Pharmacokinetics of sisomicin in patients with normal and impaired renal function; its efficacy in urinary tract infection.

Serum concentration, biological half-life, distribution space and serum clearance of sisomicin, a new aminoglycoside antibiotic, have been studied in twenty-three patients in comparison with the pharmacokinetics of 125I-labelled iothalamate, a compound only filtered by the kidney. 10 patients had normal or borderline abnormal serum creatinine (less than 1,5 mg/100 ml), 8 had various degrees of renal insufficiency (serum creatinine 1.7-9.6 mg/100 ml) and 6 were being treated by intermittent haemodialysis. After intravenous injection of sisomicin 1 mg/kg body weight in patients with normal or borderline renal function its half-life was 3.5 h, very similar to that of iothalamate, 3.2 h. The mean distribution space was 20.1% per cent of body weight; iothalamate, 23.7%. In patients with renal insufficiency there was a positive correlation between serum creatinine level and the half-life of sisomicin, and an even stronger correlation between the clearances of iothalamate and sisomicin. In patients dependent on haemodialysis, the mean serum half-life between dialysis was 40 h, compared to approximately 100 hours for iothalamate, which implies additional extrarenal clearance or tubular secretion of sisomicin. The results of pharmacokinetic studies indicated that a regime of sisomicin 1 mg/kg every 8 to 12 hours in patients with normal renal function would result in serum and urine levels sufficiently high to treat most urinary tract infections. In patients with impaired renal function the dosage interval should be increased according to the serum creatinine level, and in patients dependent on haemodialysis one standard dose at the end of each dialysis period should suffice. 9 patients with a chronic urinary tract infection severely complicated by an underlying disease were treated according to this dosage regimen with a satisfactory bacteriological and clinical result. No adverse reactions or signs of accumulation were observed.


Clinical pharmacokinetics of pancuronium bromide.

Plasma concentrations of pancuronium bromide have been studied in seven surgical patients following a 6 mg intravenous bolus injection of the drug for neuromuscular blockade. Concurrently, evoked muscle twitch response was monitored for each patient as a measure of the pharmacodynamic effect of the drug. The plasma decay curve for pancuronium was found to be biphasic and after rigorous statistical analysis the data were interpreted according to a 2-compartment open model. The half-life of the beta-phase varied between 89.5 and 161.5 min. The apparent volume of distribution of the central compartment ranged from 62.9 to145.5 ml/kg and the plasma clearance from 57.6 to 187.3 ml/min. At the first sign of recovery from neuro-muscular blockade the mean pancuronium plasma level was found to be 0.218 mcg/ml. The mean duration of action as measured from time of onset of paralysis to 20% recovery was 83.4 min with the plasma level at 20% being 0.169 mcg/ml corresponding to 45.4% of dose remaining to be eliminated from the body.


Soluble HL-A antigens in serum. I. Isolation and purification.

A new method was developed which utilizes fractional salting out, gel filtration, ion exchange chromatography and polyacrylamide gel electrophoresis for the isolation and purification of soluble HL-A antigens from serum. The method produced purified antigens possessing HL-A 5,9 specificities with yield up to 60% of that detected in the original serum. Up to 150-fold increase in specific HL-A antigenic activity above that in the starting material could be achieved as measured by the ability of the purified antigens to specifically inhibit the cytotoxic activity of HL-A alloantisera against selected target cells. Electrophoretically purified HL-A antigens with different specificities appear to have a similar molecular size, i.e. 33 000, but differ in their electric charge properties, beta2-microglobulin (beta2m) seems to be noncovalently associated with HL-A antigens present in serum, but during purification beta2m in progressively lost until at the final purification by polyacrylamide gel electrophoresis, the material is completely devoid of beta2m.


Production of migration inhibitory factor and lymphotoxin by non-T cells.

Treatment of tuberculin-sensitive guinea pig spleen or lymph node cells with a burro anti-T cell serum plus complement diminished markedly the number of functionally detectable T cells, but did not affect the amount of migration inhibitory factor (MIF) or lymphotoxin produced by the residual T cell-depleted populations.


Cell surface immunoglobulin. XII. Localization of immunoglobulin on murine lymphocytes by scanning immunoelectromicroscopy.

The localization of immunoglobulin (Ig) on the surface of murine splenic lymphocytes has been studied with the scanning electron microscope (SEM), using a rabbit F(ab')2 hybrid antibody directed against mouse Ig and tobacco mosaic virus (TMV) with TMV as a morphological marker. 35% of spleen cells and 3% of thymus cells were labeled by this method. Labeled cells were usually small lymphocytes with moderate numbers of microvilli. The Ig molecules were randomly distributed on the lymphocyte surface, appearing on microvilli as well as on the smooth surface of the cell.


A new method of radioiodine labeling of polyanionic macromolecules for radioimmunoassay.

A method is described for radiolabeling polyanionic macromolecules by the formation of an electrostatic complex with a radioiodinated tyramine conjugated DEAE-dextran. Data is presented on the use of this technique for the detection and quantitation of immunospecific binding of polynitrophenylated proteins, deoxyribonucleic acids, and cell surface antigens.


Antibody production to lipopolysaccharide in thymectomized Xenopus.

Xenopus toadlets, some of which had been thymectomized as larvae, were immunized with E. coli lipopolysaccharide. Antibody titers were similar in thymectomized and intact toadlets. Antibody was exclusively IgM. The results suggest that a division into thymus-dependent and thymus-independent anti-body responses extends at least as far back in phylogeny as the amphibians.


Participation of immunoglobulin-bearing lymphocytes in the production of macrophage migration inhibitory factor.

The role of immunoglobulin-bearing cells in the production of macrophage migration inhibitory factor (MIF) by tuberculin-stimulated lymphocytes of guinea pigs, immunized with complete Freund's adjuvant, was studied. It was found that: (1) pretreatment of lymphocytes with rabbit anti-guinea pig IgG (anti-IgG) does not block antigen-induced MIF production. (2) Passage of lymphocytes through double layer IgG-anti-IgG gelatin bead columns (the preparation of which is described) abolishes MIF formation by the eluted cells. Cells retained on the columns can be recovered and where shown to produce MIF, when stimulated by antigen. (3) Pulsing of lymphocytes with anti-IgG, for 2 h at 37degreesC, results in MIF synthesis by the cells cultured in medium, in the absence of specific antigen. These findings indicate that cells bearing Ig or Ig fragments are either able to secrete MIF themselves, upon stimulation with antigen or anti-IgG, or are required for MIF production by a different cell type.


Specific enrichment of antigen-binding receptors from sensitized murine lymphocytes.

Splenic T and B lymphocytes from sensitized mice were adsorbed to antigen-coated nylon discs and realesed from the discs  by temperature shift as described by Kiefer, H., (Eur. J. Immunol. 1973. 3: 181 and 1975. 5:624). After cell release, lymphocyte-derived antigen-binding material could be recovered from the discs. A major fraction of the activity (70 -80%) binds to anti-immunoglobulin immunosorbents. A minor fraction does not detectably cross-react with gamma, mu, alpha and kappa immunoglobulin poly-peptide chains. When filtrated through Sephadex G-200, the bulk of activity of both fractions elutes in the region of 7 S serum antibody. However, as demonstrated for (4-hydroxy-3-nitro-phenyl)acetyl (NP)-binding material from C57BL/6 lymphocytes, the average affinity for antigen of the two fractions is drastically lower than that of humoral antibody, and the average affinity of the minor fraction is lower than that of the major fraction. The fraction of lymphocyte-derived antigen-binding material that does not adsorb to insolubilized anti-immunoglobulin serum was found to be proportional to the fraction of T lymphocytes in the input cell population and may therefore represent antigen-binding T lymphocyte surface receptors.


Nature of cells binding anti-IgE in rats immunized with Nippostrongylus brasiliensis: IgE synthesis in regional nodes and concentration in mucosal mast cells.

The possibility that IgE is a secretory immunoglobulin has been examined by studying the tissue and cellular localization of IgE in rats infested with the enteric parasite, Nippostrongylus brasiliensis. The lamina propria of the small intestine and the colonic and pulmonary mucosal surfaces contained numerous anti-IgE-binding cells, but these were shown to be mast cells and not plasma cells. The major site of IgE synthesis was the regional lymph node of the small intestine, the mesenteric node, which contained large numbers of IgE-secreting plasma cells. Smaller numbers of IgE-secreting plasma cells were also found in the axillary node, which drained the site of larvae injection. Peyer's patches, the intrapulmonary bronchial lymphoid tissue and the spleen contained few, if any, IgE-secreting plasma cells. The significance of the IgE which was readily demonstrated in germinal centers of the mesenteric lymph nodes, the Peyer's patches and the axillary lymph nodes, is not known. In contrast to the infested animals, the lymphoid organs of normal rats rarely contained any IgE-CONTAINING CELLS; An unexpected observation was that mast cells in mucosal organs appear to contain intracellular IgE, differing in this respect from connective tissue mast cells. Mast cells lying between epithelial cells, the "globule leukocytes", also appear to contain intracellular IgE, and it is suggested that such cells may be responsible for the presence of IgE in exocrine secretions. This study does not support the suggestion that IgE is a secretory immunoglobulin with a physiology analogous to that of IgA.


Immunosuppressive activity of submaxillary gland extracts of the mouse. I. Effect on antibody formation in response to sheep red blood cells.

Aqueous extracts of mouse submaxillary glands yield six peaks on Sephadex G-75 columns, each containing several molecular species. Two of these fractions (III and IV) have a profound effect on the immune response. When given a day before immunization with sheep erythrocytes, the number of antibody-producing cells is reduced ten-fold or more, and the switch from IgM to IgG is virtually absent. Administration two days before or on the day of immunization results in about 50% depression; outside this range there is no effect. The dose-response curve is of sigmoid shape, with the median at about 0.5 mg of fraction III/40 g mouse, reaching an asymptote after doses of greater than or equal to 5 mg at about 5% of the response of untreated mice. Depression of the primary response does not prevent the development of immunological memory: the secondary response is typical in the early and predominant appearance of IgG-producing cells, although their numbers are somewhat lower than in the controls which received no treatment before primary immunization. Treatment on the day before boosting abrogates the secondary response. The immunotranquilizer from mouse submaxillary glands acts equally well on syngeneic and allogeneic recipients, affecting the earliest stage of the immune response. The results are compatible with the induction of a temporary block in the development of helper T cells.


