Ultrastructure of the corneal nerves in the rat.

The ultrastructure of the corneal nerves of the rat was studied in tissue fixed by immersion in and by perfusion with glutaraldehyde-containing fixatives. Of the four types of axonal terminal identified in the nerves, those with the features of adrenergic and cholinergic terminals were confined to the nerves at the limbus and were concentrated in the perivascular plexuses. The remaining two types of terminal were found on axons located in all parts of the cornea and on both intraepithelial axons located in all parts of the cornea and on both intraepithelial axons and axons in the stromal nerves. Of these, one contained the numerous mitochondria which occur in the terminals of axons associated with known mechanoreceptors and the second contained variable and often small numbers of both clear and large dense-cored vesicles. While most of the mitochondria-containing terminals were seen in nerves located near the periphery,vesicle-containing terminals were numerous in all of the nerves,and especially in those in the avascular cornea. In material fixed by immersion in glutaraldehyde-paraformaldehyde,the vesicle-containing terminals appeared to be dilated,but in material fixed by perfusion there was little evidence of any increase in the diameter of the axons in the terminal regions. The structure of the terminals was compared with that of the terminals of axons identified in the nerves of the skin and the urinary tract and the differences in the vesicle content of the terminals to those reported in other studies of the corneal nerves was related to the use of different fixation procedures. The possibility that axons possessing such terminals are identical with the beaded axons and both the cholinesterase-positive and fluorescent axons demonstrated in light microscopical studies of the corneal nerves is discussed, and the widespread distribution of the axons in the cornea is equated with the hypothesis that they are afferent in nature and represent the peripheral receptors for pain impulses.


The distribution of concanavalin A binding sites on the intestinal epithelium of the nematodes Ascaris suum and Parascaris equorum.

Receptors for Concanavalin A (Con A), were localized on the intestinal epithelium of the nematodes Ascaris suum and Parascaris equorum. Fixed tissue incubated in 3H-Con A showed labeling of the microvilli surface and basal membrane. Using Con A coupled with peroxidase, the tips of the microvilli of Ascaris suum and the tips and lateral surfaces of Parascaris equorum were stained. The basal membrane of both species was also labeld. No labeling was observed on control tissue incubated without Con A or on tissue incubated with Con A to which alpha-methyl-D-mannoside was added.


Insulin-, glucagon-, and somatostatin-immunoreactive endocrine cells in the equine pancreas.

Equine pancreas was investigated with immunohistochemical methods to study the distribution of endocrine cells immunoreactive to anti-insulin, anti-glucagon, and anti-somatostatin. A-cells demonstrable by anti-glucagon are located in the center of Langerhans islets and frequently in the duct epithelium. Few A-cells are seen associated to acini. Anti-insulin reactive B-cells form a large zone around the center of the Langerhans islets in which some B-cells lie between exocrine cells and others, although few, are located in the duct epithelium. D-cells stained with anti-somatostatin serum form a discontinuous outermost zone around the Langerhans islets. In some islets the D-cells are also observed among the B-cells or between the border of A- and B-cells. Single D-cells are seen in the duct epithelium or between acinar cells. In younger horses, endocrine cells are more frequently associated in bulges of the duct system. The histotopographic relation between these endocrine cell types is discussed with respect to its functional significance.


LHRH-like system in the brain of Xenopus laevis Daud: immunohistochemical idenfication.

Rabbit antiserum to synthetic LHRH was used with the immunofluorescence technique to identify the LHRH-secreting neurons and their axonal pathways in the brain of Xenopus laevis. Three groups of immunoreactive neurons were identified: the first, in the telencephalon, is a paired group of cells scattered near the two telencephalic ventricles; the second group lies near the preoptic recess; the third group occurs in the ventral wall of the infundibulum. Two principal neuronal pathways were observed: Fibres originating from the dorsally located telencephalic neurons converge on the cephalic median plane where they form a single bundle behind the telencephalic furrow. This bundle descends towards the anterior border of the preoptic recess where it divides into two nerve bundles which pass on either side of the preoptic recess, run above the optic chiasma then cross the infundibular floor and finally terminate in the median eminence. The second pathway is more direct. The more ventrally located telencephalic LHRH cells give rise to this second pathway. Their axons converge with the other LHRH fibres near the lateral border of the preoptic recess. Most of the LHRH nerve fibres terminate in the median eminence although some terminate near the paired pars tuberalis. No reaction was observed after the use of antiserum absorbed with synthetic antigen.


Adrenergic innervation of the mouse heart revealed at an early stage using alpha-methylnoradrenaline.

The development of noradrenergic innervation was studied in the mouse heart using fluorescence histochemistry. Following incubation of hearts with alpha-methylnoradrenaline fluorescent nerve fibres were seen as early as 13 days in utero. It is suggested that the neuronal uptake mechanism for noradrenaline is functional at an early stage.


Immuno-histochemical localization of LH-RH during different phases of estrus cycle of rat, with reference to the preoptic and arcuate neurons, and the ependymal cells.

Immunohistochemical localization of luteinizing hormone-releasing hormone (LH-RH), during different phases of the estrus cycle, in the preoptic, suprachiasmatic and arcuate nuclei, and in the OVLT of rats, with special emphasis on the ependymal cells, was studied by light, fluorescent and electron microscopy, by using rabbit anti serum to synthetic LH-RH. The LH-RH neurons in the above mentioned areas, were very active during late diestrus and early proestrus phases. Specialized ependymal cells bordering the 3rd ventricle also showed varied LH-RH positive reaction during different phases of the estrus cycle. Immunofluorescent studies showed cyclic variations in the LH-RH material in the CSF of the preoptic and infundibular recesses, as well as in the 3rd ventricle near OVLT, in that, it was maximum during late diestrus and early proestrus phases. Immediately after this, the LH-RH late proestrus was reached. We have also observed that during the proestrus phase, as the LH-RH material started declining in the CSF, it had started building up in the specialized ependyma. Estrus, metaestrus and early diestrus phases showed very weak immunofluorescent LH-RH material in the lumen of the infundibular recess and in the specialized ependyma. Our immuno-electron microscopic observations showed pleomorphic LH-RH granules in the specialized ependyma during late kiestrus and proestrus phases. All these observations lead us to believe that LH-RH is not synthesized in the ependymal cells,but is phagocytosed from the CSF of the 3rd ventricle by the specialized ependyma, which transports it to the ME portal system. In males, the fluorescent LH-RH material did not show any noticeable changes. With the present and previous work,it is concluded that the neurons in differentnuclei synthesize LH-RH and transport it to the ME portal system,primarily through the nerve fibers and secondarily by the ventricular route. It is also suggested that the ependymal transport of LH-RH to the ME portal system is cyclic and thus controls the gonadotropin secretion.


The effect of the ionophore A23187 on the ultrastructure and electrophysiological properties of frog skeletal muscle.

The divalent cation ionophore A23187 has three major effects on the thin cutaneous pectoris muscle of frog: (1) The membrane potential is depolarized, an action that is found only when the [Ca2+] of the bathing saline is very low.(2) It causes an increase in resting tension and the development of contraction. This action is produced at both normal and low values of [Ca2+]o and is, therefore, independent of Ca2+ entry and of changes in Em. The ionophore is believed to act primarily by releasing Ca2+ from intracellular stores. (3) It causes major ultrastructural damage to the muscle filaments. It is believed that this damage is the result of the action of A23187 on the sarcoplasmic reticulum and the elevation of [Ca2+]i and we suggest that the action of thisionophore may serve as a useful model for the study of certain myopathies.


Electron microscopic immunocytochemical demonstration of separate vasotocinergic and mesotocinergic nerve fibres in the neural lobe of the amphibian hypophysis.

Using the unlabeled antibody peroxidase-antiperoxidase (PAP) technique at the electron microscopic level, it was demonstrated that the hormones of the posterior pituitary of Rana temporaria are located in separate vasotocinergic and mesoticinergic nerve fibers. This observation confirms the results of our previous immunocytochemical studies at the light microscopic level.


Some observations on the ultrastructure of developing rat cerebral capillaries.

Developing blood vessels in rat cerebral cortex were studied at a number of stages between 3 and 28 days postnatal, in an attempt to obtain data on the mechanisms by which the lumen is established within cords of mesodermal cells. A combination of techniques was utilized in an attempt to elucidate these mechanisms. These were: (a) aldehyde fixation and block staining with phosphotungstic acid; (b) aldehyde perfusion followed by perfusion of a lead solution and post-fixation in osmium tetroxide; (c) conventional preparation of tissue with aldehyde and osmium fixation. Support for interendothelial lumen formation was readily forthcoming, including vessels with junctions between two or more endothelial cells cut transversely. There was some support for intraendothelial lumen formation, in the form of "seamless" endothelial cells. Other features noted included the presence of free ribosomes and vacuoles in the endothelial cells, endothelial flaps, sprouts and tendrils, intraluminal debris, endothelial degeneration and a junction with a nonendothelial cell. Large numbers of endothelial vacuoles  were noted, many of them occurring at the abluminal edge of the cells. These vacuoles may be involved in the formation of intraendothelial lumina and also in the enlargement of both types of lumina. This study provides evidence that besides the well-established interendothelial lumen formation, intraendothelial mechanisms may also be operative in rat cerebral cortex. The techniques employed in this study offer the potential for clarifying these and related issues.


Immunohistochemical and electron microscopic study of the rat hypothalamic nuclei and cell clusters under various experimental conditions. Possible sites of hormone release.

In untreated, pregnant and thirsting rats the neurosecretory hypothalamic areas were investigated by means of the immunoperoxidase technique in order to demonstrate vasopressin- and oxytocin containing elements at the light- and electron microscopic level. In addition, chromalum-hematoxylinphloxin (CHP) staining and conventional double staining of ultrathin sections were used. The areas investigated included the anterior and posterior supraoptic nuclei, the numerous accessory cell clusters in the region between the tractus opticus and the third ventricle as well as the median eminence. In all nuclei and in the accessory cell clusters, the number of vasopressin-reactive neurons exceeds that of oxytocin-reactive neurons. Compared with the anterior supraoptic nucleus, the posterior supraoptic nucleus and the accessory cell clusters react more heavily to prolonged thirst. In the median eminence the neurosecretory axons display close contacts with the portal vessels not only in its lateral portion but in thirsting animals also around the mid-line. There the internal layer is broadened and vasopressin-positive tanycytic processes reach the external zone. Parasagittally, fine vasopressin-positive material can be traced from the internal layer to small deposits at the portal vessels. In long term thirsting animals the typical feature of swollen axons exhibits a characteristic distribution in the median eminence and renders a distinct positive reaction to anti-vasopressin. The release of peptide hormones from the perikarya and from the axons within the nuclei as well as the mode of release within the the median eminence are discussed. The significance of the positive immunostaining of the ependymal tanycytes and of some perikarya of the suprachiasmatic nucleus must be reconsidered by further studies.


