Clinical trials in cancer chemotherapy.

Clinical trials in cancer chemotherapy. The success of cancer chemotherapy has increased greatly the potential areas for its inclusion in clinical trials and therefore has made the experimental design considerations more complex in these trials. New drugs still go through the three classic phases of clinical study beginning with clinical pharmacology (Phase I), efficacy screening (Phase 2) and role delineation (Phase 3). The Phase 2 and 3 trials now need to be considered within the overall therapeutic strategies which are required for each of the many diseases which are called cancer. The exigencies of a disease-oriented strategy which requires a blend of therapeutic modalities many times require a modification of what would be an ideal modality-oriented strategy geared solely to effectively testing a new agent. A new drug may need many Phase 2 and 3 trials in a variety of tumors before it can be considered adequately evaluated. Essential aspects of valid clinical trials include an adequate protocol which details the study and a design which is feasible for answering the question. Historical controls are advocated by many as a valid approach to clinical trials. Unless the numbers in the historical control are large and the prognostic variables well matches this approach should be viewed cautiously by the practicing physician.


Gastrointestinal cancer--colon (surgery-radiotherapy). The role of radiation therapy in the management of rectosigmoid cancer. The lack of an improvement in the gross survival statistics for colorectal cancer during the last decade and the negative results of randomized trials, suggests that 5-FU used as an adjuvant treatment has not increased survival. Current chemotherapeutic drug combinations, however, may prove effective in the adjuvant situation. The adjuvant use of preoperative radiation has shown improved survival in one randomized study and no change in the second study. Current preoperative radiotherapy studies using a more effective radiation dose will confirm the effect of preoperative radiation both on the primary tumor control and on metastases. Current postoperative adjuvant trials incorporating radiation, chemotherapy or a combination of the two, when completed, will provide clear indications for the physician as to the optimum treatment of Duke's B and C cancer of the colorectum. Three national randomized studies for inoperable, recurrent or residual carcinoma of the colorectum are testing the effect of radiation alone or in combination with chemotherapy or chemotherapy followed by immunotherapy.


The effects of specific antibody-complement-mediated cytotoxicity on transformed and untransformed Syrian hamster cells. Antibodies were produced against 220,000-molecular-weight proteins of Syrian hamster embryo cells. The antiserum containing these antibodies is capable of immunologically staining the surfaces and a fibrillar network around untransformed Syrian hamster embryo cells. The staining is removed by adsorption of the antiserum with Syrian hamster embryo cells and by mild trypsin treatment. Several lines of neoplastically transformed Syrian hamster embryo cells isolated and cloned after treatment with chemical carcinogens show little or no immune staining. Adsorption of the antiserum with certain transformed cells does not significantly reduce the immune staining of untransformed Syrian hamster embryo cells. The immune antiserum, in the presence of complement, is selectively cytotoxic to the untransformed Syrian hamster embryo cells. However, the transformed lines show resistance to this treatment. Analysis of the lactoperoxidase-catalyzed, iodinated cell surface proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveals that the transformed lines have either no detectable labeling or a marked reduction in the labeling of 220,000-molecular-weight proteins that are major iodinatable cell surface proteins on untransformed Syrian hamster embryo cells.


The origin of the vasopressinergic and oxytocinergic fibres of the external region of the median eminence of the rat hypophysis. The origin of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence was investigated by means of hypothalamic lesions, adrenalectomy and immunocytochemistry. The results obtained in bilaterally adrenalectomized animals with complete, or incomplete, destruction of the suprachiasmatic nuclei showed that, at least, the great majority of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence do not originate from the suprachiasmatic nuclei. From the observations obtained in bilaterally adrenalectomized animals with total or subtotal destruction of both paraventricular hypothalamic nuclei, it appears that the paraventricular nuclei must be the origin of (nearly) all the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence. The results strongly suggest that both types of fibres originate from all parts of the paraventricular nuclei.


Electron microscopic study on the innervation of the pancreas of the domestic fowl. The innervation of the pancreas of the domestic fowl was studied electron microscopically. The extrapancreatic nerve is composed mostly of unmyelinated nerve fibers with a smaller component of myelinated nerve fibers. The latter are not found in the parenchyma. The pancreas contains ganglion cells in the interlobular connective tissue. The unmyelinated nerve fibers branch off along blood vessels. Their synaptic terminals contact with the exocrine and endocrine tissues. The synaptic terminals can be divided into four types based on a combination of three kinds of synaptic vesicles. Type I synaptic terminals contain only small clear vesicles about 600 A in diameter. Type II terminals are cgaracteruzed by small clear and large dense core vesicles 1,000 A in diameter. Type III terminals contain small vesicles and small dense core vesicles 500 A in diameter. Type IV terminals are characterized by small and large dense core vesicles. The exocrine tissue receives a richer nervous supply than the endocrine tissue. Type II and IV terminals are distributed in the acinus, and they contact A and D cells of the islets. B cells and pancreatic ducts are supplied mainly by Type II terminals, the blood vessels by Type IV terminals.


Immunocytochemical study of the hypothalamo-neurohypophysial system. II. Distribution of neurophysin, vasopressin and oxytocin in the normal and osmotically stimulated rat. Antisera, with cross reactive antibodies removed by affinity chromatography, were used in the immunoperoxidase-bridge technique to study the distribution of oxytocin and vasopressin together with neurophysin in the hypothalamo-neurohypophysial system of the rat. The hormones were demonstrated in different areas of the supraoptic nucleus (SON) and paraventricular nucleus (PVN), in neurosecretory fibres of the hypothalamo-neurohypophysial tract, median eminence, and in nerve terminals of the neurohypophysis. Intact normal and rats with hereditary hypothalamic diabetes insipidus (Brattleboro strain), and rats dehydrated by the administration of oral hypertonic saline were studied. In dehydrated rats the hormone concentration in the neurons, and the number of neurons containing hormone varied according to the time of dehydration stress. The observations support the hypotheses that: 1) oxytocin and oxytocin-neurophysin, and vasopressin and vasopressin-neurophysin are synthesised in different neurons and are transported along different axons; 2) the SON and PVN are functionally indistinguishable in that neurons containing oxytocin or vasopressin are present in both nuclei; and 3) the two types of neurons respond to osmotic stimulation in a way that is qualitatively the same but quantitatively different.


Immunocytochemical study of the hypothalamo-neurohypophysial system. III. Localization of oxytocin- and vasopressin-containing neurons in the pig hypothalamus. Synthetic oxytocin and [8-arginine]-vasopressin conjugated to bovine thyroglobulin were used to induce specific antibodies in rabbits. The specificity of the anti-oxytocin serum, and the suitability of the anti-[8-arginine]-vasopressin serum for the detection of [8-lysine]-vasopressin, was evaluated by immunofluorescent studies of the respective hormones bound to Sepharose 4B particles. Oxytocin and [8-lysine]-vasopressin were specifically localized in the paraventricular (PVN) and supraoptic (SON) nuclei of the pig hypothalamus using the immunoperoxidase staining technique. After an examination of serial transverse and sagittal sections stained for either of the hormones we observed that: 1. In the rostral SON, oxytocin and vasopressin containing neurons were uniformly distributed; 2. In the caudal SON, most of the neurons contained oxytocin, but there were still a few 'vasopressin' neurons; 3. In the rostral PVN, the two hormones were evenly spread in neurons close to the third ventricle; 4. In the caudal PVN, the oxytocin and vasopressin containing neurons were differentially distributed, with 'oxytocin' neurons adjacent to the third ventricle, and 'vasopressin' neurons lateral to these and concentrated in the dorso-caudal PVN. In the cells of the PVN, there was evidence that the distribution of oxytocin and vasopressin is similar to the distribution of porcine neurophysin-II and porcine neurophysin-I respectively. This similarity is consistent with the one hormone--one neurophysin concept in the pig.


Immunofluorescence study of LRF neurons in man. Human LRF neurons were characterized by immunofluorescence, using rabbit immunesera against unconjugated synthetic LRF, previously adsorbed on polyvinylpyrrolidone. These neurons, which vary in number from one specimen to another, are mainly concentrated in the mediobasal hypothalamus (infundibular and premammillary nuclei in particular) and in the lamina terminalis and the neighbouring preoptic area. The give rise respectively to a hypothalamo-infundibular LRF tract (ending around the capillaries of the primary portal plexus of the  infundibulum) and to a preoptico-terminal tract (ending mainly around the capillaries of the primary and secondary plexuses of the vascular organ of the lamina terminalis and, in addition, between the ependymal cells lining its ventricular surface). It is suggested that these two tracts could be implicated in the tonic and cyclic control of gonadotropic secretion. Some reactive neurons are also present in the septal and pericommissural regions and in the retromammillary area and rostral mesencephalon. These neurons give rise to various extrahypophyseal LRF tracts, probably ending in the telencephalon and the brainstem. It is suggested that LRF, in addition to its major "prehypophysiotropic" action, is able to modulate the activity of certain telencephalic or mesencephalic structures.


Localisation of gonadotropin releasing hormone (GRH) in the forebrain and neurohypophysis of the trout (salmo gairdneri). An immunofluorescence study. The double antibody immunofluorescence technique was applied to serial coronal and sagittal sections of the brains of the trout, Salmo gairdneri, using an antibody against mammalian LH-RH. Immunoreactive material was found in small perikarya, situated at both sides of the ventriculus communis in the area dorsalis pars medialis of the telencephalon. The axons of these perikarya, also containing immunoreactive material, do not form a tract, but run diffusely in a caudo-ventral direction towards the pituitary stalk. The ending of these fibres has not yet been established.


Tentative immunohistochemical demonstration of melatonin in the rat pineal gland. In the present study an attempt was made to demonstrate melatonin in the rat pineal gland by means of immunohistochemistry. The anitbody used was raised against 5-methoxy-N-acetyltryptophan which is chemically similar to melatonin. Specific fluorescence was demonstrable only in pineals from rats killed during the night, when melatonin formation is high. It was restricted to parenchymal cells lying in a marginal zone of the organ. These results are discussed in relation to a subdivision of the pineal parenchyma into cortical and medullary areas.


