Research related to medical treatment of cardiovascular disease.

Medical treatment of heart disease has improved significantly in the past 30 years. The spectacular change in the natural history of rheumatic heart disease is apparent from a 1948 article that reported that 42% of children with rheumatic heart disease died of rheumatic infection or bacterial endocarditis. Antibiotics and cardiac surgery have improved the outcome from rheumatic heart disease. Cardiopulmonary resuscitation has had a major impact on the treatment of myocardial infarction and on the management of sudden death. The fundamental principle underlying the discovery of cardiopulmonary resuscitation is reviewed, and recent developments emphasizing the importance of intrathoracic pressure in the hemodynamics of cardiopulmonary resuscitation are highlighted. The important new drugs of the last 30 years include the oral diuretics, the antihypertensives and the antiarrhythmic agents. The development of the beta-blocking agents is cited as an example of the translation of basic physiological research to medical care. Finally, the role of epidemiologic techniques in the design of clinical trials to evaluate medical therapy and hence improve medical management is discussed.


Research related to surgical treatment of coronary artery disease.

In the past 20 years, basic and clinical research have provided new information on coronary artery surgery. For example, several studies have shown that coronary artery bypass grafting is more effective than medical treatment in relieving the symptoms of chronic disabling angina pectoris. However, we still do not have definitive answers to many questions. What factors in the patient, in the operation and in the care after operation determine success in surgical treatment? Does the operation prolong useful life? Is the operation affordable? These questions are difficult. Further research is needed to solve complex problems relating to surgical vs medical treatment of coronary artery disease.


Research related to surgical treatment of aortic and peripheral vascular disease.

The history of vascular surgery and the developments that made it possible are briefly traced. Approaches to the treatment of arterial lesions are considered in terms of the characteristic anatomic, pathologic and clinical patterns of arteriosclerosis or atherosclerosis, the basic underlying lesion in most aneurysmal and occlusive diseases of the aorta and major arteries. The importance of appreciating the various patterns and rates of progression of atherosclerosis is emphasized.


Competitive enzyme-linked immunoassay for Factor VIII antigen.

We describe a competitive enzyme-linked immunoassay for Factor VIII antigen. Binding of anti-factor VIII to solid-phase Factor VIII antigen is competitively inhibited by the free factor VIII antigen that is to be measured. The amount of anti-Factor VIII bound to solid-phase VIII is measured by applying in sequence a heterologous bridging antibody and a soluble antibody/enzyme immune complex. The soluble complex used was rabbit antiperoxidase/horseradish peroxidase. Peroxidase activity is inversely proportional to the Factor VIII antigen concentration in the original test plasma and is measured spectrophotometrically. The assay can be performed in as little as 4 h with only a microtiter plate, antisera, antigen, and a spectrophotometer. It is sensitive to 0.05 units of Factor VIII antigen per milliliter, and reproducibility, linearity, and normal range are similar to those reported for other techniques.


Assay of theophylline: comparison of EMIT on the ABA-100 to HPLC, GLC and UV procedures, with detailed evaluation of interferences.

The EMIT technique for theophylline measurement as adapted to the Abbott Bichromatic Analyzer (ABA-100), was compared to two other established procedures. EMIT results correlated well with those obtained with high pressure liquid chromatographic (HPLC) (r = 0.995, N = 54) and spectrophotometric (UV) (r = 0.95, N = 37) methods when the patient samples used were free of apparent interferences in the comparison methods. On samples exhibiting interferences with HPLC and/or UV methods, the EMIT assay gave the same results as a gas-liquid chromatographic procedure which showed no interferences. In addition, theophylline metabolites, xanthine analogs and many commonly administered drugs did not interfere with the EMIT theophylline assay. An interassay coefficient of variation of 3.5--6.5% was demonstrated. Our data indicate that the EMIT technique exhibits adequate precision and specificity for routine theophylline determinations. Unlike the HPLC or UV procedures, we have not encountered any interferences with the EMIT technique.


High molecular weight alkaline phosphatase: a clinical study.

High molecular weight alkaline phosphatase activities have been measured in the sera of 72 patients with a variety of forms of liver disease, 14 patients with bone disease and 8 healthy volunteers. These measurements have been compared with measurements of other indices of hepatic function in order to establish the place of this enzyme in the diagnosis of liver disease. High molecular weight alkaline phosphatase proved to be a sensitive and specific tests for detecting liver disease, particularly obstructive liver disease. It was better than all the other liver function tests in distinguishing liver metastases from other hepatobiliary diseases. It may therefore prove especially useful in the early detection of liver metastases.


The role of Escherichia coli in gastroenteritis.

During the last 10 years there has been a major break-through in understanding the pathogenic mechanisms of E. coli as a cause of acute diarrhoea. The current situation is briefly tabulated in Table 3. In the near future the field of interest is likely to centre on the investigation of adhesive factors and certainly on the possibility of using preparations of adhesive factors as vaccines. The successful use of such vaccines may have enormous benefit as intervention measures to break the acute diarrhoea--malnutrition syndrome which is so damaging to the life of infants and young children in developing countries.


Shigella infections.

Shigellosis is a complex disease involving two distinct organs and two distinctive clinical presentations of intestinal involvement. It is also associated with a wide variety of extra-intestinal manifestations. Because these may precede the onset of diarrhoea/dysentery, and confuse the diagnosis, it is safe to denote shigellosis as a protean clinical problem. The disease is readily spread by contact because so few organisms are required to establish infection. It may also be spread through contaminated food or water, related to either defective sewage or a human carrier, and explosive outbreaks may occur in closed populations. Homosexuals are also at risk of venereal transmiddion of infection. Shigellosis is treatable with effective oral antimicrobials, responding both clinically and microbiologically. When organism are sensitive, ampicillin is the current drug of choice, and when they are resistant to ampicillin, trimethoprim-sulphamethoxazole appears to be the best choice. Dehydration is not usually severe, and responds to oral rehydration therapy, which is highly recommended along with early refeeding.


Experimental immune glomerulonephritis induced in the rabbit with streptococcal vaccine.

Heavy C3 glomerular deposits were observed in rabbits injected intravenously with C5 streptococcal vaccine. Immunoglobulin deposits appeared later in a few rabbits. Although some data favour the presence of circulating immune complexes during the course of this glomerulonephritis, no evidence for their initiating role could be demonstrated. Streptococcal components are known to activate the alternative pathway of complement. It is suggested that complexes made of streptococcal components and activated C3 might deposit in glomerular tufts.


Antibodies to motor endplates demonstrated with the immunofluorescence technique.

Antibodies to acetylcholine receptors (AChR) are probably directly responsible for the pathogenesis of myasthenia gravis (MG). Methods for the demonstration of these antibodies are complicated. The present study shows that the antibodies can also be revealed by the use of the simple indirect immunofluorescence technique with rat diaphragm as substrate. Antibodies were demonstrated with FITC-labelled anti-human Ig. The location on the motor endplates was confirmed by using a TRITC-labelled anti-alpha-bungarotoxin system. Antibodies to motor endplates were only demonstrated in MG and not in either twenty-two patients with neuromuscular disorders or fifty normal subjects. Antibodies to motor endplates were found in only twelve out of fifty-seven MG patients. In fifteen of the other forty-five patients, antibodies were found of the classical anti-skeletal muscle type, 'overluminating' the anti-motor endplate antibodies.


Adjuvanticity of lactobacilli. I. Differential effects of viable and killed bacteria.

The adjuvanticity of Lactobacillus brevis and Lactobacillus plantarum was the subject of this study. The latter was the better adjuvant in both delayed hypersensivity and antibody formation to sheep red blood cells. Viable L. plantarum stimulated exclusively the delayed hypersensivity, where heat-killed bacteria had an adjuvant effect on antibody formation. For optimal adjuvant effects lactobacilli had to be injected in a dose of 10(8) into the same site as the antigen. Viable lactobacilli and to a lesser degree heat-killed bacteria induce hepato-splenomegaly, suggesting mediation of the adjuvant activity by the reticuloendothelial system. Granuloma formation with mainly mononuclear cell infiltrates could be observed after subcutaneous administration of viable lactobacilli whereas heat-killed lactobacilli induced granulomata containing about equal numbers of granulocytes and mononuclear cells. The possible clinical application of L. plantarium in the immunotherapy of tumours is suggested.


Effects of carbamazepine on valproic acid kinetics in normal subjects.

Carbamazepine and valproic acid are used together in the treatment of epilepsy. It is therefore, relevant to investigate the possibility of a carbamazepine effect on valproic acid disposition, particularly since carbamazepine is known to induce enzymes. We gave valproic acid orally to 6 normal subjects, 250 mg twice daily for 4 wk. Carbamazepine, 200 mg once daily, was begun after 4 days on valproic acid. Serum drug concentrations were measured during 4 dosing intervals, once before and 3 times after beginning carbamazepine. Minimum steady-state concentrations of valproic acid declined after carbamazepine from 34.4 +/- 5.1 to 27.1 +/- 4.4 mug/ml (p less than 0.0005). Clearance rose from 6.46 +/- 0.80 to 8.48 +/- 2.28 ml/hr/kg (p less than 0.01). The increase in clearanace and decrease in minimum steady-state levels was apparent only after 2 wk on carbamazepine. The elimination rate constant (KE) during the dosing interval did not rise during carbamazepine administration (0.0623 +/- 0.0168 hr--1 before and 0.0573 +/- 0.0168 hr--1 after, p greater than 0.25), raising the possibility of an increase in distribution volume.


Effect of imipramine on norepinephrine and blood pressure in enuretic boys.

The effect of imipramine, desmethylimipramine, and methscopolamine on blood pressure (BP) and plasma norepinephrine (NE) was measured in enuretic boys in a double-blind, placebo-controlled study. Measurements were obtained on the thirteenth day of medication (75 mg at bedtime). The tricyclic drugs induced a rise in diastolic BP as well as an increase in plasma NE but there was no significant relationshhip between the increments in plasma NE and BP. The plasma concentration of drug correlated with the drug-induced BP rise. This is the fifth study to demonstrate a hypertensive effect of tricyclic drugs in children in contrast to the systolic hypotension usually seen in adult patients. It is not clear from our data whether children have different cardiovascular compensatory reflexes or whether they experience a greater stimulant effect from the drug.


Silicone implant arthroplasty of the great toe. A review of single stem and flexible hinge implants.