Ultrastructural immunohistochemical localization of vasopressin in the hypothalamic-neurohypophysial system of three murids.

Vasopressin was immunohistochemically localized at the electron microscopic (EM) level in the hypothalamic-neurohypophysial system (HNS) of three murids. Antiserum to vasopressin was produced in rabbits injected with lysine vasopressin (LVP) conjugated to egg albumin (EA), anti-EA being precipitated prior to staining. Sternberger's unlabeled antibody peroxidase technique was employed, immunoreactivity being designated by peroxidase-anti-peroxidase (PAP) molecules and electron opacity. Immunoreactive neurosecretory granules (NSG) were found in the perikarya of the supraoptic nucleus (SON) in all three murids investigated, although far more profusely in the two wild strains. Immunoreactive axonal NSG were observed in the inner and outer zones of the median eminence (ME), and within most of the axons and terminals in the neurohypophysis. The concentration of primary serum effective for staining the SON (1:10-1:50) was far higher than that required for the ME and the neurohypophysis (1:500-1:1,200). Anti-LVP also induced electron opacity of granules in cells of the pars intermedia (PI). Discussion centers of the significance of immunoreactive NSG in the neurosecretory (NS) perikarya, on the possibility of an extragranular pool of hormone, and on speculation about the electron opacity of the PI granules.


Congenital obstructive lesions involving the major pulmonary veins, left atrium, or mitral valve: a clinical, laboratory, and morphologic survey.

The clinical, laboratory, and morphologic features of congenitally obstructive lesions causing pulmonary venous hypertension are reviewed. These lesions are responsible for considerable infant mortality and morbidity, especially in the first weeks of life, are not infrequently encountered in older children, and are occasionally seen in adults. The presence of a malformation causing pulmonary venous hypertension often can be detected clinically, but precise anatomical and physiological diagnoses, usually necessary for optimal patient management, often can be made only by detailed laboratory study. Cardiac catheterization and angiocardiography remain the prime modes of accurate diagnosis in such patients, many of whom have other significant cardiovascular malformations, and echocardiography is an extremely useful adjunct, especially in patients with mitral valvular stenosis, hypoplasia, or atresia. The only definitive treatment in any of these patients is operative relief of the obstruction, and the chances of success depend not only on the skill of the surgeon, but also on the nature of the obstructing lesion, the types of associated malformations, and the precision with which these are defined preoperatively.


A functional requirement for modification of the wobble nucleotide in tha anticodon of a T4 suppressor tRNA.

Temperature-sensitive mutants of E. coli have been isolated which restrict the growth of strains of bacteriophage T4 which are dependent upon the function of a T4-coded amber or ochre suppressor transfer RNA. One such mutant restricts the growth of certain ochre but not amber suppressor-requiring phage. Analysis of the T4 tRNAs synthesized in this host revealed that many nucleotide modifications are significantly reduced. The modifications most strongly affected are located in the anticodon regions of the tRNA'S. The T4 ochre suppressor tRNAs normally contain a modified U residue in the wobble position of the anticodon; it has been possible to correlate tha absence of this specific modification in the mutant host with the restriction of suppressor activity. Furthermore, the extent of this restriction varies dramatically with the site of the nonsense codon, indicating that the modification requirement is strongly influenced by the local context of the mRNA. An analysis of spontaneous revertants of the E. coli ts mutant indicates that temperature sensitivity, restriction of phage suppressor function, and undermodification of tRNA are the consequences of a single genetic lesion. The isolation of a class of partial revertants to temperature insensitivity which have simultaneously become sensitive to streptomycin suggests that the translational requirement for the anticodon modification can be partially overcome by a change in the structure of the ribosome.


A shift from phospholipid to triglyceride synthesis when cell division is inhibited by trans-fatty acids.

The yeast mutant Saccharomyces cerevisiae (KD 46) requires added unsaturated fatty acid for growth.  When cell growth was inhibited by the presence of trans-acids there was a marked inhibition of oleate esterification into phospholipids accompanying continued incorporation into triglycerides. Apparently some control point in phospholipid synthesis associated with the cell cycle occurs after the stage of phosphatidate biosynthesis.


Preliminary studies on the response of Plasmodium falciparum to chloroquine in the Philippines, with the in vitro technique.

Previous investigations had shown that resistance to standard regimens of chloroquine occurred in some cases of falciparum infection in the Philippines. More extensive investigations into this phenomenon were planned by the Malaria Eradication Service, by means of the in vitro technique developed by Rieckmann, in order to determine both the distribution of resistant strains throughout the country and their local incidence. Before these studies were undertaken, a series of observations were made on cases of falciparum malaria encountered in Manila and its environs, to assess the reaction of local strains of the parasite to the in vitro test. These cases were also treated with standard doses of chloroquine and some were followed up for 4 weeks to compare the predictions made as a result of the in vitro tests with the in vivo observations. Of the 34 in vitro tests carried out, 18 were followed up in vivo. In 8 cases, no recrudescence occurred, but in the other 10 recrudescences were detected during the 4-week observation period, thus indicating parasite resistance to the drug. In each of the 18 cases, the in vivo response followed the in vitro prediction.


Comparison of counter-immunoelectrophoresis with other serological tests in the diagnosis of human brucellosis.

Sera from 65 persons with clinical brucellosis were employed in a comparison of standard and rapid serological tests. The results obtained with the Rose Bengal test correlated very well with those of the standard tube agglutination test, whereas results with the rapid plate agglutination test and the Coombs (antiglobulin) test were inferior. Absorption of patients' sera with specific anti-human immunoglobulin sera showed that IgM was active in the Rose Bengal test but not in the Coombs test, whereas IgG and IgA were active in both tests. In addition to the A & M antigen, which plays the most important role in the agglutination, Rose Bengal, and Coombs tests, other antigenic fractions of Brucella were examined in precipitation tests. A protein antigen reacted with 94% of the sera in counter-immunoelectrophoresis. On the basis of the results with both groups of sera, the Rose Bengal test and counter-immunoelectrophoresis appear to be the most promising methods for diagnosing clinical brucellosis. The tests differ qualitatively since different Brucella antigens are employed.


[Bancroft filariasis in Comores archipelago (immunologic study)].

An immunological study has complemented the epidemiological survey on Bancroft filariosis in the Mayotte island (Comores) carried out by BRUNHES et al., 1971. The search for antibodies to Dipatolonema viteae and Setaria labiotopapilosa was performed by means of electrophoresis and immunofluorescence in 87 individuals and 2 hydrocele fluids. Immunoelectrophoresis confirmed the high endemicity ratio: 57 o/o of the individuals presented 1 to 7 precipitating lines; with immunofluorescence 55 o/o of the subjects had a titer of 200 to 1.600. 46 to 50 o/o of the positivities were observed in people showing clinical symptoms without microfilaremia. On the other hand, 36 to 48 o/o of the proved filarian cases yielded negative results with immunofluorescence or electrophoresis. These figures emphasize the significance and insufficiencies of immunologic techniques for the diagnostic of bancroftoses. In spite of the frequency of certain well identified precipitating systems, an immunoelectrophoretic analysis did not allow to demonstrate a constant and characteristic scheme.


George C. Griffith lecture. The role of renin in normal and pathological cardiovascular homeostasis.

Recently, the availability of a number of specific inhibitors of the renin-angiotensin system has made it possible to address certain critical questions concerning the role of angiotensin II in physiologic homeostasis and in a number of pathologic states. These studies indicate that angiotensin II does not have an obligatory role in blood pressure maintenance in the normal, sodium replete individual, but it is essential following sodium depletion. The role of angiotensin II in feedback control of renin secretion is confirmed as is its importance in aldosterone stimulation both in relation to posture and sodium depletion. Angiotensin II is responsible for the initial pressor response of experimental renovascular hypertension and appears to be important in the initiation of chronic renovascular hypertension. Converting enzyme blockers and competitive inhibitors of angiotensin II are helpful in the diagnosis of clinical renovascular hypertension and in the identification of renin dependent hypertensives. Homeostatic mechanisms leading to maintenance of blood pressure and accumulation of edema in experimental congestive heart failure appear to be dependent on angiotensin II.


Clinical efficacy and pharmacokinetics of a new orally effective antiarrhythmic, tocainide.

Tocainide, a new oral antiarrhythmic agent, was studied in man in a short-term protocol designed to evaluate the efficacy, kinetics, and toxicity of this compound. Premature ventricular contractions (PVCs) were suppressed by less than 70% in 11 of 15 patients compared with pre-drug placebo controls. For these 11 responders, there was an average PVC reduction of 91% +/- 10 (range 70 to 100%) at tocainide doses not associated with side effects. Mild transient central nervous system toxicity was observed in some patients near the time of peak concentrations during the highest dose administered. The drug was found to have linear kinetics over the dose range studied and a plasma half-life of 13.5 +/- 2 hours. Plasma concentration-response curves indicate antiarrhythmic activity over all plasma concentrations, with 70% PVC reduction above 6.0 mug/ml. This study suggests that tocainide is a safe and effective antiarrhythmic agent during short-term administration and is worthy of further clinical trials.


The VA cooperative randomized study of surgery for coronary arterial occlusive disease II. Subgroup with significant left main lesions.