Regeneration of the islets of Langerhans in the Guinea pig. The regeneration of the islets of Langerhans in the guinea pig was studied after intravenous injection of alloxan. A number of beta cells in the islets was destroyed within 24h after alloxan, but after 48h there was a rapid proliferation of the surviving cells of the islets. This depended on the dosage of the drug as well as the timing. Electron microscopy of the islet at 48h showed that the dividing cells had small electron dense granules and resembled a sub-type of normal A cells, whose function is not yet known. There were also many agranular cells in the islet. These two groups of cells seen in the regenerating islet could be precursor cells, which could differentiate into beta cells. There was no evidence for transformation of duct cells or acinar cells into islet cells. None of the guinea pigs became permanently diabetic. This was probably due to inadequate dosage which resulted in only partial degeneration of the beta cells followed by regeneration and recovery. There was also some regeneration of the liver, kidney and the adrenal cortex following alloxan.


Distribution, ontogeny and ultrastructure of the mammalian secretin cell. Immunocytochemically, secretin cells have been demonstrated to occur in the duodenum and jejunum of several mammals. Calculations on the relative frequency of such cells indicate that the bulk of secretin occurs in the jejunum, a fact supporting the view that secretin may be released by physiological stimulants other than hydrochloric acid. Electron microscopical identification of cat and pig secretin cells confirmed their identity with the ultrastructurally defined S cells, and staining experiments revealed that secretin cells were argyrophilic both with the method of Grimelius and with that of Hellerström and Hellman. Secretin cells are detected already in the 17-day old fetal rat duodenum and show a developmental pattern similar to that displayed by the gastrin cells. It is suggested that secretin may play a role in the early regulation of growth of the fetal gastrointestinal tract.


Localization of myosin and actin in ocular nonmuscle cells. Immunofluorescence-microscopic, biochemical, and electron-microscopic studies. Myosin and actin were localized by indirect immunofluorescence microscopy using specific antibodies prepared in rabbits against highly purified gizzard myosin and actin. A strong fluorescence staining with both antibodies was observed in rat corneal epithelial cells, anterior lens epithelial cells, rod inner segments, and in rat and frog pigment epithelial cells. The immunohistochemical localization of myosin in corneal epithelial cells was further supported by the electrophoretic and immunological identification of smooth muscle type myosin heavy chain in pure corneal epithelial abrasions. Electron-microscopic observations revealed a clear correlation between staining with actin antibodies and the presence of numerous thin cytoplasmic filaments (50-80 A in diameter). The functional and biochemical nature of 90-110 A filaments occurring in corneal and lens epithelial cells, as well as the ultrastructural localization of myosin in ocular nonmuscle cells under study remains obscure.


Immunofluorescent localization and ultrastructural characterization of gonadotrophe cells in the adenohypophysis of the barbary drake (Cairina moschata L.) using anti-chicken LH serum. An indirect immunofluorescence technique and an anti-chicken LH serum were used to localize cells in the adenohypophyses of drakes at different stages of their breeding cycle, after castration, and after castration combined with thyroxine treatment. Immunofluorescent cells were distributed throughout both lobes of the adenohypophyses from control and experimental birds and were shown to be alcian blue positive. PAS negative, basophiles. Immunofluorescent cells were as numerous in castrated birds as in castrated birds treated with thyroxine. Adjacent thin and semi-thin sections were used to study the cells binding anti-LH serum at light microscope and ultrastructural levels. The cells contained spherical granules with variable densities and diameters ranging between 40 and 280 nm in the rostral (=cephalic) lobe, and between 60 and 260 nm in the caudal lobe. The light microscope and ultrastructural observations showed that the anti-LH serum binds to cells which have been classified by other authors in the Pekin duck, quail and pigeon as TSH producing delta cells. The experimental technique used did not permit a distinction to be made between cells producing FSH and LH.


Somatotrophs in the human fetal anterior pituitary. An electron microscopic-immunocytochemical study. An immunocytochemical staining of ultrathin sections using the peroxidase-antiperoxidase complex was employed to localize the somatotrophs in  the anterior pituitary of the human fetus. This technique allowed a specific identification of the somatotrophs and a detailed description of their ultrastructure which was well preserved by the fixation employed. They appeared as cells with a dense cytoplasmic matrix, lamellar rough endoplasmic reticulum, and prominent Golgi apparatus. Their secretory granules had a mean diameter of 300 nm.


Innervation of the cultured fetal rat pancreas. The fetal rat pancreas, explanted at 18 days of gestation and cultured up to ten days, contains numerous acetylcholinesterase-positive neurons. These nerves usually appear in small ganglia although single nerve cells are encountered. The axons of these intrapancreatic nerves appear to terminate only in the islet tissue and not on any exocrine components of the explant. It is concluded that the fetal rat pancreas contains an islet-specific group of cholinergic neurons.


Calcium containing particles in mitochondria of heart muscle cells as shown by cryo-ultramicrotomy and X-ray microanalysis. Mitochondria of normal myocardial cells of the sand rat and the mouse as well as of the left ventricle of man, have been examined for their content of calcium. Ultrahistochemistry and X-ray microanalysis revealed two basically different inclusions: Osmiophilic mitochondrial granules and spherical mitochondrial particles. Osmiophilic mitochondrial granules were found in conventionally fixed and plastic embedded tissues as well as in cryosections of chemically fixed and sucrose infused tissues. Such granules lacked inert electron density and probably consisted mainly of unsaturated lipids. X-ray spectra obtained from these tissues revealed no peaks for calcium. Spherical mitochondrial particles were present in dry-cut cryo-sections of N2-frozen tissues not treated by fixatives and/or cryoprotectants. These particles were deeply electron dense in unstained, freeze-dried cryo-sections. They usually measured from 600 A-900 A in diameter in the normal myocardium of the sand rat and the mouse and from 250 A-400 A in diameter in the left ventricular myocardium of man. Significant calcium peaks could be identified in the X-ray spectra of these particles, whereas none occurred in the analyses of other tissue regions. Potassium was detected with about equal frequency in the particles and in other parts of the tissue. On the basis of the inert electron density of the particles and their absence in chemically fixed tissues as well as of the results of the X-ray analysis, it is concluded that they contain precipitates of extremely labile ions of mitochondrial calcium.


The araC promoter: transcription, mapping and interaction with the araBAD promoter. The start sites of the araC and araBAD gene messenger of E. coli were located by transcription in vitro from short DNA fragments, by high magnification electron microscopy and by genetic mapping. Transcription for these messengers proceeds in opposite directions from the start sites that are 150 base pairs apart. Transcription from the araBAD promoter requires araC protein plus arabinose and CAP protein plus cyclic AMP. In the experiments performed in vitro, inducing the araBAD promoter represses activity of the araC promoter.


Synthesis and assembly of the membrane proteins in E. coli. Kinetics of integration of membrane proteins were studied in E. coli to discover how membrane proteins find their final location in the functional membrane. The experiments make use of a simple and convenient method developed for isolating inner and outer membranes from a number of small-scale cultures with high recovery. Among the proteins that constitute the cell surface structures, inner membrane proteins are integrated most rapidly after synthesis, whereas outer membrane proteins delay somewhat, and periplasmic proteins delay further in reaching their destinations. Protein I, a major outer membrane protein with molecular weight of about 37,000 daltons, exhibits significantly slower rates of integration than other outer membrane proteins. The decreased fluidity of membrane lipids by temperature shiftdown of an unsaturated fatty acid auxotroph grown on elaidate results in abnormally slow assembly of the outer membrane proteins and also in an anomalous assembly of the inner membrane proteins, suggesting that the fluid state of the lipids is required for normal operation of these processes. The possible relevance of these findings to the mechanism of membrane formation is discussed.


Effect of some drugs on excision repair in E. coli cells. The intercalating dye ethidium bromide (EB), inhibits excision of pyrimidine dimers from UV-irradiated excision-proficient Escherichia coli B/r hcr+ cells. Inhibition is total at a 2.5 - 10(-4) M concentration 120 min after irradiation with a dose of 750 erg/mm2. The viability of irradiated cells diminishes in proportion to the EB concentration. Under wholly analogous conditions of cultivation and irradiation no inhibitory effect of KCN and caffeine (CFF) and only a slight effect of chloramphenicol (CAP) on dimer excision has been observed. The viability of cells is affected by these compounds but it does not appear to depend on the quantity of excised photoproducts. A change in the secondary structure of DNA induced by intercalation of EB appears to be the reason for the depression of excision of UV photoproducts.


Isosteres of natural phosphates. 6. The preparation and properties of lysophosphotidic acid. We herein report the first chemical synthesis of phosphonic acid analogues of lysophosphatidic acid. The racemic isosteric analogues, 4-acyloxy-3-hydroxybutyl-1-phosphonic acids, of lysophosphatidic acid were prepared by both catalytic and hydride reductions of the 4-acyloxy-3-oxobutyl-1-phosphonic acids, a general method for the preparation of the latter having been reported previously. The lysophosphatidic acids have been found to substrates for lysophosphatidic acid acyl transferase, and may be acylated chemically to yield phosphotidic acids. The latter reaction is of use in the preparations of differentially acylated phosphatidic acids.


Effects of heart rate augmentation on left ventricular volumes and cardiac output of the transplanted human heart. The response of the donor left ventricle to atrial pacing was measured in 11 patients 12 to 64 days after cardiac transplantation. At operation seven miniature tantalum coils were placed into the midwall of the donor left ventricle so as to outline the ventricle in a 30 degrees right anterior oblique projection. Computer aided fluoroscopic analysis of marker dynamics allowed calculation of end-diastolic and end-systolic volumes, stroke volume, cardiac output, and ventricular filling characteristics. Recipients were studied in a resting, unsedated state during a period of clinical stability that did not immediately follow or precede an episode of graft rejection. Augmentation of heart rate by an average of 40 beats per minute above sinus rate produced significant reductions in end-diastolic volume (-19%), end-systolic volume (-8%), stroke volume (-31%), and the duration of systole (-19%), and diastole (-40%). Cardiac output was invariant with rate. In all patients, end-diastolic volume reduction at augmented rates was brought about by abbreviated diastolic filling as initiation of contraction occurred at an earlier point on a common, incremental filling curve. Previous studies have shown that the normal inverse relationship between heart rate and stroke volume is not available to the transplanted heart in the immediate postoperative period. The present study shows that this intrinsic mechanism becomes available to the denervated heart in the subsequent recovery period after surgery.