One hundred sixty-five feet were reconstructed with the single stem silicone implant, including patients with hallux regidus, hallux valgus due to degenerative and osteoarthritic joint changes, hallux valgus secondary to rheumatoid arthritis and patients with previous unsuccessful "bunion" procedures. Preoperative pain and intolerance to shoe wear were relieved in the great majority of patients. The amount of correction of the hallux valgus angle was directly related to the preoperative deformity. Infection occurred in 2 patients, and inflammatory reaction to the implant occurred in one patient. One hundred five feet were reconstructed with a flexible hinge implant; 94 of the feet had joints destroyed by rheumatoid arthritis. Dramatic improvement in the hallux valgus angle resulted in these feet which had relatively severe preoperative deformities. No implant fractures were recorded. The present survey of our cases, and experiences reported by others, encourage the use of these implants by surgeons who follow the suggested indications, surgical technique, and follow up care.


Hemodynamic effects of oxprenolol and propranolol in hypertension.

Oxprenolol is a beta-adrenergic blocker with intrinsic sympathomimetic activity. Such drugs are not currently available in the United States, although they have the advantage of less negative inotropic effect than the available propranolol. In 18 patients with mild essential hypertension, oxprenolol (9 patients) or propranolol (9 patients) was added to thiazide in random double-blind fashion and continued for 7 wk during which supine heart rate, blood pressure, and noninvasively measured cardiac output (by CO2 rebreathing) were determined weekly. With thiazide dosage constant throughout, maximal dose titration to 386. +/- 52.1 (SEM) mg/day of oxprenolol and 360.0 +/- 45.4 mg/day of propranolol was achieved over the first 5 wk. Blood pressure fell with both (141.8 +/- 4.8/96.0 +/- 2.3 to 128.0 +/- 5.1/87.2 +/- 1 mm Hg on oxprenolol, p less than 0.01; 150.8 +/- 5.5/98.0 +/- 1.7 to 129.9 +/- 5.5/86.8 +/- 3.4 mm Hg on propranolol, p less than 0.01). Cardiac output fell from 6.85 +/- 0.63 to 5.77 +/- 0.45 1/min (p less than 0.01) on oxprenolol, and from 6.79 +/- 0.61 to 5.37 +/- 0.37 1/min (p less than 0.02) on propranolol. Oxpranolol. Oxprenolol reduced heart rate from 76.4 +/- 2.0 to 65.6 +/- 2.1 beats/min (p less than 0.001) and it fell from 82.0 +/- 3.8 to 65.3 +/- 3.7 beats/min (p less than 0.001) with propranolol; the fall in heart rate was less but not significantly so for oxprenolol (-14.2 +/- 1.8% and -19.8 +/- 2.8%, p less than 0.1). Thus oxprenolol is equivalent to propranolol in antihypertensive action; minor hemodynamic differences between the two drugs might reflect intrinsic sympathomimetic activity of oxprenolol. Oxprenolol should be considered as an alternative to propranolol.


[Seeing the insane: MacKenzie, Kleist, William James].

Using texts by the late 18th century Scottish writer Henry MacKenzie, the German Romantic writer Heinrich von Kleist, and the American psychologist William James, the use of visual models for the perception of the insane is illustrated. In the latter two cases, the seeming hallucinatory nature of the texts is shown to have had its roots in visual models.


[Literature and schizophrenia. Research trends and research tasks].

Research on schizophrenia and on the psychopathology of creativity is increasingly absorbing and transforming insights from literature (Büchner, Celan, Kafka, Proust) and literary aesthetics. This development is discussed against the background of the history of psychiatric research. Regarding contemporary research trends, special emphasis is placed on communication theory as presented by the Palo Alto group (double bind theory), anti-psychiatry (Cooper, Laing), French structuralism (Foucault, Lacan), and on schizoanalysis (Deleuze and Guattari). Traditional concepts of mental health and illness are called into question.


Analysis of in vitro replication of different DNAs.

The conversion of single-stranded circular DNA to duplex DNA in vitro occurs by at least three different mechanisms. These differences reside in the manner of priming of these DNAs. In contrast, the elongation of primed DNA templates is a general reaction. A number of these proteins have been isolated and further characterized. In addition, cell-free preparations capable of supporting phi X RFI DNA replication as well as the synthesis of progeny viral phi X174 single-stranded circular DNA have been prepared.


The cosmological and performative significance of a Thai cult of healing through meditation.

A cult of healing through meditation that was observed in Bangkok, Thailand in 1974 is described, and the cult is interpreted in terms of two axes, the cosmological and the performative, and the dialectical, reciprocal and complementary relations between them. The various ramifications of the cosmology are discussed--the categorization of the cosmos itself as a hierarchical scheme, the relations between man and non-human forms of existence, the ideas concerning power and its manner of acquisition and use, the relation between power and restraint, etc. The epistemological basis of the cult, which attempts cure through meditation, and the features of the ritual as they contribute to its performative efficacy are highlighted. The essay concludes by suggesting that there is a single scheme (episteme) underlying religious ideas and applications of knowledge such as meditation, medicine, alchemy, and astrology.


Maternal immunological recognition mechanisms during pregnancy.

One of the most intriguing shortcomings of modern immunology, especially transplantation immunology, is its failure to provide a satisfactory final explanation for the consistent non-rejection of immunogenetically alien conceptuses in utero, even in specifically preimmunized females. Certainly, there is no shortage of hypotheses. Over the past few years various observations have utterly refuted the simplistic notion that the much-mated but nulliparous or gravid females are immunologically unaware of the presence and activities of allogeneic cells transiently or chronically within their reproductive tracts. This evidence has engendered the belief that some kind(s) of active response(s) on the part of the female, following early recognitive events, plays an important, if not essential, role, in conjunction with adaptive hypoantigenicity of the trophoblast, not only in conferring selective benefits upon the conceptus from the time of implantation, but also in affording it protection from the possible hazards of a cellular immunity. Evidence is emerging that during mating, implantation, placentation, and gestation (1) various fetal and histocompabibility antigens are presented to the mother in a unique manner; (2) she does respond to these; and (3) her responses aid in the establishment and maintenance of a harmonious state of immunological coexistence with her fetus. There are reasonable grounds for believing that a complete understanding of the immunobiology of the maternal-fetal relationship may facilitate significant advances in both transplantation and tumour immunology.


The hormonal control of implantation.

The hormonal control of implantation in mammalian species with and without embryonic diapause is described. In a majority of species displaying the obligate form of diapause the corpora lutea appear to exhibit a low level of steroidogenic activity throughout diapause, full luteal activity being resumed just before the initiation of implantation. Fluctuations in the plasma levels of oestrogen and progesterone during diapause may serve to prime the uterus for implantation. In species exhibiting the facultative form of diapause, such as the rat and mouse, both progesterone and nidatory oestrogen are required for the induction of implantation. In species not displaying embryonic diapause implantation will take place in the presence of progesterone alone. In the light of these considerations the selection of animal models for drug-screening purposes and possible new approaches to contraception are discussed.


Hormonal control of implantation in the rat: inhibition by luteinizing hormone-releasing hormone and its analogues.

Chronic treatment of pregnant rats with luteinizing hormone-releasing hormone (LH-RH) and its analogues (Analogue I: des-Gly10-LH-RH-ethylamide; Analogue II: des-Gly10-[D-Ala6]-LH-RH-ethylamide) inhibited implantation of the ovum. Analogues I and II were 4.5 and 173 times potent than native LH-RH in inhibiting implantation respectively. On Day 8 of the treatment with Analogue II, the pituitary glands of the pregnant rats contained approximately 10% of the amount of LH and FSH found in intact pregnant rats. By contrast, serum levels of LH were significantly higher and those of FSH significantly lower in Analogue II-treated rats than in control rats. The ovaries of Analogue II-treated rats were lighter than those of control rats because of smaller corporalutea and less developed follicles. Peripheral serum concentrations of progesterone in rats treated with LH-RH and its analogues were significantly lower than those in control rats. The peripheral serum concentration of progesterone declined earlier in rats treated with LH-RH and Analogue I than it did in rats treated with Analogue II. The inhibitory affect of Analogue II was overcome by concurrent treatment with the following combinations of hormones: progesterone + oestradiol; progesterone + human chorionic gonadotropin (hCG); prolactin + oestradiol; and prolactin + hCG. Treatment with progesterone or prolactin alone did not overcome the effect of Analogue II. These results indicate that Analogue II stimulates the production and release of LH and release of FSH to cause high ratios of LH/prolactin and LH/FSH. The induced imbalance of gonadotropins suppresses the development of ovarian follicles and corpora lutea and reduces secretion of both oestrogen and progesterone. The resulting low levels of ovarian steroids cause a delay in implantation.


Ecto 5'-nucleotidase deficiency in primary hypogammaglobulinaemia.

The activity of the lymphocyte ectoenzyme 5'-nucleotidase is very low in the majority of patients with primary 'common variable' hypogammaglobulinaemia. In order to test whether this can be explained by lymphocyte subpopulation deficiencies we measured 5'-nucleotidase activity, using both biochemical and histochemical techniques, in purified T and B cells from patients and healthy subjects. Purified B cells from normal subjects have about four times the activity of T cells. This explains why the levels of lymphocyte 5'-nucleotidase activity are at the lower limit of the normal range in patients with X-linked hypogammaglobulinaemia who lack B cells. The low levels in the 'common variable' group can be explained by low activity in their T lymphocytes associated with either low activity in their B cells or depletion of B cells. The finding that inhibition of the enzyme does not interfere with in vitro lymphocyte transformation or immunoglobulin production in normal subjects indicates that the enzyme deficiency is not directly responsible for the hypogammaglobulinaemia. These and other studies suggest that this enzyme appears on lymphocytes at a certain stage of development and that both T and B lymphocytes in some patients with 'common variable' hypogammaglobulinaemia are developmentally immature.


Allosteric regulation of calf thymus ribonucleotide reductase.

Ribonucleotide reductase was purified 3400-fold from calf thymus. The enzyme preparation was essentially free of kinases and phosphatases and therefore allowed a conclusive study of the allosteric regulation of a eukaryotic ribonucleotide reductase to be made for the first time. Comparable maximal activities were obtained for the reduction of all four ribonucleotide substrates in the presence of their optimal stimulatory effectors. These and other results strongly argue for the existence of only one ribonucleotide reductase in mammalian cells. No reduction was observed in the absence of effector. The reduction of CDP and UDP both required ATP, with no stimulatory effect of any other nucleoside triphosphate. The only activator of GDP reduction was dTTP and the only activator of ADP reduction was dGTP. Reduction of the purine ribonucleotides was further stimulated by ATP but only in combination with dTTP or dGTP. The reduction of all four ribonucleotides was strongly inhibited by dATP, the inhibition being partly released by ATP. The data can be integrated into a scheme which links ribonucleotide reduction to DNA synthesis.