From a large cooperative prospective randomized study, data relating to a subgroup of 113 patients with angina pectoris and a significant lesion of the left main coronary artery were analyzed. Of these patients, 53 had been randomly allocated to a medical treatment group and 60 to a surgical treatment group. The former group received conventional medical treatment, while the surgical treatment group received one or more aortocoronary saphenous vein bypass grafts. Important risk factors were approximately uniformly distributed between the two groups. Both are being followed up to 60 months (average follow-up, 30 months). To date, 12 of 60 surgical patients (20%) and 19 of 53 medical patients (36%) died (P less than 0.06). The operative (30-day) mortality declined from a rate of 25% for the first 2 years of the study to 7% for the last 3 years. Of patients randomized in the latter 3 years of the study, 12 of 41 medical patients (29%) and three of 42 surgical patients (7%) died (P less than 0.01). The average follow-up period in this group was 24 months. The proportion surviving 24 months was clearly larger in the surgically treated group (P less than 0.02). The difference in the proportion of patients surviving after surgery as compared with medical treatment was greatest in patients with additional significant disease involving the right coronary artery, with or without left ventricular dysfunction. Relief of angina as assessed by an "anginal score" was also better in surgical patients to a significant degree. Graft-patency rates correlated well with relief of angina, but objective studies including treadmill testing are not yet available.


Modification by sex steroids of LHRH response in the polycystic ovary syndrome.

Exogenously administered oestradiol or progesterone was found to induce augmentation of LH and FSH release in response to LHRH administration in patients with the polycystic ovary (PCO) syndrome. The effect of oestradiol upon LH release in the patients was significantly less than the augmented release induced in normal women (P less than 0-02). In contrast, progesterone induced a significantly greater LH release in PCO patients than normal women studied during the early follicular phase (P less than 0-001) but less than that in normal women studied during the mid-follicular phase of the cycle (P less than 0-01). The effects on FSH release in the PCO patients were less marked but similar to those in normal women with comparable basal steroid levels. The results suggest that both oestradiol and progesterone can modify the hypothalamic-pituitary axis in the PCO syndrome in a manner similar to that observed in normal women and a failure of these 'positive feedback' mechanisms does not appear to be the cause of the ovulatory deficiency in this syndrome.


The control of gonadotrophin release in women with hyperprolactinaemic amenorrhoea: effect of oestrogen and progesterone on the LH and FSH response to LHRH.

The effect of the administration of oestradiol benzoate and of progesterone on the subsequent response to LHRH has been investigated in women with hyperprolactinaemia. There was an amplification in the release of LH in four out of ten patients and of FSH in one out of ten patients at 44 h after the administration of 2-5 mg oestradiol benzoate. The average amount of LH released before and after oestrogen did not change, but there was a significant decrease in the amount of FSH released. There was no correlation between the LH released and the oestradiol concentration in serum at the time of the LHRH tests but there was a negative correlation between the FSH released and the oestradiol concentration (r = 0-507;P less than 0-05). These results contrast with those obtained in normal subjects in the follicular phase of the cycle when there is a positive correlation of oestrogen concentrations and the amount of LH and FSH released. As in normal subjects, however, a significant suppression of basal FSH concentrations, persisting until 44 h, was produced by the oestrogen (P less than 0-01). Seven out of eleven patients showed an amplification of LH response and six out of eleven an FSH response 20 h after the administration of 25 mg progesterone. The mean amplifications are not significantly different from those of normal subjects tested in the early follicular phase of the cycle, but are significantly less than those tested in the mid follicular phase of the cycle (LH P less than 0-001; FSH P less than 0-01). This may be related to the serum concentrations of oestradiol which in patients with hyperprolactinaemia are significantly less than those found in the mid follicular phase of the cycle (P less than 0-05). These results indicate that in women with hyperprolactinaemia oestrogen negative feedback, necessary for cycle initiation, is normal: failure of ovulation may be related to failure of positive feedback to oestroen. Oestrogen-negative feedback is unopposed and this may explain the follicullar development and lack of oestrogen in the mid-follicular phase.


Synthesis of immunoglobulin and secretory component by gastrointestinal mucosa in patients with hypogammaglobulinaemia or IgA deficiency.

Biopsies of intestinal mucosa from patients with adult hypogammaglobulinaemia or selective IgA deficiency have been studied for the ability to synthesize immunoglobulins and secretory component. Tissue fragments were cultured in vitro in medium containing 14C-labelled amino acids and newly snythesized proteins were detected by radioimmunoelectrophoresis. Synthesis of IgA, and in some cases IgG and IgM, was found in intestinal mucosal biopsies from hypogammaglobulinaemics and IgA-deficient subjects. Biopsies from all the patients also synthesized secretory component, but evidence was obtained which indicated that secretory component does not combine normally with IgA. Tissue sections of these biopsies have also been studied by immunofluorescence and immunoglobulin bearing cells have been demonstrated. The present findings demonstrate that immunoglobulin synthesizing cells are present in the intestinal mucosa of immunoglobulin-deficient individuals. Local immunoglobulin synthesis may partially explain why these patients do not often have major problems with intestinal infections.


An extractable nuclear antigen not attaching to tannic acid-treated erythrocytes.

One of the antigens present in isotonic phosphate buffer extract of calf thymus nuclei against which antibodies are produced in collagen diseases, was shown not to attach to tanned sheep erythrocytes. It is, however, well exposed on slides coated with smears of ENA-extract and fixed with methanol-acetone. Reactions with the antigen can be demonstrated by the mixed haemadsorption technique. Preliminary results suggest that it may be a ribonucloprotein closely related to the 'soluble ribonucleoprotein" (sRNP) attachable to tanned erythrocytes. It may also have a similar relationship to clinical symptoms.


Immunodepression during Trypanosoma brucei gambiense infections in the field vole, Microtus montanus.

The effects of Trypanosoma brucei gambiense infections upon immune responses were examined in an outbred laboratory colony of field voles. Microtus montanus. Antibody levels to challenge with heterologous erythrocytes and bovine serum albumin were significantly depressed in infected animals. Trypanosome infections impaired both primary and secondary humoral responses, although previously established specific antibody levels were not affected by infection. Specific antibody-producing capabilities of previously infected, trypanoicidal drug-treated voles were found to be comparable to uninfected controls, within 3 days following chemotherapy. Cell-mediated hypersensitivity responses to oxazolone were also significantly depressed by trypanosome infection; responses to the initial sensitization of oxazolone and to secondary challenge treatments were depressed compared to uninfected controls. Possible mechanisms of trypanosome-induced immunodepression are discussed.


A simple immunoenzymatic method for measuring IgE in human sera.

A reliable and relatively simple method for the estimation of serum IgE by single radial immunodiffusion is described. The method requires glucose oxidase-labelled antibodies. The method permits the measurement of IgE concentrations ranging from 20 to 700 i.u./ml. The values obtained in unknown samples were in good agreement with those obtained by radioimmunoassay (correlation coefficient r=0-9557).


Thyroiditis in T cell-depleted rats: suppression of the autoallergic response by reconstitution with normal lymphoid cells.

Qualititive, quantitative and functional differences were found in lymphoid cells of female thymectomized and irradiated (Tx-X) PVG/c strain rats as compared to normal females of the same strain. Tx-X rats were lymphopenic and had reduced numbers of cells within spleen and cervical lymph nodes, depressed transformation responses of peripheral blood lymphocytes to PHA and lower percentage killing of their spleen cells by anti-T-cell serum and complement. There was an increased percentage of immunoglobulin-bearing cells in the lymph nodes. Reconstitution of Tx-X rats by the intravenous route using syngeneic lymph node cells, spleen cells or thymocytes abrogated the autoimmune responses to thyroid components generally observed in this state. Lymph node and spleen cells, but not thymocytes, also prevented thyroid changes when given intraperitoneally. In contrast, bone marrow cells appeared to give enhanced responses. Quntitative studies showed that the relative proportions of the suppressor or autoregulatory cells in various lymphoid tissues were lymph node greater than spleen greater than thymus. Complete abrogation of the autoimmune responses was possible only when cells were administered within a short time of final dose of irradiation and moderate thyroid change was again seen if transfer was delayed for 14 days post-irradiation. At 28 days reconstitution had no influence on the development of the autoimmune responses. Preliminary characterization studies using an anti-T-cell serum and fractionation of lymph node cells on a linear Ficoll gradient suggested that autoregulatory cell is a large T cell.


Human sperm antigens and antisperm antibodies. II. Age-related incidence of antisperm antibodies.

Naturally occurring human antisperm antibodies (ASA) detectable by immunofluorescence, were found to have a peak indicence of 90% in both sexes before puberty. Thereafter, the incidence declined to about 60% and persisted through life. This age-related incidence resembled that for foreign and not self antigens. The natural incidence of the six antibodies under investigation varied: the most frequent were antibodies to acrosomal antigen Acl and equatorial antigen; the least frequent were antibodies to acrosomal antigen Ac2 and sperm nuclear protamine; and between these were antibodies to the mainpiece of tail and post-acrosomal region. Irrespective of their natural incidence, these antibodies increased at a comparable rate in men following vasectomy indicating these six sperm antigens have comparable immunogenicity. These results provide evidence for the lack of immunologic tolerance in man toward many sperm antigens. The variation in the natural incidence of individual ASA is explanable by differences in prevalence of crossreaction between each sperm antigen with exogenous antigens. Thus, antibodies to acrosomal antigen Acl and equatorial antigen, which occurred most frequently, appeared to crossreact with ubiquitous microorganisms.


Cross-reaction of antibodies present in sera of vasectomized mice with human swollen spermheads.

Vasectomy induced anti-sperm antibodies in mice were seen to cross react with human spermatozoa. Similarly anti-sperm antibodies in an infertile human male subject were found not only to react with human but also with bovine, porcine and murine sperm. Both observations made with preparations of swollen spermatozoa demonstrate the cross-reactivity of anit-sperm protamine activity of different species. Therefore in spite of known species dependent differences in amino acid composition and sequence of protamines, they appear to have common antigenic determinants.


Lymphocyte transformation test in leprosy; correlation of the response with inflammation of lesions.