Determination of B lymphocyte population using antibody-coated polyacrylamide beads. The present study describes a method for the determination of the human B cell population by the use of polyacrylamide beads which are coated with antibody specific for each of the immunoglobulin classes. This method provides a simpler determination of the B cell count than the EA or EAC rosette tests. The antibodycoated beads also have the potential of being used to quantify the subpopulations of B cells bearing specific classes of immunoglobulins.


Linear growth and pubertal development in treated congenital adrenal hyperplasia due to 21-hydroxylase deficiency. 21 years experience with management of seventeen cases of congenital adrenal hyperplasia due to 21-hydroxylase deficiency has been analysed with respect to growth, bone maturation and related events at puberty: age at menarche and the occurrence of menstrual irregularities, this study showed that growth retardation is still a problem; that irregular treatment and prolonged exposure to adrenal androgens or oestrogens, may lead to disturbance in hypothalamo-pituitary-gonadal function and may be the cause fo delayed menarche, or menstrual irregularities in the case of the female. In males the start of puberty and its completion was within the normal range.


The effects of halothane on plasma vasopressin during cardio-pulmonary bypass. Plasma vasopressin concentrations were estimated in twelve patients undergoing cardio-pulmonary bypass for open heart surgery. In six patients anaesthesia was maintained with 66% nitrous oxide in oxygen, whilst the remaining six additionally received halothan as a vasodilator during the bypass period. Induction of anaesthesia had little effect on plasma vasopressin concentrations, whilst marked increases were seen during surgery and bypass in both groups of patients. However, in those patients receiving halothane, significantly higher concentrations were reached, a maximun b36.1 +/- 8.9 (SEM) muu/ml being seen in contrase to 15.4 +/- 2.2 muu/ml in the group receiving nitrous oxide in oxygen alone.


The interaction of fructose, dextrose and ethanol on human performance. 1. The effects of orally administered fructose (1.2 g.kg-1) and dextrose (1.2 g.kg-1) on the performance decrement induced by ethanol (1.0 g.kg-1) in a series of perceptual, cognitive and motor tests were studied in twelve healthy, paid University student volunteers using a double-blind cross-over design. Ethanol was consumed at a constant rate over 20 min and the hexoses or placebo were given approximately 40 min later. 2. The peak blood ethanol concentration was attained earlier when the subjects also received either hexose than after placebo but although its rate of fall was unaffected, significantly lower ethanol concentrations were encountered in the hexose-treated groups in the latter part of the experiment. 3. In general, the subjects were impaired to a slightly lesser degree when they received a combination of hexose and ethanol than after ethanol alone. There were no differences between the effects of fructose and dextrose. 4. The results do not support contentions that fructose is able to accelerate a return to sobriety and observed differences between hexose and placebo-treated groups were attributed to an effect on the absorption of ethanol rather than its metabolism.


Allograft survival in patients receiving anticonvulsant medications. The use of phenobarbital and diphenylhydantoin in transplant recipients is associated with reduced cadaver allograft survival when compared to transplant recipients not receiving anticonvulsant medication. An attempt was made to overcome the adserve effect on allograft survival by either discontinuing the anticonvulsants (2 patients) or maintaining the dose of prednisone at greater than 1 mg/kg/day (2 patients). Discontinuation of phenobarbital was successful after transplantation, with subsequent stabilization of allograft function. Of the 2 patients receiving greater than 1 mg/kg/day of prednisone, renal function is normal in one patient; however, the second is undergoing chronic rejection. Neither patient appears cushingoid. Anticonvulsant medication should be discontinued prior to transplantation. If the patient has an active seizure disorder, prednisone dosage should be maintained at a higher level.


Paralytic instability of the hip in myelomengoceles. The results of a Colonna type arthroplasty with several modifications were evaluated in 29 hips of 22 myelomeningocele children with subluxations or dislocations. The operations were performed only in children with high neurosegmental levels where no muscle transfers were possible or when previous surgical procedures failed to maintain hip stability. Improvement in radiographic and clinical stability was demonstrated in 79 per cent of the hips. Eighty-three per cent of the hips lost motion in the flexion-extension arch with a mean loss of 30 degrees. The arthroplasty improved hip stability but at the expense of some loss of flexion-extension. Favorable results are more likely when the neurosegmental level is T-12 or above, the hip has had no previous operative procedure and the arthroplasty is accomplished without capsular interposition. Improvement of ambulatory status was impossible to evaluate because in any one case, the level of ambulation was determined by influences and physical factors other than the operation.


Phenytoin in the treatment of diabetic symmetrical polyneuropathy. The efficacy of phenytoin in the treatment of the symptoms of diabetic symmetrical polyneuropathy has been tested in a double-blind, crossover study. Symptoms were evaluated daily by linear analogue self-assessment, while control of blood glucose and plasma phenytoin level were monitored on a weekly basis. There was no significant improvement in symptoms on phenytoin, whether analyzed in aggregate, as pain alone, or on the last 3 days of each treatment week. Blood glucose, however, was elevated in diabetic patients taking phenytoin, and the incidence of  undesirable side effects was increased. It is concluded that phenytoin has no role in the treatment of diabetic symmetrical polyneuropathy.


Influence of sex on drug kinetics in man. Very few investigations in man have been specifically devoted to the influence of sex on drug kinetics. Although the potential role of various factors such as the phase of the menstrual cycle, hormonal steroid contraceptive use, menopause or andropause is virtually never taken into account, sex-linked differences in drug kinetics do exist for a limited number of substances such as certain antibiotics, a few tricyclic antidepressants, lithium and aspirin. These sex-linked differences are relatively minor from a quantitative standpoint and appear to be due chiefly to differences in body composition and activity of circulating enzymes and hormones. On the basis of present knowledge, in general these differences in drug kinetics do not warrant modifying dosage as a function of sex.


Prolongation of gastric emptying by oral propantheline. The present study shows that a single oral recommended dose of propantheline bromide normally doubles the mean gastric half-emptying time in man. In a prospective, double-blind, randomized crossover design 13 normal subjects were given 30 mg propantheline or placebo 90 min before taking a 113m-indium-labeled liquid test meal, the volume of which was adjusted to body weight. The disappearance of radioisotope from the area of the stomach was determined by external gamma counting. After placebo the mean half-emptying time was 68 min and after propantheline it was 135 min (p  less than 0.005). Although salivary flow decreased and pulse rate increased there were no visual disturbances. In studies already reported maximally tolerated oral doses of quaternary ammonium anticholinergic drugs have not consistently retarded gastric emptying in man.


Drug kinetics in pregnancy.

Drug kinetics in pregnancy. Any drug given to a pregnant woman must be considered as possibly harmful in the fetus, since all drugs administered to the mother cross the placental membrane, although at different rates. Important physiological changes occur in pregnancy, which may influence the kinetics of drugs. Differences in  gastrointestinal function are likely to alter drug absorption rates in the stomach or gut. Ventilatory alterations modify pulmonary drug absorption or elimination. Important changes in haemodynamics and alterations in body water compartments influence drug distribution and elimination. Renal drug elimination is generally enhanced, whereas hepatic drug elimination may be modified in different ways. Computerised pharmacokinetic models representing the compartmental aspects of the fetal-maternal unit and fetal-maternal drug interrelationships may be used to predict kinetic consequences of fetal drug exposure. Such information may be a useful guide for the clinical use of drugs during pregnancy, particularly for treatment of fetal disease.


Pharmacokinetics and response to diazoxide in renal failure. Diazoxide is given by rapid intravenous injection for the urgent reduction of high blood pressure in patients with all grades of renal function. Oral diazoxide produces less consistent effects. Protein binding of diazoxide is reduced in renal failure and this can be related to reduction of albumin concentration. There is a relation between impairment of renal function and the hypotensive effect of rapidly injected diazoxide. This is explicable in terms of the greater concentration of free (unbound) drug achieved after rapid injection in patients with renal failure. Renal clearance of diazoxide and its metabolites is impaired in renal failure but this is unlikely to affect its activity.


Protein binding of cardiac glycosides in disease states. Digitoxin in 97% bound to serum albumin and digoxin only to the extent of 24%. Hypoalbuminaemia significantly changes the  protein binding of digoxin in Kwashorkor serum and the binding of digitoxin in patients with chronic active hepatitis and the nephrotic syndrome. Sprue patiens with normal albumin values have normal binding of digitoxin. Preliminary data in patients with thyrotoxicosis and myxoedema show digitoxin binding within the normal range. The effect of uraemia per se on digitoxin binding is controversial as both normal and slightly decreased values have been reported. In uraemic patients on treatment with haemodialysis, heparin administration has been shown to be a powerful serum binding displacing agent for both digitoxin and digoxin, the mechanism probably being a heparin-induced release of free fatty acids. Patients with a significant decrease in serum protein binding of digitoxin or digoxin should be maintained on a total serum concentration lower than usually considered within the therapeutic range.


Plasma calcitonin in chronic renal failure: relation to other factors of importance in bivalent ion metabolism. 1. Plasma concentrations of human calcitonin were measured in groups of patients with chronic renal failure, treated either conservatively or by haemodialysis, and compared with a normal group of persons. 2. Plasma calcitonin was statistically significantly elevated in both groups with renal failure. 3. When the data from the three groups were pooled, plasma calcitonin was found to be inversely correlated with total calcium and directly correlated with plasma creatinine.


Two main features of T-cell development: thymus traffic and postthymic maturation. It appears from the results presented in Table V and discussed in the preceding pages that a precursor population of immunologically competent T cells can be characterized and defined by multiple biological criteria as being probably of postthymic origin and substantially different from either the immunologically competent T lymphocyte or the prethymic stem cells. On the basis of recent data on functional characteristics of of different subsets of T cells, which suggest parallel lines of differentiation (see Chapter 2), it is possibile that the postthymic precursor compartment may also show heterogeneity; however, this cannot be presently asserted from our results.


Roentgenographic assessment of causes of cardiac pacemaker failure and complications. Implantation of a permanent pacemaker to stimulate the heart is a well established procedure of great benefit. However, a malfunctioning pacemaker is a hazard to well-bing and to life. Once a pacemaker has been installed the physician's attention is directed not only to the patient's illness but also the unique complications of cardiac pacemakers. Roentgenograms are a valuable resource in the assessment of patients with pacemakers. It is essential that the radiologist thorougly understand the extent to which radiologic studies can depict the functioning of the complications associated with pacemakers. The roentgenographic assessment of the causes of pacemaker failure and the complications associated with permanent pacemakers are reviewed and the role of the roentgenogram is defined.