Purine metabolism in adenosine deaminase deficiency.

Deoxyadenosine was identified in the urine of a second child with almost undetectable levels of adenosine deaminase (ADA) in erythrocyte lysates. Deoxyadenosine excretion thus appears to be characteristic of ADA deficiency: the acid lability of deoxyadenosine (responsible for the frequent confusion of this abnormal urinary metabolite with adenine) may be used in screening for this defect by isotachophoresis. The deoxynucleotides dATP, dADP and dAMP found initially in the child's erythrocytes (in comparable amounts to ATP, ADP and AMP) disappeared after a successful marrow graft from an unrelated donor, as did the urinary deoxy metabolites. Erythrocyte ADA activity decreased after the marrow graft but was still greater than 10% of normal congruent to 10 weeks after the last red cell transfusion.


The sodium-potassium pump in volume expanded hypertension.

Decreased arterial Na+-K+ pump and cardiac Na+, K-+ATPase activities have now been demonstrated in several types of experimental volume expanded hypertension. The changes are not secondary to elevated pressure since they also occur in veins and right ventricle where the pressure is not elevated. Decreased arterial Na+-K+ pump activity can be reproduced by acute volume expansion of the normal rat and plasma extracts from this rat suppress pump activity when applied to arteries from another rat. Suppression of Na+-K+ pump activity in arteries, veins and heart, with ouabain for example, leads to increased contractile activity. Thus the volume expansion, reduced pump activity, and hypertension appear to be causally related through an ouabain-like humoral agent. Certain other evidence suggests that the pump defect extends to the sympathetic nerve endings, thereby reducing the efficiency of neural compensatory mechanisms.


Amoxycillin injectable: a review of its antibacterial spectrum, pharmacokinetics and therapeutic use.

Amoxycillin, an acid stable semisynthetic penicillin shown to be effective against a wide range of infections when given orally, is now available for intramuscular and intravenous injection. Amoxycillin has an antibacterial spectrum and level of activity essentially the same as for ampicillin. Amoxycillin has been shown to have more rapid and complete bactericidal action than ampicillin against E. coli in vitro and in animal models of infection, but the clinical importance of this difference has not yet been determined. Amoxycillin is present in therapeutic amounts in the cerebrospinal fluid of meningitis patients given the drug intravenously and parenteral amoxycillin has been successfully used in the treatment of meningitis, and in urinary tract infections, septicaemia, upper and lower respiratory tract infections and a variety of other infections caused by Gram-negative and Gram-positive aerobic bacteria. However, the extent of experience is limited compared with ampicillin, and thus further studies are needed to more clearly delineate its relative therapeutic role. Parenteral amoxycillin is generally well tolerated. Pain at the site of intramuscular injection occurs in about one-third of patients, but can be minimised by the use of lignocaine or procaine hydrochloride.


Naproxen up to date: a review of its pharmacological properties and therapeutic efficacy and use in rheumatic diseases and pain states.

Naproxen is a propionic acid derivative with analgesic and anti-inflammatory activity which has been widely used in the treatment of rheumatic diseases. Naproxen has been well studied in rheumatoid arthritis and is as effective as aspirin but better tolerated, thus enabling more patients to continue with treatment. For this reason some clinicians now prefer to try propionic acid derivatives, such as naproxen, before aspirin in arthritic patients. In comparative studies with other non-steroidal anti-inflammatory drugs, such as indomethacin, ibuprofen, fenoprofen and others, all drugs were usually of similar overall efficacy although naproxen was sometimes preferred: but as with other non-steroidal anti-inflammatory agents, not all patients will respond to naproxen and in such cases other agents should also be tried until the most satisfactory drug is found for each patient. Naproxen is also effective in degenerative joint diseases of the hip and knee, although further well designed studies are needed to more clearly define its relative place compared with newer drugs such as diclofenac or diflunisal. Results of other comparative studies have shown that naproxen is a suitable alternative to phenylbutazone or indomethacin in ankylosing spondylitis and to aspirin in juvenile rheumatoid arthritis. Naproxen appears to be effective in reducing pain and swelling in acute gout and is an effective analgesic in patients with pain following surgery or trauma and in pain of dysmenorrhoea. Naproxen has generally been better tolerated than aspirin or indomethacin at the dosages used. Because of its relatively long plasma half-life, naproxen can with convenieice be given twice daily, and there is some evidence that once daily dosage is as effective in rheumatoid arthritis.


Drugs and vitamin deficiency.

During the past 10 to 20 years it has become increasingly clear that a certain number of drugs may lead to increased vitamin requirements. However, it is unusual for symptomatic avitaminosis to develop, and then only when circumstances are present which in themselves increase the risk of vitamin deficiency. Therapeutic doses of drugs will interfere with the vitamin status only to a restricted degree, provided they are administered for brief periods and to patients receiving a normal supply of vitamins. Most cases of vitamin deficiency have in fact been described in connection with drugs usually taken for a longer period of time by patients who were already in negative vitamin balance as a result of disease or marginal supply of the necessary vitamins. This review describes some of the more important articles covering this topic.


Drug interactions with alcohol.

Ethanol and drugs can affect each other's absorption, distribution, metabolism, and excretion. When ingested together, ethanol can increase drug absorption by enhancing the gastric solubility of drugs and by increasing gastrointestinal blood flow. However, high concentrations of ethanol induce gastric irritation causing a pyloric spasm which in turn may delay drug absorption and/or reduce bioavailability. The 'quality' of the alcoholic beverage, independent of its ethanol content, can contribute to altered absorption of a drug. Ethanol is not bound to plasma proteins extensively enough to modify drug distribution. However, serum albumin levels in chronic alcoholics may be abnormally low so that some drugs, e.g. diazepam, have an increased volume of distribution. In addition to the amount ingested, the duration of regular intake determines the effect of ethanol on drug metabolism. Acute intake of ethanol inhibits the metabolism of many drugs but long term intake of ethanol at a high level (greater than 200g of pure ethanol per day) can induce liver enzymes to metabolise drugs more efficiently. At the present time there are no accurate means, with the possible exception of liver biopsy, to clinically predict the capacity of an alcoholic to metabolise drugs. Several drugs can inhibit the metabolism of ethanol at the level of alcohol dehydrogenase. Individual predisposition determines the severity of this drug-ethanol interaction. During its absorption phase, ethanol inhibits the secretion of antidiuretic hormone and is also able to induce increased excretion of a drug through the kidneys. However, chronic alcoholics with water retention may show reduced excretion of drugs via this route. At the pharmacodynamic level, ethanol can enhance the deleterious effects of sedatives, certain anxiolytics, sedative antidepressants and antipsychotics and anticholinergic agents, on performance. Mechanisms of lethal interactions between moderate overdoses of ethanol and anxiolytics/opiates/sedatives are poorly understood. On the other hand, certain peptides, 'nonspecific' stimulants, dopaminergic agents and opiate antagonists can antagonise alcohol-induced inebriation to a significant degree.


Mild hypertension: to treat or not to treat?

In the second half of this century, morbidity and mortality from cardiovascular disease has reached epidemic proportions. The major risk factor associated with cardiovascular disease has been found, from various epidemiological studies, to be elevated blood pressure. Therefore, a lot of energy has been expended in prospective therapeutic trials in attempting to detect whether treatment of high blood pressure is beneficial. In severe hypertension, where the risk to the individual is considerable, this benefit has been demonstrated with relative ease. However, it has not yet proven possible to show conclusive benefit of treatment in the mild group of hypertensives, which form the vast majority of patients seen in general practice. A different approach has been advocated where account is taken of other coincidental cardiovascular risk factors; their presence or absence should help the clinician decide whether to treat or not to treat in the individual case.


Neurofilament protein in clonal lines of mouse neuroblastoma.

As indicated by immunofluorescence with neurofilament antiserum and by electron microscopy two neuroblastoma clones in suspension culture contained a pool of neurofilament subunits which could be induced to assemble into filaments following exposure to vinblastine and colchicine. Under the same culture conditions neuroblastoma cells treated with aluminum extended thick processes with many filaments. The processes were much smaller in nontreated cells but still contained bundles of filaments. These filaments persisted after inhibition of protein synthesis by cycloheximide. In the long neurites formed by cells attached to plastic immunofluorescence with neurofilament antiserum was particularly intense at the growth cone.


[Effect of leucogen on the proliferation of antibody-synthesizing cells].

The authors examined the effect of leucogen on cells synthesizing antibodis in spleens of white mice during immune response to sheep erythrocytes. They established a strong stimulating action on the proliferation of immune competent lymphocytes-cells, producing hemolysin. Therefore the stimulating effect of leucogen was not limited only to its action on peripheral leucocytes, but also on the number of cells, forming antibodies.


Effect of caerulein on exocrine and endocrine pancreas in the rat.

The secretion of insulin, glucagon, pancreatic juice, and amylase in response to a 20-min iv infusion of synthetic caerulein were studied simultaneously in the anesthetized rat. Caerulein, a chemical analogue of cholecystokinin, was used in doses of 1-1000 ng/kg.min. The maximum stimulatory effect of caerulein on pancreatic juice volume and amylase output was obtained with doses of 10 ng/kg.min. With increasing doses, the effect decreased progressively. On the other hand, the release of insulin and glucagon was stimulated only by supramaximal doses of caerulein, which had little or no effect on pancreatic exocrine secretions. These results raised the question of whether, under physiological conditions, cholecystokinin regulates the secretory activity of the endocrine pancreas.


Isolation and characterization of somatostatin from anglerfish pancreatic islet.

Somatostatin was purified from anglerfish pancreatic islets using acetic acid extraction, gel filtration (Bio-Gel P-10), ion exchange chromatography (CM Bio-Gel A), and reversed phase high pressure liquid chromatography. The resulting peptide was characterized by RIA, bioassay, and determination of amino acid composition. Anglerfish islet somatostatin was found to possess an amino acid composition and immunological and biological activities equivalent to synthetic somatostatin. Sequence analyses revealed that the primary structure was H-Ala-Gly-cyclo-[Cys-Lys-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys]-OH. These results demonstrate that anglerfish islet somatostatin has the same primary structure as somatostatin from all other sources characterized to date.