A plaque assay for all cells secreting Ig of a given type or class.

A modification of the hemolytic plaque assay using protein A-coated red cells is described which makes use of the fact that the Fc portion of IgG binds to protein A. A number of murine plasmacytomas secreting different classes of Ig have been tested for plaque formation with these indicator red cells. In the presence of complement-binding antibodies specific for the corresponding class of secreted Ig, between 10 and 70% of all plated myeloma cells formed plaques. The assay shows a prozone effect in excess of antibody, suggesting that complexes of antibody and secreted Ig effect lysis of the target cells. This assay can be used to enumerate cells secreting any molecules for which complement-binding antibodies are available.


Thymus dependency of bone marrow stem cell proliferation in response to certain antigens.

The activity of mouse bone marrow CFU's following systemic injection of antigen has been investigated. In normal mice quiescent CFU's may be triggered into cycle after antigenic challenge. Prior adult thymectomy prevents stimulation of CFU if thymus-dependent antigens are injected but not when thymus-independent antigens or non-specific stimuli are used. Similar results were obtained in congenitally athymic mice. This seems to indicate an independent pathway for CFU triggering for each cell lineage and for T and B cells.


Comparative survey on enzyme localization, ultrastructural arrangement and functional organization in the optic tectum of non-mammalian vertegrates.

The histochemical localization of some enzymatic activities is surveyed in the optic tectum of vertebrates from cyclostomes to birds. These data are compared with results arising from ultrastructural and experimental works in order to outline some possible connections between enzyme localization and functional organization of the optic tectum. The most interesting result derives from acetylcholinesterase which, in the majority of vertebrate species, is localized in tectal layers in which visual and other sensitive afferents discharge. Such a situation, together with some experimental and developmental results, suggests that cholinergic mechanisms play an important role in the function of the optic tectum and that these mechanisms are worthy of further and more detailed investigations.


Calorimetric characterization of microorganisms.

A short survey is given on recent calorimetric studies of microbial growth, which raised the question whether calorimetrically obtained thermograms can be used for identification of different organisms. Advantages and the efficiency of the method are critically discussed.


Capping of concanavalin A- or ricin-binding sites does not influence phagocytosis in polymorphonuclear leukocytes.

Rabbit polymorphonuclear leukocytes (PMNs) were capped with territin-conjugated concanavalin A or ricin, and then allowed to phagocytose yeast cells. Phagocytic activity and lectin distribution were determined by ultrastructural morphometry. Capped PMNs were found to phagocytose as efficiently as control PMNs, and always to ingest the particles with a lectin-free portion of their plasma membrane. This clearly indicates that concanavalin A- and ricin-binding sites of the PMN membrane are not involved in the recognition and phagocytosis of yeast particles.


The relationship between the pituitary response to luteinizing hormone-releasing hormone and the ovulatory response to clomiphene citrate.

Luteinizing hormone-releasing hormone (LH-RH), at a dose level of 10 mug, was administered intravenously to 31 patients, 9 with oligomenorrhea and 22 with secondary amenorrhea. A serum luteinizing hormone level higher than 45 mIU/ml at 30 or 45 minutes after injection was considered to be a "positive response" to LH-RH. All 9 patients in the oligomenorrheic group and 11 of the 22 patients with secondary amenorrhea had a "positive" response. The patients were then treated with increasing doses of clomiphene citrate until an ovulatory dose was reached. Eight of the nine patients with oligomenorrhea ovulated with doses of up to 150 mg of clomiphene daily for 5 days. Of the 11 patients in the group with secondary amenorrhea who had a "positive" response to LH-RH, 7 ovulated at a dose of 150 mg (or less) of clomiphene citrate, while only 1 of the 11 patients with a "negative" response ovulated while receiving a similar dosage of clomiphene. These results suggest that a simplified LH-RH test might be of value in selecting the level of therapy for ovulation induction.


Prolonged elevation of luteinizing hormone (LH) after intranasal administration of an analog of LH-releasing hormone.

[D-Leu-6,desGly-NH2-10]-LH-RH-ethylamide, an analog of luteinizing hormone-releasing hormone (LH-RH), was administered by intranasal spray or tube to 18 healthy men. A significant release of LH occurred within 15 minutes and was sustained for as long as 12 hours in what appeared to be a diphasic response. Both the spray and rhinyl tube were shown to be effective methods for releasing LH by intranasal administration of the LH-RH analog.


Behavior of the lysosomal system during organ perfusion. An inquiry into the mechanism of hepatic proteolysis.

In this chapter we have attempted to provide some insight into a mechanism(s) of protein degradation in liver which we believe is implicated in the physiological regulation of free amino acid pools. The association we have observed between regulatory effects of insulin and amino acids in the perfused rat liver and physical alterations of the lysosomal system in similarly treated livers suggests that the final stage of the process is localized within the lysosomal-vacuolar system. The present studies show that perfusion of the isolated rat liver is in itself a potent stimulus to general proteolysis, an effect which is associated with increases in lysosomal size, density and sensitivity to osmotic shock. Since the osmotic and density alterations can be prevented or reversed by the provision of insulin or amino acid mixtures, we believe these agents normally act to restrain some inherent proteolytic process. Although the attendant lysosomal alterations are similar in many respects to those induced by glucagon, they differ sharply from the latter in having no sequestered organelles or evident membrane structures and the fact that they rise in the absence of any increase in the tissue level of cyclic AMP. Aside from the obvious problem of evaluating the quantitative importance of the lysosomal system in hepatic proteolysis, three interesting problems remain to be solved: (1) the nature of the proteins involved, (2) their mode of acquisition by the lysosome, and (3) the relationship of this inherent proteolytic process to glucagon or cyclic AMP-induced cellular autophagy. With regard to the former it seems reasonably clear from our studies and those of others that protein substrate does not gain access to lysosomal proteases by the direct penetration of lysosomal membranes, but more likely by fusion of a preexisting lysosome with some, as yet undefined, membrane-bound element. Whether such an element is formed in a manner analogous to that of the precursor vacuoles in autophagy (Ericsson 1969) or by some other process is not known. Closely related to this is the question of the nature of the substrate and the factors determining its uptake by the lysosome. Space does not permit us to review evidence from a number of laboratories which suggests that the turnover of intracellular protein may be a function of size (Dehlinger and Schimke 1971; Dice et al. 1973), charge (Dice and Goldberg 1975), or conformational stability (Li and Knox 1972; Ballard et al. 1974). We have no idea at the moment whether the mechanism we have outlined will eventually be shown to encompass a wide range of turnover of intracellular proteins or, alternatively, whether it will be limited to a certain class of proteins that turnover as a unit. While there is undoubtedly far more specificity in the regulation of intracellular proteins than our present data indicate, we do not view this as an incompatibility, but rather as another aspect of a complex problem we are only beginning to understand.


Heavy metals and lysosomes.

Much can be gained by reassessing the processes which determine the ability of lysosomes to take up or exclude, sequester and mobilize heavy metals. To achieve a better understanding of these events, the chemical forms, intracellular pathways and modes of delivery of metals to lysosomes, as well as the specific physiologic ligands and molecular targets susceptible to metal toxicity have to be identified. None of these can be derived from measurements of metal contents of whole lysosomal fractions because the metal's "effective concentration" at a specific target site may be affected by the binding properties of the lysosomal ligand as well as by those of cation carrier proteins present in the cytosol (e.g., metallothionein), and by interactions with and competitions by other cellular organelles. Therefore, the possibility of such events diminishing or enhancing a metal's direct effect observable in in vitro systems has to be considered before extrapolating to the in vivo situation. Another pitfall to be wary of is the equation of an organelle's relative affinity for a metal in vitro with its susceptibility to the metal's toxic effects. This is evident, albeit at a tissue level rather than at that of organelles, from the discordance between the low affinity of nervous tissue for lead and this metal's pronounced encephalopathic effect. The answers to some of the questions raised in this review may possibly lead to pharmacologic applications, particularly to the development of effective agents for the removal from or the inactivation of toxic metals deposited in lysosomes. At present, considerable uncertainty exists regarding the possible interaction of therapeutic chelating agents with lysosomes in vivo. We do not know, for example, whether the contrasts between the remarkable effectiveness of penicillamine in mobilizing copper from tissues and the limited effectiveness of desferioxamine in removing excess iron stores can be accounted for by differences in accessibility of these two chelators to lysosomes. Or, alternatively, can these differences in effectiveness be related to different ligands or macromolecules interacting with each metal? At least part of the lysosomal iron is bound to ferritin molecules which may not be susceptible to the action of chelating agents after incorporation. Such speculation is not without foundation since ferritin molecules are heterogeneous. However, whether this heterogeneity, which is reflected in different organ-specific patterns of distribution (Powell et al. 1973), is the result of differing affinities of the isoferritins for specific subcellular organelles has not been established. It is conceivable that ferritin molecules present in the cytoplasm may be subtly different from those taken up by lysosomes, implying that the latter are endowed with capabilities for selection of specific macromolecules...


Secretion of lysosomal enzymes induced by immune complexes and complement.

Immune complexes react with neutrophils and macrophages by way of surface membrane Fc receptors. Alternatively, after fixation of complement, interaction may be with membrane C3b receptors. As a result, phagocytosis is induced. A consequence of this process is the liberation to the extracellular medium of lysosomal constituents. In neutrophils, fusion of granules with phagocytic vacuoles liberates the contents to the outside because some of the vacuoles, for various reasons, are inefficiently closed off. Alternatively, reaction of neutrophils in vitro or in vivo with immune complexes and/or activated complement components (C3b, C3a, C5a, C567) on surfaces too large to be phagocytosed, induces direct exocytosis of granules with consequent release of constituents. In both cases the liberation of lysosomal constituents is a secretory process, does not involve cell lysis, and resembles closely the secretory reactions of other widely differing cell types.


[Studies on the secretion of gonadotropins in patients with collagen diseases].