Lymphocyte transformation tests (LTT) using 'whole washed" and 'sonicated" preparations of Mycobacterium leprae (M. leprae) as antigen were studied in eighty-one patients with borderline leprosy. The results were correlated with the histological and the clinical pictures. There was a good correlation with the histological spectrum, LTT responses generally being higher in the borderline tuberculoid leprosy patients and lower in the borderline lepromatous. However, considerable variation was noted in each group of the borderline leprosy spectrum, and it was found that this was due in part to the degree of inflammation in the skin. Thus those with 'inflamed" skin lesions had higher responses than those with 'silent" lesions, and even those with borderline lepromatous leprosy with inflamed lesions had higher responses than those with borderline tuberculoid leprosy whose lesions were silent. Those who had reversal reactions, where inflammation is very marked, had very high LTT responses which fell with treatment of the reaction with steroids. It thus appears that the LTT in leprosy is influenced by the occurrence of hypersensitivity reactions as well as by the patient's ability to resist bacillary multiplication.


In vitro reversal of cellular unresponsiveness induced by levamisole.

Mononuclear cells from twenty-one patients with depressed cellular reactivity were assessed for the ability to produce leucocyte inhibitory factor (LIF) and to transform after PHA stimulation, in the presence or absence of levamisole. Cells from nineteen patients failed to produce significant lymphokines when stimulated with PHA alone, but after a prior 1-hr levamisole pulse normal amounts of LIF were produced. Unstimulated mononuclear cell supernatants from six patients showed LIF-like activity, which could be abolished or decreased in five of the six when the cells were initially treated with levamisole. Mononuclear cells from seven of twelve patients which failed to incorporate [3H]thymidine after PHA activation, showed an increased response after a 1-hr levamisole pulse. Unstimulated mononuclear supernatants from six patients inhibited the lymphoproliferative response of normal cells to PHA. After treatment with levamisole, however, the suppressive effect of these supernatants was decreased or abolished. In vitro levamisole treatment, therefore, not only restores cellular responsiveness in anergic patients but also restricts the uncontrolled release of inhibitory factors.


Local adoptive transfer to mice of human delayed hypersensitivity: reactions by the radioisotopic footpad assay.

Human delayed hypersensitivity to living BCG organisms and/or Varidase was adoptively transferred to mice and assayed in mice by a radioisotope footpad assay (FPA). A mixture of the antigen and peripheral blood lymphocytes from patients (positive skin reactions to PPD and/or Varidase) or healthy volunteers was inoculated into the footpads of lethally X-irradiated mice. A positive footpad reaction was accompanied by an increased leakage of the radiolabelled serum protein from the blood stream into the intercellular space at the site of inoculation, and was measured by the 'foot-count ratio', or radioactivity in the test foot divided by radioactivity in the contralateral foot. The intensity of the footpad reaction correlated directly with the skin test response of the human lymphocyte donors.


T-cell helper response to antigens of Schistosoma mansoni in CBA mice.

It is believed that a T-cell helper response against the schistosome surface is a necessary prerequisite for the development of protective immunity in schistosomiasis. Accordingly, the carrier effect has been used to assay eleven antigenic preparations of Schistosoma mansoni for their helper T-cell priming against surface components of the schistosomula. Three weeks after i.v. injection of the preparation, CBA mice were injected with schistosomula coated with trinitrophenol (TNP) and 4 days later, their spleens were assayed for plaque-forming cells (PFC) against TNP. Formalin-fixed schistosomula and crude adult worm tegumental membrane induced the highest response; only thirty schistosomula or 10mug of crude membrane protein were needed to generate a T-cell helper response equivalent to that induced by a living infection. All other antigenic preparations, including formalin-fixed cercariae, live miracidia, eggs and adult worm culture fluid, generated some response indicating the presence of shared carrier components.


Specific antibody production against a soluble antigen in the Harderian gland of the domestic fowl.

Three groups of chickens were immunized with bovine serum albumen (BSA). One group received the antigen i.v., the second had BSA dropped into the orbit and the third group was immunized both systemically and by topical application of BSA. Agar-gel diffusion studies showed antibodies to BSA to be present in the tears and sera of the first and third groups but only in the tears of the second group. Immunofluorescence revealed positive anti-BSA staining in lymphoid cells of the spleens of the first and third groups and in plasma cells of Harderian glands of all three groups. It is concluded that systemic and/or topical applications of BSA affect the tears and Harderian gland whereas only systemic injections produce an effect in the spleen and serum. The Harderian gland may have an important part to play in the local immunologic mechanism of the eye and upper respiratory tract of the fowl.


The effect of recurrent early homograft rejection on subsequent patient and renal graft survival. Two hundred renal transplants performed in Newcastle between 1968 and 1974 and followed up for at least 6 months are reviewed. There was a significant fall ingraft survival at 1 year in those patients who suffered a rejection episode during the first two months post-transplant and this fall became greater with each successive rejection episode. The rise in patient mortality with increasing numbers of rejections showed a similar trend but was less (40-50% at 1 year in those suffering 3 or 4 rejection episodes) and did not reach statistical significance beyond the first episode. We, therefore, conclude that in patients not suitable for home dialysis and in whom, because of uncommon tissue type, a second transplant is not likely to be offered under prevailing conditions of kidney donor shortage, it is justifiable to treat third and fourth rejections occurring during the first two months.


Renal transplantation in children: a psychological survey.

Psychological tests were carried out on 24 children with renal disease during the time they were hemodialyzed and after a minimum of one year after their transplant. These investigations included regular interviews with children and families, drawings and their commentaries, and projective techniques: Rohrschach tests. The results of these investigations indicated that the psychological status of the transplanted child could not be singled out and isolated from the effects of being ill, the different therapeutic procedures, and the personal history. Beyond the appearance adolescents give of a satisfactory social adjustment and beyond the explosion of vitality frequent amongst younger children, an impoverishment of their personality and major difficulties manifest themselves in coping with personal problems and identity crises especialy as far as agressiveness and sexuality are concerned. This neurotic mechanism appears mainly due to the traumatic experiences these children have experienced and their lack of support in mastering them through the verbalization process.


Etiologic and prognostic significance of isolated acute renal hypoxic change (so-called tubular necrosis or shock kidney) in kidney transplants.

20 biopsies were studied in order to determine whether or not the pathologist is able to predict on morphological grounds the etiology and prognosis of acute renal hypoxic change in renal transplants. We concluded that: 1) Intertubular blood stasis seems to be pathognomonic of peripheral vascular rejection and/or renal artery thrombosis. 2) A focal distribution of acute tubular luminal dilatation suggests peripheral vascular rejection. 3) The finding of lymphatic casts suggests the presence of ureteric obstruction. 4) Linear fluorescent staining of the peripheral glomerular basement membrane appears to be due to leakage of plasma proteins in the presence of endothelial damage. 5) The histological distinction between camouflaged rejection and genuine shock kidney is impossible in a biopsy. 6) The precise cause of acute renal hypoxic change and the prognosis of kidney transplants with this alteration often cannot be determined.


Influence of cannabidiol on secobarbital effects and plasma kinetics.

To investigate the possible metabolic interaction between cannabidiol (CBD) and secobarbital, 6 male volunteers received 150 mg/70 kg sodium secobarbital orally immediately after smoking a marihuana cigarette prepared to deliver 0, 150, or 500 mug/kg CBD. The study was performed in a double-blind manner with each of the three treatments being assigned to every subject. Clinical effects and plasma secobarbital concentrations were recorded at periodic intervals. CBD did not alter the summary parameters which describe the secobarbital plasma concentration time curve. Secobarbital half-life, peak concentration, time of peak concentration, and area under the curve were unchanged by the coadministration of CBD. Clinical effects of secobarbital were also unaltered by CBD pretreatment. Thus at the doses administered, CBD does not appear to inhibit secobarbital metabolism in man.


Antihypertensive efficacy of a single bedtime dose of methyldopa.

To compare the antihypertensive efficacy of methyldopa administered once at bedtime with the same total dose given three times daily, a double-blind crossover study was performed in 14 patients previously well controlled on methyldopa. Each patient received a total daily dose of 0.37 gm, 0.75 gm, or 1.5 gm of methyldopa, depending on the dose of drug that had previously been successful in that individual. The trial design included either 12 wk of methyldopa three times daily (TID) followed by 12 wk of single daily bedtime (HS) doses of methyldopa or administration of drug in the reverse order. Supine and erect blood pressures were recorded 4 times daily (8 a.m., 12 noon, 4 p.m., and 8 p.m.) every 4 wk throughout the study. Blood pressure control was excellent in all patients whether the drug was administered three times daily or at bedtime. Systolic pressures were slightly lower at 8 a.m., when methyldopa was given at bedtime than on doses three times daily, and systolic and diastolic pressures were slightly higher at 8 p.m. that at 8 a.m. on the bedtime regimen.


Absorption and metabolism of phenytoin from tablets and capsules.

Crossover studies were conducted to compare plasma levels of phenytoin and urinary throughput of the major metabolite, 5-(p-hydroxyphenyl)-5-phenylhydantoin (HPPH), after ingestion of single 250-mg oral doses as tablets or as capsules. Plasma levels of phenytoin did not differ significantly for the 2 formulations except at 8 and 12 hr after ingestion of drug, when the mean values for the tablets were higher than for the capsules. The normalized area under the curve, however, did not differ significantly for either preparation, indicating that equivalent amounts of each preparation were absorbed. Urinary throughput of HPPH did not differ significantly between the 2 preparations. The results suggest that the 2 preparations provide equivalent therapeutic effects.


Patient dissatisfaction and attempted suicide.

This paper deals with the presence of suicidal behavior in a mental health clinic and psychiatric emergency room, and its relationships to what has been described in the literature as the "dependent-dissatisfied" personality. Procedures included a suicidal potential scale, projective figure drawings, and a brief interview about past and present suicidal behavior. The scale used significantly differentiated between high and low potentially suicidal groups. The high suicidal group showed a significantly greater dissatisfaction with treatment both past and present. The implications for both evaluation and therapy of suicidal patients are discussed.


Antibiotic-associated pseudomembranous colitis.

This review defines the entity pseudomembranous colitis and briefly outlines the supposed etiologic causes of pseudomembranous colitis including antibiotics. The incidence, mortality rate, and natural history of antibiotic-related pseudomembranous colitis is contrasted with other forms and causes of pseudomembranous colitis. The clinical spectrum of antibiotic-related pseudomembranous colitis, ranging from a nonbloody, watery diarrheal state to a life-threatening condition mimicking an acute surgical abdomen, is reviewed. The classic proctoscopic and pathologic findings, as well as common problems encountered in interpretation, are discussed. A complete review of the spectrum of radiographic findings is presented from the nonspecific to the quite characteristic radiographic findings, including both plain film and contrast studies of the colon. These findings are contrasted with the X-ray features of other inflammatory and ischemic colitides and a differential diagnosis is discussed. A section dealing with the treatment of antibiotic-associated pseudomembranous colitis will be included. This section will review the various modes of therapy that have been employed. Finally, a brief section will speculate on the possible etiologic role that antibiotics play in pseudomembranous colitis, including the alteration of the bacterial flora and possible effect on bile salt metabolism.


