to be avoided as no raw healing surfaces are obtained. Results in 60 eyelids with a minimum follow-up of one year are presented.


Mass culture of human corneal endothelial cells. Human corneal endothelial cells have been grown in mass culture for the first time to our knowledge. Cultures from donors who were under 20 years of age grew well. Cells from older donors were difficult to grow or did not grow at all. Initial outgrowth usually began one week after explantation. After an initial lag phase, mass increased by 15% weekly, and confluency was approached in eight to nine weeks. The lag phase of growth correlated with the age of the donor. A basal lamina was secreted by human corneal endothelial cells in culture. Chromosomal counts demonstrated a modal number of 46. With this technique, sufficient human corneal endothelial cells may be obtained for biochemical investigation.


Origin of the retrocorneal membrane in the rabbit. Electron microscopic studies have offered presumptive evidence that the origin of both cellular and extracellular components of the retrocorneal membrane (RCM) derive from corneal endothelial cells. In an attempt to demonstrate conclusively the origin of the cell in the RCM, we performed 8-mm exchange, penetrating keratoplasties between male and female rabbits. After allowing for complete healing and using only clear corneas, we produced central RCM by freezing the central cornea after inducing intraocular inflammation. Care was taken to ensure that the membrane was surrounded by normal donor endothelium and Descemet's membrane was intact. Sex chromatin counts of the cells in the RCM of a female host with a male graft showed an average of 2% Barr bodies. Cells in the membrane of a male host with a female graft showed an average of 40% Barr bodies. The experimental and control sex chromatin counts were almost identical. This study provides conclusive evidence that central retrocorneal, fibroblast-like cells and membranes in rabbits are derived from corneal endothelial cells.


The maxillo-septal syndrome and temporomandibular joint dysfunction. The results of twenty patients treated for the maxillo-septal syndrome and associated temporomandibular joint dysfunction are reported as well as the method of treatment. The treatment plan included rapid maxillary expansion with either a fixed or removable appliance, retention of the increased maxillary width with a Hawley retainer, simultaneous increase in mandibular width with a Crozat appliance. Finally the occlusion was equilibrated. Certain cases are discussed in detail and maxillary expansion was usually restricted to between 1 . 0 and 2 . 0 mm.


Protection of mice against plasmodium and babesia infections: attempts to raise host-protective sera. In an attempt to generate large numbers of mice resistant to Plasmodium berghei and Babesia rodhaini to be used as donors of antibody-secreting cells for hybridoma production, various methods of inducing resistance to repeated challenge with infected blood cells have been explored. Although results of independent experiments varied markedly, prior injection of CBA/M mice with BCG, and prior infection of BALB/c mice with Plasmodium yoelii, were found to be manipulations capable of inducing resistance to P. berghei. A single dose of serum, harvested from resistant mice challenged several times with P. berghei, could transfer resistance against P. berghei to a proportion of naive CBA/H recipients. Although resistance to multiple B. rodhaini challenge could be induced in mice, in no situation was a host protective effect of a single high dose of serum demonstrated in naive recipients.


Transfer of mouse IgG2 production by IgM-bearing spleen cells separated by a fluorescence-activated cell sorter. The purpose of the present study was to determine whether at least some splenic B lymphocytes can switch from the synthesis of one isotype of immunoglobulin to another during B cell differentiation. The experimental system invovled the transfer of characterized cell suspensions between allotype congenic strains of mice followed by analysis in the recipient for donor type immunoglobulin production. Donor splenic lymphocytes were incubated with specific fluorescent labelled anti-mu antiserum and passed through the Los Alamos fluorescence-activated cell sorter; mu-depleted cell suspensions were transferred into sublethally irradiated congenic recipients and the amount of donor type immunoglobulin of IgG2 type was measured at weekly intervals. The results demonstrated taht at least some cell bearing membrane bound IgM can differentiate in vivo into IgG2-secreting cells, although not all IgG2-secreting cells have been recently derived from IgM positive precursors.


Induction of long-term tolerance in the quokka (Setonix brachyurus) by thymus and skin allografts into early pouch young. Intact quokkas (Setonix brachyurus) were grafted with thymus and skin as neonates. Fifty seven percent of primary thymus grafts persisted when donors were less than 32 days of age, compared with only 20% when the dornors were juveniles. This suggested that neonatal tissue was more readily accepted than tissue from adult animals. When the recipients had left the pouch, induced tolerance was tested by grafting thymus and skin from the original donors. Second thymus grafts were generally unsuccessful; however, second skin grafts were accepted and remained intact over the three-year period of observation, provided that the hosts were less than about 40 days of age at the time the first graft were placed.


Antibacterial activity of murine spleen cell extract. Exposure to a peptide extract from murine spleen reduced the viable count of E. coli and E. coli spheroplasts. The reduction in viable count was reflected by reduced incorporation of radiolabelled thymidine and was apparently due to a loss of the ability of treated ogranisms to divide.