Simple method for the collection of pancreatic islets by the use of Ficoll-Conray gradient.

Simple method for the collection of large numbers of viable pancreatic islets from normal rats is described. This method involves collagenase digestion of the pancreas, followed by the use of Ficoll-Conray discontinuous gradient for the separation of isolated islets from unwanted acinal debris. Ficoll-Conray A solution was prepared by mixing undialyzed 12.5% Ficoll and 33.4% Conray in the ratio 2 : 1, to make a specific gravity of 1.095 and osmolarity of 396 mOsm/l. The solution B, C, and D, with specific gravities 1.084, 1.072, and 1.048, respectively were obtained by diluting A solution with distilled water. Using Ficoll-Conray gradient, about 200 viable islets which maintained excellently their morphology and function, were collected consistently from a young adult rat pancreas.


The gonadotropin releasing hormone stimulation and the clomiphene tests in female patients with anorexia nervosa.

In 9 female patients suffering from acute anorexia nervosa (a.n.) and in two patients in whom this disease had reached the remission phase, the response of the gonadotropin-producing cells in the adenohypophysis was checked by administration of Gn-RH and the degree to which the hypothalamic-hypophyseal axis could be stimulated was checked by administration of clomiphene (5 x 100 mg). Hormonal screening examinations (cervical score after Insler and hormonal vaginal cytology) were used to assay the basal estrogen production. LH and FSH concentrations in the serum were determined radioimmunologically using the principles of the double antibody technique. The gonadotropin-producing cells did not respond to Gn-RH administration in 8 of the 9 patients with acute a.n. The response was disturbed in one of these patients. The response to Gn-RH stimulation was normal in the two patients in the remission phase. Clomiphene had no stimulatory effect on the hypothalamic-hypophyseal axis during either the acute or remission phase. Hormonal treatment during the acute phase of a.n. is not indicated since, after recovery of a normal body weight, the symptoms recede and the cycle normalises spontaneously.


[Partial effects of estrogens and 19-nortestoron derivatives].

In the present review the extragenital effects of steroidal estrogens and 19-norandrogen derivatives on mammalian organs and tissues are described. In detail experimental and clinical findings concerning the influence of these compounds on carbohydrate-, lipid- and protein metabolism, on the cardio-vascular system, the blood coagulation, the bone-tissue, the connective-tissue, the central nervous system and the immune system are summarised. These effects are mainly discussed as possible side effects and also as base for new therapeutic concepts.


Linear relationship between plasma concentration and dosage of sodium valproate.

Plasma valproate concentration was studied in 7 hospitalized nonepileptic patients who received sodium valproate at daily doses of up to 60 mg/kg. A linear relationship between dose and plasma concentration of valproate (r = 0.81--0.99) was found in each patient, although the slopes of the regression lines (reflecting clearance rates) varied twofold. This suggests that if the valproate plasma concentrations at two different doses are known and the clearance of valproate is not altered by concomitant administration of other drugs, it should be possible to predict the plasma valproate concentration that will result from further dose increments.


Meiotic nondisjunction in the mouse: methodology for genetic testing and comparison with other methods.

Since trisomies produce adverse effects relatively late in development or even postnatally, they are an important component of the array of genetic damages that might be caused by environmental agents. Whole-chromosome aneuploidy (as opposed to breakage-derived aneuploidy) might come about secondarily from crossover depression, or could follow damage to the meiotic spindle or to kinetochores. For simplicity, the event-by whichever of the mechanisms-is referred to as meiotic nondisjunction (ND). A genetic method has been devised which is based on the facts that ND involving the sex chromosomes produces mostly viable mice, and that such exceptional animals can be externally recognized by the use of appropriate markers. The method is compared with the following other ND indicators: univalent and/or chiasma frequencies at M I; number of dyads at M II; extra sex chromosomes in spermatids; karyotypes in cleavage, morula, or blastocyst metaphases; and chromosome constitution of mid-gestation embryos. Some of the cytological endpoints are found to be unreliable. Various biological variables (germ-cell stage, sex, age) are examined with a view toward maximizing the chances for detecting induced nondisjunction. While experimental evidence on this question is equivocal, a consideration of the probable ND mechanisms suggests that the early spermatocyte (in stages including the premeiotic S phase) may be a favorable test object. The numerical sex-chromosome anomaly (NSA) method is useful not only in the study of ND but also in detecting breakage-derived chromosome losses induced in females, where the dominant lethal test is not easily applicable.


Testing for nondisjunction in the mouse.

Tests for nondisjunction have been carried out in male and female mice. Ten-day fetal progeny of control and treated adults have been karyotyped to establish spontaneous and induced levels of aneuploidy. In males, the effects of 100 rad x-rays on type A spermatogonia and early primary spermatocytes, and the effects of Mitomycin C (2 mg/kg) on early primary spermatocytes, have been tested. The results show insensitivity of primary spermatocytes to both agents, but a 3.5-fold increase in nondisjunction following spermatogonial irradiation. In females, comparisons have been made between young controls, young x-rayed (5 rad), aged controls and aged x-rayed (5 rad) animals. The "ageing effect" on nondisjunction is observed, but too few fetuses have been analyzed to reach conclusions regarding enhancement of nondisjunction levels by low doses of x-rays.


Analysis of anaphase in cell culture: an adequate test system for the distinction between compounds which selectively alter the chromosome structure or the mitotic apparatus.

A system of anaphase analysis is presented as an adequate test to detect aberrations in chromosome structure and alterations in the mitotic apparatus. Particular emphasis is placed on those data which suggest that proteins may be the preferred targets of chemical compounds that induce aneuploidy.


Analysis of aneuploidy in first-cleavage mouse embryos fertilized in vitro and in vivo.

First-cleavage mouse embryos, fertilized in vitro and in vivo, provide ideal material for chromosomal analysis. With the appropriate incubation in a mitotic inhibitor, syngamy is prevented and the sperm- and egg-derived chromosomes remain as separate clusters. Because the latter chromosomes undergo condensation sooner than those from the spermatozoon, the parental source of chromosome sets can be identified even without a marker chromosome. Thus these embryos can be analyzed both for the primary incidence and the parental source of a number of chromosomal anomalies, including aneuploidy. By using fertilization in vitro to obtain the embryos, the synchrony of fertilization and nuclear development is such that 80% or more of the chromosomal preparations are suitable for analysis, compared with about 50% for embryos fertilized in vivo.


Microtus oeconomus (Rodentia), a useful mammal for studying the induction of sex-chromosome nondisjunction and diploid gametes in male germ cells.

Preliminary data indicate that chemicals can also increase the frequency of sex-chromosome nondisjunction. Positive results--which certainly need further confirmation--have been obtained for MMS, p-fluorophenylalanine, vincristine, procarbazine, carbendazim, and bleomycin. Nocodazole, benomyl, colcemic, 6-mercaptopurine, and halothane were all negative at the concentrations tested. For the induction of diploid spermatids positive results were only obtained for MMS and parafluorophenylalanine. In view of the results obtained, the Microtus system is considered a very useful tool for analyzing factors contributing to the high frequency of aneuploidy and triploidy among abortuses and of aneuploidy in liveborn infants of men. A method is described for the detection of sex-chromosome nondisjunction and diploid spermatids in male germ cells of the field vole Microtus oeconomus. The method is based on the unique distribution pattern of heterochromatin in Microtus cells, which makes it possible to identify X and Y chromosomes in early spermatids with a simple C-banding procedure. Slide preparation is easy. Scoring of early spermatids for extra sex-chromosomes is simple and 2000-4000 cells per hour can be examined. With the Microtus system it has now been demonstrated that radiation of spermatocyte stages with doses of 50, 100 and 200 R results in a higher frequency of sex chromosome nondisjunction and of diploid gametes. Both types of aberrant gametes can be produced during the first and second meiotic division.


Detection of nondisjunction in mammals.

Methods have been developed in the past to assess spontaneous and induced chromosomal aneuploidy in germ cells and in early pre- and postimplantation mammalian embryos. Some of these methods yield still more information when combined with chromosome banding techniques. Various chemicals and x-rays have been tested in mammalian oogenesis and x-rays in spermatogenesis. The inference may be drawn from these studies that spontaneous nondisjunction is considered to occur only rarely in mouse and hamster oogenesis and spermatogenesis. X-rays induce nondisjunction during male and femlae meiosis, thus giving rise to significantly more aneuploid oocytes and F1 embryos. The alkylating agents trenimone and cyclophosphamide induce chromosomal missegregation in oocytes; the incidence depends on the dose injected. Hormones used as oral contraceptives did cause aneuploidy in oocytes, but only after daily treatment with high doses. Hormones used for stimulated ovulation did not interfere with chromosome segregation in the mouse and Chinese and Syrian hamsters. The following problems may be considered in futre studies: the problem of a species-specificity for induced nondisjunction; the question of a stage sensitivity (transplacental treatment); what happens after chronic exposure, also at low doses; the presence of a threshold; the existence of a dose-effect relation; the nature of cellular target(s) responsible for induced nondisjunction (spindle, regulatory proteins for polymerization of microtubules and ther depolymerization, centrioles, centromeres, RNA, or gene expression); whether DNA is involved and whether repair capacity plays a role.


Detection of aneuploidy in human sperm.

Adequate methods for monitoring any type of gametic mutation directly in man are virtually nonexistent. A method is presented by which one can monitor Y chromosomal nondisjunction directly in the male gamete by quantifying the number of spermatozoa with two fluorescent bodies (YFF) in 1000 sperm counted. Dried semen slides are stained with quinacrine dihydrochloride and examined under a fluorescent microscope with dark field illumination. This method eliminates the biopsy required for other meiotic studies and further eliminates bias in gametogenic selection by evaluating ejaculated mature spermatozoa. Since chromosomal numerical errors are found in 0.4% of term births and 35% of miscarriages, it is evident that chromosomal aneuploidy constitutes the major mutagenic load in man. In view of the increases observed in the incidence of YFF sperm in patients receiving antineoplastic therapy and in the DBCP-exposed workers, it may be prudent for men who have a history of exposure to mutagens and who are contemplating reproduction to be evaluated by this method prior to attempting conception. Further, this procedure could also be applied to the clinical phase of new drug testing to evaluate the effects of that agent with respect to aneuploidy since the increases in Y chromosomal nondisjunction may well act as a barometer for increases in overall autosomal nondisjunction.