Transverse axial tomography of the spine.

Transerse axial tomography is a technique by which the spine is viewed radiographically in cross section. The tomographic apparatus, technique of examination, and radiation exposure will be discussed. Examples of normal and pathologic anatomy of the cervical, thoracic, and lumbar spine will be presented. In the lumbar spine cross-sectional views of stenotic canals due to facetal hypertrophy, congenital stenosis, spondylolithesis, and ventral overgrowth of spine fusions will be demonstrated. Measurements of the sagittal and interpedicular distances on conventional x-rays have correlated poorly with the anatomic state in lumbar stenosis. A high degree of correlation has been demonstrated by plotting the cross-sectional area configuration of the bony canal, as well as a new measurement, the interfacet distance, on axial tomograms. Transverse axial tomography is an exciting, new approach in neuroradiology for the evaluation of spinal disorders which have primarily axial distortion.


Essential knowledge about pitfalls in tomography.

Tomographic cut is not synonymous to a roentgenogram of an anatomic cut. Blurring is inherent in the tomogram, and the edges do not appear as sharp as in the conventional roentgenogram. Part of the contrast is lost in tomography. Therefore, when tissue contrast is low, or when we intend to see an entire structure in one cut, zoneography is preferred. On the other hand, when many details are crowded in a small area and/or when the tissue contrast is high, a thin section tomography is indicated. Proper knowledge of the level of area of interest (focal plane) and proper spacing of the cuts decrease radiation and expenses. The Law of Tangent, namely, "The edge of a structure should be momentarily parallel to the beam at the focal plane, in order to show on the film," is an essential part of every tomographic study. The Law of Tangent applies equally to spheres and other curved surfaces. The effective tomographic angle is the angle described by the tube during exposure. The thickness of the cut depends on the effective tomographic angle. Prior viewing of conventional roentgenogram and proper knowledge of the mechanism of parasitic and phantom images help us to avoid mistaking these images as real structures.


A comparative study of clemastine ('Tavegil') and chlorpheniramine maleate in the treatment of hay fever.

A double-blind randomised parallel group study was carried out in 46 patinets to compare the effectiveness of clemastine with that of chlorpheniramine maleate in the treatment of hay fever. The results showed that both drugs were equally effective in controlling the symptoms of hay fever without side-efects or other untoward reactions. It is suggested that clemastine is a logical first choice antihistamine as well as being an effective alternative where tolerance to other antihistamines develops.


Double-blind trial of prostaglandin F2alpha and oxytocin in the induction of labour.

In a random double-blind trial in 28 pregnant women at term, labour was induced with either prostaglandin F2alpha or oxytocin given by intravenous infusion. The results showed that prostaglandin F2alpha was as active as oxytocin and induction was successfully carried out in 24 patients; 3 patients had a caesarean section because of an obstructed labour and 1 patient on oxytocin did not have regular contractions after 10-hours' infusion although she delivered spontaneously 2 days later. No difference was found in the induction--delivery interval in the 8 patients in each group who had early amniotomy (first 90 minutes). No adverse effects were noted in either mother or the child. The authors recommend that to avoid side-effects the dosage of 20 mug prostaglandin F2alpha per minute should not be exceeded and that, although no cardiotocographic abnormalities were noted, this method of control should be used.


Double-blind comparison of the antirheumatic activity and gastric side-effects of indomethacin used alone or in combination with a gastro-protective agent, proglumide.

A double-blind placebo controlled trial was carried out in 32 hospitalized patients with various rheumatic disorders to assess the therapeutic effectiveness and gastric tolerance of indomethacin (150 mg/day) used alone or in combination with proglumide (900 mg/day). Patients were treated for periods ranging from 20 to 28 days. The therapeutic response to both treatments, as assessed by objective criteria and final overall evaluation, was not significantly different. There was, however, a marked difference in gastric disturbance between the two groups. Uropepsinogen levels increased significantly during treatment in patients receiving indomethacin plus placebo but decreased significantly after Day 10 in those receiving proglumide. Although stomach X-ray or faecal occult blood investigations showed litle change from normal in either group, no gastric side-effects were reported in the proglumide patients whereas only 2 of the 16 patients receiving indomethacin plus placebo were entirely free of such side-effects throughout the trial, and most had developed gastric symptoms by the 9th day of treatment.


Comparison of azapropazone and naproxen in rheumatoid arthritis.

A double-blind crossover trial was carried out in 15 patients with definite rheumatoid arthritis to compare the effectiveness of 1200 mg azapropazone daily and 750 mg naproxen daily, each drug being given for 2 weeks. Patients also received placebo therapy before and between the two treatment periods. The results of subjective and objective assessments showed that both drugs were significantly superior to placebo, but no significant difference could be demonstrated between the two drugs.


Replacement therapy and piperazine oestrone sulphate ('Harmogen') and its effect on memory.

A formal memory test was administered to 18 female patients with signs or symptoms of oestrogen deficiency taking part in a double-blind study of piperazine oestrone sulphate. A significant improvement in memory was seen in the treated group compared with the placebo group. The findings are discussed.


Case report: lithium in reactive depression.

A 25-year old woman suffering from severe reactive depression was treated with lithium carbonate 1500 mg per day. On this regime she showed a very good improvement, and after 4 weeks of in-patient treatment was free of symptoms. At 6-months' follow-up there was no recurrence of symptoms; her serum lithium levels were within therapeutic range (0.6 to 1.2 mEq/1).


[Bridging extensive peritoneal defects with terminal mesoileum].

Extended intestinal operations such as right hemicolectomies and operations in the pouch of Douglas can cause considerable difficulties in restoring the missing peritoneum. In the authors' four cases the problems were solved anatomically and functionally by suturing only the mesenterium of the terminal ileum to the edge of the previously cut peritoneum. With this technique of peritonealization the mobility and function of the gut was left intact. The resulting retroperitoneal space was drained separately.


[Choice of surgical procedure on the trachea].

The operative possibilities: fenestration and transsection of the trachea with closure of the defect by autoplasty, alloplasty, or end-to-end-anastomosis are described with a review of the literature. The choice of surgical procedure depends on the localization, type and extension of the tracheal lesion. Two case histories - one patient with a cylindroma and another with an inflammatory stenosis - are demonstrated. In both cases tracheal fenestration and closure of the defect by a pleural graft including support with a free transplanted autologous ribbone was applied. The reasons for this procedure are described. Of particular interest is the demonstration of the advantages and limitations of this method.


Iron-binding proteins and other factors in milk responsible for resistance to Escherichia coli.

Human milk contains large amounts of the iron-binding protein lactoferrin. This is normally unsaturated with iron. It also contains large amounts of IgA and small amounts of IgG and IgM. A combination of lactoferrin and specific antibody has a powerful bacteriostatic effect on Escherichia coli. In sucking infants the milk proteins probably reach the small intestine intact. Experiments with sucking guinea pigs show that milk suppresses E. coli in the gut and that the unsaturated iron-binding protein plays an essential role in the bacteriostatic reaction. Inhibited E. coli appear to be acutely iron deficient. E. coli growing slowly in iron-deficient media show abnormal forms of certain aminoacyl tRNAs. In bacteria inhibited by colostrum the proportion of abnormal tRNA is as high as 90%. These abnormal tRNAs are converted to the normal form by the addition of iron. This occurs in the absence of further RNA synthesis and is accompanied by renewed bacterial growth. The normal flora of the gut also plays an important role in resistance. Human milk has a low buffering capacity and bacterial fermentation of lactose produces a low pH.e. coli is inhibited by acetic acid/acetate buffer at pH 4.8-5.6, whereas these conditions allow normal growth of Lactobacillus bifidus. The faeces of babies fed on breast milk have a low pH, low counts of E. coli and high counts of L. bifidus. Artificially fed babies have more alkaline faeces which contain few L. bifidus and large numbers of E. coli.


The problem of bacterial diarrhoea.

The reported incidence of "pathogenic" bacteria, as judged by serotype, in the stools of children with acute diarrhoea has varied from 4 to 33% over the last twenty years. Techniques such as tissue culture provide a means for detecting enterotoxin-producing strains of bacteria, strains which often do not possess "pathogenic" serotypes. "Pathogenicity" requires redefinition, and the aetiological importance of bacteria in diarrhoea is probably considerably greater than previous reports have indicated. Colonization of the bowel by a pathogen will result in structural and/or mucosal abnormalities, and will depend on a series of complex interactions between the external environment, the pathogen, and the host and its resident bacterial flora. Enteropathogenic bacteria may be broadly classified as (i) invasive (e.g. Shigella, Salmonella and some Escherichia coli) which predominantly affect the distal bowel, or (ii) non-invasive (e.g. Vibrio cholerae and E. coli) which affect the proximal bowel. V. cholerae and E. coli elaborate heat-labile enterotoxins which activate adenylate cyclase and induce small intestinal secretion; the secretory effects of heat-stable E. coli and heat-labile Shigella dysenteriae enterotoxins are not accompanied by cyclase activation. The two major complications of acute diarrhoea are (i) hypernatraemic dehydration with its attendant neurological, renal and vascular lesions, and (ii) protracted diarrhoea which may lead to severe malnutrition. Deconjugation of bile salts and colonization of the small bowel with toxigenic strains of E. coli may be important in the pathophysiology of the protracted diarrhoea syndrome. The control of bacterial diarrhoea requires a corrdinated political, educational, social, public health and scientific attack. Bacterial diarrhoea is a major health problem throughout the world, and carries an appreciable morbidity and mortality. This is particularly the case during infancy, and in those developing parts of the world where malnutrition is common. This paper is concerned mainly with acute bacterial diarrhoea, and reviews the problem as a whole.


Taking science where the diarrhoea is.