Convolution analysis of transcription by yeast DNA-dependent ribonucleic acid polymerase A. A mathematical method for studying ribonucleic acid chain elongation. The rate of initiation of RNA synthesis catalysed by yeast RNA polymerase A on native calf thymus DNA decayed exponentially with a half-life of about 4.3 min. The rate constant for initiation was unaffected by preincubating the enzyme with DNA, or by decreasing the concentration of GTP 4-fold. The rate of RNA synthesis was constant for 15--20 min and then decreased. Each enzyme molecule made no more than one RNA molecule. In this situation, initiation, elongation and total RNA synthesis are related by a convolution integral. Solution of the convolution integral revealed that the rate of elongation was apparently biphasic. Analysis of the size of the RNA product showed that this biphasic profile arose because most but not all of the enzyme stopped RNA synthesis soon after initiation.


Effects of collagen, ionophore A23187 and prostaglandin E1 on the phosphorylation of specific proteins in blood platelets. Human platelets that had been preincubated with 5-hydroxy[(3)H]tryptamine and [(32)P]P(i) were stirred with various agents; the secretion of 5-hydroxy[(3)H]tryptamine from platelet granules and the radioactivity of platelet [(32)P]phosphopolypeptides separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis were then measured. Exposure of the platelets to collagen fibres or ionophore A23187 selectively increased the phosphorylation of polypeptides with apparent mol.wts. of 47000 (P47) and 20000 (P20) by approx. 3-fold, in association with the release of 5-hydroxy[(3)H]tryptamine. The 47000-mol.wt. phosphopolypeptide (P47) was clearly separated from platelet actin by the electrophoresis system used. Prostaglandin E(1), which inhibits platelet function by increasing platelet cyclic AMP, decreased the phosphorylation of polypeptides caused by collagen as well as the release of 5-hydroxy[(3)H]tryptamine. Prostaglandin E(1) also selectively increased the phosphorylation of distinct polypeptides with apparent mol.wts. of 24000 (P24) and 22000 (P22) by approx. 2-fold. As the phosphorylation reactions caused by collagen are probably mediated by an increase in Ca(2+) concentration in the platelet cytosol and may have a role in the release reaction [Haslam & Lynham (1977) Biochem. Biophys. Res. Commun.77, 714-722; (1978) Thromb. Res.12, 619-628], we suggest that a cyclic AMP-dependent phosphorylation of the 24000- and/or 22000-mol.wt. polypeptides caused by prostaglandin E(1) may initiate processes that decrease the Ca(2+) concentration in the cytosol, so inhibiting both the Ca(2+)-dependent phosphorylation reactions and the release reaction. Treatment of platelets with prostaglandin E(1) did not inhibit the increased phosphorylation of polypeptides with apparent mol.wts. of 47000 and 20000 (P47 and P20) caused by ionophore A23187, which may therefore short-circuit cyclic AMP-dependent mechanisms that decrease the Ca(2+) concentration in the platelet cytosol. As prostaglandin E(1) did inhibit the release of 5-hydroxy[(3)H]tryptamine by ionophore A23187, cyclic AMP may also inhibit the release reaction by additional mechanisms.


Association of IgG anti-brain antibodies with central nervous system dysfunction in systemic lupus erythematosus. Sera from 20 patients with systemic lupus erythematosus (SLE) and active central nervous system (CNS) dysfunction were examined by indirect immunofluorescence for antibodies to neuronal membrane determinants. Warm-reactive IgG antibodies were demonstrable in 82% (9/11) of patients with clinical evidence for seizures or diffuse CNS disease, but these antibodies generally were absent in non-CNS SLE sera or when focal neurologic deficit or psychosis was the primary CNS manifestation. Cold-reactive antibodies of the IgM class were equally prevalent in patients with or without CNS disease and appeared to be more directly correlated with extra-CNS systemic illness. Absorption experiments with lymphocytes, brain homogenate, and various other tissues suggested a predominant brain-specificity for IgG antibodies and partial lymphocyte cross-reactivity for IgM antibodies. Interpretations of this special association between IgG anti-brain antibodies and diffuse CNS dysfunction in SLE are discussed.


Circulating immune complexes in mixed connective tissue disease. Circulating immune complexes were measured by three assays (monoclonal rheumatoid factor radioimmunoassay, C1q-binding assay, Raji cell radioassay) in sera from patients with mixed connective tissue disease. Evidence for circulating immune complexes was found by at least one method in 94% of sera. The highest frequency of positive results was noted by the Raji cell radioassay (88%). However, a significant number of patients also had immune complexes measured by the C1q-binding assay (50%) and monoclonal rheumatoid factor radioimmunoassay (45%). Further, the levels of immune complexes are measured by the Raji cell radioassay, and to a lesser extent the C1q-binding assay, correlated with disease activity and represent a potentially useful clinical tool. The pattern of reactivity in mixed connective tissue disease sera differed from rheumatoid arthritis, scleroderma, and polymyositis. This suggests that a spectrum of immune complexes is found in these rheumatic diseases and supports the concept that mixed connective tissue disease is distinct from these diseases.