Meiotic aneuploidy: its origins and induction following chemical treatment in Sordaria brevicollis.

A system suitable for the detection of meiotic aneuploidy is described in which various different origins of the aneuploidy can be distinguished. Aneuploid meiotic products are detected as black disomic spores held in asci containing all the products of a single meiosis. Aneuploidy may result from nondisjunction or from a meiosis in which an extra replica of one of the chromosomes has been generated in some other way, e.g., extra replication. By using this system it has been shown that pFPA treatment increase aneuploidy, primarily through an effect on nondisjunction. Preliminary results with trifluralin have indicated that this compound, too, may increase aneuploidy. There is a good possibility that the system can be further developed to permit a more rapid screening using a random plating method; this will allow a more efficient two-part analysis of the effects of compounds under test.


Neurospora prototroph selection system for studying aneuploid production.

Various environmental agents have been tested for their ability to produce aneuploid products of meiosis in a Neurospora cross. The cross was between two multiply marked strains designed specifically for this purpose. These parental strains were heterozygous for four auxotrophic mutations on chromosome 1. Prototrophic disomics could be selected by plating ascospores on minimal medium. Out of 48 agents tested, at least 10 increased the aneuploid frequency significantly above control levels. Some of these positive-testing agents have also been found active in other aneuploid detection systems. It is suggested that if it should become necessary to perform widespread testing of agents in the human environment for their ability to generate aneuploids at meiosis, the Neurospora system could be successfully applied for this purpose.


Analysis of mitotic nondisjunction with Aspergillus nidulans.

Two methods to detect the induction of nondisjunction with a diploid stable strain of A. nidulans are described. The first method gives only qualitative results, while the second method is quantitative and dose-effect curves can be done. Some physiological parameters affecting the induction of nondisjunction can also be studied, because either quiescent or germinating conidia can be treated with the drug under test. Some agents inducing nondisjunction were also tested for the induction of point mutation and somatic crossing-over with these comparative analysis. Two classes of agents inducing nondisjunction may be detected: the first causes all possible types of genetic damage either on quiescent or germinating conidia (a representative of this class is MMS) and acts presumably on the DNA level; the second acts only on germinating conidia and does not produce point mutation or crossing over. A representative of this class is Benomyl which interferes with spindle microtubules. A list of compounds tests is included.


Detection of mitotic and meiotic aneuploidy in the yeast Saccharomyces cerevisiae.

A number of genetic systems are described which involve the use of the yeast Saccharomyces cerevisiae. The systems may be used to detect the production of aneuploid cells produced during both mitotic and meiotic cell division in the presence of genetically active chemicals. During mitotic cell division, monosomic colonies (2n - 1) may be detected by plating upon selective medium. Increases in such monosomic colonies are produced by exposure of cells to a number of chemical mutagens such as ethyl methane-sulfonate and mitomycin C. More importantly, monosomic colonies are also induced by nonmutagens such as sulfacetamide and saccharin, which suggests that such chemicals are capable of inducing aneuploidy (aneugenic) in the absence of mutagenic activity. Genetic analysis of aneuploid colonies produced on nonselective medium indicate that at least a proportion of the monosomic colonies were the result of mitotic nondisjunction. During meiotic cell division, disomic cells (n + 1) produced by chromosome nondisjunction may be detected by plating on selective media. The frequency of disomic cells has been shown to increase after exposure to p-fluorophenylalanine.


Creatine kinase isoenzyme BB in serum of patients undergoing chronic hemodialysis and with kidney transplant.

The activity of creatine kinase isoenzyme BB (CK-BB) was determined in serum of healthy adults and in patients undergoing maintenance hemodialysis and with kidney transplant. In the healthy adults examined, an activity of 0.56 +/- 0.16 U/l (mean +/- SD) was found. The arithmetic mean of CK-BB activity in patients with renal insufficiency under hemodialysis was 1.42 +/- 0.87 U/l and differed from that of the healthy group. The CK-BB activity in patients with kidney transplant was not different from that of the control group. The occurrence of CK-BB in serum is discussed from diagnostic and methodological point of view.


Evidence for an enhanced de novo synthesis of alkaline phosphatase in cholestatic rat liver using immunotitration and precursor incorporation techniques.

An antikidney phosphatase serum was produced. This showed a cross-reaction with liver phosphatase and precipitated the latter enzyme specifically in the double antibody method. An U-[14C] protein hydrolysate was injected intraperitoneally into rats, which had previously undergone bile duct ligation. Liver alkaline phosphatase was partially purified and immunoprecipitated. By determination of phosphatase labelling the extent of de novo synthesis of the phosphatase protein was evaluated. Comparing livers from control and cholestatic rats, it could be shown that 12 h after beginning of cholestasis the de novo synthesis of alkaline phosphatase was increased up to 4-fold and that is remained at a 2-fold increased level for at least 2 days.


Uptake and conversion of the antibiotic albomycin by Escherichia coli K-12.

The antibiotic albomycin is transported into cells of Escherichia coli K-12 by the same uptake system as the iron-supplying ferrichrome complex. The iron-complexing hydroxamate moieties of albomycin and ferrichrome are structurally similar. During the phase of rapid iron uptake the chelators were not found in the cells. In order to understand the antibiotic activity of albomycin, it was labeled in the hydroxamate with tritium and in the presumed antibiotically active area with radioactive sulfur. While the tritium label was not retained by the cells, part of the sulfur label was taken up and concentrated 500-fold within the cell. The sulfur was not incorporated into proteins or nucleic acids since it was recovered as a low molecular weight component. Gel filtration on Bio-Gel P-2 revealed one tritium-labeled and two sulfur-labeled cleavage products in the incubation medium. We conclude that albomycin is actively transported via its ferrichrome-like portion into the cells and that the growth-inhibitory moiety is released by hydrolysis intracellularly and remains there.


Controlled sequential pacing in isolated perfused auxiliary rat hearts.

An important question after clinical auxiliary heart transplantation is whether controlled pacing of both hearts is desirable in order to protect the diseased recipient heart. A comparison of the effects of spontaneous beating with three types of pacing (random, simultaneous and sequential) was undertaken in an isolated perfused working double rat heart. The two hearts were attached to each other by the atria, in a way comparable to the clinical situation. During controlled sequential pacing of both normal hearts, the peak systolic pressure fell and diastolic pressure rose, but the combined cardiac output did not changes. In contrast, during simultaneous pacing, systolic pressure rose, diastolic pressure  fell and coronary flow was less than with sequential pacing, but the oxygen uptake was higher. Hence the effects of sequential pacing could be expected to be beneficial when function of one of the hearts was depressed by left main coronary artery ligation. During sequential pacing the cardiac output was significantly improved from 10 to 30 ml/min (P less than 0.001) and coronary flow rose. Myocardial oxygen consumption increased from 80 to 110 microliters/g/min (P less than 0.05). Thus sequential pacing could improve the severely depressed hemodynamics of the coronary-ligated heart in the presence of an auxiliary heart. The increased myocardial oxygen consumption is viewed as a beneficial effect of increased diastolic perfusion pressure and was not associated with increased enzyme release. It is concluded that the effects of sequential pacing warrant assessment in clinical auxiliary heart transplantation.


Comparison of pirbuterol and pirbuterol and hydroxyzine in patients with reversible airway obstruction.

The effects of single oral doses of pirbuterol 10 mg plus hydroxyzine 10 mg, and of pirbuterol 10 mg plus placebo, were compared in a single-blind cross-over trial in 17 adult patients with reversible airway obstruction. The initial bronchodilator response after the two treatments was similar, but mean specific airway conductance at 3, 4, 6 and 8 h, and mean FEV1 and FVC after 3 and 4 hours, were significantly higher after pirbuterol + hydroxyzine than after pirbuterol + placebo. The mean thoracic gas volume measured at 3 and 4 h was lower after pirbuterol + hydroxyzine. Heart rate, blood pressure and a continuous ECG record did not differ after the two treatments. The findings suggest that the combination of pirbuterol and hydroxyzine brings about more prolonged bronchodilatation in patients with reversible airway obstruction than does pirbuterol alone.


Clinical trial of brinase and anticoagulants as a method of treatment for advanced limb ischemia.

A randomized, single-blind trial of repeated intravenous infusion of Brinase was carried out in 70 petients with severe chronic limb ischemia, who were candidates for lumbar sympathectomy or amputation. The enzyme caused s significant increase in calf and ankle pressure index. At six months follow-up, the clinical results were statistically significant in favour of Brinase when all patients were considered, but not if 10 patients with Buerger's disease were omitted from the analysis. Patients treated with a combination of Brinase and coumarins had a better clinical outcome than patients receiving either treatment on its own.


The enumeration of rat IgE-secreting cells using a reverse plaque-forming cell assay.

A reverse hemolytic plaque assay utilizing protein A-coated sheep red cells and a specific rabbit anti-rat IgE preparation has been adapted for the enumeration of rat IgE-secreting cells derived from the IR-162 rat plasmacytoma and from rats infected with Nippostrongylus brasiliensis. Under optimal conditions, approximately 10-15% of the viable plasmacytoma cells were scored as plaque-forming cells. In rats infected with 5000 Nippostrongylus brasiliensis larvae, a maximum of 2 x 10(6) IgE-secreting cells were found in the mesenteric lymph nodes, and no IgE plaque-forming cells in their spleens. The kinetics of the mesenteric lymph node plaque-forming cell responses closely coincided with total serum IgE levels, with maximum responses occurring 15-16 days after infection. There was a high degree of correlation between the mesenteric lymph node IgE plaque-forming cell responses and total serum IgE levels of individuals rats. It was concluded that the IgE-secreting cells in the mesenteric lymph nodes contributed, in a large way, to the elevated levels of IgE found in the circulation of these rats.


Detection of albumin-secreting hepatocytes at the single cell level by a protein A plaque assay.

In a complement-dependent assay, using protein A-coupled erythrocytes as target cells and anti-albumin immunoglobulin (Ig) as a developing serum, it was possible to detect single albumin-secreting hepatocytes as plaque-forming cells. Plasmacytoma cells were detected using anti-Ig antibodies and did not form plaques with anti-albumin Ig. This method can potentially be used for detecting any cell-secreting products in sufficient amounts.