With attack rates exceeding two episodes per year in the young diarrhoea with attendant dehydration is by far the major single killer in the developing world. An invariable accompaniment of the more insidious and chronic protein-energy malnutrition (PEM), diarrhoea is itself an acute form of malnutrition: fluid-electrolyte malnutrition (FEM). Scientific attention to FEM has focused heavily on mechanisms of pathogenesis and disordered physiology, often to the neglect of preventive and effective control measures. A notable exception was the huge step from the short-circuit chamber to the cholera ward which carried the science of coupled transport to the field. Glucose-electrolyte solutions provide effective prevention and treatment of dehydration and, where combined with early proper feeding, an interruption of the FEM-PEM cycle. Wider use of this simple technology awaits greater understanding and interact-on with the social systems that determine the ecology of diarrhoeal disease.


Neonatal Escherichia coli infections in domestic mammals: transmissibility of pathogenic characteristics.

Apart from the fact that different serotypes are involved, natural and experimental Escherichia coli infection in domestic mammals closely resembles natural E. coli infection in human beings. Some of the important characteristics of E. coli strains that cause disease in domestic mammals are determined by transmissible plasmids. These include enterotoxin, haemolysin and K88 antigen in piglet enteropathogenic strains and enterotoxin and K99 antigen production in calf and lamb enteropathogenic strains; most strains that cause generalized infections in young domestic mammals, i.e. invasive strains, also produce plasmid-determined colicine V. These are all good reasons for employing young domestic mammals as the animal model for studying certain aspects of E. coli infection in human beings. Exploiting the fact that plasmids can be introduced into bacterial cells by conjugation and can be removed from them by "curing", bacterial strains were created that differed from each other, as far as could be determined, only by the presence or absence of one or more of these plasmid-determined properties. These strains, or cell-free preparations of them, were then given by mouth to piglets, calves, lambs and baby rabbits. The results showed that the K88 antigen, probably on account of its adhesive properties, permitted pig enteropathogenic strains of E. coli to proliferate in the small intestine of piglets; the K99 antigen performed a similar function in calf and lamb enteropathogenic strains. The enterotoxin produced by the proliferating organisms was then chiefly responsible for the subsequent movement of fluid from the body into the small intestine and the consequent diarrhoea. Possession of the Col V plasmid contributed significantly to the virulence of invasive strains of E. coli by enabling them to resist more successfully the defence mechanisms of the host.


The nature and action of cholera toxin.

Cholera diarrhoea is due to the action of a toxin that acts on all animal cells by stimulating the enzyme adenylate cyclase, which catalyses the production oc cyclic AMP from ATP. In intestinal brush border cells raised cyclic AMP levels result in increased secretion of chloride ions, leading to fluid accumulation in the gut. Escherichia coli produces a similar toxin. The receptor for cholera toxin on the cell membrane appears to be a complex containing the ganglioside GGnSLC (or GM1). Cholera toxin is a protein composed of two different kinds of subunits linked non-covalently. Each toxin molecule has one subunit A and four or more subunits B. Subunit B is inactive but binds to the ganglioside GGnSLC on the cell surface. Subunit A does not bind to cell membranes or gangioside and is slightly toxic to intact cells but strongly and instantly active in lysed cells. The binding of whole toxin through the B subunit to the cell is followed by a lag before subunit A penetrates the cell membrane (leaving subunit B on the surface) and stimulates the adenylate cyclase. The stimulation of adenylate cyclase depends on the presence of NAD and other co-factors present in the cell sap.


[Propranolol and pindolol in chronic obstructive lung disease (author's transl)].

The effects of 5 mg pindolol and 100 mg propranolol on the airway resistance were compared with a placebo in an inter-individual double blind trial in eight patients with chronic obstructive lung disease. Six hours after drug administration the patients inhaled two puffs of fenoterol (0.4 mg) from a metered aerosol to test the reversibility of the bronchoconstrictive effect. Airway resistance and intrathoracic gas volume were measured by body plethysmography. Propranolol increased the airway resistance significantly (P less than 0.01). The highest increase with a mean of 44% was measured two hours after ingestion. After six hours the mean increase was still 18%. Pindolol showed a barely detectable, statistically insignificant bronchoconstrictive effect in comparison with the placebo. With regard to the circulatory effects 5 mg pindolol and 100 mg propranolol were aequipotent, propranolol had a more pronounced effect in lowering the pulse rate, pindolol in lowering the blood pressure. Fenoterol can reduce airway resistance even after beta-blockade, but the effect was more profound after the placebo than after the beta-blocker.


[Kinematics and load distribution in braced free-end dentures. III. Influence of saddle surfaces on saddle kinematics and on the load-distribution abutiments-edentulous prosthesis saddle. Discussion and results].

In two upper-jaw free-end dentures with proximal rigid anchorage and equal average saddle width, the relationship between resilience of the mucous membrane of the alveolar ridge, the saddle area and the distribution of the load between abutment teeth and edentulous denture-bearing base was studied.


[Studies on the stability of total prostheses].

1. The load capacity of complete upper dentures objectively proved greater than that of lower dentures. 2. The load capacity and, consequently, the stability of position of the lateral area of upper dentures objectively diminished with advancing time of wearing, but increased in the lower jaw. 3. The stability of complete dentures is not determined exclusively by the quality of the denture bearing tissue. 4. The measured stability of position of complete dentures did not correspond with the subjective impression of full function; in general, this was stated to be as good as ever by all patients.


[Zinc oxide-eugenol as dental material (1)].

Zinc oxide-eugenol cements are considerably better tolerated by tissue than other dental materials. As they alleviate pain and are bacteriostatic and antiseptic, they are well tolerated by patients. The cements are good insulators and possess better sealing properties than zinc phosphate cements. Because of their poor mechanic properties, the conventional zinc oxide-eugenol cements are mainly used as temporary fixing contents and filling materials, for gingival dressings and together with filling materials as impression materials. Recently, reinforced zinc oxide-eugenol cements and cements containing ethoxy benzoic acid (EBA) have been developed. These new cements have considerably better mechanic properties and are therefore used for cement bases, indirect capping, long-term temporary fillings and in selected cases as definite fixing cements.


[Changes in shape of complete dentures in the mouth].

Form changes of complete dentures were measured in 60 patients. Some of the dentures were made of Paladon 65 and the others of PMMA by the "casting method". The first check was made on the day when the denture was inserted and the last up to 90 days thereafter. In the time the denture was worn, the form changes were recorded and evaluated by means of special markings. It was found that, under masticatory load, the dentures were slightly expanded transversally by 0.2 to 0.4% in the regions of the tuberosity and the anterior teeth. It was not possible to demonstrate a clear difference in the form stability of the base material Paladon 65 and that of PMMA (using the "casting method") during the three month in which the dentures were worn.


[Quantitative studies on the retention mechanism of complete dentures. I. Studies on a phantom model].

In four full maxillary dentures, the physical mechanisms of retention were examined and quantitatively determined by inductive pressure and motion recorders, in tests performed on a model and on the patients. In addition to capillary or viscous forces, the pressure difference existing in the space between denture and palate on the one hand and the oral environment on the other and resulting during pull-off, provides the greatest retaining effect.


Long-term organ culture of rat anterior pituitary glands.

An organ culture system for neonatal rat anterior pituitary glands has been developed in which cellular integrity and responsiveness to LH releasing hormone (LHRH) are maintained for at least 4 days. Anterior pituitary glands from Sprague-Dawley rats were cultured individually at 37 C in an atmosphere of 95% O2-5% CO2 in BGJb culture medium, Fitton-Jackson modification. Histological examination of 5-day-old rat pituitary glands cultured for 96 h in control medium revealed no evidence of tissue necrosis. By contrast, 40-day-old-rat anterior pituitary glands were centrally necrotic after only 24 h of incubation under the same conditions. The neonatal glands were treated for 24 h with synthetic LHRH, and medium and pituitary LH FSH concentrations were determined by double antibody radioimmunoassay. On the first day of culture, LHRH caused a dose-related release of LH with the minimal effective dose between 10(-10) and 10(-9)M. Near-maximal release appeared to be attained with 10(-6)M LHRH, which induced a 12-fold elevation in medium LH over controls. Release was evident within 3 h following stimulation with 10(-9)M LHRH. Pituitary glands were responsive to this dose at 0,24 and 72 h of culture with progressively increasing ratios between LHRH-stimulated and control LH levels of 3,13, and 24, respectively. Although FSH release by LHRH was not observed on the first day of culture, on the second day LHRH caused a dose-related secretion of this hormone. Specificity of the pituitary response was demonstrated for LH and TSH release following addition of LHRH and thyrotropin releasing hormone (TRH). The simplicity, sensitivity, reproducibility, and long-term viability of this neonatal rat anterior pituitary organ culture system make it a valuable technique for the study of gonadotropin secretion.


Neurosecretory pathways in the mallard duck (Anas platyrhynchos) brain: localization by aldehyde fuchsin and immunoperoxidase techniques for neurophysin (NP) and gonadotrophin releasing hormone (Gn-RH).

Localization studies of the hypothalamohypophysial and tuberoinfundibular neurosecretory systems were performed in the adult male mallard duck with an immunoperoxidase techinque for the demonstration of neurophysin (NP) and gonado-tropin-releasing hormone (Gn-RH) and with aldehyde fuchsin for the staining of neuosecretory material (NSM). A comparison was made between the distribution of NSM stained with aldehyde fuchsin and NP seen by immunocytochemistry. The magnocellular perikarya of the supraoptic (SON) and paraventricular (PVN) nuclei, the zona externa of the anterior median eminence (ME), the fiber layer of both the anterior and posterior ME, and small neurons in the tractus quintofrontalis were stained by both the immunoperoxidase method for NP and by the aldehyde fuchsin stain. In contrast, the parvocellular neurons of the PVN, extra-hypothalamic neurosecretory fibers dorsal to the anterior commissure in the septal region and tanycytes lining the ventral 1/3 of the third ventricle at the level of the anterior ME, were stained only by the immunocytochemical procedure for NP. These observations indicate that immunocytochemistry is more sensitive than aldehyde fuchsin staining for detecting low concentrations of NP in cells and tissues, but the two techniques produce comparable results where the concentration of the NP is relatively high. Two populations of beaded axons containing Gn-RH were distributed throughout the zone externa of both the anterior and posterior ME. One group of fibers paralleled the hypothalamo-hypophysial neurosecretory tract whereas the other was distributed in the contact zone of the ME. Immunoreactive Gn-RH was found in the cytoplasm of a sparse population of cell bodies in the dorsolateral portion of the arcuate nucleus as well as in the axons that project from this nucleus ventrally towards the ME.