Cardiopulmonary manifestations of progressive systemic sclerosis: associations with circulating immune complexes and fluorescent antinuclear antibodies. Sixteen patients with progressive systemic sclerosis were evaluated for cardiopulmonary manifestations of their disease and for serologic immune abnormalities. Twenty-five percent of patients had abnormal echocardiograms, and 81% had a significant reduction in pulmonary function by spirometry. Circulating immune complexes (IC) were detected in 44% of patients by using a fluorescent Raji cell assay, and these patients were more likely to have an abnormal echocardiogram (P less than 0.02). Seventy-five percent of the patients had fluorescent antinuclear antibodies (FANA) and these patients were more likely to have pulmonary disease (P less than 0.01).


Scleroderma after cosmetic surgery: four cases of human adjuvant disease. In 9 cases of human adjuvant disease, 4 cases of scleroderma (3 progressive systemic sclerosis and 1 localized morphea) were observed. Seven to 19 years after injection of foreign substances into the breasts or nose for cosmetic purposes, some patients developed human adjuvant disease. In one case the foreign substances were removed by bilateral mastectomy with no discernible effects on the patient's clinical course. Histopathologic findings of the removed breasts demonstrated foreign body granulomas with calcification. The injected substance was identified as a mixture of liquid and solid paraffin. Human adjuvant disease might be caused by prolonged hypersensitization activated by the injected foreign materials which act as an adjuvant.


[Examination of leg volume of patients for determining the therapeutic effect of 1-phenylephrine hydrochloride (author's transl)]. Distorsions and other injuries of a more or less grave kind are often found in combination with exudative processes. The anti-exudative effect of adequate drugs has so far been tested only on the rat paw oedema. As results gained from animal tests cannot immediately be applied to man, tests with human subjects were carried out. With a device by Diebschlag it has become possible to fulfil the demands of a simple but still exact measuring process for quantification of leg volumes. 12 subjects with a distorsion of the ankle joint were treated with a gel (active substance: 1-phenylephrine HCl) which reduces the volue of the leg and with a similar placebo gel, and the effects of both treatments were compared and examined. The analysis of data yielded statistically highly significant values for the effectivity of the active substance gel.


[Prolongation of the mitotic life span of diploid human glia cells in a quantitative cell culture system by centrophenoxine (author's transl)]. The accumulation of lipofuscin in postmitotic and reversible postmitotic cells of animals and man is an age correlated process. The mechanism of the lipofuscin accumulation and the function of lipofuscin in the aging cell is not fully understood. The accumulation of lipofuscin in vivo and in vitro can be slowed down by the action of centrophenoxine (Helfergin). Diploid cells are the only reversible postmitotic cells of man that have a genetically determined limited cell division capacity and accumulate lipofuscin in the process of the cellular aging in a quantitative cell culture system in vitro. The treatment of diploid human glia cells with centrophenoxine results in increasing the cell division capacity by 30--40% in vitro. The data demonstrate that the centrophenoxine induced inhibition of lipofuscin accumulation has a positive influence on the cell metabolism and the mitotic division capacity and causes a delay of the cellular aging of the human glia cells in vitro.


Sisomicin in surgical infections. Laboratory and clinical studies. A clinical and bacteriological study to evaluate the effectiveness and toxicity of sisomicin in the treatment of surgical infections is described. Fifteen patients received i.m. injections of 150 or 225 mg/day for seven days. The daily doses was divided into three applications. The mean serum levels at the first hour were 5.26 +/- 2.41 microgram/ml and at the seventh hour 1.0 +/- 1.19 microgram/ml. Similar values were determined on the seventh day of treatment with sisomicin: 6.36 +/- 2.36 microgram/ml one hour, 1.17 +/- 1.67 microgram/ml seven hours after i.m. injection. All the microorganisms isolated had MICs below the sisomicin levels measured. The clinical results were assessed as "excellent" in 11 patients. Therapeutic results were considered as "fair" in three patients. The clinical conditions of one patient affected by osteomyelitis following osteosynthesis of the femur was not improved by treatment. No side effects were observed.


[Long-term study on a new beta2-sympathicomimetic agent and a combination of an oral theophylline-ethylenediamine with retard effect and hexoprenaline (author's transl)]. The effects of a treatment with a new beta2-adrenergic agent (hexoprenaline, Etoscol) as well as a combined therapy of theophylline (Euphylin retard) with the Etoscol on the total resistance were studied on two groups of patients. The therapy resulted in a significant decrease of the total resistance and the combined therapy showed an additional decrease of the resistance values.