It is well known that majority of patients with collagen diseases are women and that collagen diseases take turn for the worse or the better when the secretion of sex hormones changes greatly at menarche, pregnancy, delivery or menopause. These facts suggest that sex hormones are involved in the pathophysiology of collagen diseases. In the present study, the responses of plasma luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels following the intravenous injection of 100 mug luteinizing hormone-releasing hormone (LR-RH) were investigated in 34 patients with systemic lupus erythematosus (SLE) and in 15 patients with rheumatoid arthritis (RA). The results obtained were as follows: 1) The magnitude of plasma LH response to LH-RH in 29 mature female patients with SLE was significantly greater than that in normal subjects. 2) On the other hand, the magnitude of plasma FSH response to LH-RH in patients with SLE was comparable to that in control subjects.  3) In patients with RA who have normal menstrual cycles, the magnitude of increase in plasma LH and FSH levels after the injection of LH-RH was the almost same as that in normal subjects. Increased responses in plasma LH and FSH levels to LH-RH were observed in 7 patients with RA who were 3 menopausal females and 4 aged males. These findings suggest that the secretion of LH, but not FSH, in response to LH-RH might augment in patients with SLE. On the other hand, in RA patients the function of the pituitary-gonadal axis might maintain within normal limits. For that reason, I guess the following possibilities: 1) In patients with SLE the pathological changes of the disease reached to the ovary and ovarial function was slightly suppressed and then a hypersecretion of LH was observed, 2) the hypothalamus was attacked with the disease and then an unknown mechanism caused the hypersecretion of LH.


Pituitary and adrenal control of pancreatic endocrine function in the duck.

Recent clinical and immunological data suggest that the classical concept of "idiopathic autoimmune diseases" is to be revised. In a normal population, autoimmunity reactions against thyroid gland, gastric mucosa and adrenals develop slowly with increasing age and are found more frequently in women than in men (at least so far as thyroid antibodies are concerned) as is lowering of the functional activity of T lymphocytes. Diabetes takes its place among a series of factors diminishing immunocyte reactivity, and thus enhancing the development of the autoimmunity process. This may perhaps be promoted in some way by genetic factors, perhaps those which also play a definite though as yet ill-defined role in determining the emergence of diabetes. For the present, diabetes mellitus itself must only rarely be considered a consequence of an autoimmune process and then only in certain insulin-dependent cases. By contrast, diabetes appears frequently to be an activator of autoimmune phenomena against tissues other than pancreas, namely thyroid gland, adrenals and gastric mucosa. Awareness of these associations should encourage physicians to seek latent humoral or cellular evidence of autoimmune phenomena in diabetics; this would favour the early recognition of clinically important abnormalities which may accompany the diabetes.


Study of glucagon secretion using in vitro perifusion of rat pancreas pieces.

The in vitro perifusion technique of rat pancreatic tissue has been studied in respect of sensitivity and reproducibility regarding alpha-2-cell function. Convenient and quickly performed, it allows the kinetic analysis of humoral factors of glucagon release when the blood vessels are too small to be cannulated, for instance in case of fetal pancreas and of tumor tissue. A2 cells in the perifused pancreatic tissue release glucagon in response to arginine, norepinephrine and the decrease of glucose concentration in the medium. The sensitivity of this preparation is lower than that of the perfused pancreas: this may be related to unphysiological conditions of fluid circulation, of the degradation of the stimuli and degradation of the hormone released. Satisfactory conditions of perifusion flow rate and amount of panceatic tissue have been defined and several protective agents have been tested.


[Participation of catecholamines in the inhibition of insulin secretion and stimulation of glucagon release during exercise and stress (author's transl)].

The participation of catecholamines in the inhibition of insulin secretion and stimulation of glucagon release during exercise and stress is reviewed. In rats injected with guinea-pig anti-insulin serum and either compelled to swim or exposed to electrical shocks, the rate of insulin secretion is markedly inhibited, mimicking the situation found after administration of exogenous epinephrine. Prior injection of phentolamine abolishes the exercise-induced inhibition of insulin release. These findings support the concept that such an inhibition is due to activation of the alpha-adrenergic receptors of the beta-cell by endogenously released catecholamines. In addition to inhibiting insulin secretion, epinephrine is shown to stimulate glucagon release in vitro, using pieces of pancreas from duct-ligaturated rats incubated at both low and high glucose concentrations. Augmentation of glucagon release is also observed in vivo during exercise or stress. It is concluded that the hormonal regulation under the latter situations is opptimally oriented for the mobilization of energetic substrates such as glucose and fatty acids.dicale (INSERM), Hòpital des Enfants Malades, Paris.


Feed-back inhibition of insulin secretion in subjects with high and low insulin response to glucose.

The plasma insulin response to two successive stimulations by glucose was measured in 12 healthy subjects, 5 of whom demonstrated subnormal insulin responses to glucose infusion (low insulin responders). 15 g of glucose were injected rapidly i.v. at 0 and 20 min, and blood glucose and plasma insulin levels determined at intervals of 2-10 min. In both groups, the peak elevation of plasma insulin, and the peak insulinogenic index, were significantly inhibited at the second stimulation. The degree of inhibition was smaller in subjects with low insulin response. The integrated insulin response and the integrated insulinogenic index during the 20 min following glucose administration were also significantly inhibited at the second glucose challenge in subjects with low insulin responders. The degree of inhibition of the second insulin response (20-40 min) shows significant correlation with the insulinogenic index of the first response (0-20 min) in the total material. It is suggested that this inhibition of insulin release on repeating the stimulus, (a) may offer an explanation sustained stimulation by glucose; (b) is probably generated by the phenomenon of insulin secretion, and (c) might thus represent a negative feed-back loop in insulin secretion. (d) Decreased insulin secretion in low responders is not due to the presence of increased inhibition.


[Genetics of diabetes mellitus (author's transl)].

Since Pincus and White's claim in 1933 that diabetes mellitus is an inherited disease, the precise mode of inheritance remains a matter of dispute. The reason for the controversy is that the geneticist is confronted with a number of impediments to genetic analysis. As pointed by Neel, "diabetes mellitus is in many respects a geneticists nightmare". The obstacles are : 1) a precise definition of diabetes is difficult to establish, 2) the frequency of the disease which is sex and age dependent is not well known, 3) the probability of genetic heterogeneity is great but whether early onset and late onset diabetes are different genetic diseases or the same one remains controversial, 4) the basic defect (s) is unknown, 5) environmental factors (e.g. nutritional status) influence the frequency of the disease. Despite these problems many studies have been devoted to the mode of inheritance of diabetes mellitus. Many authors favour an autosomal recessive mechanism. However, low penetrance (25 %) is necessary to support this mode of inheritance. Simple autosomal dominant mode of inheritance has also been suggested, but this pattern fits only few families. The majority of geneticists think, at the present time, that diabetes has a multifactorial mode of inhritance. The heritability which express the extent to which the phenotypes exhibited by parents are transmitted to their offspring is in the neighbourhood of 50%. Many arguments favour this mode of inheritance: 1) low penetrance is necessary to aistinct genetic diseases, and especially in chronic glaucoma, which also have a multifactorial mode of inheritance;  in particular, one must note the association between glucose intolerance and ocular hypertension induced by dexamethasone, 3) the association between diabetes and antigen A of the ABO system and antigens HL-A8 and W 15 of the HL-A system.


Inhibitory and augmentative effects of oestradiol on LH-RH-induced release of LH by anterior pituitary glands from intact female rats in vitro.

Anterior pituitary glands from intact dioestrous female rats were incubated in vitro and the LH released into the medium was measured at hourly intervals. The effect of a supramaximally active dose of LH-RH was first inhibited and later augmented by oestradiol added to the medium. The augmented response also occurred when LH-RH was absent during the preceding phase of the incubation. Both effects of oestradiol could be blocked by cycloheximide, which agent, however, also partly inhibited the response to LH-RH.


Luteinizing release hormone tests in impotent diabetic males.

Assessment of pituitary-gonadal function was made in impotent diabetic males using luteinizing hormone-release hormone tests. Serum testosterone, sex-hormone-binding globulin capacity, and basal and incremental gonadotrophin concentrations in the impotent diabetics were similar to those in control diabetics, suggesting a primary neurologic rather than an endocrine defect as the cause of the impotence.


Anencephalus in the Oxford Record Linkage Study area.

The incidence of anencephalus in the largely rural areas of Oxfordshire and west Berkshire from 1965 to 1972 was ascertained from the files of the Oxford Record Linkage Study as 1-49 per 1000 total births. There was little variation within the area, but there was a marked increase in incidence over the eight years of the study. There was a slight seasonal variation, with an excess of conceptions in the first two quarters of the year, a slight excess of births to primiparous women and to women aged between 20 and 24 years. There was an increase in incidence with falling social class, with a significant excess of fathers who were printers, painters and decorators, transport drivers or in the armed forces. Among 115 siblings, 11 (9-6 per cent) had defects of the central nervous system, and the incidence of anencephalus among twins was twice as high as expected.


Issues and problems in remedial education.

Children with reading and other education problems present for help to a wide variety of professionals from different disciplines. At present there appears to be a wide gulf between sophisticated assessments and effective remedial intervention. This paper examines some of the issues involed in providing and evaluating remedial education, and the issues of definition, measurement and prevalence; it is concluded that as many as 20 per cent of 10-year-olds in inner cities in the United Kingdom may require extra help with reading. The evidence suggests that, over-all, remedial help is not at present very effective. A concerted research programme is needed to identify effective approaches. Constructive suggestions are advanced whereby such a research programme would make a  significant contribution to services for children with educational problems.


Proinsulin synthesis in islets of Langerhans from spiny mice (Acomys cahirinus). Comparison with rats and mice.

Proinsulin synthesis in response to different concentrations of glucose has been studied in islets of Langerhans of acomys, rat and mouse. The response to increased glucose was small in acomys when compared with rat or mouse. The time course of the effect of glucose on proinsulin synthesis was also studied in sequential 15 min periods up to 60 min. With 5.5 mM glucose the rate of 3H-leucine incorporation for acomys islets were greater over the periods 15-30, 30-45 and 45-60 min after the addition of glucose to the islets than at 0-15 min. By contrast rat islets exhibited a greater glucose-induced stimulation between 15-30 and 30-45 min relative to the 0-15 min period and a further increase of the incorporation rate between 45 and 60 min. With 27.5 mM glucose acceleration of incorporation relative to the first 15 min period occurred at 15-30 min; it was then stable up to 60 min for islets both from acomys and from rat. Incorporation into total protein was much greater in acomys than in rat or mouse: thus in 2.75 mM glucose, acomys islets incorporated 5 and 19 times more 3H-leucine into total protein than did islets from rat or mouse respectively. In 27.5 mM glucose the incorporation into total protein of the islets from acomys was 4 and 13 times greater. Islet content of insulin was similar for all three species. No significant changes in ATP content were observed in response to changes in glucose concentration from 2.75 to 27.5 mM. The results demonstrate a decreased responsiveness of proinsulin synthesis to glucose in acomys and are discussed in terms of the known decreased sensitivity of insulin release in this animal.