The coronary collaterals: their development, morphology, function, and classification. Coronary collaterals are important in evaluations made for bypass coronary artery surgery because of their role in the preservation of myocardial function. This paper attempts to define collaterals, explain how they develop, and describe what role they play in the normal heart. It presents a model of the different types of coronary collaterals, emphasizing the intracoronary and intercoronary anastomoses. The controversial role of the function of collaterals in coronary artery disease is discussed extensively, with special attention given to more recent angiographic in vivo studies. Recent investigations indicate that collaterals may limit the size of myocardial infarction as they help to preserve myocardial functions; they assist in the evaluation of both runoff and function for patients being considered for bypass surgery. As more is learned about collaterals, undoubtedly we shall gain a greater appreciation of their significance.


Renal osteodystrophy--a radiological review. Renal osteodystrophy is an amalgam of a number of distinct pathological conditions, in particular, hyperparathyroidism and osteomalacia. In addition, there may be a change in the guantity of bone, i.e., osteopenia (osteoporosis) or osteosclerosis. While bone biopsy may be the most reliable method for detecting these lesions, it is not yet a routine procedure in many centers. Radiological assessment of the bones, therefore, is the most widely used method for assessing the type and severity of the bone lesions in patients with chronic renal failure. This article reviews the world literature and pays attention to conventional radiological techniques as well as macroradiography. In addition, studies in which radiological appearances are correlated with histological appearances are described. Mention is also made of the effects on radiological bone disease of dialysis and transplantation. Consideration is also given to the manifestations of soft-tissue calcification, both of the vascular and subcutaneous type, and to the effects of treatment.


Radionuclide techniques in the evaluation of cerebrospinal fluid shunts. Currently, radionuclide studies offer the simplest and most accurate means of determining the patency of CSF shunts. The selection of the best test procedure depends upon the type of hydrocephalus and the type of shunt which has been inserted. This review documents the development of radionuclide shunt patency tests and suggests the optimum test procedure for a given clinical problem.


Comparison between simple nebulization and intermittent positive-pressure in asthmatic children with severe bronchospasm. Tweinty-three asthmatic children had severe sudden bronchospasm due to numerous factors. Baseline values for the peak expiratory flow rate were less than 25 percent of predicted. Utilizing an analysis of variance, three methods of administering isoproterenol hydrochloride (hand-held Freon-propelled nebulization, continuous nebulization, and intermittent positive-pressure breathing [IPPB]) were compared and found to be similar in reversing the bronchospasm (F = 1.56; degrees of freedom, 2/44; P = 0.22). There were no patients whose condition consistently improved with IPPB over the methods of therapy using simple nebulization. Therapy with IPPB did not offer any advantage over simple nebulization in patients with severe, reversible airway obstruction.


Exfoliative cytology in the rapid diagnosis of pulmonary blastomycosis. Four patients with pulmonary blastomycosis are reported. Their bronchial washings, submitted for cytologic evaluation and stained by the standard Papanicolau technique, yielded the diagnosis, subsequently confirmed by cultural identification of the fungus. In three additional cases, retrospective evaluation of cytologic material also revealed the organism, even though the diagnostic significance was not appreciated originally. Since cytologic techniques are simple, readily available, and rapid, they can be helpful in differentiating pulmonary neoplasms from pulmonary blastomycosis, thus reducing the need for diagnostic thoracotomies.


Saddle embolism of the innominate artery. A case of saddle embolism to the innominate artery in a patient with congestive myocardiopathy is presented. Embolectomy was successfully performed under local anesthesia using a combined carotid and axillary approach.


Interactions stabilizing DNA tertiary structure in the Escherichia coli chromosome investigated with ionizing radiation. The structure of the bacterial chromosome was investigated after introducing breaks in the DNA with gamma irradiation. It is demonstrated that irradiation of the chromosome in the cell prior to isolation results in partial unfolding of the isolated condensed DNA, while irradiation of the chromosome after it is released from the cell has no demonstrable effect on DNA folding. The results indicate that RNA/DNA interactions which stabilize DNA folds are unstable when breaks are introduced in the DNA prior to isolation of the chromosome. It is suggested that the supercoiled state of the DNA is required for the initial stabilization of some of the critical RNA/DNA interaction in the isolated nucleoid. However, some of these interactions are not affected by irradiation of the cells. Remnant supercoiling in partially relaxed chromosomes containing a limited number of DNA breaks has the same superhelical density as the unirradiated chromosome. This suggests that restraints on rotation of the packaged DNA are formed prior to the physical unwinding which occurs at the sites of the radiation induced DNA breads. - Analysis of the in vitro irradiated chromosomes shows that there are 100 +/- 30 domains of supercoiling per genome equivalent of DNA. The introduction of up to 50 double-strand breaks per nucleoid does not influence rotor speed effects of the sedimentation coefficient of the chromosome.


Chromosomes and DNA of Microtus. III. Heterochromatin rearrangements in M. agrestis bone marrow clones. Newborn Microtus agrestis were given single acute whole-body gamma-irradiation (350, 500, or 750R). C-banded bone marrow preparations showed cells with radiation-induced rearrangements of constitutive heterochromatin of the sex chromosomes, usually the consequence of single events, encompassing a wide spectrum of deletions and translocations. These cells persisted in bone marrow for more than a year after irradiation; however, many cells showing the same redistribution of heterochromatin constituted clones of both deletions and translocations. These results indicate that deletion or rearrangement of constitutive heterochromatin does not impair the capacity of bone marrow cells for further proliferation.


Parenteral haloperidol in psychiatric emergencies. Double-blind comparison with chlorpromazine. In a double-blind study of 58 acutely disturbed men and women brought to an emergency psychiatric unit, parenteral haloperidol was generally more useful than parenteral chlorpromazine in the control of disruptive signs and symptoms of psychosis. Half the patients (15/30) who received one injection of haloperidol 5 mg were calmed, cooperative and alert, and another fourth (8/30) were improved. By contrast, only 3 of 28 patients who received chlorpromazine were controlled successfully, and 11 of 28 were partly controlled. Covariant analysis of data from the Brief Psychiatric Rating Scale showed that haloperidol was superior (p less than .05) to chlorpromazine in five of the signs and symptoms evaluated, notably hostility and excitement. None of the statistical comparisons favored chlorpromazine. There were no adverse reactions in any of the patients. Results of this study and of work reported elsewhere indicate that haloperidol is the drug of choice for the control of disruptive symptoms and signs of psychosis in patients who require emergency treatment with an antipsychotic agent.


Amantadine and a fixed combination of levodopa and carbidopa in the treatment of Parkinson's disease. Forty-two patients with Parkinson's disease were given amantadine HC1 (Symmetrel) and placebo in an 18 week double-blind cross-over study to determine if amantadine provided additional benefit when combined with levodopa and carbidopa (Sinemet). Analysis of our results showed that amantadine effected a 92% improvement over baseline in symptom scores and a 95% improvement over baseline in activity impairment scores, compared with corresponding values of 4% and 18% for placebo. The difference between amantadine and placebo was statistically significant. Except for one case of mild livedo reticularis and two of blurred vision in the amantadine group, side effects were generally similar for amantadine and placebo in type and frequency. This study provides new evidence of the importance of combinations of antiparkinson drugs to achieve maximum therapeutic benefit.


The indoleamine hypothesis of depression: an overview and pilot study. This paper reviews the evidence for a specific indoleamine deficiency in depression and the attempts to correct this suspected deficiency with serotonin precursors. It also presents the clinical and biochemical data on six patients with depression treated with L-5-HTP in a nonrandom, double-blind protocol. The oral administration of L5-HTP was associated with a rise in CSF5-HIAA, but only two of six patients studied had any decrease in depression ratings. 5-HTP was also shown to decrease urinary excretion of 17 hydroxycorticosteroids in twodepressed patients and three normal controls suggesting an interrelationship between serotonin metabolism and the pituitary adrenal system. This leads to the suggestion that in a postulated subgroup of depressed patients with pituitary adrenal hyperactivity and evidence of serotonin deficiency, L5-HTP deserves a further trial as an experimental treatment.


Computer-aided teaching in ophthalmology. Computer-aided teaching is of a growing importance in ophthalmology. The following problems are particularly reviewed: sequential, ramified and tutorial teaching, exercises in simulation and the checking of knowledge. Most of the technical obstacles have now been resolved. The aim of this paper is to overcome some psychological obstacles due to partial ignorance of the subject by most lecturers in our field.


The use of mini- and micro-computers in eye clinical practice. In an ophthalmological department there are many sectors in which the use of computers can give useful support. We have singled out three major areas of interest: 1. the rationalization of the flow of patients; 2. the management of clinical records; 3. the automatization of ophthalmological devices. Instead of using different dedicated computational systems we preferred to focus our attention on a unique system based on a minicomputer driving several parallel micro-processors. Preliminary results are given.


The visual field examination and its automation. An automated system for examination of the visual fields is described. The entire campimetric examination was simulated in order to enable an in-depth analysis of actual examinations. The program runs on a HP2100 computer with a 16K core. The main advantages of the system are: greater speed, less patient fatigue, greater reliability, repeatability and accuracy.


Automatic testing of the visual field using electro-oculographic potentials. The central visual field is tested using static perimetry on a tangent screen located 1 m from the patient. During the test electro-oculographic (EOG) potentials, associated with eye movements, are recorded. For this purpose two pairs of electrodes are placed in such a way that one pair records the vertical and the other the horizontal component of the EOG potential. The EOG signals that are produced when the tested eye moves toward the light target on the screen are digitized and fed into a computer programmed to relate each pair of signals to the specific lamp on the tangent screen. The results of the test appear on a computer-printed chart that shows the distribution of the lamps on the testing board together with the information whether the lamp was seen by the subject. This method of visual field testing is both objective and automatic.