Correlation of the pituitary response to synthetic LH-RH to the estradiol levels in anovulatory and ovulatory women.

This paper describes the LH and FSH responses to synthetic LH-RH in 81 anovulatory women and 37 ovulatory women at various estradiol (Ed) levels. Besides, effects of mestranol of releases of LH and FSH in response to LHrh were examined in 4 anovalatory women. Subjects were divided into 3 groups: low Ed group, moderate Ed group and high Ed group. No significant correlation was found in ovulatory women, but in anovllatory women a positive significant correlation was found between the LH/FSH ratio after LHrh injection and the circulating Ed level. In ovulatory women, a mean LH/FSH after administration of LHrh was not significantly different from LH/FSH at the LH peak in the normal menstrual cycle. But in anovulatory women, a mean LH/FSH after the adminitration in the low Ed group was significantly lower than LH/FSH and mean LH/FSH after the administration in two other Ed groups were significantly higher than that at the LH peak in the normal menstrual cycle. Among the low Ed group, four patients with primary ovarian failure  and elevated gonadotropin levels in serum were treated with mestranol. With increased doses of mestranol, the LH/FSH after administration of LH-RH was increased and reached to the same level as the normal ovulatory peak in these 4 patients. Our results demonstrated that responses of pituitary gonadotropins to LH-RH in anovulatory women were significantly different in the LH/FSH from those in ovulatory women. Abnormality of these responses in anovulatory women seems to be unfavorable to ovulation.


Inhibition of insulin release by synthetic rat C-peptide II in isolated rat islets.

The effect of synthetic rat C-peptide on glucose-induced insulin release was studied in isolated islets of rat pancreas. Addition of rat C-peptide II to the medium inhibited glucose-induced insulin release from islets by 30 to 40 percent. Rat C-peptide I, human connecting peptide and bovine, porcine and canine C-peptides had no significant effects on insulin release from islets of rat pancreas.


The action pattern of amylomaltase from Escherichia coli.

Amylomaltase, the inducible 4-alpha-glucanotransferase of Escherichia coli strain ML, has been purified to homogeneity. Its specific activity with a commercial maltose substrate was 500 mkat/kg protein (30 mumol glucose formed min-1 mg protein-1). The purified enzyme, dependent on buffer concentration, exists in interconvertible low-molecular-weight (apparent molecular weight 71000) and high-molecular-weight (apparent molecular weight 370000) forms. The specificity of amylomaltase has been redefined. Hitherto, the enzyme was thought to be a glucosyltransferase, catalysing the transfer of single glucosyl units, and maltose has been regarded as its most important substrate. Amylomaltase is now shown to exhibit both glucosyl-transfer and 4-alpha-glucanosyl-transfer specificity. 4-alpha-Glucanosyl chains containing up to at least nine glucosyl units can be transferred. However, it is concluded that the transfer reaction by which amylomaltase action was originally expressed, does not take place, i.e., Maltose + maltose in equilibrium Maltotriose + glucose and that maltose has a restricted role as a substrate. This may be due to the inability of maltose to function as a donor substrate, serving only as an acceptor substrate. It is confirmed that when a maltodextrin serves as a donor, that portion of the molecule transfered by the enzyme is that containing the nonreducing-end-group. Enzyme action on chromatographically pure maltose is characterized by a lag phase in the time course of glucose release. The lag pahse is overcome by addition of 'priming' (catalytic) concentrations of maltotriose or higher maltodextrins. An autocatalytic reaction mechanism involving the generation of primer molecules is proposed to explain the action of the enzyme on maltose. The redefined action pattern of amylomaltase is consistent with the redefined role of the enzyme in the utilization of exogenous and endogenous 1,4-alpha-glucans by E. coli.


Protease I from Escherichia coli. Some physicochemical properties and substrate specificity.

Protease I, a periplasmic endopeptidase from Escherichia coli has been further purified by a modified procedure. While the purified protein consists of a single polypeptide chain of about 21000 daltons, its molecular weight in dilute salt solution was estimated to be near 43000, suggesting that the enzyme has a marked tendency to dimerize. It has only one disulphide bond and is very sensitive to urea. In agreement with previous evidence of a chymotrypsin-like specificity, hydrolytic assays of various p-nitrophenyl esters of N-substituted amino acids showed that phenylalanine and tyrosine derivatives are the best substrates for the enzyme. The Km(app) for N-benzoyloxycarbonyl-L-tyrosin-p-nitrophenyl ester at pH 7.5 In 100 mM sodium phosphate buffer at 25 degrees C was found to be 0.2 mM. In contrast to chymotrypsin, protease I is unable to hydrolyse N-acetyl-L-phenylalanine ethyl ester and its tyrosine analogue. Moreover, the enzyme appears devoid of amidase activity and exhibits a low activity upon polypeptides. At 37 degrees C, it cleaves the carboxymethylated B-chain of bovine insulin at four points: Phe25-Tyr26, Phe24-Phe25, Leu15-Tyr16 and Ser9-His10. From a detailed study of peptides bonds hydrolyzed, it was concluded that protease I has a stringent requirement for both residues forming the scissile bond, and appears to possess an extended hydrophobic binding site.


Bovine neurophysins I, II and C: new methods for their purification and for the production of specific antibodies.

1. Bovine neurophysins were prepared by a modified method, in which a Biogel P-60 column was used. This yielded two neurophysin fractions, the first containing neurophysin I and small quantities of the other neurophysins,the second containing neurophysin II and C, and only traces of neurophysin I. 2. Antibodies against neurophysin I, II and C were prepared by an original method, 5 mug in 100 mul water of each of the two fractions were applied on a gel slab and separated by iso-electric focusing in a pH gradient 4--6. The separated bands were visualized with 8-aniline-1-naphthalene sulfonic acid, magnesium salt and strips respectively containing neurophysin I, II or C were cut out. The neurophysin-containing strips were homogenized in complete Freund's adjuvant and injected into rabbits. 3. The specificity of the antisera were tested by immunocytochemistry and by radioimmunoassay. By this latter method, it was determined that cross-reactivity was less than 1%. The cross-reaction, observed with the immunohistochemical method could be eliminated by differential absorption. 4. It was found that neurophysin C antisera were undistinguishable from the neurophysin II antisera, while showing little cross-reactivity with the neurophysin I antisera. This suggests that in vivo neurophysin C is not a real neurophysin, or at least, that it is very similar to neurophysin II. 5. Highly purified bovic focusing method. By modifying a LKB Uniphor electrophoresis apparatus, the elute the proteins without switching off the voltage. The resolution of the technique is close to that offered by analytical gel iso-electric focusing.


Activities of protein-deficient particles derived from 50-S ribosomal subunits by NH4Cl/ethanol treatment.

Protein-deficient ribosomal particles obtained by treatment of 50-S subunits from Escherichia coli ribosomes with 1 M NH4Cl and 50% ethanol contain less than 3% of proteins L7 and L12 and about 7% of proteins L10 and L11. Proteins L1, L5, L8/9 and L25 are also released during the treatment but in amounts accounting for less than 40%. The particles are able to form peptide bonds in different systems, such as 'fragment reaction', puromycin reaction and formation of dipeptides. They also bind N-acetylphenylalanyl-tRNA and phenylalanyl-tRNA non-enzymically but are unable to support any of the elongation-factor-dependent reactions tested. However, when methanol is present, they display up to 20% of the control EF-G-dependent GTP activities such as GTP hydrolysis and formation of the ternary complex EF-G-GuoPP(CH2)P-ribosome. The first activity is totally sensitive to the antibiotic thiostrepton while the formation of the ternary complex is unaffected by the drug. When measured by equilibrium dialysis the core particles are shown to be able to bind radioactive thiostrepton. The results show that protein L11 is not an absolute requirement either for peptidyl transferase activity or for the binding of thiostrepton, although in the last case the protein strongly enhances the ribosome affinity for the antibiotic.


Accumulation of free ribosomal proteins S1, L7, and L12 in Escherichia coli.

The total content of free ribosomal proteins in the cells of Escherichia coli was determined to study the nature of intracellular accumulation during growth. Labeled ribosomes and post-ribosomal supernatant were prepared from exponentially growing and stationary-phase cultures. The fraction of free ribosomal protein in the supernatant was estimated by resolving both the acidic and basic proteins separately with two different techniques of two-dimensional gel electrophoresis. Free ribosomal proteins in the cell sap were identified on the basis of coelectrophoresis with authentic ribosomal protein markers, molecular weights and amino acid composition. Among the acidic proteins, S1, L7, and L12 were identified and examined in detail. All three proteins accumulated to significant levels in these cultures. Stationary-phase cells contained 2-4 times more free S1, L7, and L12 than midlogarithmic phase cells. Moreover, free S1, L7, and L12 and ribosome-bound forms were stable during exponential and post-exponential growth of cultures. At this growth transition, non-ribosomal proteins in the supernatant and those associated with the ribosomes showed different characteristics of accumulation. The ratio of L12:L7 in the supernatant did not exhibit a remarkable shift during the growth cycle like the ratio of L12:L7 in ribosomes. In addition, free L12 in the supernatant was not acetylated, although there was a rapid acetylation in the cells.


Active transport by membrane vesicles from anaerobically grown Escherichia coli energized by electron transfer to ferricyanide and chlorate.