The antithrombotic activity of BAY g 6575. The activity of 1-[2-(beta-naphthyloxy)ethyl]-3-methyl-2-pyrazolin-5-one (= BAY g 6575) was evaluated in models of experimental thrombosis caused by traumatically induced damage of vessel segments. After prophylactic administration of BAY g 6575 (0.3 mg/kg p.o.) to rats the thrombus formation was significantly reduced in the carotid artery as well as in the jugular vein. The thrombus formation in the femoral arteries of rabbits is inhibited at a minimal effective dose of 1 mg/kg p.o. The incidence of occlusive thrombi is not influenced. BAY g 6575 is 10 times more potent than acetylsalicylic acid (ASA). In the arterial system the thrombus formation is frequently completely abolished.


Effect of non-steroidal antiinflammatory drugs on some biological activities dependent on complement activation. The paper presents the results of a study on the action of five non-steroidal antiinflammatory agents (phenylbutazone, indometacin, acetylsalicylic acid, niflumic and flufenamic acids) on two biological activities which are dependent on complement activation: opsonization of bacteria and membrane damage, the latter evaluated both with the classic immunohemolytic system and with a bactericidal assay. The three biological assays differ for complement sequences involved. Flufenamic and niflumic acids showed high inhibitory activity in the lytic assays. Human complement was more sensitive to inhibition than guinea-pig complement. Phagocytic test confirmed the inhibitory activity of flufenamic acid on complement dependent opsonization.


Mutagenicity of inhalation anaesthetics: trichloroethylene, divinyl ether, nitrous oxide and cyclopropane. The mutagenic potential of trichloroethylene, divinyl ether, nitrous oxide and cyclopropane was assessed in vitro by microbial assay employing two histidine-dependent strains of Salmonella typhimurium, TA1535 and TA100. Anaesthetic agents in various concentrations were incubated with bacteria in the presence or absence of an enzyme system prepared from enzyme-induced rat liver. Nitrous oxide and cyclopropane were not mutagenic, whereas divinyl ether gave a strongly positive response. Results for trichloroethylene were equivocal. These and previous studies with the salmonella system, together with mutagenicity studies using different test systems, indicate that modern inhalation anaesthetic agents are unlikely to be mutagenic.


Systemic air embolism following induction of artificial pneumothorax under anaesthesia, with successful management. Systemic air embolism occurred in a patient during general anaesthesia, with positive pressure ventilation, following induction of artificial pneumothorax to assist in the diagnosis of a mediastinal mass. A sudden change in vital signs together with neurological abnormalities suggested involvement of both coronary and cerebral arteries. A trace of blood was noticed in the syringe which the surgeon had used to create the artificial pneumothorax. The patient was treated with hyperbaric oxygen and recovered satisfactorily, despite a 10-h interval between the air embolus and the institution of definitive therapy.


The effect of assay methods on plasma levels and pharmacokinetics of theophylline: HPLC and EIA. 1 The effects of two assay methods, HPLC and EIA, on theophylline plasma levels (n = 254) and pharmacokinetics (n = 6) were examined on blood samples obtained from asthmatic patients and normal adult male volunteers. 2 The standard curves obtained and the plasma level values during plasma level monitoring of theophylline measured with two methods are well correlated each other within concentration range of 2.5 to 20.0 microgram/ml (P less than 0.001). EIA, however in general, appears to give a lower value than HPLC. 3 The values of plasma theophylline half-life and clearance analyzed using a two-compartment open system model were found different between the two assay methods and dissimilar to those previously reported by using ultraviolet method. 4 The sampling end-point and/or assay sensitivity particularly to measure a lower level of theophylline (less than 2.5 microgram/ml) can influence the pharmacokinetics for a potent bronchodilator with a narrow therapeutic margin, theophylline.


Atenolol and chlorthalidone in combination for hypertension. 1 The hypotensive effect of single daily dosing with atenolol 100 mg and chlorthalidone 25 mg given alone or in combination has been assessed in a double-blind, crossover, placebo controlled trial in fifteen hypertensive patients. 2 Average lying blood pressures were: Placebo 155.4/103.9 mm Hg, atenolol 134.6/85.8 mm Hg, chlorthalidone 139.5/90.1 mm Hg, combination 127.7/82.5 mm Hg. 3 The effect of the combination therapy in reducing lying diastolic pressure compared with placebo (a fall of 21.4 mm Hg) was significantly less than the 31.9 mm Hg fall predicted from the sum of the individual effects (P = 0.01). 4 Observations on blood pressure at rest and under mental, isometric and bicycle ergometer stress were made pre-dose and post-dose for a 12 h period at the end of the last treatment period. 5 Lying blood pressure declined from the zero hour (pre-dose) reading on all treatments to a low at 15.00--18.00 h and then rose again. 6 The rise in systolic blood pressure after isometric exercise and mental stress was of a similar magnitude with all four treatment regimes. 7 Atenolol, alone and in combination with chlorthalidone, reduced the blood pressure and the pulse rate increase on exercise 2 h post-dose when compared with readings 24 h post-dose. 8 Once daily dosing with a combination of atenolol and chlorthalidone produced a fall in supine blood pressure over a 24 h period but the effect on exercise induced changes was not uniform over this period.