The inhibition of insulin secretion from the perfused rat pancreas after thyroxine treatment.

Thyroxine treatment did not significantly affect the immediate insulin secretory response of the perfused rat pancreas, but it inhibited the late phase of D-glucose-induced insulin secretion. Thyroxine treatment did not inhibit D-glyceraldehyde-, D-mannose-, and tolbutamide-induced insulin release from the perfused pancreas. An increase in the D-glucose concentration of the perfusion medium as well as feeding of the rats did not restore insulin secretion after thyroxine treatment. The inhibition of D-glucose-induced insulin release in response to thyroxine treatment was reversed after addition of either D-glyceraldehyde, dihydroxyacetone, DL-glyceric acid, pyruvate, or alpha-ketobutyrate to the perfusion medium. Tolbutamide, L-glucose, D-fructose, D-mannose, L-lactate, and propionic acid were not able to overcome the inhibition of D-glucose-induced insulin secretion. Except for alpha-ketobutyrate all substances which were effective in reversing the inhibition of D-glucose-induced insulin release were glycolytic intermediates. Comparing the glycolytic alpha-ketoacid pyruvate and the non-glycolytic ketoacid alpha-ketobutyrate, the only part common to both substances was the ketoacid moiety. It is concluded from these findings that the ketoacid moiety of the alpha-ketoacids plays an important role in reversing the effect of thyroxine on D-glucose-induced insulin release.


Diagnosis of carcinoma of the papilla of Vater by duodenofiberscopy: simultaneous attempt on endoscopic observation, aspiration cytology, retrograde pancreatocholangiography and biopsy.

Clinical features and validity of a duodenofiberscopic examination for the diagnosis of carcinoma of the papilla of Vater were evaluated on 13 cases preoperatively diagnosed endoscopically. Followings are the conclusions derived. 1) Average age was 59.8 years. Incidence was equal among men and women. 2) As initial symptoms, general fatigue and easy fatigability or symptoms of cholangitis should be emphasized. 3) Other important symptoms include jaundice, fever with chills and weight loss. Abdominal pain, when present, was noted as one of symptoms of cholangitis in most cases. 4) Important laboratory findings include elevated serum bilirubin and alkaline phosphatase, elevated erythrocyte sedimentation rate, positive occult blood in stool. 5) Carcinomas of the papilla of Vater were divided into 3 types (type I, II and III) according to endoscopical and pathological findings. Clinical features and laboratory findings were discussed in relation to the type of lesions. Characteristic endoscopic findings of each type of the lesions were described. Validity of aspiration cytology, pancreatocholangiography and biopsy under duodeno-fiberscopic observation was also discussed. As a result, duodenofiberscopy was considered to be the most useful method for the diagnosis of carcinoma of the papilla of Vater because it provides us with an opportunity to perform simultaneously an endoscopic observation, aspiration cytology, pancreatocholangiography and biopsy.


Ultrastructural studies of human acute pancreatitis.

The pancreatic tissue from a patient with acute hemorrhagic necrotizing pancreatitis was studied by routine electron microscopic observation. The remarkable change was the destruction of pancreatic acinar units. 1) The acinar lumen was dilated with filling of fibrillar materials, which occasionally contained degenerated cellular components or neutrophils. 2) Accumulations of fibrillar materials were present at the periphery in the acinar cells, especially depositted thickly at the basal portion. 3) In some acinar cells, the accumulation of fibrillar materials occupied the entire acinar cell, accompanied by disappearance of luminal margin, and the intracellular fibrillar materials were mixed with acinar lumen contents. 4) The acinar units which showed above described changes had almost intact basal lamina.


Comparison of dose-response curves between acid and pepsin to betazole hydrochloride in the dog with gastric fistula.

Secretory responses to subcutaneously or intravenously administered betazole hydrochloride of graded doses were compared between acid and pepsin in the dog with gastric fistula. Although the dose related increments in acid and pepsin outputs were parallel in the range of lower doses than 2.0 to 3.0 mg/kg body weight of betazole hydrochloride, higher doses caused in decrease in pepsin output, but did not in acid output. It was pointed out that the dose of betazole hydrochloride that stimulates acid secretion maximally dose not correspond to the maximal stimulation for pepsin secretion in the dog.


Clinical course of peptic ulcer and the effect of pharmacotherapy:--multi-institutional double-blind controlled study--.

Comparative clinical trials were conducted by the double-blind technique in a series of 302 cases of gastric ulcer at fifteen medical institutions in order to investigate the clinical course of peptic ulceration and the effect of pharmacotherapy on it, using N-acetyl-L-glutamine aluminum complex (KW) as test drug and basic aluminum sucrose sulfate (UL) as reference drug as well as an inactive placebo of lactose (PL) with the results as follows: In 12 weeks of treatment with inactive placebo, a higher cure rates were noted in inpatients than in outpatients, and in patients with ulceration in the corpus ventriculi than in those with ulceration at the angulus. The overall evaluation of therapeutic responses indicated that, both KW and UL were significantly superior in efficacy to placebo in 12 weeks of therapy. These two medicaments displayed different modes of efficacy, possibly reflecting their different mechanisms of pharmacologic action. The efficacy of the medicaments was particularly superior in outpatients and patients with peptic ulceration at the gastric angle both of which showed lower rates of response to placebo.


Malignant lymphoma of large intestine --15-year experience and review of literature.

From our experience of malignant lymphoma in the large intestine and reviewing the Japanese literature, it might be concluded that this particular lesion was extremely rare, and was found predominantly in the right side of the colon. In contrast to the other malignant neoplasms in the large intestine, malignant lymphoma was found fairly often in children, which was accompanied by intussusception not infrequently.


Natural histamines.

Twelve histamines have been so far identified in living animal tissues. The two reactive sites of the histamine molecule involved in the production of these derivatives are represented by the imino nitrogen of the nucleus and the amino group of the lateral chain. The ring imino nitrogen may be methylated, to form 3-methylhistamine, or conjugated with ribose to give origin to histamine riboside, ribotide and dinucleotide. The amino group of thelateral chain, in its turn, may be methylated (N'-methyl-histamine, N',N'-dimethylhistamine), conjugated with aliphatic acids (N'-imidazoleproprionylhistamine, N'-urocanylhistamine) or amino acids (beta-alanylhistamine or carcinine), and finally linked, through a methyl group, to the imidazole nucleus to give origin to imidazo-c-pyridine derivatives or cyclized histamines. The biosynthesis of the above natural histamines and their physiological significance are discussed.


[Effect of adenine, adenosine and adenylic nucleotides on the mutagenic effect of hydroxylamine].

The effect of some adenyl precursors of DNA synthesis on the mutagenic activity of hydroxylamine (HA) is studied. It is shown that the addition of adenine to a suspension of Escherichia coli B cells increases the yield of mutants by more than two times as compared with HA alone. The effects of adenosine, AMP and dAMP are somewhat different. It is suggested that the increase of the HA mutagenic effect produced by the addition of adenine may be due to: 1) the excess of the amount of adenylic precursors of DNA synthesis over guanilic ones, which promotes the erroneous base-pairing during the replication of the HA modified template; 2) the modification of adenylic precursors by HA into N6-oxy-dATP, and their incorporation into DNA. The mutagenic effect of N6-hydroxyadenosine, the product of the adenine modification by HA, in E. coli B pur- was studied. The experiments showed that N6-hydroxyadenosine induced about 1% of mutations, a relatively low lethal effect (the cell survival was 80%), and provided a high mutagenic action of this compound.


[Differences in the mutagenic effectiveness of N-nitrosomethyl- and N-nitrosoethylurea acting on bacterial DNA in vivo].

Chemically induced lethals and reversion frequencies of two mutants of Salmonella typhimurium LT-2 1530 and 1532 were examined. Prototrophic mutants to histidine independence occurred by two different mechanisms and were induced by nitrous acid, hydroxylamine, N-nitrosomethylurea (NMU), N-nitrosoethylurea (NEU) and N, N, nitrosodimethylurea (NDMU). The ability of the mutants to revert was affected by the chemical structure (in the row of N-nitrosoalkylurea - by alkyl group) of the mutagen. NEU is found to induce in bacterial DNA only base pair substitutions; NMU and NDMU - deletions and base pair substitutions. An original model of deletion mechanism with Goldwait endonuclease II participation is proposed.


Coeliac disease, gluten-free diet, and malignancy.

Two hundred and two patients with coeliac disease or idiopathic steatorrhoea previously reported from this unit have been reviewed after a further 10 years of follow-up. Of 20 more deaths, 10 were due to malignancy. A separate study was made of all patients with histologically confirmed coeliac disease seen to the end of 1972 and followed up to the end of 1974. Twenty-one of the 43 deaths in this series were due to malignant tumours, of which 13 were reticulum cell sarcomas. There was no evidence that patients showing sub-optimal clinical response to gluten withdrawal or persisting falt jejunal biopsies were more prone to die of cancer. A gluten-free diet did not appear to be effective in preventing malignant complications, but a longer follow-up of patients will be necessary to provide a final answer to this question.


Transfer of computer-aided diagnosis of dyspepsia from one geographical area to another.

This paper reports a comparison in Airedale District General Hospital between computer-aided diagnosis of dyspepsia and endoscopy in a prospective unselected series of 165 patients. Patients were interviewed immediately before endoscopy and the findings analysed by a small desk-top computer-aided system. Each 'new' patient was compared by the computer with a group of 360 similar patients from Leeds (25 miles away). Overall, 83% of the positive lesions found at endoscopy were correctly predicted by the computer, including all but three of the 22 cases of gastric cancer. It is suggested (1) that there is little loss of accuracy in transferring the computer-aided system from one locality to another; and (2) that a computer-aided analysis of the patient interview may be of value in selecting 'high-risk' patients for intensive investigation.


Tranexamic acid and upper gastrointestinal haemorrhage--a double-blind trial.