[Tissue distribution of native and enzyme-treated human immunoglobulin (author's transl)]. The tissue distribution of intravenously injected native and enzyme-treated human gammaglobulin has been investigated in the small intestine, kidney, striated muscle and the salivary gland of the mouse. Peroxidase-as well as FITC-labeled antibodies to human gammaglobulin were used to locate the injected antibody preparation. With this assay, the enzyme-treated immunoglobulin was found intracellularly, whereas the intact molecule seemed not to penetrate the cell wall and could mostly be found intra-and perivascularly. Since highly purified tetanus toxoid was fixed by thin sections of the intestine tissue containing intracellularly located enzyme-treated gammaglobulin it is concluded that the antibodies penetrating the cell wall retain their biological activity.


[Surgical aspects of secondary hyperparathyroidism (author's transl)]. During long-term dialysis of patients in chronic renal failure disorders of mineral metabolism in the sense of renal osteodystrophy occur increasingly with time. Quite separate from these patients with renal osteopathy (osteomalacia and osteopenia) as a result of dialysis there is a group of patients in whom, despite appropriate dialysate composition, a full-blown picture of renal osteodystrophy due to secondary hyperparathyroidism develops. Subtotal parathyroidectomy will significantly improve them. Experience in 12 patients has demonstrated that, in addition to the clinical signs, biochemical findings and radiological changes, the quantitative evaluation of iliac crest biopsy and determination of parathormone and ionised serum-calcium fraction are decisive as indications for surgery. In these patients subtotal parathyroidectomy gave satisfactory results, but the treatment of renal osteopathy in the sense of osteopenia is often unsatisfactory and the full clinical picture can often be improved only by early renal transplantation.


[Filling of the neck of the tooth by means of acid etching]. The technique of acid cauterisation is only effective in the area of the enamel. The technique of preparation has to be modified if fillings go beyond the area of the enamel. Those modifications and solutions are discussed which combine the anchoring possibilities of conventional preparations with the technique of acid cauterisation.


Stimulation of glucagon release by addition of anti-stomatostatin serum to islets of Langerhans in vitro. Addition of anti-somatostatin serum to rat islets of Langerhans which had been previously maintained in organ culture for 48h, resulted in a marked stimulation of glucagon release with no change in the rate of insulin secretion during the entire 90 min period during which the secretion of both hormones was followed. Prior saturation of the anti-somatostatin serum with somatostatin prevented the stimulatory acting on glucagon release. These data strongly suggest a physiological role for endogenous somatostatin in the control of glucagon secretion.


The influence of glycyl residues on the flexibility of peptide hormones in solution. A 13C-nuclear-magnetic-resonance study of luteinizing hormone-releasing hormone (luliberin) and its des-glycinamide10 N-ethylamide analog. 13C nuclear magnetic resonance spectroscopy in used to gain information on the flexibility of the backbone in peptide hormones and peptide hormone analogs. 13C spin-lattice relaxation times (T1) were measured on luliberin, the luteinizing-hormone-releasing hormone and des(Gly-NH2)10-luliberin-N-ethylamide in aqueous solution at 25.2 and 67.9 MHz at temperatures of 32 degrees, 40 degrees and 55 degrees C. The 13C spin-lattice relaxation times indicate increased flexibility of the peptide backbone in the immediate environment of glycyl residues in luliberin (less than Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) and the hormone analog des(Gly-NH2)10-luliberin-N-ethylamide (less than Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-NH-CH2CH3) in aqueous solutions. 13 C NMR spectroscopy is shown to be a sensitive technique for monitoring the time-averaged conformational flexibility of peptides in solution. Activation energies (Ea) of about 25 kJ/mol were obtained for rotational reorientation of non-terminal alpha-carbons in the peptide backbone. Rotation of methyl groups was characterized by an Ea of 9.6 kJ/mol whereas reorientation of the N-terminal pyroglutamyl residue showed an Ea value of 14.6 kJ/mol. The Ea values of individual carbons in the side-chains of prolyl, arginyl and leucyl residues in the peptides were similar to those obtained for the alpha-carbon of the same amino acid residue in the peptide backbone of the hormones.


Energy-dependent transport of manganese into yeast cells and distribution of accumulated ions. Manganese transport into yeast cells is energy-dependent. It is dependent on endogenous sources of energy and is inhibited by olygomycin (12.5-25 microgramg/ml), 2,4-dinitrophenol (1 mM), 2-deoxyglucose (1-50 mM) and sodium azide (1-10 mM), but is stimulated by cyanide and glucose. The stimulating effect of glucose is eliminated by N-ethylmaleimide and iodoacetate, which apparently inhibit the transport of glucose itself. About 75% of the manganese accumulated in the presence of glucose is found in yeast protoplasts and nearly 25% in the cell walls. A major portion of the accumulated manganese is found in vacuoles. The concentration of osmotically free manganese in the cytosol did not exceed 2 mM, but the concentration in vacuoles was up to 14 mM. The tonoplast is assumed to have a transport system for divalent cations, thereby regulating their concentration in the cytosol.


Tyroslyl-tRNA synthetase from baker's yeast. Rapid isolation by affinity elution, molecular weight of the enzyme, and determination of essential sulfhydryl groups. Tyrosyl-tRNA synthetase (EC 6.1.1.1) has been isolated from baker's yeast with an overall purification factor of more than 5000. After opening the cells, pH 4.8 precipitation, ammonium sulfate fractionation, removal of the nucleic acids with DEAE-cellulose and chromatography on CM-Sephadex, the critical purification step is the elution of the cation-exchanger-bound tyrosyl-tRNA synthetase with tRNATyr. The homogeneous enzyme exhibits a molecular weight of 40 000 as estimated by sedimentation equilibrium centrifugation and dodecylsulfate-gel electrophoresis under reducing and non-reducing conditions. Gel filtration experiments show a molecular weight of about 100 000 indicating the existence of an active dimeric form. The possibility of proteolytic cleavage of the enzyme is excluded. The reaction of tyrosyl-tRNA synthetase with p-chloromercuribenzoate and N-ethylmaleimide reveals two repidly reacting sulfhydryl groups per subunit of molecular weight 40 000, as demonstrated by the inhibition of aminoacylation and the isolation of enzyme-inhibitor complexes. In addition an efficient purification method is described for isolating tRNATyr from soluble ribonucleic acid from baker's yeast in three chromatographic steps in a yield of 28%.


L-Ornithine carbamoyltransferase from Saccharomyces cerevisiae: steady-state kinetic analysis. Ornithine carbamoyltransferase of Saccharomyces cerevisiae is subjected to an enzymatic regulation of its anabolic activity when it is bound to the inducible catabolic arginase as described earlier. This regulatory ornithine carbamoyltransferase essentially catalyzes the synthesis of citrulline, but the reverse reaction could be demonstrated using arsenate instead of phosphate. Steady-state initial velocity studies of the reverse reaction indicate that the mechanism is consistent with a rapid-equilibrium random model (in which all steps are in equilibrium, except that concerned with the interconversion of the central ternary complexes) involving the formation of enzyme - ornithine - arsenate and enzyme - citrulline - phosphate dead-end complexes. In the forward direction, although the mechanism also appears to be random, the results are in better agreement with a preferred ordered binding of substrates, with carbamoylphosphate adding first. This degenerate form of the random mechanism is discussed.


Subunit structure of the methionine-repressible aspartokinase II--homoserine dehydrogenase II from Escherichia coli K12. The quaternary structure of Escherichia coli K12 aspartokinase II--homoserine dehydrogenase II has been examined. This multifunctional protein has a molecular weight Mr = 176000. It is composed of two subunits having the same molecular weight and the same charge. The results obtained from the examination of tryptic maps, the number and amino acid composition of cysteine-containing peptides and the uniqueness of the N-terminal sequence, strongly suggest that the 2 subunits are identical. The properties of aspartokinase II--homoserine dehydrogenase II can be compared to those of the much better known protein aspartokinase I--homoserine dehydrogenase I.


On the stereochemistry of activation of phenylalanine by phenylalanyl-tRNA synthetase from baker's yeast. Because of its chiralic alpha-phosphorus atom adenosine 5'-O-(1-thiotriphosphate) (ATPalphaS) exists in two diastereomeric forms, arbitrarily named (A) and (B). For phenylalanyl-tRNA synthetase ATPalphaS (A) is a substrate whereas ATPalphaS (B) is neither a substrate nor an inhibitor. During the ATPalphaS (A)/PPi exchange reaction with phenylalanyl-tRNA synthetase the configuration at the alpha-phosphorus is retained. The mechanistic implications of these findings are discussed. Preliminary investigations with several other aminoacyl-tRNA synthetases show that the stereochemical requirement with respect to the alpha-phosphorus of ATP is not identical for all aminoacyl-tRNA synthetases.


Isolation of mutants conditionally blocked in the biosynthesis of the 3-deoxy-D-manno-octulosonic-acid--lipid-A part of lipopolysaccharides derived from Salmonella typhimurium. A procedure is described for the selection of conditional 3-deoxy-D-manno-octulosonic-acid--Lipid A mutants which depends on temperature sensitivity for both synthesis of complete lipopolysaccharide and for growth. Using this procedure new types of mutants were isolated which cease growth and accumulate lipid A precursors following a shift to nonpermissive temperatures. All precursor molecules differ in their charge as judged by DEAE-cellulose chromatography. While they all contain glucosamine, phosphate and 3-hydroxymyristic acid, they lack detectable 3-deoxy-D-manno-octulosonic acid (dOclA) as well as the nonhydroxylated fatty acids of the complete lipid A structure. Three mutants proved to be conditionally defective in dOclA metabolism, whereas one seems to be blocked at a relatively early step in lipid A synthesis. The phenotypes of all these mutants appear to be due to single mutations by reversion analysis and by characterization of the temperature-resistant revertants. Studies of these mutants may shed light on the essential role of the complete dOclA--lipid A part of lipopolysaccharides in membrane function.


A trypsin-resistant fragment from complexes of ribosomal protein S4 with 16-S RNA of Escherichia coli and from the uncomplexed protein. A fragment of ribosomal protein S4 was prepared by limited trypsin degestion of a specific complex between protein S4 and 16-S RNA. It was characterised for amino acid sequence and the N-terminal 46 amino acids were found to be absent. An intermediate fragment, cut at Arg-43, was also observed at low trypsin concentrations. Evidence is presented that the protected fragment constitutes the primary RNA-binding region of the protein. No smaller protein fragments were found that rebound to the RNA. A mechanism for the degradation of the N-terminal region of the protein is proposed and two probable functions of the excised region are given. Under milder trypsin digestion conditions than for the complex, the same fragment, cut at Arg-46, was also prepared from the free protein. This result, together with that from a control experiment, indicates that at least within this local region, the protein conformation is conserved in both the free protein and the protein-RNA complex. This is the first direct evidence for the conservation of conformation in a protein when both complexed and uncomplexed with a ribosomal RNA.