Antihypertensive effect of a non-selective (propranolol) and a cardioselective (metoprolol) beta-adrenoceptor blocking agent at rest and during exercise. 1 The antihypertensive effects at rest and during physical exercise of the non-selective beta-adrenoceptor blocker propranolol and the cardioselective beta-adrenoceptor blocker metoprolol were compared in a double-blind cross-over study. 2 Eighteen patients with mild hypertension entered the trial. One patient was withdrawn from the study due to side effects on both drugs. 3 The two beta-adrenoceptor blockers were compared using doses earlier shown to have the same beta-adrenoceptor blocking potency, as measured by their effect on exercise tachycardia in healthy men. 4 Arterial blood pressure was reduced to the same extent by propranolol and metoprolol at rest as well as during submaximal work. 5 It is concluded that the antihypertensive effect of beta-adrenoceptor blockers is mainly mediated through blockade of the beta 1-adrenoceptors.


Modification of the immunogenicity and antigenicity of rat hepatoma cells. II. Mild heat treatment. I.p. immunization with gamma-irradiated hepatoma cells induces resistance to s.c. tumour-cell challenge in syngeneic WAB/Not rats. Mild heat treatment of these cells (greater than 41 degrees C for 30 min) destroyed this immunoprotective effect, but did not abolish tumour-specific antibody production in treated rats. The binding of syngeneic and alloantibodies to surface antigens expressed on hepatoma cells was unaffected by heat treatment. Thus, heat-treated gamma-irradiated hepatoma cells retain a serologically defined tumour-specific antigen but are unable to elicit immunoprotection. By examining the incorporation of radioactive precursors into DNA, RNA and protein in heat-treated cells, it was determined that above 41 degrees C there was a significant decrease in metabolic activity. It is postulated that for the effective induction of transplantation immunity to tumours, tumour-specific antigens should be present on the surface membranes of a metabolically active cell. This hypothesis accounts for the absence or marked reduction of immunoprotection induced by inviable or glutaraldehyde-treated cells, isolated cell membranes and soluble tumour extracts which retain serologically defined tumour-specific antigens.


Radiosensitization of E. coli B/r by the cytotoxic agent procarbazine: a hypoxic cell sensitizer preferentially toxic to aerobic cells and easily oxidized. Procarbazine has been shown to be a hypoxic cell sensitizer of moderate ability in E. coli B/r, with an achievable enhancement ratio of 1.4 at subtoxic concentrations. The drug appears to act in a manner similar to the expected with the electron-affinic radiosensitizers. However, procarbazine and the electron-affinic sensitizers differ in two important respects. Unlike the electron-affinic sensitizers, procarbazine is not easily reduced, but is easily oxidized. It is more toxic to aerobic than to hypoxic cells. At the drug dosages in present clinical use, procarbazine is likely to be only a weak radiosensitizer. The possible implications of the data for the further development of a new class of sensitizers and combination therapy are discussed.


Systemic and in-situ natural killer activity in tumour-bearing rats. Single-cell suspensions prepared by enzymatic disaggregation of an immunogenic 3-methylcholanthrene-induced sarcoma (Mc40A) contain a significant proportion of infiltrating leucocytes (approximately 42%), comprising T lymphocytes, macrophages and non-phagocytic FcR+ lymphoid-like cells. Tumour-infiltrating lymphocytes (TIL) were isolated and purified by successive passage over Sephadex G-10 columns and their cytotoxic activity in vitro compared with that of lymphoid cells from normal rats and from tumour-bearers at different times after implantation. For this purpose, surviving target cells were quantified by incorporation of the gamma-emitting analogue of methionine, 75Sel-methionine, in a 48-h assay which detected both cytotoxic and cytostatic effects. The reactivity of TIL, which was consistently demonstrable from 11 days after tumour transplantation, was essentially similar to that of normal splenic lymphocytes in magnitude and specificity. Reciprocal cytotoxicity tests using TIL and cultured targets from an antigenically unrelated tumour of similar aetiology (Mc57) showed that the manifestation of TIL cytotoxicity was determined, not by the tumour of origin, but by the susceptibility of the target cells. Evidence that the effector function of TIL was mediated in part by natural killer (NK) cells was derived from concurrent experiments using human myeloid cells (K562) as targets in an 18h 51Cr-release assay. In this system the level of NK activity was critically dependent on the numbers of tumour cells in the TIL population; contamination in excess of 2% gave rise to dose-dependent inhibition of NK function. The results show that within a progressively growing tumour known to possess rejection antigens, NK reactivity was detected in the absence of a demonstrable tumour-specific cytotoxic component.