Lipopolysaccharide and lipid A-induced human blood lymphocyte activation as detected by a protein A plaque assay.

Various purified cell wall lipopolysaccharides (LPS) from gram-negative bacteria and derivatives of these LPS were tested for their stimulatory capacity for human peripheral blood cells. Immunoglobulin (Ig) production was tested by an indirect plaque-forming cell assay using Staphylococcus aureus protein A-coupled erythrocytes and specific anti-Ig as developing serum. This method allows the detection of the majority of cells secreting Ig of a single class, and the number of plaque-forming cells detected are approximately 100-1000 times the amount obtained using normal sheep red cells as targets. LPS containing the O antigen-specific chain, as well as mutant products only containing lipid A and ketodeoxyoctonate trisaccharide, could induce cell division and antibody synthesis. The polypeptide antibiotic polymyxin B was found to inhibit LPS-induced activation. Furthermore, purified lipid A, complexed with bovine serum albumin, was also found to activate human peripheral blood B cells. These findings demonstrate that human peripheral blood lymphocytes can be activated by LPS and also indicate that lipid A is the active part of these molecules.


Wheat germ agglutinin activates macrophages for collaboration with B cells.

Wheat germ agglutinin (WGA), added to Mishell-Dutton-type cultures of nude spleen cells challenged with erythrocyte antigens, allows a primary specific plaque-forming cell (PFC) response. WGA, however, does not display direct mitogenicity to either T or B lymphocytes. The activity of WGA in the specific PFC response is macrophage-dependent and parallels a nonspecific PFC response in the same cultures.


Prostaglandin-like substances in Propionibacterium acnes. III. Differential contractile effects on smooth muscle layers of the human utero-tubal junction.

The biological activity of a lipid fraction extracted from P. acnes was tested on isolated smooth muscle strips from the human utero-tubal junction. The bioassay experiments support the concept that prostaglandin-like substances (PLS) occur in P. acnes. However, in the bioassay system used, the effect of PLS was different from that of PGF2 alpha and PGI2 but similar, although not identical, to that of arachidonic acid and PGE2.


Systemic candidiasis from Candida albicans colonizing the gastrointestinal tract of mice.

Reproducible induction of systemic Candida infection was achieved by treating mice in which Candida colonization had been established in the gastrointestional tract by aminobenzylpenicillin treatment. Systemic candidiasis was induced in these mice by X-ray irradiation followed by immunosuppressive doses of dexamethasone or X-ray irradiation followed by immunosuppressive doses of trypan blue. Macrophages seem to play an important role in thie systemic infection.


Octopamine levels during the moult cycle and adult development in the migratory locust, Locusta migratoria.

Octopamine content of the head of the locust Locusta migratoria has been determined during the last larval stage, moulting and adult life of 3 groups of insects: female and male gregarious, solitary and CO2 solitarized. An important difference was found between these 3 groups. Octopamine contents increased in the middle of the larval life and during the adult life. The moulting time is characterized by a sharp decrease of the octopamine content which becomes identical in the 3 groups of insects. The relation between octopamine content, hormone cycles and motility is discussed.


Colony stimulating activity in acute and chronic endotoxinemia in man.

Blood granulocyte-macrophage colony stimulating activity (GM CSF) was measured in 6 normal individuals challenged with low-dose endotoxin and in 63 unselected patients with nonhaematological disorders. 5/63 patients were febrile and 5 other patients whoed detectable endotoxin levels, as measured by the Limulus assay. CSA levels showed a rapid increase in normal individuals following endotoxin administration, but were in the normal range in patients with chronic endotoxinemia or in those with febrile disorders. Thus, unlike acute endotoxinemia, chronic endotoxinemia is not associated with elevated activity that promotes growth of myeloid commited stem cells. In addition, fever per se did not coincide with elevated blood CSA levels.


Relationship of endogenous to dietary sorbitol. A study in normal and diabetic rats.

The relationship of endogenous to dietary sorbitol was investigated by measuring the sorbitol content of plasma, lens and kidney in normal an streptozotocin-injected rats fed for two-three weeks a control diet or a diet rich in either sorbitol or sucrose. The insulin content and secretory capacity of the pancreatic islets removed from these rats was also measured. In normal rats, the carbohydrate-rich diet increased the concentration of sorbitol in plasma but not lens or kidney. The carbohydrate-rich diet also increased the secretory responsiveness of the B-cell to glucose. Accumulation of sorbitol in lens and kidney occurred in diabetic rats. When the diabetic rats were treated with insulin, the sorbitol content of plasma and lens was less elevated than in untreated diabetic animals, despite poor control of the glycemia. The insulin treatment failed to prevent sorbitol accumulation in the kidney. There was no obvious difference between dietary sorbitol and sucrose respectively, in terms of sorbitol accumulation in tissues or stimulation of B-cell secretory potential.


Metformin: a review of its pharmacological properties and therapeutic use.

In a survey, the pharmacological and clinical documentation of metformin is presented and discussed, and the present state of knowledge relating to metformin-associated lactic acidosis is reviewed. The use of metformin in the treatment of diabetes is based on clinical experience over twenty years. It has been well documented that metformin is effective in maturity-onset diabetes both as monotherapy and in combination with a sulphonylurea. An advantage of metformin treatment is the tendency to weight reduction and the absence of significant hypoglycaemia; blood glucose levels are reduced only to normal. The disadvantages are the gastro-intestinal side effects and the potential risk of vitamin B 12 and folic acid deficiency during long-term use. Metformin-associated lactic acidosis is a very rare complication, which has mainly occured in patients with serious renal insufficiency or other contra-indications to the use of metformin. The association between phenformin and lactic acidosis has led to withdrawal of this biguanide in several countries. Metformin differs from phenformin in certain important respects, and the normal use of metformin does not involve the risk of side effects disproportionate to the intended effect. Further experimental studies are required to substantiate pharmacokinetics and metabolic effects of metformin in man.


[Treatment of diabetic retinopathy with laser photocoagulation (author's transl)].

Laser photocoagulation has changed the visual prognosis of diabetic patients affected by retinopathy. It aims to lower the risk of blindness of diabetic patients. The goal of photocoagulation is to reduce the tissue damage of microangiopathic origin expressed by nonperfusion areas and permeability abnormalities that are responsible for retinal ischemia and oedema respectively. Loss of visual acuity in the diabetic is due mainly to two causes : first, vitreous hemorrhage with its dramatic loss of vision; secondly, macular cystoid oedema, occuring more commonly and with progressive loss of central vision. The efficiency of pan-retinal photocoagulation in reducing the risk of vitreous hemorrhage and consequent blindness in patients with disc or preretinal newly formed vessels, has been ascertained by American and British randomised studies. The indications, technics and results of photocoagulation in non-proliferative diabetic retinopathy are the subject of many studies. Only photocoagulation for macular oedema due to intra-retinal microvascular abnormality has shown to be of benefit.


Comparative trial of a new hypnotic (Finorgal) with nitrazepam and triclofos sodium.

Thirty patients complaining of insomnia were studied in a double-blind trial with crossover of Finorgal, nitrazepam and triclofos sodium. Finorgal given as a hypnotic produced similar results to nitrazepam and triclofos sodium in terms of induction of sleep, duration and quality of sleep, dream recall and morning 'hangover'. Reported side-effects were not serious and occurred less frequently in association with Finorgal treatment than with nitrazepam or triclofos sodium. Laboratory investigations gave no indication of the development of any drug toxicity during the three-week period of the trial.


Comparison of a new hypnotic (Finorgal) with placebo in a double-blind trial.

The hypnotic effects of Finorgal (ethchlorvynol with diphenhydramine) were compared with those of placebo in a double-blind study with crossover of treatments in thirty-five hospital in-patients. During the four-week period of Finorgal treatment there was a significant reduction in the mean time elapsing between the administration of the hypnotic and the onset of sleep, and a significant increase in the duration of sleep, compared with the four weeks of placebo treatment. There was also a significant increase in the proportion of nights when the patients felt they had slept well, and in the incidence of morning 'hangover' and nocturnal confusion during the Finorgal treatment periods. Patients had to be actively woken in the morning significantly more often following Finorgal administration. In patients experiencing pain in the night there was a significant reduction in the occurrence of pain during the nights when Finorgal had been given.


Neurobiochemical aspects of maprotiline (Ludiomil) action.

In the first part of the paper a short review of the neurobiochemical effects of the antidepressant drug maprotiline is given. Its most obvious effect is the inhibition of noradrenaline uptake in peripheral and central neurones. The peculiarity of this action consists in its high degree of selectivity, as no inhibition of serotonin uptake could be demonstrated in vivo. In the second part, the results of new experiments are described. These show that serotonin uptake is not diminished in rat mid-brain synaptosomes even after treatment with very high doses of maprotiline (600 mg/kg p.o.). In addition, the influence of the antidepressant on noradrenaline and serotnin uptake was studied in rat cerebral cortex, cerebellum, hypothalamus and pons-medulla. Dopamine and serotonin uptake were measured in the corpus striatum. Again, only the uptake of noradrenaline was found to be inhibited. There was not even a slight tendency towards inhibition of serotonin uptake. This high degree of selectivity distinguishes maprotiline from the tricyclic antidepressants and thus makes it an intersting 'extreme-type' uptake inhibitor.


Comparison of diazepam and flurazepam in the treatment of insomnia in general practice.

In a double-blind crossover study in general practice, flurazepam was shown to be significantly better (p less than 0.001) than diazepam in treating sleep disturbance. Fewer patients reported side-effects on flurazepam.


A double-blind controlled trial of betamethasone dipropionate 0.05% (Diproderm) in comparison with fluocinolone acetonide 0.025% (Synalar) in psoriasis and other steroid-responsive dermatoses.

In a double-blind randomized study, sixty-two patients suffering from psoriasis or eczema were treated, twice daily for three weeks, either with 0.05% betamethasone 17, 21-dipropionate cream or with 0.025% fluocinolone acetonide cream. The results showed that both topical corticosteroid preparations were effective, well tolerated and cosmetically acceptable. Fifty-seven per cent of the patients treated with betamethasone dipropionate were rated as being 'much better' in the overall assessment of response at the end of the trial period compared to only 25% of those in the fluocinolone acetonide group.


A clinical trial of choline theophyllinate versus salbutamol with placebo control.