Active transport of amino acids by membrane vesicles from Escherichia coli, grown anaerobically on glucose in the presence of nitrate, can be energized under anaerobic conditions by electron transfer in the nitrate respiration system with formate as electron donor and nitrate as acceptor. A high rate of amino acid transport is also obtained under anaerobic conditions by electron transfer from formate to the nitrate analogue chlorate or to the membrane-impermeable electron acceptor ferricyanide. Electron transfer from formate to nitrate results in the generation of an electrical potential as is indicated by the uptake of the lipophilic cation triphenylmethylphosphonium. Ferricyanide accpets electrons from at least two sites of the nitrate respiration system. One of these sites appears to be nitrate reductase, because cytochrome b, reduced by formate, is completely reoxidized by ferricyanide and glutamate transport energized by formate plus ferricyanide and formate plus nitrate are affected by the same electron transfer inhibitors. A second site of electron transfer to ferricyanide appears to be located prior to nitrate reductase in the nitrate respiration system, since formate is oxidized at a higher rate in the presence of ferricyanide than with nitrate while formate/ferricyanide energizes transport of amino acids at a lower rate than formate/nitrate. Moreover, electron transfer inhibitors block electron transfer from formate to nitrate to a significantly higher extent than from formate to ferricyanide. The effects of irradiation of the membrane vesicles with near ultra-violet light suggest that quinones play an essential role in the electron transfer from formate to nitrate or ferricyanide. Irradiation blocks completely formate-dependent nitrate and ferricyanide reduction and active transport driven by formate/nitrate and formate/ferricyanide, but has hardly any effect on the activity of formate dehydrogenase and on ascorbate/phenazine methosulphate/oxygen-driven transport. Similar effects of ferricyanide have been observed in membrane vesicles from E. coli, grown anaerobically in the presence of fumarate. In these membrane vesicles a high rate of lactose and triphenylmethylphosphonium uptake under anaerobic conditions is obtained by electron transfer from glycerol 1-phosphate to fumarate and also to ferricyanide and evidence has been presented for the involvement of cytochromes in these electron transfers.


Heart failure in experimental refractory shock.

Severe haemorrhagic shock was studied in a series of animal experiments. Reduction in myocardial oxygen and substrate utilisation in association with cardiac failure was demonstrated when the duration of shock was prolonged. The probable aetiological factors included ischaemia and the presence in the blood of certain vasoactive and metabolic depressant substances. It was concluded that the process leading to irreversible shock was multi-factorial in origin but failure appeared to play an important role.


The effect of a progressive decrease in the circulating blood volume of the dog on the transthoracic impedance.

The correlations between the haemodynamic and transthoracic electrial impedance changes resulting from a progressive reduction in the circulating blood volume were studied in four intact mongrel dogs artificially ventilated with a mixture of halothane in nitrous oxide-oxygen. The cardiac output of the dogs was measured by both the electrical impedance and the fibre optic dye dilution techniques. It was found that significant correlations existed between the blood lloss and the arterial blood pressure, the maximum first derivative of the transthoracic impedance, the Heather Index, the transthoracic impedance, the maximum rate of change of aortic pressure and the cardiac stroke work. There was also a good correlation between the dye and impedance cardiac output values, the impedance value always being higher than the corresponding dye value. The correlation between the Heather Index and the PEP/LVET ratio and 1/PEP2 varied markedly from dog to dog.


The use of nasal CPAP in newborns with respiratory distress syndrome.

The efficiency of applying continuous positive airway pressure (CPAP) by the nasal route was retrospectively nalyzed in 32 newborns with RDS (23 uncomplicated HMD with additional cardiac or pulmonary complications and 7 RDS of non-hyaline membrane etiology) who underwent nasal CPAP treatment at the Kinderspital Zurich from 1972--1974. 16 of the 23 infants with uncomplicated HMD were successfully treated with CPAP. They showed a significant rise in PaO2 as well as a significant drop in respiratory frequency during nasal CPAP application, the PaCO2 did not change significantly. The remaining 7 infants in this group (7/23) had to be intubated and mechanically ventilated owing to a persistent high FIO2 (4 infants), technical difficulties (1) or nasal hypersecretion (2). Two of these 23 infants died, one of meningitis, one of cerebral hemorrhage. The two infants with HMD and additional cardiac or pulmonary complications and 3 of 7 infants with RDS of non-hyaline membrane etiology had to be intubated and mechanically ventilated after failure of nasal CPAP. All 9 infants in these two groups survived. The nasal CPAP system as described is a simple, inexpensive and effective method of applying CPTPP in newborns with uncomplicated HMD, except radiological stage IV. In HMD with additional cardiac or pulmonary complications and in RDS of non-hyaline membrane etiology the results of nasal CPAP treatment were not convincing.


Correlation of PPD and BCG-induced leukocyte migration inhibition, delayed cutaneous hypersensitivity, lymphocyte transformation in vitro and humoral antibodies to PPD in man.

In a study of healthy human individuals a complete lack of correlation between the results of the agarose leukocyte migration inhibition (LMI) test, using purified protein derivative (of tuberculin) (PPD) and Bacillus Calmette Guérin (BCG) as antigens, and delayed cutaneous hypersensitivity and lymphocyte transformation in vitro to PPD was found. There was a reasonable correlation between PPD- and BCG-induced LMI. Antibodies to PPD proved to have no influence on PPD-induced LMI. Purified polymorphonuclear leukocytes, whether derived from donors sensitive to PPD in the agarose LMI test or from nonsensitive donors, did not show migration inhibition to PPD. It was concluded that polymorphonuclear leukocytes and lymphocytes need to be present simultaneously for migration inhibition of peripheral blood leukocytes by PPD. Furthermore, because a consistent relation with conventional parameters of cell-mediated immunity was lacking, it is doubtful whether the agarose LMI test can be considered as an alternative parameter of this kind of immunity.


Selective breeding of mice for antibody responsiveness to flagellar and somatic antigens of salmonellae.

Selective breeding has been performed in mice for their high or low antibody responsiveness to Salmonella antigens (Ag). Two noncross-reacting Salmonellae (S. typhimurium and S. oranienburg) have been alternately used to immunize successive generations in order to avoid interference of maternally transmitted antibody. Both Salmonellae carry two independent antigens: flagellar (f) and somatic (s). Two two-way selections were carried out: one for agglutinin response to fAg and the other for agglutinin response to sAg (Selection Ags). The agglutinin response to the other independent Ag was also measured (Associated Ags). The phenotypic character chosen for selection is the maximal agglutinin titer in response to optimal immunization. In both selections, high and low responder lines diverged progressively. There was a 27-fold interline difference in the F13 generation of selection for fAg response and a 38-fold one in the F11 generation of selection for sAg response. This divergence and the continuous distribution of phenotypes in the foundation population indicate that the quantitative character investigated is subject to polygenic regulation. The realized heritability (h2) of the character, measured from the interline divergence, was 0.18 +/- 0.06 in both f and s selections. This means that about 20% of the phenotypic variance of the foundation population is additive. The modification of immune responsiveness to the selection Ag produced by selective breeding was accompanied by a parallel and equivalent effect on immune responsiveness to the noncross-reacting Associated Ag. This effect is therefore entirely nonspecific. A small but definite sex effect in favor of female antibody responsiveness was demonstrated. The results are discussed in comparison with other selection experiments made in mice for quantitative antibody response to other antigens.


Dual pathway of B lymphocyte differentiation in vitro.

Direct visualization of the events resulting from LPS stimulation of mouse spleen cells in vitro was achieved by characterizing the cells during four days of culture for morphology, Ig and theta surface markers and autoradiography after [3H] thymidine uptake. The changes observed were related to biochemical parameters such as incorporation of [3H] thymidine into DNA, Ig biosynthesis and secretion. Two pathways of B lymphocyte differentiation were observed: a) the generation of a large number of small B lymphocytes with high density of surface Ig but no internal pool detectable by immunofluorescence, and b) the maturation of a very small proportion of cells with a large intracellular pool and the ability to secrete Ig. Both cell types arise from dividing blast cells, either physically separated or traced by pulse chase experiments with [3H] thymidine. We discuss whether this duality is caused by the triggering of different B cell subpopulations at different developmental stages, preprogramed to one or the other pathway or whether the final direction of development depends on the microenvironment of individual dividing cells.


Control mechanism of lymphocyte traffic. Altered distribution of 51Cr-labeled mouse lymph node cells pretreated in vitro with lipopolysaccharide.

The effect of in vitro treatment of mouse lymph node cell populations with lipopolysaccharide (LPS) on their migration in syngeneic recipients was studied. LPS-exposed cells had a decreased lymph node localization and an increased spleen distribution when compared to control untreated cells, after intravenous injection into syngeneic recipients. This effect of LPS on cell traffic was maximal at 24 h and could not be detected at either 1 or 72 h after cell transfer. These changes induced on cell distribution by LPS treatment are interpreted as the consequence of a modified interaction between the circulating lymphocytes and the resident cells within the spleen of the recipients.


Reduction of trapping of aggregated human IgG in germinal centers by mitogens.

The effect of nonspecific mitogens on the trapping of 125I-labeled aggregated human IgG (125I-AHGG) in germinal centers (GC) of mouse spleens has been investigated by both radioactivity uptake and immunofluorescence. Phytohemagglutinin and concanavalin A (Con A) significantly decreased trapping. Lipopolysaccharide produced less inhibition, and pokeweed mitogen had no significant effect. The maximum inhibition occurred with 250--500 mug Con A. This had no effect on 125I-AHGG uptake in liver kidney and blood. No differences were found between i.p. and i.v. routes of Con A injection. The effect of mitogens on the 125I-AHGG trapping in GC is due more likely to modification of the migratory properties of lymphocytes brought about by surface binding, than to their mitogenic properties, since Con A decreased 125I-AHGG localization in thymectomized, x-irradiated and bone marrow-reconstituted animals.


Effects of morphine on the turnover of brain catecholamines and serotonin in rats-acute morphine administration.

Morphine increased the rate of brain dopamine (DA) depletion when given before alpha-methyl-p-tyrosine (AMT) or alpha-propyl-dopacetamide, but not when given after AMT. No effect of morphine was found on the rate of depletion of brain noradrenaline (NA) or serotonin (5-HT) after the two synthesis inhibitors. The accumulation of homovanillic acid and 5-hydroxy-indoleacetic acid induced by probenecid was significantly increased by morphine pretreatment, whereas the accumulation of 3-methoxy-4-hydroxy-phenylglycol sulphate was not changed. These findings can be best explained by the hypothesis that morphine increases the non-functional intraneuronal catabolism of newly synthesized DA and 5-HT, without much effect on the monoamines already taken up in the synaptic vesicles. NA turnover does not seem to be changed by acute morphine administration.