The LH and FSH responses to LH-releasing hormone (LH-RH) in girls with true precocious puberty treated with cyproterone acetate. Ten girls with precocious puberty ranging in age from 7 to 10 7/12 years who were treated with oral cyproterone acetate on a long term basis, were subjected to LH-RH tests, prior to and 3 to 16 months after the institution of therapy. Cyproterone acetate was given in doses from 60 to 153 mg/m2, which proved to be clinically effective, as evidenced by the slowing down of sexual maturation. The basal levels of LH were found to be unaffected by therapy and corresponded to the pubertal stages of the individual girls. The peak increment of LH after LH-RH stimulation was markedly suppressed by the therapy. FSH secretion and its responsiveness to LH-RH was not affected by cyprotereone acetate. The basal levels of FSH were higher during therapy than before, but the peak FSH increment remained  the same. An escape phenomenon in the LH peak response was evident in 2 patients upon retesting after prolonged therapy. It is possible that the antigonadotrophic action of cyproterone acetate is due to its progestational nature.


The importance of continuous pump suction in TUR hydraulic hemostasis. An intravesicoprostatic hydrostatic pressure below 10 mm Hg maintains normal anatomy and physiology of the muscular and vascular prostatic structures by continuous pump suction. An increase of the hydrostatic intravesicoprostatic pressure above 10 mm Hg produces a distortion in the musculature of the prostate, especially at the true capsule, opening the cut vessels, making possible the absorption of the irrigant free of electrolytes (TUR syndrome). A low hydrostatic intravesicoprostatic pressure permits the compression of cut vessels by the inflow hydraulic pressure of 90 cm H2O achieving hydraulic hemostasis during TURP reducing the bleeding and operative time by more than 50% and making hemostasis easier. In our last 400 TURs, blood transfusions have been unnecessary. Less electrocoagulation is required, resulting in a more rapid recovery.


Renal function after warm ischaemia. I. Effect of initial perfusate and its temperature. The left kidneys of anaesthetized, heparinized rats were subjected to a 60-min period of warm ischaemia in vivo and perfused in situ with either isotonic saline, human plasma protein fraction, Perfudex (Pharmacia), 20% mannitol or Sacks' solution at 37 degrees C. A second similar series were perfused at 4 degrees C. One further group had autologous arterial blood perfusion at body temperature. Immediate contralateral nephrectomy was done and function was assessed by regular plasma creatinine estimation and survival (life supporting function). Control groups consisted of unilateral nephrectomy alone, 45 or 60 min of ischaemia with immediate contralateral nephrectomy. The only significant improvement in function in the experimental groups when compared to the 60-min ischaemia control group was seen following perfusion with isotonic saline at 37 degrees C (p less than 0.05) and with autologous arterial blood perfusion (p less than 0.005). Renal function following this period of warm ischaemia was not significantly improved by any of the other perfusates tested.


Interactions between cardioactive drugs and antidepressants. The most important cardiovascular interactions between cardioactive drugs and monoamino-oxidase inhibitors or tricyclic antidepressants are reviewed. Post-ganglion blocking agents (e.g., guanethidine), clonidine, reserpine, and alpha-methyl-dopa should not be used in patients needing antide-present therapy. For hypertension, diuretics, beta-blokkers, and vasodilators should be used. In patients with cardiac disease negative interactions between membrane-stabilizing antiarrhythmic agents and tricyclic antidepressants may be observed. Caution is recommended when using both drugs. On the other hand, the use of digitalis is relatively free of negative interactions. Some of the mechanisms possibly involved are discussed. The practical implications of such effects are presented and some clinically useful guidelines are suggested.


[Studies on the resistance of a congenic resistant strain of mice (DKIR) against infection with Salmonella enteritidis (author's transl)]. A congenic strain of mice (DKIR) having a relatively resistant gene for mouse typhoid was established by the beckcross mating between C3H/He and DKI strains. DKI strain is highly and uniformly susceptible to the infection with Salmonella enteritidis and C3H/He is relatively resistant to that infection. The present paper reported the consistency of resistance to Salmonella infection of DKIR strain throughout generations after 10 backcross matings. Difference of the number of infected organisms in the peritoneal fluid and organs between DKIR and DKI or C3H/He strains was also described. The newly established DKIR strain seems to be a suitable experimental animal for the study of genetical resistance for mouse typhoid when compared with its original DKI strain.


Relationship between myocardial contractility and the effects of digitalis on ionic exchange. Therapeutic concentrations of the digitalis glycosides that produce significant positive inotropy cannot be dissociated from certain effects on ionic exchange. These effects produce an increase in cellular Na and an augmentation of Ca influx. An increase of K efflux can be dissociated from the therapeutic action of the glycosides. The increase of Ca influx does not produce an increase of a component of slow inward current that is usually ascribed to transmembranous movement of Ca with voltage clamp technique. It is proposed that the primary action of the glycosides is to produce an elevation of intracellular Na--secondary to inhibition of the cellular Na-K pump. The increase of cellular Na results in augmentation of the activity of an electroneutral Na-Ca carrier system with stimulation of outward Na and inward Ca movement. The enhanced inward Ca movement is directly responsible for the positive inotropic effect of the drug.


The effects of a "low-estrogen" oral contraceptive on carbohydrate metabolism during six months of treatment: a preliminary report of blood glucose and plasma insulin values. A prospective study of carbohydrate metabolism was carried out on seven women by using an oral glucose tolerance stimulation test and measuring blood glucose and plasma insulin values over a 3-hour time period. The test was performed before and after 6 months of treatment with a "low-estrogen" oral contraceptive containing 0.4 mg of norethindrone and 35 micrograms of ethinylestradiol. There was no change in the weight or diastolic blood pressure of the women, but there was a slight elevation of the systolic blood pressure with treatment. There was no significant change in the plasma insulin values or in four of five blood glucose values. The 1-hour blood glucose value was significantly lower after 6 months of treatment. These results suggest that the "low-estrogen" oral contraceptives may reduce the secondary effects on carbohydrate metabolism seen with regular oral contraceptives.


Surface immunoglobulins on stimulated lymphocytes. The effect of phytohaemagglutinin, lipopolysaccharide, lipid A and streptococcal erythrogenic toxin on the expression of surface immunoglobulins on nerborn piglet spleen lymphocytes was studied, using iodinated anti-pig Ig, L-, mu- or gamma-antisera. It has been concluded that only IgM is the surface immunoglobulin on non-stimulated as well as LPS-stimulated piglet lymphoid cells. An increase in the numberof labelled cells was observed after cultivation of piglet spleen cells with all the mitogens tested.


In vitro and in viro effects of the fifth thymosin fraction. Using the 5th fraction of thymosin, it was possible to enhance the capacities of peripheral blood lymphocytes to form rosettes with sheep erythrocytes and to respond to PHA in those cases of cancer or sarcoidosis in which these functions were depressed. Moreover, the 5th thymosin fraction increased the resistance of CBA mice to infection with virulent H37Rv mycobacteria and augmented their ability to acquire delayed hypersensitivity.


Active rosette test in cutaneous lymphoproliferative disorders. T and B peripheral blood lymphocytes were evaluated in 17 patients with clinical and histological evidence of mycosis fungoides and in ten patients with benign lymphoplasias of the skin. The T subpopulation (T-Ea) characterized by a rapid rosette formation with sheep erythrocytes was also measured in these patients. While both T-total population (T-Et) and T-Ea subpopulation were within the normal range in pseudolymphoma group, a statistically significant decrease was observed in T-Ea and T-Et populations in the mycosis fungoides group. Moreover, the decrease in T-Ea subpopulation was found in all cases under investigation, even when T-Et population was still at the normal levels. A close relation between T-Ea values and clinical status was observed. B and null cells were substantially unchanged in all patients.


A double-blind comparative study of the inhibitory effect of intraduodenally administered hyoscine-N-butylbromide on human duodenal motility. In a double-blind crossover study hyoscine-N-butylbromide (HBB) was compared with placebo in its inhibitory effect on duodenal motility stimulated by metoclopramide. Measurements were made both by the balloon technique and the open-tipped tube method with pressure transducer. 20 mg or 50 mg HBB or placebo were instilled in random sequence into the intestinal tract of 12 healthy volunteers. The frequency of Type II waves was determined in each experiment after administration of the test substance following prior metoclopramide stimulation. This frequency was used as the inhibitory index for each drug. The test substance was administered 15 minutes before a second stimulation. A four-way analysis of variance (mixed model) was applied to the calculated number of waves per period. Subjects were required to respond equally to stimulation with increased motility on both trial days as evidenced by a minimal difference in the averages following the first stimulation on each trial day and before application of the test substance. The frequency of Type II waves after 20 mg and 50 mg HBB was reduced in a statistically significant manner in each case when compared with placebo. The probabilities of error were p less than 0-05 and p less than 0-025.


Penbutolol in hypertension: a pilot study with single daily doses. A pilot single-blind placebo controlled crossover within-patient study was undertaken in essential hypertension. In ten patients single daily doses of 25 mg and 50 mg and in two patients 25 mg, 50 mg and 100 mg were used. Satisfactory reductions in both systolic and diastolic blood pressure in the supine and erect postures were observed. Reduction in heart rate was of the order of 6-32%, there being no correlation between reductions in blood pressure and decrements in heart rate. Three patients were dropped from the final analyses. Seventy-eight per cent (7/9) of patients had a final diastolic pressure (lying) of 90 mm Hg or less. Single doses of penbutolol controlled blood pressure for at least twenty-four hours. At the end of two weeks on placebo medication, following nine weeks of active drug medication, blood pressure had reverted to near pre-treatment levels. Penbutolol was well tolerated.