Using a randomized double-blind procedure, seventeen patients with chronic airways obstruction completed a placebo-controlled study to compare bronchodilator effects of single doses of choline theophyllinate 400 mg and salbutamol 4 mg. As determined by a range of spirometric measurements, the effects of the two drugs were remarkably similar and there were no statistically significant differences in any of the parameters measured. Both drugs produced highly significant differences in comparison with placebo response. Side-effects were mainly complaints of digital tremor; this was seen in four of the patients after salbutamol and in less pronounced form in two patients after choline theophyllinate.


Hydrocortisone alcoholic solution in eczematous external otitis.

A double-blind, randomized study is described involving a total of fifty-five patients suffering from eczematous external otitis comparing for efficacy hydrocortisone alcoholic solution and hydrocortisone-17-alpha-butyrate. The trial duration was seven days. No difference in efficacy could be demonstrated between the two preparations. No side-effects were observed.


Clinical effects of trithiozine, a newer gastric anti-secretory agent.

Trithiozine is a new product which in animal studies proved to have a marked anti-secretory and anti-ulcer effect, along with a very low acute and chronic toxicity and no ganglioplegic, anticholinergic and anti-H2 activity. Clinical trials with trithiozine have been performed in some European countries. The results of these trials, most of which were conducted on a double-blind basis, and including already several hundreds of patients, have shown that oral trithiozine: exerts a very significant action on both basal and stimulated gastric acid secretion, without a rebound hypersecretion; promotes, in most patients, a complete endoscopic healing of peptic ulcer, in addition to an early symptomatic relief; has a mild sedative action; does not affect the pancreatic secretion; is well tolerated even for long-term (up to 10 months) treatments.


Isolation and characterization of preproinsulin mRNA from fetal bovine pancreatic islets.

Total polyadenylated RNA prepared from isolated islets of fetal bovine pancreas was physically characterized. Incubation of this poly A+ RNA with a cell-free protein-synthesizing system derived from wheat germ results in the synthesis of insulin-immunoreactive polypeptide identical in size to that described earlier, 11 200 daltons (Lomedico et al. (1977) J. Biol. Chem. 252, 7971--7978). This material comprised approximately 22% of the total 3H-labeled translation products. Compared to poly A+ RNA from the total pancreas, we conclude that islet mRNA is enriched in proinsulin mRNA.


Somatostatin in the pancreas and hypothalamus of obese mice.

The pancreatic content of somatostatin, insulin, and glucagon and the hypothalamic content of somatostatin were examined in ob/ob mice at various ages and in goldthioglucose-obese mice. The total pancreatic content of somatostatin was increased in ob/ob mice compared to controls: 92 ng vs 75 ng (a 22% increase) at 2 months of age; 208 ng vs 131 ng (a 60% increase) at 6 months of age; and 184 ng vs 118 ng (a 60% increase) at 8 months of age. The total pancreatic content of glucagon in ob/ob mice was already enhanced by 70% over controls at 2 months of age (301 ng vs 173 ng) and did not increase further at later stages, whereas that of insulin progressively rose with age. In goldthioglucose-obese mice the pancreatic content of insulin was also increased but to a lesser extent than in ob/ob mice; the pancreatic levels of somatostatin and glucagon were unaltered. In both ob/ob mice (regardless of age) and goldthioglucose-obese mice, there was no significant change in the hypothalamic content of somatostatin compared with that of lean controls.


Bone loss in diabetes: effects of metabolic state.

The significance of different risk factors for the development of bone loss in diabetes mellitus was evaluated in a cross sectional study of 215 insulin treated diabetic outpatients. Bone mineral content in the forearms was measured by photon absorptiometry and the metabolic status was evaluated by three indices: residual B-cell function, insulin dosage and fasting blood glucose. The mean bone mineral content was reduced to 90.2% of sex- and age-matched normal mean values (P less than 0.001). Stratification of the patients showed that bone mineral content was 99.3% of that found in sex- and age matched normal subjects in the group with residual B-cell function, low insulin dosage and low fasting blood glucose; it was only 79.3% of normal in the group with no detectable insulin secretion, high insulin dosage and more severe hyperglycaemia. Thus, residual insulin secretion and the quality of metabolic control are major factors in determining bone mineral content in insulin treated diabetic patients.


Diabetic autonomic neuropathy.

This review attempts to outline the present understanding of diabetic autonomic neuropathy. The clinical features have been increasinly recognised but knowledge of the localization and morphology of the lesions and their pathogenesis remains fragmentary. A metabolic causation as postulated in somatic nerves accords best with clinical observations. Most bodily systems, particularly the cardiovascular, gastrointestinal and urogenital, are involved with added disturbances of thermoregulatory function and pupillary reflexes. Possible effects on neuroendocrine and peptidergic secretion and respiratory control await definition. Current interest centres around the development of a new generation of tests of autonomic nerve function that are simple, non-invasive, reproducible and allow precision in diagnosis and accurate quantitation. Most are based on cardiovascular reflexes and abnormality in them is assumed to reflect autonomic damage elsewhere. Probably no single test suffices and a battery of tests reflecting both parasympathetic and sympathetic function is preferable. Little is known of the natural history. The prevalence may be greater than previously suspected and although symptoms are mild in the majority, a few develop florid features. The relation of control and duration of diabetes to the onset and progression of autonomic neuropathy is not clearly established. Once tests of autonomic function become abnormal they usually remain abnormal. Symptomatic autonomic neuropathy carries a greatly increased mortality rate possibly due to indirect mechanisms such as renal failure and direct mechanisms such as cardio-resiratory arrest. Improved treatment of some of the more disabling symptoms has been possible in recent years.


Ultrastructural and morphometric studies of delta cells in pancreatic islets from C57BL/Ks diabetes mice.

An ultrastructural and immunocytochemical study was undertaken to elucidate the temporal and quantitative aspects of the changes occurring in the delta cells in the pancreatic islets of C57BL/KsJ db/db (diabetes) mice. Electron microscopy revealed that prior to the major topographical redistribution of delta cells from their peripheral location to the islet interior, long delta cell filopodial extensions penetrated into the islet, greatly increasing the area of surface contact between delta cells and hypersecretory beta cells. Coincident with delta cells redistribution in islets of 8 to 10 week diabetes mice, the mean number of delta cells per islet had increased significantly. In contrast, their volume density had decreased, indicating incomplete compensation for beta cell hyperplasia which had commenced approximately 4 weeks earlier. In the 14 week mutants, numbers of delta cells per islet and islet volume reached maximum values while delta cell volume density had been restored to a control level. Delta cell volume density exhibited a 2-fold increase in the mutants at 20 weeks that coincided with massive beta cell necrosis. However, a decline in the number of delta cells per islet (173.6 +/- 20.9 at 14 weeks versus 91.2 +/- 20 weeks) suggests that islet degeneration in terminal stages of the syndrome also includes some loss of these cells.


Histogenesis of adenomatous polyps in the human large intestine.

Detailed histologic analyses have been performed in 75, minute, colorectal polyps from familial polyposis patients. All polyps were composed of typical adenomatous tissue, with, in addition, some normal-looking glands still to be found among the neoplastic elements. Bifurcating glands were not observed, and branching patterns were present in 27% of the polyps only. In 86% of the lesions, the number of gland openings along the polyp surface was larger than the number of gland bases observed along the muscularis mucosae. This difference increased with polyp size (r = 0.9043). Those data, together with previous radioautographic observations, suggest that formation of new adenomatous glands mainly results from an infolding of the surface epithelium between normal, preexisting glands. This mechanism sharply contrasts with villous polyps in which papillary projections arise upwards from the mucosal surface.


Effects of prostacyclin and a stable analogue, 6-beta-PGI1, on gastric acid secretion, mucosal blood flow, and blood pressure in conscious dogs.

We studied the effect of prostacyclin, PGI2, its chemical decomposition product, 6-oxo-PGF1 alpha, and a stable 5-6-dihydro analogue, 6-beta-PGI1, on gastric acid secretion, mucosal blood flow (14C-aminopyrine clearance), and mean arterial pressure in unanesthetized dogs. During submaximal acid secretion from a gastric fistula induced by intravenous histamine dihydrochloride (20 microgram kg-1 h-1), prostacyclin and its stable analogue, 6-beta-PGI1, reduced acid output with ID50s (dose causing 50% inhibition) of about 0.2 and 3.0 microgram kg-1 min-1 i.v., respectively, whereas 6-oxo-PGF1 alpha was inactive at 100 times the effective dose of prostacyclin. The ratio of mucosal blood flow to acid output remained unchanged during prostacyclin administration and was significantly elevated during 6-beta-PGI1 infusion, suggesting that with both compounds the changes in mucosal blood flow were not the cause of the antisecretory action. For doses causing equivalent antisecretory action, 6-beta-PGI1 lowered systemic arterial blood pressure much less than prostacyclin, indicating selectivity of action. Prostacyclin is unlikely to be a circulating antisecretory agent, but may play a role as a local humoral modulator of secretion and blood flow in the gastric mucosa.


Histidine mutants requiring adenine: selection of mutants with reduced hisG expression in Salmonella typhimurium.

A method is described for the selection of Salmonella typhimurium mutants with reduced levels of hisG enzyme activity. This method is based on the fact that the hisG enzyme catalyzes the consumption of ATP in the first step of histidine biosynthesis. Normally, this reaction is closely regulated, both by feedback inhibition and by repression of the operon. However, conditions can be set up that result in the uncontrolled use of adenine in histidine biosynthesis. Cells grown under these conditions become phenotypic adenine auxotrophs. Some revertant clones that no longer require adenine contain mutations in hisG, hisE, or the his-control region. The hisG mutations are of all types (nonsense, frameshift, missense, deletion and leady types), and they map throughout the hisG gene.


A refined map of the hisG gene of Salmonella typhimurium.

The hisG gene is the most operator-proximal structural gene of the histidine operon; it encodes the feedback-inhibitable first enzyme of the biosynthetic pathway. Previously, hisG mutants were mapped into seven intervals defined by the availble deletion mutations having endpoints in the hisG gene. The map has been refined using over 60 new deletion mutants. The new map divides the gene into 40 deletion intervals, which average approximately 30 base pairs in length. The map has been used to analyze the distribution of insertion sites for the transposable element Tn10 and has permitted conclusions on the diistribution of duplication endpoints. The map promises to be useful in analysis of his regulation and, more particularly, in the determination of the possible role of the hisG enzyme in this mechanism.


Mutagenesis by cytostatic alkylating agents in yeast strains of differing repair capacities.