Predominance of T lymphocytes in the dermal infiltrate of atopic dermatitis. Tissue sections from diseased skin of sixteen patients with atopic dermatitis were investigated with haematoxylin-eosin and toluidine-blue staining, with direct immunofluorescence staining using FITC-conjugated antisera against IgG F(ab')2 and IgM, and with the indirect immunofluorescence method utilizing specific rabbit anti-human T lymphocyte antiserum with FITC-conjugated goat anti-rabbit Ig antiserum as the second layer. Furthermore, cryostat sections were investigated in a closed chamber immune adherence method using aminoethylisothiouronium bromide (AET) treated sheep red blood cells to detect E receptors on T lymphocytes, and with various types of coated sheep red blood cells to detect cells with IgG Fc receptors and complement factor C3b receptors. All sections presented dermal perivascular infiltrates of mononuclear cells as judged by haematoxylin-eosin staining. Staining with toluidine-blue demonstrated varying numbers of mast-cells, but in no case pathological increased number. The majority of the infiltrating cells presented rim-like membrane fluorescence with the anti-T antiserum, and the AET treated sheep red blood cells (SRBC) adhered to the infiltrates, thus indicating a predominance of T lymphocytes in the skin infiltrates of atopic dermatitis.


Evidence for a conformational change in the Escherichia coli maltose receptor by excited-state fluorescence lifetime data. The initial signaling event during maltose chemoreception in Escherichia coli is identified with a delocalized liqand-induced conformational change in the maltose binding protein. Substantiation for the conformational change involves a new application of the "distant reporter group technique" [Zukin, R.S., Hartig, P.R., & Koshland, D.E., Jr. (1977a) Proc. Natl. Acad. Sci. U.S.A. 74, 1932-1936] utilizing excited-state fluorescence lifetime measurements. Binding of maltose to its receptor results in changes in the microenvironment of the two tryptophan residues of the receptor protein and of an experimentally attached reporter group, 5-(iodoacetamido) fluorescein. The minimum distance between the two typtophans from efficiency of fluorescence energy transfer theory is 17 A; the minimum distance from the farther tryptophan to the fluorescein is 50 A. Thus, the maltose receptor is shown to undergo molecular rearrangements at distant sites upon ligand binding. The general feature of conformational change as the initial signaling event during chemoreception in the enteric bacteria is discussed.


Antibiotic effects on the photoinduced affinity labeling of Escherichia coli ribosomes by puromycin. The effect of ribosomal antibiotics on the photoinduced affinity labeling of Escherichia coli ribosomes by puromycin [Cooperman, B.S., Jaynes, E.N., Brunswick, D.J., & Luddy, M.A. (1975) Proc. Natl. Acad. Sci. U.S.A. 72, 1974; Jaynes, E.N. Jr., Grant, P.G., Giangrande, G., Wieder, R., & Cooperman, B.S. (1978) Biochemistry 17, 561] has been studied. Although blasticidin S, sparsomycin, lincomycin, and erythromycin are essentially without effect, major changes are seen on addition of either chloramphenicol or tetracycline. The products of photoincorporation have been characterized by one- and two-dimensional gel electrophoresis and by specific immunoprecipitation with antibodies to ribosomal proteins. In the presence of chloramphenicol, protein S14 becomes the major labeled protein. In the presence of tetracycline, L23 remains the major labeled protein, but the yield of labeled ribosomes is enormously increased, and the labeling is more specific for L23. These results are discussed in terms of the known modes of action of these antibiotics and the photoreactivity of tetracycline.