The efficacy of antifibrinolytic therapy in the management of acute upper gastrointestinal haemorrhage has been investigated in a double-blind clinical trial. Two-hundred patients were studied using tranexamic acid, a potent antifibrinolytic agent. Of these, 103 were in the treatment group and 97 in the control group. Patients were analysed to determine severity of initial blood loss, transfusion requirements, together with the incidence of recurrent bleeding, surgical intervention, and death. Final diagnosis as to the site of bleeding was arrived at using endoscopy, barium studies, and the findings at operation and necropsy. The groups were well matched as regards severity of initial haemorrhage, age, sex, aetiological diagnosis, and precipitating factors. A significant difference was observed in the requirement for surgical intervention to control continuing or recurrent haemorrhage. Twenty-three of 97 in the control group and seven of 103 in the treatment group required surgery. There appeared to be a reduction in the transfusion rate after the first three days of hospitalization in the treatment group. There were no significant differences in mortality or in side-effects between the two groups.


The changing pattern of pyogenic infections of the hand.

Four hundred cases of pyogenic hand infection treated in 1947 are compared with a similar number of cases treated in 1974.


Can HMOs improve health care for the poor?

Routine and preventive health services are underutilized by low-income groups, who make greater use of inpatient and emergency services than the population at large. Studies reported on in this article suggest that the low-income population might be more effectively served, and at less expense, by health maintenance organizations that provide service on a prepaid, capitation basis.


Treating sexual dysfunction in dialysis and transplant patients.

Sexual dysfunction can be a significant problem for patients with end-stage renal disease and other chronic or disabling conditions. To a large extent, however, social workers and other professionals have overlooked the special needs of patients whose sexual capacities have been affected by illness or injury. This article presents an assessment and treatment model for helping such patients.


The Peckham Experiment revisited: cultivating health.

The Peckham Experiment, begun in London in 1935, was a combination health and recreation center that embodied a visionary, ecological approach to health care. In the face of the continued and costly failure of today's episodic medical services, the author invites us to re-examine Peckham and perhaps take a cue from the past for the future.


[Increase of brain serotonin and its metabolite in rats caused by meclofenoxate].

Influence of meclofenoxate (MF) on the 5-HT and 5-HIAA contents in the cortex, diencephalon and brain stem of the rat was studied with the following results. MF caused a dose-dependent elevation of 5-HIAA level in the three different brain regions and particularly in the brain stem. An increase of 5-HT was also noted in the brain stem, whereas 5-HT in the diencephalon tended to decrease. The increase of 5-HT after pargyline was accelerated and the pargyline-induced decrease of 5-HIAA was equally inhibited by MF and probenecid, which suggests that MF blocks the efflux of 5-HIAA from the brain. The effects of MF on the brain 5-HT and 5-HIAA contents were identical with those of its hydrolysate, p-chlorophenoxyacetic acid.


Selection of auxotrophic mutants in Saccharomyces cerebisiae by a snail enzyme digestion method.

Yeast cells in the stationary phase of growth are relatively resistant to snail enzyme digestion. This resistance was shown to decrease abruptly in the course of only 3-5 duplications, when stationary cells were allowed to grow in a fresh medium. The selective digestion of growing prototrophs from a mutagenized culture in minimal medium by snail enzyme was applied to increase the proportion of auxotrophs which remained relatively resistant.


The broth effect and mutation frequency decline in cells of Escherichia coli after irradiation with UV-light.

Dependence of the broth effect and the phenomenon of mutation frequency decline on dose of the applied UV radiation was investigated in the strain Escherichia coli B/r Hcr+ thy trp. Reversions to Trp+ were followed. The degree of the broth effect and the mutation frequency decline is minimal within the range of UV doses corresponding to a survival of cells lower than 10(-1). In connection with the two effects, excision of thymine dimers, initiation of synthesis, synthesis and degradation of DNA were also investigated. It was found that stimulation or inhibition of an inaccurate postreplication repair mechanism, rather than inhibition or stimulation of excision of thymine dimers, are responsible for the broth effect and the mutation frequency decline, respectively.


[Elements of Spanish psychiatry].

The development of a psychiatric science in Spain took place on various levels. To this end an examination was made as to the cultural-historical side with the Christian and stoical tradition as well as to the ethnologic-psychological side, especially with the aspect of inferiority and pride taking into consideration the reflection of these problems in the so-called "generation of 1898". Furthermore, linguistic aspects were taken into consideration as the example of the Spanish term ""gana" and the expressions of the Valencian dialect in the Middle Ages show. The "forerunner"-psychiatry as well as the significance of the first European Christian mental hospital in 1409 were described. The examination comes to the conclusion that whilst the Spanish psychiatry, after comparatively late maturity, followed the German phenomenological school, it  still succeeded in enriching this school with conceptions of its own, such as the vital fear (López Ibor) and physiodynamic factors.


[Clinical studies on Frubienzyme in a controlled double-blind trial].

In a controlled clinical trial Frubienzym (throat lozenges with 5 mg lysozyme, 2 mg papaine and 200 I.U. bacitracin) or placebo have been given to 100 patients with pharyngitis and/or tonsillitis for 4 days. Under treatment with Frubienzym reddening, swelling, matter and mucus in the throat, coughing, swelling and pain of lymphatic ganglions and pain of swallowing vanished more quickly than under placebo. The differences were significant (p less than 0,05, p less than 0,001 or even p less than 0,001; U-test of Wilcoxon, Man and Whitney). There were no side effects which could be attributed to Frubienzym.


Evidence for a hypophyseal factor that stimulates insulin secretion by the pancreas (insulotrophine?)

The continuous infusion of glucose (1 mg/kg/min) via the carotid artery in anesthetized dogs produced a biphasic pattern of insulin secretion. The first peak reached a maximum 3 min after glucose infusion and decreased to basal level at 7 min. As long as glucose infusion persisted a slow and maintained increase in insulin level in the pancreatico-duodenal vein was observed. The same amount of glucose infused in to the carotid arteries of hypophysectomized dogs, failed to induce any change in plasma insulin level. Plasma sample obtained from the jugular vein of dogs receiving glucose via the carotid arteries were infused into a second dog via the pancreatico-duodenal artery. One minute after the onset of infusion a rise in insulin was observed in the pancreatico-duodenal vein. The stimulating effect was not due to the high blood glucose level present in the jugular vein of dogs undergoing the cephalic glucose infusion. Infusion through the pancreatico-duodenal artery of a glucose solution at a concentration equal to the highest blood glucose level observed in the jugular vein did not evoke insulin secretion. Plasma samples obtained from the jugular vein of dogs receiving saline via the carotid arteries did not evoke insulin secretion when receiving into the pancreatico-duodenal arteries of dogs. Pancreatic infusion of plasma obtained from the jugular vein of hypophysectomized dogs infused glucose through the carotid arteries did not evoke any pancreatic response. These findings are demonstrative of the presence of a hypophyseal humoral insulin stimulating factor in the jugular vein of dogs receiving a cephalic glucose load.


Ionophore A23187-induced insulin secretion in the isolated, perfused dog pancreas.

Perfusion of ionophore A23187 (10 muM) in the isolated dog pancreas resulted in a monophasic release of insulin. Ionophore A23187 (10 muM) failed, however, to elicit insulin secretion when added to calcium deficient (0.1 mmoles/L) perfusate. Simultaneous reintroduction of calcium (1.27 mmoles/L) and discontinuance of ionophore A23187 following calcium deficient periods caused a monophasic secretion of insulin which was quantitatively very similar (41,400 +/- 13,800 muU) to that stimulated by ionophore during normocalcemic perfusion. With reference to the current literature, these results suggest that ionophore A23187 elicits insulin secretion in the perfused dog pancreas preparation by increasing the level of free intracellular calcium, a process which is dependent upon a normal extracellular ionic calcium concentration.


Effects of iodoacetate and fluoride on islate respiration and insulin biosynthesis.

Fluoride and iodoacetate inhibited the oxidation of glucose by islets of Langerhans isolated from the rat pancreas. Fifty % inhibition occurred with either 17 mM fluoride or 0.5 mM iodoacetate. The rate of insulin biosynthesis was more strongly inhibited by these inhibitors, especially fluoride. Fifty % inhibition occurred with approximately 1.5 mM fluoride. At high concentrations of iodoacetate and fluoride, the inhibitory effect on insulin synthesis was not reversed to a significant degree by the addition of pyruvate in the incubation medium. In addition to inhibiting the glycolysis and depriving islets of energy essential for the biosynthesis of insulin, fluoride probably exerts a direct inhibitory influence on the biosynthetic mechanism. A separate experiment with [6-14C]glucose indicated that 0.2 mM iodoacetate does not inhibit glycolysis completely.


[32P] Orthophosphate efflux from pancreatic islets: graded response to glucose stimulation.

The dose-response relationships of the glucose-induced rapid transient efflux of [32P] orthophosphate from prelabeled pancreatic islets ("phosphate flush") have been investigated. Threshold levels for eliciting a "phosphate flush" were between 0.5 and 1.0 mg/ml glucose, the apparent "Km" for this event was 1.0-1.5 mg per ml and the apparent "Vmax" was reached at ambient glucose concentrations between 1.5 and 2.0 mg per ml. This dose-response curve is somewhat shifted to the left in comparison with previously published data for glucose-induced insulin release. Thus, the "phosphate flush" appears to display a more narrow dose-response curve to ambient glucose concentrations than the actual release of insulin. It is proposed that this may constitute further evidence that the "phosphate flush" reflects an early step in stimulus-secretion coupling and that there may be some loss of sensitivity as the glucose signal is transmitted from the site of recognition to the final site of hormone release.


Autoreactivity developing spontaneously in cultured mouse spleen cells. II. Comparison of cytotoxicity of cultured male and female spleen cells.

Male and female spleen cells were compared before and after culture for their cytotoxicity to autologous-embryo fibroblasts. Cultures of male cells developed significantly greater reactivity than cultures of female cells. Moreover, while the cytotoxicity derived from cultures of male cells was totally abolished by treatment of effector cells with a mouse anti-T cell antiserum, such an antiserum had less affect on the effector cells of female mouse cultures.


Comparative studies on the actions of antigen and polyclonal B-cell activator in differentiation and proliferation of B-cells and B memory cells.