Reversion of two nulcear ochre nonsense alleles and cell inactivation induced by mono-, bi-, and tri-functional alkylating agents and by UV has been investigated in stationary-phase haploid cells of yeast strains with differing capacities for DNA repair. The ability to survive alkylation damage is correlated with UV repair capacity, a UV-resistant and UV-mutable strain (RAD REV) being least and a UV-sensitive and UV-nonmutable strain (radi rev3) most sensitive. Mutagenicity of alkylating agents is highest in the former and is abolished in the latter strain. Deficiency in excision repair (rad1 rad2) or in the RAD18 function does not lead to enhanced mutability. Mutagenesis by the various agents is characterized by a common pattern of induction of locus-specific revertants and suppressor mutants. Induction kinetics are mostly linear, but UV-induced reversion in the RAD REV strain follows higher-than-linear (probably "quadratic") kinetics. The alkylating agent cyclophosphamide, usually considered inactive without metabolic conversion, reduces colony-forming ability and induces revertants in a manner similar but not identical to the other chemicals tested. These findings are taken to support the concept of mutagenesis by misrepair after alkylation, which albeit sharing common features with the mechanism of UV-induced reversion, can be distinguished therefrom.


[Acceptance and transfer of plasmid Rts1 by bacteria of the genus Erwinia].

The ability of 13 Erwinia strains to accept, to inherit and to transmit the Rts1 factor by conjugation was studied. 11 strains accepted the Rts1 factor from Escherichia coli K-12 CSH-2 with the frequency of about 10(-7)--10(-3). The Rts1 factor was genetically stable in the Erwinia cells and was not eliminated by acriflavine and under the temperature of 37 and 42 degrees C. All the R+ exconjugants were characterized with more high degree of the resistance of kanamycin than E. coli cells harbouring the same R factor. Erwinia strains harbouring the Rts1 plasmid transferred it by conjugation into homologic (Erwinia) and heterologic (E. coli) bacteria. The study of kinetics of the transfer of the Rts1 factor in different mating systems showed that the transfer of this plasmid from R+ Erwinia into R- Erwinia and R- E. coli--in the liquid medium. It is concluded that Erwinia can be the host and the donor of the Rts1 factor.


[Functional interactions of the genomes of Shigella sonnei phages and Escherichia coli phage T4 in mixed infection].

A comparative study of Shigella sonnei phages U and G and Escherichia coli phage T4 has shown that enzymes coded for by the Sh. sonnei phages can functionally substitute for some T4-coded products. This finding in indicative of an evolutionary relationship between T-even phages and disenteric phages U and G. The U phage is uncapable to compensate amber mutants for the genes that control the conversion of cytosine into 5-hydroxymethyl cytosine (5-HMC) and the glucosylation of the latter, which agrees with our earlier finding that the U phage DNA contains no 5-HMC. U and G phages are also found to exclude the T4 phage in the course of mixed infection.


Preparation and characterization of large amounts of restriction fragments containing the E. coli lac control elements.

Large quantities of pure DNA fragments (789, 203 and 95 bp in length) containing the Escherichia coli lac controlling elements (operator, promoter, CRP binding site) were prepared from appropriate recombinant plasmids. High pressure liquid chromatography on RPC-5 or preparative sucrose gradient centrifugation was used to fractionate the pVH51 vector from the inserts. The fragments had few, if any, nicks or depurinated sites, and the majority of the fragment ends were intact. Absorbance-temperature profiles on the fragments showed multiphasic transitions.


Thermo-inducible expression of cloned early genes of bacteriophage Mu.

An EcoRI fragment, containing approx. 5100 base pairs (bp) of the immunity-end of bacteriophage Mu, was inserted into the multicopy plasmid pMB9 by in vitro recombination. The expression of early Mu genes, located on the cloned fragment, is thermo-inducible because of the presence of the ts mutation in gene c. The isolation of a transformant harbouring the recombinant plasmid, pGP1, was possible only when expression of Mu genes was prevented. pGP1 can be maintained at 28 degrees C at high copy number, but at 42 degrees C the pGP1 containing cells are killed due to the expression of the kil gene of Mu. The following Mu genes are present on pGP1: the ner gene, the integration and replication genes A and B, the cim gene, and the kil gene. pGP1 containing cells do not show Gam and Sot activity at 42 degrees C, therefore the leftmost EcoRI site on the Mu DNA is located between genes kil and gam or sot, or within the gam or sot gene.


Effect of alpha-glucosidehydrolase inhibition and intestinal absorption of sucrose, water, and sodium in man.

The effect of a new complex oligosaccharide exhibiting potent inhibitory action on alpha-glucoside hydrolases on intestinal absorption of sucrose in man was tested by constant in vivo perfusion of the jejunum. At concentrations of 4.65 or 15.5 X 10(-6)M the alpha-glucosidehydrolase inhibitor (alpha-GHI) markedly inhibited absorption of glucose from sucrose and absorption of sodium and water. Oral administration of the alpha-GHI resulted as well in depression of solute, sodium, and water absorption. This new compound can serve as an interesting tool to induce carbohydrate malabsorption by inhibition of final digestion and may possibly be of beneficial therapeutic effect in dietary control of diabetes or obesity.


Arterial and portal blood supply in cirrhosis: a functional evaluation.

The uptake of (125)I albumin microaggregates (U-(125)I-AMA) from portal blood, during a single passage through the hepatic reticuloendothelial system, has been found to be generally decreased in cirrhosis. To investigate if a similar phenomenon occurs for the colloid flowing through the hepatic artery, the U-(125)I-AMA was first calculated in normal dogs after injection of a mixture of (51)Cr red blood cells ((51)Cr-RBC) and (125)I-AMA into the hepatic artery by comparing hepatic indicator dilution curves (IDC) obtained with both indicators. In nine dogs, the U-(125)I-AMA from hepatic artery blood was generally over 90%, as previously reported for the same colloid flowing through the portal vein in another group of normal dogs. This approach was then applied in nine patients with alcoholic cirrhosis who underwent combined umbilicoportal vein, hepatic vein, and hepatic artery catheterisation because of severe portal hypertension. Hepatic indicator dilution curves were obtained in the nine patients after injection of a mixture of (51)Cr-RBC and (125)I-AMA into the portal vein and the hepatic artery. The U-(125)I-AMA from portal and hepatic artery blood was measured by comparing (51)Cr-RBC and (125)I-AMA hepatic IDC. U-(125)I-AMA varied between 5.2 and 90.5% after portal vein injection and between 13.7 and 90.1% after hepatic artery injection; not difference was found between paired values. In all patients the extraction of indocyanine green (E-ICG) was calculated during a continuous infusion and significant correlations were found between E-ICG and U-(125)I-AMA from portal blood (r=0.931; p <0.001) or from hepatic artery blood (r=0.861; p <0.005). The decreased uptakes can be related to intrahepatic shunts or sinusoidal changes responsible for ineffective phagocytosis and restricted access of dye to parenchymal cells. These data indicate that in cirrhosis the hepatic artery and portal vein blood is cleared of colloid and ICG in a similar fashion and suggest nearly identical blood supply to the regenerative nodules by the hepatic artery and portal vein. Thus U-(125)I-AMA from hepatic artery or portal vein blood, as well as the E-ICG, may be used to estimate the functional hepatic blood supply in cirrhosis; this may prove to be useful in the prognosis of patients before portacaval shunts.


Chemotherapy in advanced gastric cancer: a controlled, prospective, randomised multi-centre study.

Seventy-six patients with advanced gastric adenocarcinoma were studied in a prospecitive, randmoised, controlled trial using vincristine, methotrexate, cyclophosphamide, and 5-fluorouracil in an initiation course and mitomycin-C with 5-fluorouracil as maintenance therapy. Thirty-seven patients were inoperable and 39 had the primary tumour resected with histological evidence of residual disease. Survival in the inoperable group was short and showed no significant difference between treated and control patients. The median survival times for treated and control groups were 9.5 and 9.0 weeks respectively. In the resected patients there was no difference in ultimate overall survival between the groups but up to 20 weeks there was a suggestion that the probability of survival in treated patients was higher (P = 0.06). The patients were well-matched and it is concluded that chemotherapy has had an early effect but that a further trial with more detailed stratification, particularly of staging and histological grade, is needed. No patient received treatment for longer than two years and unacceptable toxicity occurred in only two patients. Nausea occurred more frequently in the treated group but was short-lived and clinically manageable.


Effect of massive bowel resection on enteroinsular axis.

Pancreatic endocrine function was studied in 13 patients who had undergone massive bowel resection. The patients were divided into two groups: one month after operation (short-term group), and three or more months after operation (long-term group). Oral administration of glucose caused a persistent low insulin secretion in almost all the patients and the glucose tolerance curve showed a diabetic pattern in four. In contrast, the insulin response to intravenously infused arginine was impaired in the short-term group, but was at an approximately normal level in the long-term group. Pancreatic glucagon response to intravenous arginine, however, remained unchanged in both groups. Interruption of the enteroinsular axis as the result of massive bowel resection probably plays a major part in this discrepancy of the behaviour of insulin in the long-term group.


The inhibitory effect of PGI2 (prostacyclin) on white platelet arterial thrombus formation.

Prostaglandins play an important role in the platelet-vessel wall interactions. Superfusion with PGI2 of arterial segments prepared in an in vivo model for thrombus induction resulted in a very marked dose-effect-related decrease of white arterial thrombus formation.


[Extension of graft survival time by means of a modification of the treatment with autologous blood].

A modification of the therapy with patients' own blood was achieved by the application of blood components together with an adjuvant. In a defined experimental rat model it was to be clarified whether it is possible to sensitize animals with an antibody directed against skin together with an adjuvant, in order to produce an anti-antibody, which is able to protect a consecutive skin graft from rejection. In a similar way animals were sensitized with cells from rats previously immunized against skin from the donor. By this treatment it is possible to improve survival time of skin grafts. Whereas the prolongation of survival time after pretreatment with skin antibodies can be explained by an anti-antibody, which was found in the serum of the animals, such an anti-antibody could not be detected in the serum of animals, pretreated with "sensitized" cells. Possibly the treatment with cells induces an unspecific effect, which can be achieved also with normal cells. It is conceivable that this principle of the prolongation of skin graft survival time by immunization with certain components of the blood is not only useful in transplantation immunology, but also in other diseases, i.e. autoimmune diseases or allergic reactions.