Interaction of phosphate analogues with glyceraldehyde-3-phosphate dehydrogenase. The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase catalyzes the oxidative phosphorylation of D-glyceraldehyde 3-phosphate. A variety of phosphonates have been shown to substitute for phosphate in this reaction [Gardner, J. H., & Byers, L. D., (1977) J. Biol. Chem. 252, 5925--5927]. The dependence of the logarithm of the equilibrium constant for the reaction on the pKa2 value of the phosphonate is characterized by a Brłnsted coefficient, betaeq, of approximately 1. This represents the sensitivity of the transfer of the phosphoglyceroyl group between the active-site sulfhydryl residue (in the acyl-enzyme intermediate) and the acyl acceptor on the basicity of the acyl acceptor. Molybdate (MoO42-) can also serve as an acyl acceptor in the glyceraldehyde-3-phosphate dehydrogenase catalyzed reaction. The second-order rate constant for the reaction with molybdate is only approximately 12 times lower than the reaction with phosphate even though the pKa2 of molybdate is 3.1 units lower than the pKa2 of phosphate. The immediate product of the molybdate reaction is the acyl molybdate, 1-molybdo-3-phosphoglycerate. The acyl molybdate, like the acyl arsenate (the immediate product of the reaction when arsenate is the acyl acceptor), is kinetically unstable. At pH 7.3 (25 degrees C), the half-life for hydrolysis of the acyl molybdate, or the acyl arsenate, is less than 2.5 s. Thus, hydrolysis of 1-molybdo- and 1-arseno-3-phosphoglycerate is at least 2000 times faster than hydrolysis of 1,3-diphosphoglycerate under the same conditions. Glyceraldehyde-3-phosphate dehydrogenase has a fairly broad specificity for acyl acceptors. Most tetrahedral oxy anions tested are substrates for the enzyme (except SO4(2-) and SeO4(2-)). Tetrahedral monoanions such as ReO4- and GeO(OH)3- are not substrates but do bind to the enzyme. These results suggest the requirement of at least one anionic site on the acyl acceptor required for binding and another anionic group on the acyl receptor required for nucleophilic attack on the acyl enzyme.


Shapes of proteins L1, L9, L25, and L30 from the 50S subunit of the Escherichia coli ribosome, determined by hydrodynamic studies. Proteins L1, L9, L25, and L30, purified by a nondenaturing method from the 50S ribosomal subunit of Escherichia coli A19, have been characterized. The four proteins were studied under conditions which resemble those used for reconstitution experiments. These proteins have S020,W values of 2.0 S, 1.8 S, 1.8 S, and 1.0 S and D20,W values of 8.4 X 10(-7), 9.0 X 10(-7), 14.0 X 10(-7), and 15.0 X 10(-7) cm2/S. Apparent specific volumes at 20 degrees C are 0.738, 0.733, 0.700, and 0.735 mL/g for the four proteins. The respective molecular weights determined by sedimentation equilibrium are 25 000, 17 300, 12 000, and 6500. The intrinsic viscosity values for the four proteins are 4.0, 5.5, 3.6, and 3.2 mL/g. From these hydrodynamic parameters L1 and L9 appear to have globular or at most only slightly elongated shapes, whereas L25 and L30 appear to be definitely globular.


Biosynthesis of the pyrimidine moiety of thiamin in Escherichia coli: incorporation of stable isotope-labeled glycines. Methods are described for the cleavage, extraction, and subsequent gas chromatographic-mass spectrometric analysis of the pyrimidine moiety of thiamin as 2-methyl-4-amino-5-[(ethylthio)methyl]pyrimidine. The methods are of a general nature and can be applied to any system. Using these methods to evaluate the incorporation of 13C-, 15N-, and 2H-labeled glycines into the pyrimidine moiety of thiamin by Escherichia coli, we established that the nitrogen and carbon atoms of glycine are incorporated as a unit into the pyrimidine. 13C- and 15N-labeled glycines are incorporated at greater than 60% but deuterium from [2-(2)H2]glycine was incorporated at only 18%. A detailed analysis of the mass fragmentation pattern of the pyrimidine derivative has established that the glycine nitrogen atom supplies the N-1 of the pyrimidine and that the C-1 and C-2 of the glycine supplies the C-4 and C-6 of the pyrimidine, respectively. This evidence is consistent with the substitution of a C2 unit between the C-5 and C-4 of the 4-aminoimidazole ribonucleotide precursor during the biosynthesis of the pyrimidine moiety of thiamin in E. coli.


Light-induced calcium release in isolated intact cattle rod outer segments upon photoexcitation of rhodopsin. By applying flash-spectrophotometry with the calcium-indicating dye arsenazo III rapid light-triggered calcium release in various cattle rod outer segment preparations was studied. It is shown that light-induced calcium signals can be unambiguously discriminated from underlying absorption changes due to photolysis of rhodopsin and apparent absorption changes resulting from lightscattering transients. The following results have been obtained: 1. Calcium-induced arsenazo III responses can be quantitatively and kinetically resolved within the time domain of the visual transduction process. 2. Photoexcitation of rhodopsin results in calcium release from intradiscal binding sites. 3. Calcium released does not appear in the cytoplasmic space unless the disc membrane is made permeable to calcium ions by an ionophore. 4. The shortest observed half-rise time of calcium release (300 ms) is possibly limited by the ionophore. 5. The stoichiometric ratio of calcium released/rhodopsin bleached is 0.5 at a free calcium concentration of 2 microM. The amount of calcium released is proportional to the precentage of rhodopsin bleaching (from 1--10%). 6. Upon disruption of the disc stack by lysis of intact rod outer segments the light-induced calcium release is greatly altered. The results are discussed in relation to previous reports on a light-induced calcium release from retinal discs and in terms of the proposed role of calcium as an intracellular transmitter in vertebrate photoreceptors.