Free calcium in full grown Xenopus laevis oocyte following treatment with ionophore A 23187 or progesterone. Free intracellular Ca2+ was monitored in isolated Xenopus laevis oocyte during induced maturation using the Ca2+ -sensitive luminescent protein, aequorin. Internal free Ca2+ was not precisely measured but data suggest it was quite low (in the micromolar range). No change in internal free Ca2+ was detected during maturation induced either by progesterone or by p-chloromercuribenzoate. By contrast, the ionophore A 23187 gave an increase in the free Ca2+ level when there was a raised external Ca2+ (10 mM), conditions which also induce oocyte maturation. About 3 h after progesterone or p-chloromercuribenzoate stimulation, the oocyte membrane potential decreased by about 50 mV while the membrane resistance increased transitorily.


Identification of four cell types in the human endocrine pancreas by immunoelectron microscopy. In order to correlate the different cell types of the human endocrine pancreas to a specific secretion product, an immunoelectron microscopic localization of the hormones whose production had been attributed to pancreatic islets was conducted. Glucagon and insulin were respectively localized in the typical A- and B-cells, whereas no subclasses of A-cells could be identified. With antibodies that reacted with the gastrin cells in the human gastric mucosa, it was not possible to detect gastrin in any of the islet cell types. In confirmation of recent results obtained by light microscopy, somatostatin was found in all the typical D-cells containing large, weakly electron-dense secretory granules. The human pancreatic polypeptide (HPP), a newly postulated hormone, was clearly associated with a fourth cell type, which is characterized by the presence of small secretory granules (100-150 nm.). These results suggest that each of the four cell types that are easily identifiable by ultrastructural observations is responsible for the production of a specific secretory product.


[The effect of several sexual steroids, 2-bromo-ergokryptin and lisurid-hydrogenmaleate an the postpartum concentration of serum prolactin and lactation (author's transl)]. The effect of Ablacton, Estrovis 4000, 2 Brom-alpha-ergokryptin (Cb 154) and of Lisurid-hydrogenmaleate (LHM) on lactation and the serum concentration of prolactin postpartum was studied. 10 normal nursing postpartum patients served as control. LHM was tested in a double blind study compared to placebo. The serum prolactin (PRL) was determined daily for the first 10 postpartum days by radioimmunoassay. Inhibition of lactation was consistent with Cb 154 (89% of the cases). Within two to three days after the onset of treatment with Cb 154 the changes of pregnancy in the breasts had completely subsided. The sexual steroids resulted in inhibition of lactation in 60% of the postpartum patients but mastodynia continued. LHM showed no difference from group taking placebo. The serum prolactin levels postpartum remained as high as a nursing mother's or rose with the administration of steroids. The administration of Cb 154 resulted in a drop of the serum prolactin to non-pregnant levels with in 2 days. The serum prolactin levels under treatment with LHM were not different from the group slowly. In nursing mothers, the stimulus of suckling maintain the pituitary secretion of prolactin as showed by higher serum prolactin levels and a slower decreased to normals than in the non-nursing mothers.


[Diabetes mellitus and pregnancy (author's transl)]. In present day, recognized factors which influence the carbohydrate metabolism of a diabetic during pregnancy have been reviewed. Possible effects on the fetus are discussed. Based on the good results obtained by Roversi and Canussio with the "Maximal tolerated dose of insulin", a state of glycemia within the normal limits was sought; this aim is attained most readily with a 3-4 times daily injection of soluble insulin. With satisfactory control of metabolism and regular surveillance of fetal parameters (ultrasonics, CTG, oestriol, HPL), the time for delivery can approach the calculated date: thus, the danger of fetal respiratory distress is reduced. Also, fetal hyperinsulinism can be avoided by stabilising the diabetic condition as early as possible within the normal limits of glycemia. Spontaneous birth is attempted with preparations made for caesarean section.


Effects of the mitotic cell-cycle mutation cdc4 on yeast meiosis. The mitotic cell-cycle mutation cdc4 has been reported to block the initiation of nuclear DNA replication and the separation of spindle plaques after their replication. Meiosis in cdc4/cdc4 diploids is normal at the permissive temperature (25 degrees) and is arrested at the first division (one-nucleus stage) at the restrictive temperature (34 degrees or 36 degrees). Arrested cells at 34 degrees show a high degree of commitment to recombination (at least 50% of the controls) but no haploidization, while cells arrested at 36 degrees are not committed to recombination. Meiotic cells arrested at 34 degrees show a delayed and reduced synthesis of DNA (at most 40% of the control), at least half of which is probably mitochondrial. It is suggested that recombination commitment does not depend on the completion of nuclear premeiotic DNA replication in sporulation medium.--Transfer of cdc4/cdc4 cells to the restrictive temperature at the onset of sporulation produces a uniform phenotype of arrest at a 1-nucleus morphology. On the other hand, shifts of the meiotic cells to the restrictive temperature at later times produce two additional phenotypes of arrest, thus suggesting that the function of cdc4 is required at several points in meiosis (at least at three different times).


Inheritance of the 2 micrometer m DNA plasmid from Saccharomyces. A variety of Saccharomyces strains were examined for the presence of 2micro DNA and, if present, for the pattern of fragments produced by its digestion with site-specific (restriction) endonucleases. Two strains were found that did not contain detectable levels of 2micro DNA, and two strains contained 2micro DNA molecules having only one EcoRI restriction endonuclease recognition site rather than the usual two.-A haploid containing 2micro DNA with one EcoRI restriction site was mated with a haploid containing 2micro DNA with two EcoRI restriction sites and the resulting diploid maintained both types during vegetative growth. Sporulation of the diploid produced four spores, and the clones from these spores contained both types.-A haploid lacking 2micro DNA was mated with a haploid containing 2micro DNA and the resulting diploid contained 2micro DNA. The four clones derived from the haploid spores after sporulation of this diploid all contained 2micro DNA. A rho(-) strain without 2micro DNA was mated to a rho(+) strain with 2micro DNA, and heteroplasmons were selected that had received the nucleus from the strain without 2micro DNA and the mitochondria from the strain with 2micro DNA. Twelve of twenty-four such clones contained 2micro DNA.-I conclude that: (1) the different types of 2micro DNA identified in these strains do not restrict one another, (2) the different types are inherited extrachromosomally, (3) lack of 2micro DNA in two strains is not due to the absence of genes needed for maintenance and (4) the approximately 100 copies of 2micro DNA contained within a single cell are probably clustered within one or a few cytoplasmic organelles.


Mutants of formyltetrahydrofolate interconversion pathway of Saccharomyces cerevisiae. Thirteen mutants of Saccharomyces cerevisiae that lack one or more of the three enzyme activities of the pathway for interconversion of tetrahydrofolate coenzymes at the formate level of oxidation have been isolated. They do not require adenine. All fail to complement mutations in the ade3 locus. Mutations that greatly reduce activity for the other two interconversion enzymes. The three enzyme activities cochromatograph on TEAE-cellulose columns. A mutation that eliminates synthetase activity also alters the chromatographic behavior of the remaining cyclohydrolase and dehydrogenase activities. It is suggested that the three activities reside in an enzyme complex encoded by the ade3 locus.


Ochre suppression in Salmonella typhimurium. A bacterial strain was constructed which permitted positive selection for ochre suppressor mutations as well as for the loss of suppressor function. A derivative bearing an ochre suppressor mutation was selected following mutagenesis with N-methyl-N-nitroso-N'-nitroguanidine. The suppressor-bearing strain was treated with nitrous acid to eliminate suppressor function by mutation, and a strain lacking suppressor activity was selected. The selected strain which had lost suppressor function was then subjected to mutagenesis to induce a second suppressor mutation. The alternating sequence (induction of an ochre suppressor mutation leads to induction of a mutation eliminating ochre suppressor activity) was repeated 29 and one-half times in a single strain. Some of the suppressor mutations were tentatively mapped at four locations on the chromosome. The first suppressor mutation selected maps at about minute 30 on the chromosome. The second suppressor selected maps at approximately minute 60, while the third suppressor maps nearby, possibly as far as minute 72. Among the subsequently selected suppressor mutations, all eleven which were mapped were cotransducible with the gal and nic loci near minute 36 on the chromosome and may represent more than one suppressor gene. Deletions were selected which inactivate two of the ochre suppressor alleles mapping near the gal-nic region, suggesting that one or more such genes are dispensable. Some evidence also suggests that the occurrence of either deletion mutations or transduction-mediated recombination events in the gal-nic region can cause instability of nearby suppressor alleles.


The effects of genotype frequency and population density on fitness differentials in Escherichia coli. Two strains of Escherichia coli K-12, a lac+ wild type and a lac- auxotroph, were grown both as pure and mixed cultures, using a serial transfer procedure. Four different growth media were employed, consisting of the same minimal salts solution, but different total concentrations of the sugars lactose, arabinose, and glucose (in proportions 5:4:1). Population densities and genotypic frequencies were assayed every 48 hours, at the time of transfer. Population density of the pure lac+ culture was greater than that of the pure lac- culture for all media; this was expected, since the latter cannot utilize lactose. Mixed cultures quickly approached the same density as the corresponding lac+ controls, and the frequency of the lac+ genotype increased steadily for all media. Trajectories of lambda = log (P divided by Q) were strictly nonlinear, indicating a dependence of the selective differential on population density and genotypic frequency. The rate of substitution decreased slightly with increasing sugar concentration, contrary to theoretical expectation. It was speculated that either the generation interval was longer for denser cultures (higher substrate concentrations) of that buildup of organic by-products reduced the selective differential in denser cultures. For a single medium, however, the behavior of completing genotypic strains was reasonably well predicted by theoretical models of frequency and density-dependent selection, the parameters of which may be related to the experimental inputs.


Clotting factors and platelets. Immunofluorescence evidence that fibrinogen and factor VIII are present in human washed platelets, whereas prothrombin complex factors and factor XIII are lacking. Factor I (fibrinogen) and factor VIII were shown, by an indirect immunofluorescence technique, to be present in human washed platelets. In the case of fibrinogen, the immunofluorescent pattern had a 'clod distribution' up to a 1:128 dilution of the antiserum. Factor VIII showed a 'speckled' pattern up to the 1:64 dilution. Cross-absorption studies confirmed the presence of such factors. Factor II, VII, IX, X and XIII, however, were not found in washed platelets. A diffuse fluorescence was noted using normal rabbit sera, but this was due to a cross-reaction, since it disappeared using a 1:32 diluted sera.