Using the capsular polysaccharide of Klebsiella pneumoniae (CPS-K) as a polyclonal B-cell activator (PBA) and sheep red blood cells (SRBC) as a T cell-dependent antigen, we compared the ability of PBA and antigen to differentiate (generate antibody-forming cells, AFC) and proliferate (generate immunological memory) virgin B cells and B memory cells. In vitro CPS-K induced the differentiation of IgM virgin B cells, IgM B memory cells and IgG B memory cells to AFC, as well as or better than SRBC. The differentiation of B memory cells to AFC by CPS-K did not require the participation of macrophages or T cells, whereas the action of SRBC depended strictly upon the helper actions of these cells. The responsiveness to CPS-K and SRBC of normal and antigen-primed spleen cells as judged by anti-SRBC PFC responses in vitro was markedly decreased after stimulation of virgin B cells and B memory cells in vivo by CPS-K injection into normal or primed mice but greatly increased after the injection of SRBC. The decrease in the responsiveness to CPS-K of spleen cells from mice treated with CPS-K appeared principally due to exhaustion of the functions of B cells and B memory cells. From the present data it has been concluded that the signals required for the differentiation and proliferation of B cells of B memory cells are different from each other, the signal for differentiation being provided by either antigen (SRBC) or PBA (CPS-K), while the signal for proliferation only by antigen.


Immunological studies of human placentae: subclass and fragment specificity of binding of aggregated IgG by placental endothelial cells.

Fluorescein-conjugated heat-aggregated human IgG binds to endothelial cells of foetal stem vessels in cryostat sections of normal, full-term human placentae. No binding was observed using native human IgG of heat-aggregated human albumin, IgM or IgA2. No inhibition of binding of heat-aggregated human IgG was observed by pre-treatment of placental tissue sections with native IgG or non-aggregated Fc fragments. The binding was blocked using heat-aggregated Fc fragments prepared from IgG1, IgG2, IgG3 and IgG4 myeloma proteins, but not with heat-aggregated human light chains, Fab and F(ab)2 fragments of human IgG, or with heat-aggregated human IgM and IgA2. It is suggested that the placental endothelial cell receptor for aggregated IgG may function to keep immune complexes from entering the foetal circulation.


The use of 1-fluoro-2,4-dinitrobenzene as an affinity label for the antigen receptor of delayed hypersensitivity.

The passive transfer of delayed contact sensitivity to dinitrophenyl can be specifically inhibited by brief treatment of lymph node cells (LNC) from sensitized guinea-pigs with 1-fluoro-2,4-dinitrobenzene (DNFB). Analysis of KC1 extracts of LNC previously treated with (3H)DNFB, using polyacrylamide gel electrophoresis, revealed a peak of radioactivity present in both sensitive and normal cell extracts as well as a peak found only in sensitive cell extracts. Cell-free extracts appeared capable of transmitting delayed-onset skin reactivity to non-sensitized recipients.


Nonspecific resistance against infection with Salmonella typhi and Salmonella typhimurium induced in mice by cord factor (trehalose-6,6'-dimycolate) and its analogues.

Mice pretreated intraperitoneally with trehalose-6,6'-dimycolate (cord factor) were protected against an intraperitoneal challenge with Salmonella typhi strain Ty2 or Salmonella typhimurium strain SR 11. The nonspecific resistance to S. typhi and S. typhimurium was still detectable 7 and 14 days, respectively, after administration of cord factor. The effect of cord factor was local. Synthetic analogues of cord factor--trehalose-6,6'-dipalmitate and trehalose monopalmitate--also induced nonspecific resistance to the above virulent bacteria. The results are discussed.


Comparative evaluation of dexamethasone with cyclophosphamide, chloramphenicol and cyproheptadine hydrochloride as immunosuppressive agents on skin homografts in rats and rabbits.

An experimental study on immunosuppressive action of cyclophosphamide, chloramphenicol and cyproheptadine hydrochloride in comparison with the control and dexamethasone was undertaken on skin homografts in rabbits and rats. The survival period of skin homografts in cyclophosphamide treated group was significantly increased (P less than 0.01) in rabbits, but there was insignificant increase in rats (P greater than 0.01). With chloramphenicol there was significant increase in survival period in both the species (P less than 0.01). Cyproheptadine did not show significant result in any of the species. The probable mechanisms of immunosuppressive action have been discussed.


Isolation of a low-molecular-weight antibacterial system from human amniotic fluid.

A low-molecular-weight antibacterial system has been isolated from human amniotic fluid. The bacterial inhibitor requires the metal cation zinc and a peptide with a molecular weight of 630. The peptide component was purified using ultrafiltration, gel filtration, and ion-exchange chromatography. It can be inactivated by digestion with carboxypeptidase. The amino acid composition of the peptide is: 3 glutamine-glutamic acid, 2 glycine, and 1 lysine. Removal of zinc from the peptide has been shown to remove bacterial inhibitory activity.


Electron microscopic and radioisotopic studies on cap formation in Toxoplasma gondii.

Trophozoites of Toxoplasma gondii from mouse peritoneal exudate are capable of shedding antibodies with which they have been previously coated. The antibodies are first moved towards the anterior pole of the parasite, at which they form a "cap". From the pole they are shed into the environment in the form of antigen-antibody complexes. An internalization of the accumulated material has never been encountered. Most probably, the observed phenomenon reflects a mechanism by which the parasite evades the host's immune response.


Persistence, pathogenesis, and morphology of an L-form of Streptococcus pyogenes adapted to physiological isotonic conditions when in immunosuppressed mice.

Evidence obtained using nonimmunosuppressed and newborn mice suggests that the immune response of the host plays a role in the rapid removal of a physiologically isotonic L-form of Streptococcus pyogenes, since its inability to persist in vivo was not due to osmotic lysis. With mice immunosuppressed with methylprednisolone sodium succinate, viability and detection of this L-form by fluorescent antibody was prolonged for at least 2 weeks, the approximate duration of immunosuppression in these mice. However, heat-killed L-form cells only persisted for 3 days in such mice. Therefore, persistence of a viable L-form in these treated mice was not simply due to the lack of removal of L-forms by a compromised host. At no time was there any indication of illness in nonimmunosuppressed or immunosuppressed mice after L-form injection, and all internal organs, when examined macroscopically, remained normal. Thus, overt pathogenesis was not a characteristic of this L-form in a suitable host even when its immune response had been compromised. The microscopic morphology of the L-form after isolation from immunosuppressed mice changed drastically. It was typically micrococcal in appearance and exemplified the cellular variability achievable by this organism in vivo. Also, streptolysin S production was increased markedly by passage of the L-form in immunosuppressed mice. However, M protein, as a cellular component, was not detected serologically, nor was any reformation of a rigid cell wall apparent by electron microscopy after isolation of this streptococcal L-form from such mice.


Limulus amebocyte lysate reaction with streptococcal pyrogenic exotoxin.

Three immunologically distinct pyrogenic exotoxins derived from group A streptococci were found to cause coagulation of limulus amebocyte lysate. These exotoxins were shown to be several thousand times less active for lysate coagulation than gram-negative endotoxin.


[Bacteriological studies in outpatients with acute urinary tract infections with particular reference to the resistance spectrum (author's transl)].

Bacteriological investigation of urinary samples from 1,926 non-hospitalised patients with documented or suspected acute urinary tract infection revealed organisms pathogenic for the urinary tract in 56.4% of the patients, who came from various parts of West Germany. Prevalent pathogens were E. coli (69%) and Proteus mirabilis (14%). E. coli and P. mirabilis demonstrated a low rate of resistance against ampicillin, the cephalosporines, gentamicin, tobramycin, and also against nitrofurantoin, nalidixine acid and trimethoprimsulfamethoxazole. The situation was more unfavourable in the case of Klebsiella and indolpositive Proteus species however, there being a noticeably high proportion of strains resistant to gentamicin and tobramycin.


[Antimicrobial effectiveness of sisomicin. I: In vitro activity of sisomicin compared with gentamicin, tobramycin, amikacin and kanamycin (author's transl)].

The aminoglycosides sisomicin, gentamicin, tobramycin, amikacin and kanamycin are highly active against staphylococci including the penicillinase-positive strains. Sisomicin is more effective than amikacin and kanamycin. Mixed infections with staphylococci and Enterobacteriaceae or Pseudomonas aeruginosa are thus on indication for treatment with sisomicin or other aminoglycosides. Infections with E. coli, Enterobacter, susceptible Klebsiella, and susceptible Pseudomonas strains can be treated with sisomicin, gentamicin or tobramycin. In such cases sisomicin is the most effective antibiotic because of its high antimicrobial activity. In infections with these organisms amikacin can also be used for treatment especially if there is resistance to other aminoglycosides. In hospital-acquired infections with Serratia marcescens amikacin and sisomicin are the drugs of choice. Both aminoglycosides have to be given in high doses in infections with Serratia because of the high inhibitory concentration for Serratia. Sisomicin demonstrates a high antimicrobial activity particularly against indole-positive Proteus species such as Proteus vulgaris and Proteus morganii, Enterobacter, and gentamicin-sensitive Pseudomonas strains. In infections with Pseudomonas aeruginosa tobramycin is the most effective bactericidal antibiotic. Amikacin is the drug of choice against gentamicin-resistant Pseudomonas strains which are also not infrequently resistant to other aminoglycosides. The low proportion of resistance to sisomicin of 7,6% in 370 organisms is only exceeded by amikacin with a rate of 0,6% (resistance to tobramycin 11,4%, gentamicin, 13,2% and kanamycin 42,4%). The low rate of resistance and the high antimicrobial activity are essential advantages of sisomicin.


[Changes in the antibiotic sensitivity of urinary tract pathogens (author's transl)].

The resistance of 2,314 bacterial strains from 1968 to 1972 was investigated drawing on case histories from the Munich University Urological Clinic. Using the disc-diffusion method 12,160 sensitivity tests were conducted with these strains against gentamicin, co-trimoxazole, ampicillin, tetracycline and chloramphenicol. The material was compiled from 35,000 individual data by means of modern electronic data processing and evaluated according to variable criteria. As expected the incidence of resistance in hospital-acquired pathogens was considerably higher than that in community-acquired pathogens; however, the resistance pattern of hosptial-acquired pathogens remained essentially unchanged from 1968 to 1972, whereas a significant increase of resistance in community-acquired infections was recorded. It can thus be concluded that the methods used in this clinic for combatting hospital infection and a more critical attitude towards anti-bacterial therapy have had positive results. An express warning is issued against unspecific antibacterial therapy which is often administered outside the clinic on the pretext of long-term therapy. In the light of the resistance situation of urinary tract pathogens, antibiotics should be used sparingly, keeping in mind the acidification of urine and the wash-out principle by means of high fluid intake.


Effect of active and passive immunizations with lipid A and salmonella minnesota Re 595 on gram-negative infections in mice.