Yeast plasma membrane ghosts. An analysis of proteins by two-dimensional gel electrophoresis. We have examined yeast cell ghost preparations to assess their value in obtaining plasma membrane proteins. Ghosts prepared by two methods involving stabilization of spheroplast envelopes had similar protein patterns by two-dimensional gel electrophoresis, and approximately 200 proteins were resolved. Spheroplasts were lactoperoxidase iodinated, and recovery of label in ghost preparations was greater than 60%. Spheroplasts appeared to be impermeable to the lactoperoxidase reagents as judged by an examination of two-dimensional gel electrophoretic patterns of ghost proteins that had been iodinated in spheroplasts or in unsealed ghosts. Spheroplasts were also impermeable to pronase proteases. Surface iodination and surface proteolysis allowed us to identify exposed ghost proteins; the major ghost glycoprotein was exposed in spheroplasts. Two-dimensional patterns of ghost proteins were not heavily contaminated (less than or equal to 25% of all proteins) by proteins present in soluble or promitochondrial fractions, and estimates of surface label and total cell protein recovery suggested that the ghost fraction represents a cell envelope enrichment of 8--10 fold over whole cells. Resolution of ghost proteins by two-dimensional gel electrophoresis appears to be a powerful aid toward identifying membrane proteins.


Nature of the regions involved in the insertion of newly synthesized protein into the outer membrane of Escherichia coli. Outer membrane proteins are synthesized by cytoplasmic membrane-bound polysomes, and inserted at insertion sites which cover about 10% of the total outer membrane when cells grow with a generation time of 1 h. A membrane fraction enriched in outer membrane insertion regions was isolated and partly characterized. The rat at which newly inserted proteins are transferred from such insertion regions into the rest of the outer membrane was found to be very fast; the new protein content of insertion regions and that of the remaining outer membrane equilibrate completely within about 20 s at 25 degrees C. Given the rather rigid structure of the outer membrane and the multiple interactions between outer membrane components and the murein layer, lateral diffusion of newly inserted proteins from insertion sites to the remaining outer membrane is not likely to explain this rapid equilibration. Instead, the data support a model in which insertion regions move along the cell surface, leaving behind stationary, newly inserted outer membrane proteins.


Physical and coding properties of poly(5-methoxyuridylic) acid. The synthesis of poly(mo5U) requires a high concentration (2.7 mg/ml) of polynucleotide phosphorylase as well as a long reaction time (48 h). The resulting polynucleotide has a chain length of approximately 100 nucleotides. It shows no indication of a stable secondary structure. When poly(mo5U) is mixed with poly(A), a triple-stranded complex poly(A) . 2poly(mo5U) is formed. This complex has a melting temperature of 68.5 +/- 0.5 degrees C at 150 mMNa+ and exhibits a hysteresis loop between melting and reformation of the complex having a delta Tm of 11.5 degrees C. Poly-5-methoxyuridylic acid stimulates the binding of Phe-tRNA to 70-S ribosomes but is inactive in directing poly(Phe) synthesis.


Interaction of Cibacron Blue F3GA with Escherichia coli DNA polymerase I and with T4 DNA polymerase. Individual rapid procedures for the enrichment of Escherichia coli DNA polymerase I and of bacteriophage T4 DNA polymerase free of endonuclease activity are described using Blue dextran-Sepharose chromatography. The blue dye of Blue dextran-Sepharose selectively binds to the deoxynucleoside triphosphate substrate site of the E. coli but not the T4 enzyme indicating that the catalytic sites of these two enzymes which catalyze the same polymerization reaction in vitro are quite distinct.


Product inhibition studies of yeast phosphoglycerate kinase evaluating properties of multiple substrate binding sites. Product inhibition studies on yeast phosphoglycerate kinase (ATP:3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3) have been performed with 1,3-P2-glycerate. The results indicate that: 1. The catalytic reaction can be affected via four substrate binding sites, two for MgATP2- and two for 3-P-glycerate. 2. There is one catalytic centre per enzyme molecule. 3. The catalytic reaction primarily occurs at the 'first' or 'high affinity' MgATP2- and 3-P-glycerate binding sites. The 'second' set of sub-sites for these substrates are located in a region for regulation of the catalytic reaction. 4. The products of the reaction, 1,3-P2-glycerate and ADP, are preferentially bound to the regulatory region. 5. MgATP2- and 1,3-P2-glycerate are able to bind simultaneously to this region. When liganded with MgATP2- the apparent Ki value for 1,3-P2-glycerate increases from 3 microM to 20 microM.