[Metabolic fate of carteolol hydrochloride (OPC-1085), a new beta-adrenergic blocking agent. (6) Pharmacokinetics of carteolol in the rat, dog, rabbit and man]. The kinetics (absorption, distribution and excretion) of carteolol were investigated after oral and intravenous administration to man, rats, Beagle dogs and rabbits. The half-life of carteolol in plasma was 1.22 approximately 1.45 hr in rats, 1.73 approximately 2.08 hr in dogs and 1.42 approximately 1.43 hr in rabbits, and was independent of the route of administration. The absorption rate constants, obtained from log(C1-C) approximately time plot, after oral administration were 1.89 hr-1 in rats, 1.04 hr-1 in dogs and 1.54 hr-1 in rabbits. There were no differences between tablet and film coated tablet in the pharmacokinetic parameters of carteolol in man after oral 30 mg (tablet or film coated tablet) administration [half life (t1/2)=4.50 hr (tablet), 4.30 hr (film coated tablet), elimination rate constant (k2) equals 0.154 hr-1 (tablet), 0.161 hr-1 (film coated tablet)]. The elimination rate constant, obtained from Sigma-minus plot after 2, 5 and 10 mg oral administration, was 0.137 approximately 0.160 hr-1.


Inhibition of beta-galactosidase synthesis in Escherichia coli after infection with different DNA and RNA phages. Infection of Escherichia cooi with T1, T2r+, T3 and T4 phages leads to an immediate inhibition of beta-galactosidase synthesis. Similar results were obtained with the virulent mutant of phage lambda. The degree of inhibition of beta-galactosidase synthesis depends on the time delay between the addition of the inducer and the phage particles, and on the amount of phage DNA, which has penetrated into the host cell. RNA phage MS2 exhibited no inhibitory effect on enzyme synthesis.


Replication of DNA in UV-irradiated Escherichia coli in the absence of amino acids. Since pyrimidine dimers are considered to be the cause of the synthesis of short DNA segments, normalization of DNA replication after UV irradiation should be in a temporal correlation with their removal. This correlation holds in exponentially growing excision-proficient Escherichia coli cells. However, when these cells are preincubated and postincubated without amino acids, synthesis of short segments continues although dimers are efficiently excised.


[T- and B-lymphocyte determination in infectious mononucleosis]. In 12 children with infectious mononucleosis the peripheral blood T- and B-lymphocytes were studied and compared to a control group of 26 children, aged 2 to 13 years, without any infection. Spontaneous sheep red blood cell rosettes were used as a T-cell marker, demonstration of surface immunoglobulins by immunofluorescence technique was used as a B-cell marker. Patients with infectious mononucleosis showed a marked increase of T-lymphocytes (percentage and absolute numbers) in comparison with the control group; the difference was statistically significant. Since only B-lymphocytes are the target cells fo Epstein-Barr virus, the increase of T-lymphocytes in infectious mononucleosis probably represents the response of cell mediated immunity against B-lymphocytes which are altered in their antigenic properties by Epstein-Barr virus.


[First rank symptoms of schizophrenia (Kurt Schneider): Are they still of any importance? (author's transl)]. The general importance of paying attention to symptomatology in psychiatry is stressed. Research in this area has been advanced by modern psychopathometric methods. A renewal of general interest in the "classical" symptomatology of the older German authors is also shown by research involving Kurt Schneider's first rank symptoms of schizophrenia in England and the USA. Important papers concerned with this latter trend are discussed.


Maize silage: incidence of moulds during conservation. The basic principle of silage making--preservation under anaerobiosis--is recognized. With this environment the substrate can harbour a spontaneous and active microflora that can be divided into two sequential phases; a bacterial phase followed by a yeast phase. The imperfections of the techniques used in making silage, however, frequently result in invasion by fungi. The moulds that are responsible for serious changes (loss of nutrients, mycotoxins) are classified into a dozen characteristic divisions (species or groups of ecologically related species). These appear in a definite chronological order and form a relatively simple model of the development of the flora. This dynamic transformation can also be utilized profitably to determine the quality of the fodder and estimate the storage life. A simple mycological puncture analysis reveals in fact the stage reached in the fixed sequence, and the results enable extrapolations to be made regarding the history and the future of the material. It is also hoped that a better knowledge of the biotope will lead to complete control of the floral equilibrium, good conservation and high quality foodstuffs.


Malaria and hereditary ovalocytosis.

Malaria and hereditary ovalocytosis. Hereditary ovalocytosis in Papua New Guinea is restricted to areas of endemic malaria and may confer increased resistance to the disease. The incidence of malaria was investigated in 1616 Melanesiams of known red cell morphology and severity of infection determined in a smaller subsample. Ovalocytics tended to be more resistant to severe malarial infections than normocytics. The ratio of parasitaemia in 112 ovalocytics compared with 741 normocytic children was 1.05 for P. falciparum; 0.90 for P. vivax; 0.54 for P. malariae, and 0.91 for infection with any species. The difficulties in conclusively demonstrating any selective advantage of the condition are discussed.


[Technical advances in demonstration of treponema-specific 19S-IgM antibodies in patients with latent or late-latent syphilis (author's transl)]. In patients with latent or late-latent syphilis as well as neurosyphilis the demonstration of treponema-specific 19S-IgM antibodies is not regularly possible. In this publication it is shown that high IgG antibodies are the reason for a competitive IgM inhibition in the indirect immunofluorescence tests. It is demonstrated that in the above mentioned states of the infection the IgM-FTA-19S test seemed to be the method of choice to realize the existence of 19S-IgM antibodies. In sera with competitive inhibited IgM it is possible to calculate the IgM antibody activity with the aid of the socalled F factor.


[Delayed type hypersensitivity and Immunity (author's transl)]. Delayed type hypersensitivity is a T-cell-mediated phenomenon. It is characterized by the following features: a) development within 24-72 h after antigen challenge in sensitized individuals, b) perivascular infiltration by mononuclear cells at the inflammatory site, c) transfer by lymphocytes, but not by serum. Delayed type hypersensitivity occurs during infections with facultatively intracellular bacteria and certain viruses, tumor diseases and rejection of transplanted foreign tissue. This article samples evidence that delayed type hypersensitivity is an outward phenomenon indicating the presence of specific T-cells and mononuclear phagocytes, both acting together to produce local immunity which results in the elimination or containment of the noxious agent.


The influence of thymosin on affinity binding in the E-rosette assay. Human peripheral blood lymphocytes (PBL) were analyzed in the E rosette assay with respect to (1) the effect of SRBC/lymphocyte ratio on rosette number; (2) the effect of this ratio on the presence of lymphocytes with multiple binding sites for SRBC; and (3) the influence of thymosin on increasing the number of lymphocyte receptors for SRBC. An increase in the SRBC/lymphocyte ratio was associated with an increase in rosette number. The number of multiple binding rosettes was also dependent on the SRBC/lymphocyte ratio. At saturing SRBC concentrations, greater than 30% of the rosettes were in the morula form. Treatment of PBL with thymosin was associated with a slight increase in the number of total rosettes and a large increase in number of morulas. It is suggested that the morula represents a mature form of T cell in contrast to the 2 - 3 SRBC rosette, which may be induced by thymosin to become a morula form.


Effects of the new anti-lymphocytic peptide cyclosporin A in animals. The fungus metabolite cyclosporin A is a small cyclic peptide acting as a novel antilymphocytic agent. It is effective following either parenteral or oral administration in mice, rats and guinea-pigs. The suppressive effect after short and prolonged treatment on plaque-forming cells, the inhibition of the secondary humoral response and the reversibility of its effect on haemagglutinin formation is demonstrated. Cyclosporin A inhibits delayed hypersensitivity skin reaction to oxazolone (primary and secondary responses) in mice and to tuberculin in guinea-pigs. Its failure to suppress antibody synthesis to lipopolysaccharide antigens in nude mice suggests a selective effect on T cells. High doses of the compound affect the haemopoietic tissues very weakly as shown by the bone marrow and stem cell numbers in mice, which finding markedly contrasts with most other immunosuppressive and cytostatic drugs.


Leucocyte migration inhibitory activity in supernatants of cultured human mono-nuclear cells stimulated with mitogens or C3- and Fc-receptor reactions. We studied whether reactions at the lymphocyte membrane receptors for complement, immunoglobulin or mitogens would induce lymphokine production. Human peripheral blood mono-nuclear cells were stimulated by C3- or Fc-receptor reactions and with phytohaemagglutinin (PHA) or lipopolysaccharide (LPS) and cultured for 3 or 4 days. C3- and Fc-receptor reactions were brought about by rosette formation with red cells coated with human complement or antibody. Culture supernatants were assayed for migration inhibitory activity with human leucocytes by the leucocyte-migration-in-agarose test. On the average, no migration inhibitory activity could be detected in cultures stimulated by C3- or Fc-rosette formation, mean migration indices (MI) being 99-2 and 96-1, respectively. Of the mitogens, PHA induced distinct lymphokine synthesis (mean MI 60-2) while the mean MI with LPS varied from 97-9-69-8, depending on the mitogen preparation used and the conditions of culture. We conclude that PHA and LPS are able to activate human lymphocytes into elaboration of migration inhibitory factors whilst reactions at C3- or Fc-receptors fail to do so.


The immunological response of CBA mice to P. yoelii. I. General characteristics, the effects of T-cell deprivation and reconstitution with thymus grafts. Experimental infection of normal CBA mice with the parasite Plasmodium berghei yoelii (P. yoelii) resulted in a mild, non-fatal and self-limiting infection which lasted for 15-17 days. Animals which recovered from the primary infection were immune to reinfection though parasites could be detected in the kidneys of such mice 4 weeks after recovery from infection. (No plasmodia were demonstrated in the peripheral blood and other tissues examined.) In T cell-deprived mice, P. yoelii infections resulted in a progressive parasitaemia and proved fatal in 35-40 days. Studies of fluorescent antibody levels and morphological changes in the spleens of infected normal and T cell-deprived mice showed that while normal mice produced high levels of IgG1, IgG2 and IgM antiplasmodial antibodies and developed a strong and sustained germinal centre response, in T cell-deprived animals the production of IgG1 antibodies was almost completely abolished and the germinal centre response severely impaired. Reconstitution of T cell-deprived mice with syngeneic thymus grafts resulted in partial restoration of immunological responsiveness. P. yoelii infections in these reconstituted animals ran a self-limiting course akin to that seen in normal CBA mice; the level of protective immunity and the germinal centre response correlated with the degree of reconstitution achieved.