The capacity of lipid A, a structure common to the lipopolysaccharide cores of all gram-negative bacteria, to serve as an active immunizing agent in mice and to protect these animals against gram-negative infections was investigated. Active immunization experiments were also performed with the Re mutant of Salmonella minnesota 595 which carries a lipopolysaccharide composed of lipid A and three residues of ketodeoxyoctonic acid. Single injections of lipid A complexed to acid-hydrolyzed bacteria as carriers failed to induce specific protection against subsequent challenge infections with E. coli O4 and S. breslau. Repeated injections of lipid A resulted in good protection against intraperitoneal challenge with S. breslau and partial protection against intravenous challenge with the same organism but did not alter the sensitivity of mice to challenge infections with E. coli or Pasteurella multocida. Whole antisera or serum fractions from rabbits in which high titers against lipid A had been attained by repeated intravenous injections of the antigen did not protect mice against challenge infections with E. coli O4. In contrast a single injection of the Re mutant of S. minnesota antigen in combination with incomplete Freund's adjuvant provided substantial protection against an otherwise lethal intraperitoneal infection with S. breslau over a period of at least 45 days. Repeated application of the Re antigen resulted in partial protection against experimental infections with E. coli O4, S. breslau and Pasteurella multocida. Injections of S. minnesota Re 595 antiserum provided better protection against an E. coli O4 infection than lipid A sera or antibodies of the IgG or IgM type directed against this antigen.


Colonization of caesarean section babies by Excherichia coli.

Eight babies all delivered by Caesarean section were observed for evidence of colonization by E. coli from birth until discharge from hospital 10 to 14 days later. Faecal specimens were cultured daily for E. coli as were swabs from the rectum, groin, umbilicus, head, hands und mouth. Different serotypes of E. coli were isolated from four babies on day 1, one baby on day 2, one baby on day 7 and two babies on day 9. Three babies excreted more than one serotype of E. coli. Serotypes of E. coli isolated from the faeces and rectum were usually isolated from the groin but less often from the umbilicus. Other sides were rarely colonized. In all instances except one the primary site of colonization appeared to be the gastrointestinal tract. Colonization by E. coli was influenced by antibiotic therapy and possibly by the method of feeding.


Escherichia coli isolated from babies delivered by caesarean section and their environment.

The sources from which eight Caesarean section babies acquired E. coli are described and the probable routes by which the organisms reached the babies are outlined. Suggestions are made concerning the control of the spread of E. coli in premature nurseries and during outbreaks of E. coli gastroenteritis.


Escherichia coli in a maternity ward.

The Escherichia coli isolated from faecal and nasal swabs from all babies present in a maternity ward were serotyped and their anbitiotic resistance patterns and fermentation characteristics with six carbohydrates were determined. These were compared with E. coli isolated from hands and clothing of attendants and samples of ward air. The results suggest that mothers may be the primary source of E. coli in the ward but the colonized babies themselves provide the main reservoir of nursery E. coli. As most of these serotypes of E. coli were found on the attendant's hands or clothing or in the air, it is difficult to assess transmission routes for E. coli, particularly as some strains appear to possess a greater inherent ability to colonize babies than others. By the use of markers other than O antigens it was possible to characterize strains to a far greater extent.


[Antibody binding bacteria in the urine in chronic pyelonephritis].

Using the direct immunofluorescence technique antibody-coated bacteria were demonstrated in urine samples from 49 of 57 patients with the clinical diagnosis of chronic pyelonephritis, but were not observed in urines from 11 patients with cystitis. A correlation rate was found between the presence of antibody-coated bacteria in the urine seidment and elevated serum antibody titers in the pyelonephritis group as determined by the indirect immunofluorescence technique. Patients with elevated serum antibody titers without antibody-coated bacteria in the urine, and vice versa, were also found. Patients with cystitis did not have elevated serum antibody titers against the homologous strain isolated from the urine. The clinical diagnosis of chronic pyelonephritis could be confirmed in 86.5% of patients taking into account the presence of antibody-coated bacteria and/or elevated serum antibody titers.


[The liver concentration of cephradin and cephacetril and their elimination in the bile].

Liver biopsies and serum samples were collected after intravenous application of 2 g cephradin (n = 13) or 2 g cephacetril (n = 11) during surgery. There was no difference in the serum levels of cephradin and cephacetril. 30 min. after i.v. application of cephradin the liver tissue concentration was 72.62 mcg/g. 30 min. after i.v. cephacetril the liver tissue concentration was 5.83 mcg/g. The quotient of liver tissue concentration to serum concentration for cephradin was between 0.36 and 0.83, and for cephacetril between 0.02 and 0.16. The excretion of cephradin and cephacetril in human bile was studied by collecting bile samples from the common bile duct via T-tube drainage (n = 17). Cholecystomized patients were given 2 g of antibiotics intravenously. Serum levels of cephradin were 263 mcg/ml 5 min after application, and 22 mcg/ml after 240 min. Serum levels of cephradin were 263 mcg/ml 5 min after application, and 22 mcg/ml after 240 min. Serum levels of cephacetril were 193 mcg/ml 5 min after application, and 27 mcg/ml after 240 min. The highest levels of cephradin in the bile were found 75 min after injection at a concentration of 86.4 mcg/ml; the highest level for cephacetril was 21.8 mcg/ml at 15 min. In patients with hyperbilirubinaemia cephradin reached a mean maximum concentration of 29.6 mcg/ml in bile samples, in comparison to 117.4 mcg/ml in normal patients, while no difference was seen with cephacetril. After intravenous administration of 2 g cephradin biliary concentration are achieved which may be sufficiently high to be effective not only against the very sensitive gram-positive organisms, but also against most strains of E. coli, Klebsiella and indol-negative Proteus. Cephradin is effective in the treatment of cholangitis and intrahepatic abscesses, as was observed in 18 patients. A free bile-flow is essential.


The acquisition of Escherichia coli by new-born babies.

In discussing the background of these studies the importance of faecal carriage of Gram negative organisms by hospital patients is stressed. In many instances it was shown that transmission is by an oral route. This discussion leads on to an assessment of the dose required for Escherichia coli to implant in the bowel. The difficulties of studying the spread of E. coli within a faecal specimen are discussed. A number of papers are quoted which show that E. coli are present in the vagina of women and that the acquisition of these E. coli by babies is related to the length of time that the birth takes, and that there is a relationship between the E. coli found in the faeces of the mothers, the mucus swallowed by the babies at birth and subsequently in the faeces of the babies. Most of the eralier studies quoted deal predominantly with enteropathogenic serotypes, but it was later shown that other serotypes can be similarly acquired by the babies. Although this appears to be the general method by which babies acquire their faecal E. coli, it is well established that they can also be obtained from the environment, hence ward outbreaks of infantile gastroenteritis. Studies on normally delivered babies show that generally two thirds obtain their faecal E. coli from their mothers while the rest appear to pick up environmental strains. Very detailed biochemical and serological studies need to be performed to assess this. Caesarian section babies are generally not likely to become colonized by their mothers' faecal E. coli and studies are described which show that the babies faeces or rectal swabs are usually the first areas colonized and that the E. coli are spread from there. Extensive environmental studies suggest that contaminated hands and uniforms of the nursing staff may be the main vector for transmitting E. coli. There is a wide variety of E. coli serotypes present in a maternity ward at any time and these are related to the presence of the babies excreting them. A variation in the ability of strains to spread was noted.


Chemotherapy of typhoid fever: a review of literature.

Until the last few years, chloramphenicol was recognized positively as the drug of choice in the treatment of acute typhoid fever. Its hematoxicity, as well as the recently observed epidemic and the present endemic occurrence of S. typhi strains with R-factor-mediated resistance to chloramphenicol in Mexico, India and South-EAst Asia, render the clinical evaluation of new antibacterial agents extremely important. By means of a literature review on controlled comparative trials, the value of thiamphenicol, ampicillin, amoxycillin, furazolidone and co-trimoxazole as alternative drugs for the treatment of acute typhoid fever is examined. Co-trimoxazole seems to be the drug of choice in the treatment of acute typhoid fever. For the treatment of the chronic typhoid carrier ampicillin is most frequently used, but amoxycillin and co-trimoxazole seem to be just as effective.


Properties of the K562 cell line, derived from a patient with chronic myeloid leukemia.

The K562 cell line derived from a CML patient in blast crisis was examined for properties of B and T lymphocytes and cell lines. K562 lacks the B markers of immunoglobulins, Epstein-Barr virus (EBV) genome and associated nuclear antigen, and receptors for EBV. A low proportion of cells from rosettes with sheep erythrocytes, the frequency of which is considerably increased after neuraminidase treatment. Unlike B lines but like T lines, K562 cells are lysed rapidly by C'/Fc receptor-positive human blood leukocytes and do not stimulate MLC reactions. On the other hand, K562 lacks T antigen, high radiosensitivity and sensitivity to growth inhibition by thymidine. The cells do not contain N-APase, an enzyme found in all lines derived from lymphoid cells and in lymphoproliferative diseases. By scanning electron microscopy, K562 cells were seen to be rounded and relatively smooth, with small numbers of short microvilli resembling undifferentiated leukemic cells. A few cells had narrow ridge-like profiles and small ruffles similar to granulocytic leukemic cells. K562 is strongly positive for immunoglobuln Fc receptors and pinocytosis, but does not phagocytose or mediate antibody-dependent phagocytosis or cytolysis. Among histochemical stains, K562 is positive for esterase, lipid, and acid phosphatase. There seems to be no doubt that K562 is not a B cell line. While it has some T cell properties, these are not exclusive. Some of its characteristics indicate that it is probably not lymphoid. Due to its low level of differentiation, its nature cannot be stated with certainty. On the basis of the possible presence of the cellular marker of chronic myeloid leukemia, the Ph chromosome, it may be regarded as belonging to the granulocytic series of cells.


Emergence of a subpopulation of lymphocytes bearing theta antigen and complement receptor during tumor growth.

Spleens from BALB/c mice transplanted with mammary tumors display a significant increase in the percentage of lymphocytes bearing complement receptors (CRL) while the percentage of cells bearing surface immunoglobulins (SIg) remains unchanged in comparison with that in spleens from normal animals. Upon separation on nylon columns, the increase in CRL in the spleens from tumor-bearing mice was limited to the non-adherent cell population. Simultaneous maker analysis disclosed that these CRL lacked detectable SIg. Indirect immunofluorescence indicated that these cells possessed theta antigen. Absence of SIg and the presence of theta antigen coupled with the restricted occurrence of these cells in the nylon-non-adherent population which responded to PHA and Con A but not to LPS indicate that these cells may constitute a subset of T cells. The paucity of this subpopulation of spleen lymphocytes in normal spleen suggests that their emergence is related to tumor growth.


Sequential changes in the numbers of B and T lymphocytes and other leukocytes in the blood in Marek's disease.

Numbers of B, T and total lymphocytes, monocytes, heterophils, eosinophils and basophils have been examined in the peripheral blood of chickens between 2 and 42 days after infection with Marek's disease virus. During the stage of the acute restrictively productive virus infection of lymphoid tissues at 2-9 days after infection, absolute numbers of B cells, T cells, total lymphocytes and heterophils were increased, those of monocytes and eosinophils were decreased, and those of basophils were unchanged. The lymphoproliferative phase of the disease, from 21-42 days after infection leading to lymphoma formation, was accompanied by an increase in T cells, total lymphocytes and possibly eosinophils, and a decrease in B cells, monocytes, heterophils and basophils. The T-cell increase following infection occurred only in female birds, and there were more lymphomas in females than in males. The increase in lymphocytes in the blood of six birds with leukemia was mainly due to an increase in T cells, but in one bird B cells were also increased. Blast cells and atypical lymphoid cells were increased in leukemic birds. Regression coefficients were calculated between different pairs of leukocytes in infected and uninfected birds at different stages of the disease. Particularly noteworthy were the associations between B and T cell numbers, which indicated constant proportions of these cells irrespective of total numbers, possibly due to a common control mechanism.


Results obtained with fractionated ifosfamide massive-dose treatment in generalized malignant tumours.

In 409 sufferers from various malignant tumours, we used the cytostatic Ifosfamide (ASTAZ4942) in fractionated doses. The total i.v. dose averaging 300 mg/kg bodyweight, was either spread over 5 consecutive days (5 X 60 mg/kg i.v.) or over 10 consecutive days (10 X 30 mg/kg). At the same time, most patients were irradiated, the radiation dose usually being only one tenth the antitumour dose. Infections and electrolyte imbalance were first treated before Ifosfamide therapy was instituted. Cases of advanced cerebral sclerosis, thrombopenia below 75,000/cmm, cerebral metastases, impaired renal function and inadequate cooperation of the patient were excluded from the studies. To prevent and control side effects, various premedications and adjuvants are required: Antiemetics, prevention of cystitis and infections, cardiovascular agents etc. Corticosteroids are contraindicated. Out of 360 assessable patients 101 had a full remission, 150 a partial remission, 79 were failures; 30 cases were not evaluated. Good results were seen especially in ovarian carcinoma, mammary carcinoma and microcellular bronchial carcinoma. Particularly striking is the drug's effectiveness in testicular tumours including teratomas, osteosarcomas, chondrosarcomas and myosarcomas as well as in some adenocarcinomas of the gastro-intestinal tract, particularly pancreatic carcinoma. In lymphoreticular tumours and haemoblastoses, its potency is less pronounced. The side effects of Ifosfamide are the same as those of other alkylating agents. They are reversible and can usually be controlled or even avoided by adequate preventive measures. In the order of incidence we observed: Alopecia, leukopenia, fall in haemoglobin, cystitis, intercurrent infections, nausea and vomiting as well as cerebral disorders. Since haemorrhagic cystitis considerably interferes with Ifosfamide treatment, its prevention is of essential importance. Because of possible complications and specific premedication and adjuvant measures for their control, this type of treatment should for the present only be carried out by oncologists or special cancer centres.


[The number of subjects or patients in clinical pharmacological studies].

A technique permitting without preliminary informations the assessment of the minimal number of test persons or patients to be studied in clinicopharmacological investigations dealing with multiple variables (vital functions, laboratory data) is reported. The procedure is applicable to investigations using data recorded at different time intervals and concomitant variables (basic values at the start of the investigation, age, and body weight). The minimal number of test persons or patients is estimated from the results of a formal examination. In this examination is tested whether a multivariate analysis of a therapeutic effect using all data only once and simultaneously is based on sufficient degrees of freedom of the error variance.


Radiometric studies of Mycobacterium lepraemurium.

The radiometric method has been applied for studying the metabolism of M. lepraemurium and the conditions which might force or inhibit its metabolic activity in vitro. These organisms assimilate and oxidize (U-14C) glycerol, and (U-14C) acetate, but are unable to oxidize (U-14C) glucose, (U-14C) pyruvate, (U-14C) glycine and 14C-formate. When incubated at 30 degrees C M. lepraemurium oxidizes (U-14C) acetate to 14CO2 faster than 37 degrees C. The smae effect was observed with increasing concentrations of polysorbate 80 (Tween 80), or the 14C-substrate. No change in metabolic rate was observed when the organisms were kept at -20 degrees C for 12 days. Although tried several times, it was not possible to demonstrate any "inhibitors" of bacterial metabolism in the reaction system. The radiometric method seems to be an important tool for studying metabolic pathways and the influence of physical and biochemical factors on the metabolism of M. lepraemurium in vitro.


Separation of M. leprae from human leproma and the development of a cytoplasmic skin test antigen from purified bacilli.

A successful method for purification of M. leprae from human leproma without subjection to heat has been developed. The "floater" phenomenon has been described which consists of bacillary tendency to float in the supernate when bacilli which are not autoclaved are separated from tissues by enzymatic digestion. A method for preparing cytoplasmic fractions from purified M. leprae has been developed for the production  of a skin test antigen for leprosy. The cytoplasmic fraction of M. leprae elicited positive skin test responses in people with tuberculoid leprosy and negative responses in lepromatous leprosy. Cytoplasmic preparations from purified M. leprae had little cross-reactive relationship with the organism BCG. The small particulate fraction elicited positive reactions in PPD-S negative as well as BCG vaccinated individuals.


Double-blind comparison of phlebitis associated with cefazolin and cephalothin.

In a double-blind, crossover study in 28 patients having infections susceptible to cephalosporins (mostly Staphylococcus or Streptococcus), phlebitis developed in twice as many patients (46.4%) receiving intravenous cephalothin within 48 hours, as in patients (21.4%) receiving intravenous cefazolin over the same time period. Likewise, drug therapy had to be discontinued more frequently with cephalothin (39%) than with cefazolin (21%), and the percentage of patients reporting pain or discomfort scores of 1+ or higher was greater with cephalothin (46.4%) than with cefazolin (21.4%). In our study, the intravenous administration of cefazolin was associated with a lower incidence of phlebitis than was cephalothin.


Macrophage function in leprosy.

The macrophage function in patients with leprosy was assessed by estimating histochemically the acid phosphatase activity in skin biopsies and by assessment of phagocytic and lytic capability of in vitro cultured macrophages derived from peripheral blood monocytes, challenged with live M. leprae. Acid phosphatase was demonstrated in skin biopsies of different groups of leprosy patients classified according to the Ridley and Jopling scale. The degree of acid phosphatase positivity was correlated with clinical spectrum, Bacterial and Morphologic Indices and treatment status. Peripheral blood monocytes from patients with leprosy, either tuberculoid or lepromatous, were cultured in monolayers and challenged with M. leprae. The phagocytosis and lysis of mycobacteria by macrophages was observed at different time intervals from the 1st to the 28th day. The morphology of the macrophages in different types of leprosy was also studied. The results suggest that macrophages from patients with either tuberculoid or lepromatous leprosy are not by themselves capable of lysing live M. leprae. Live M. leprae injected into the foot pad of Wistar strain of rats evoked similar responses on the tenth day, in normal and protein deficient animals.


The genetic control of liquid-holding recovery and U.V.-induced repair resistance in the yeast, Saccharomyces cerevisiae.

Liquid-holding treatment in non-nutrient solutions after U.V.-exposure results in an increased resistance of wild-type (RAD) yeast cells to a second U.V.-treatment (repair resistance). U.V.-sensitive (rad) mutants of yeast which show variation in their response to liquid-holding treatment and a second U.V.-dose-range have been classified into two groups. Mutants of Group 1 show increased viability after post-U.V.-liquid-holding treatment and show repair resistance. In contrast, mutants of Group 2 which show reduced viability during post-U.V.-liquid-holding treatment have the same U.V.-sensitivity, both before and after liquid-holding treatment. Genetic analysis of crosses of the rad mutants to wild-type cultures indicate that the phenotype of repair resistance to U.V.-treatment is under genetic control and depends on the presence of alleles of radiation sensitivity genes, which also confer the property of liquid-holding recovery.


The analysis of single-strand breaks in E. coli using a curve-fitting procedure.

A curve-fitting technique has been used to analyse the activity profiles occurring when the tritium labelled DNA of irradiated E. coli is sedimented in an alkaline sucrose gradient. The fitted curve is a modification of an expression derived elsewhere, which assumes that DNA of unit length undergoes scissions at random positions on the DNA strand. The theoretical curves fit the data well, even after considerable repair or excision of the radiation damage has been effected by cellular enzymes. The profile resulting from sedimentation of unirradiated DNA may be fitted with the same theoretical expression provided that the velocity of centrifugation is not too high. The number of single-strand breaks derived from the analysis is almost unaffected by the presence of extraneous activity at the top or bottom of the sucrose gradient. The method is therefore more reliable than many other methods used to analyse sedimentation data.


The myocardium.

A comparison of some of the mechanical properties of cardiac with other types of muscle has been made, showing that, except for the speed of some responses, cardiac muscle is similar to other types of muscle. Furthermore, the techniques used in both living and glycerol-extracted insect fibrillar and vertebrate skeletal muscle are applicable to heart muscle, where the focus of the technique is now on cross-bridge mechanics and energetics. It is particularly encouraging to see many well known phenomena such as inactivation with shortening, stress related increases in active force, and the Fenn effect begin to find some more specific relation to cross-bridge mechanical and chemical activity. The high compliance of cardiac preparations still clouds the interpretation of data obtained from whole muscle preparations; however, the reduced compliance of glycerol-extracted cardiac muscle offers some hope of obviating some series compliance. Indeed, the correspondence in mechanical responses of living and glycerol-extracted preparations shows that glycerol preparations are of great utility since the time dependence of activation also can be removed in these studies. A more complete analysis of muscle models, in which the cross-bridge contribution to muscle elasticity is more realistically evaluated, should help in relating muscle measurements to cross-bridge activity. Furthermore, studies on both living and glycerol-extracted cardiac muscle, particularly if sarcomere length can be controlled, offer new hope of closing the perpetual gap in our understanding of cardiac muscle physiology relative to skeletal muscle.