[Experiences with selective proximal vagotomy in the treatment of chronic duodenal ulcers]. After 138 selective proximal vagotomies--130 of which were performed because of chronic duodenal ulcer--the rate of the postoperative complications was 3,8%, which is very low; the primary lethality was 0,76%. At re-examination 68% of the patients were without complaints; 8% had a recurrent ulcer. Epigastric fulness (6%), dumping (1%) and diarrhea (1%) were rare. Because of our results we believe, that selective proximal vagotomy is the optimal method for the surgical treatment of duodenal ulcer.


[Contact lenses: indications and limits]. Contact lens wear may be indicated for cosmetic, professional, optical or therapeutic reasons. For each of these indications the arguments are given, why and when contact lenses are better than spectacle correction and which possibilities there are. Contact lens wear is contraindicated in certain eye diseases and in the case of problems of handling and hygiene. The author's preferences concerning the different contact lens materials are summarized briefly.


[B-scan echography in eye and orbital diseases]. After a short description of ultrasound A- and B-scan-techniques, the ultrasonic findings in the different diseases of the eye and orbit are discussed. The results, as presented in this paper, show that ultrasound proves to be a useful aid in the evaluation and differential diagnosis of ocular pathology.


[Metabolic changes in the multiply injured. Biochemical studies in multiply injured patients with reference to the severity of the injury]. Leading physical sign of "trauma-disease" in polytraumatized patients is hypovolemic shock. Changes in metabolism have been described previously. At present there are no reports concerning the time at which these changes occur. The early posttraumatic phase is characterized by normosodiemia and initial transient hypopotassemia, which is based on the renin-angiotensin-mechanism. The metabolic acidosis depending on trauma causes decreased oxygen perfusion of tissue, which possibly is found even before alterations of circulation are detected. Already at the site of the accident hyperglycemia and hyperglucagonemia with normal values of insulin were found. The increase of blood glucose was correlated to the values of adrenalin and to the insulin antagonist glucagon. The catabolism of proteins closely related to the degree of trauma continued as long as 5 days after injury. Analysis of the blood clotting system showed that in the early phase after trauma there are changes in hemostatic potential consisting in hemostatic defect. In several essential points the early posttraumatic phase of metabolism is different from that in a later phase. These facts should be kept in mind if planning therapy.


[Erythropoietic protoporphyria].

[Erythropoietic protoporphyria]. Erythropoietic protoporphyria (EEP) is the most frequently found erythropoietic porphyria in men. This inborn error of heme metabolism with autosomal dominant mode of inheritance is based on a deficiency of ferrochelatase. This defect leads to an increase for protoporphyrins predominantly in the red cells. Under the influence of sun light, especially UV-A, photohemolysis occurs and the content of protoporphyrin in the tissue increases. This is associated with acute sun burn like skin lesions, which usually clear up completely. Persistent skin lesions are seen in form of hyalinosis like infiltrations of the most intensively light exposed skin areas in some patients. Associated symptoms might be: gallstones or rarely a cirrhosis of the liver. Treatment with photoprotective ointments and especially carotinoids (beta-carotene, canthaxanthine) is very effective and only in some rare cases this symptomatic therapy fails. Prognosis of the disease is good. Only a few patients died of the associated hepatic failure.


[Pre- and postoperative clearance studies in the individual kidneys in juvenile kidney surgery]. In 1978 43 patients have been operated in the Department of Pediatric Surgery, Krankenhaus München-Schwabing, because of recurrent urinary-tract infections with ureterovesical reflux or obstructive urinary-tract diseases. In all patients a split whole body clearance-study with a method modified after the method of Mühle with 131J-Hippuran was carried out pre- or postoperatively, respectively. In 78,5% of these cases the clearance-results were corresponding to the results of x-ray examination; 6 of the remaining patients had a worse clearance result and 3 had a better result than the x-ray findings would have suggested.


Response of plasma somatostatin-like immunoreactivity to the administration of alloxan in dogs. The present study was designed to examine the effects of intravenously injected alloxan (75 mg/kg) upon plasma somatostatin-like immunoreactivity (SLI), glucagon (IRG), insulin (IRI) and glucose levels in 6 dogs. Within 2 hours of the injection of alloxan, SLI and IRI levels decreased significantly below their respective baselines, while IRG and plasma glucose concentrations increased. At 8 hours SLI levels had increased significantly by 55 pg/ml, together with a rise in IRI and a decrease in IRG and glucose concentrations. After 24 hours, marked hyperglycemia and hyperglucagonemia had developed whereas SLI levels were not different from preinjection values.


In vitro effects of fatty acids on the actions of serum on rat and pig cartilage. It has been reported that fatty acids preferentially inhibit serum-stimulated incorporation of sulfate by embryonic chick cartilage, suggesting that they may interfere with the effects of a proposed mediator (serum somatomedin) of the actions of growth hormone (GH). This was studied further in mammalian cartilage. Butyrate and octanoate at concentrations of 0.5 to 5 mM produced a concentration-dependent inhibition of both basal and serum-stimulated sulfate and thymidine incorporation by costal cartilage from hypophysectomized rats. Butyrate also inhibited basal and serum-stimulated sulfate incorporation in cartilage from normal pigs and normal sucklings rats. In all 3 test systems, oleate (0.2--5 mM) bound to serum albumin (4 g/dl) was ineffective. There was no evidence that fatty acids preferentially inhibited the stimulation of sulfate incorporation produced by serum.


Effect of metabolic control on urinary excretion and plasma levels of catecholamines in diabetics. Urinary excretion and plasma levels of catecholamines were determined in 20 normal and 39 diabetic subjects to evaluate the sympathetic activity. Diabetic patients were divided into 4 groups according to the metabolic control. Sympathetic activity showed no differences between normal and subjects with chemical diabetes (group I, n = 5). In insulin-treated diabetics in good metabolic control (group II, n = 11) only urinary excretion of free norepinephrine was significantly higher than normals (p less than .05). In insulin-treated diabetics in poor metabolic control (group III, n = 16) urinary excretion and plasma levels of norepinephrine showed a marked increase over groups I and II (p less than .001). In insulin-treated diabetics with ketosis (group IV, n = 7) urinary excretion and plasma levels of both norepinephrine and epinephrine showed the highest values (p less than .001 and less than .1). Finally, in groups III and IV, after achieving improved metabolic control, a significant decrease of urinary excretion and plasma levels of catecholamines was observed. The results confirm that there is an increased rate of catecholamine release in poorly controlled diabeties and suggest a close correlation between sympathetic activity and metabolic derangement in diabetes.


Cholesterol and triglycerides of the liver after administration of a chromatographic fraction of thymus: variations in tissue and blood. In rats fed on a fat diet and injected with a chromatographic thymus fraction, intrahepatic distribution of cholesterol and triglycerides have been studied. The results show that the hematic decrease of these compounds, always observed in previous works, is accompanied by a smaller infiltration in hepatocytes and the lipidic drops are more finely dispersed. These results lead us to suppose that liver is a target organ of the thymus fraction.


rT3 production in normal man, assessed from variations in serum rT3 during short-term rT3 infusion. The metabolic clearance rate (MCR) of 3,3'5'-triiodothyronine (reverse T3, rT3) was estimated in normal human subjects by a modified noncompartmental method using the integrated increase in serum rT3 following intravenous infusion of 0.10 nmol/min rT3 for 4 hr. The MCR-rT3 was calculated to be 102.8 +/- 17.01/day and the daily rT3 disposal to be 33.0 +/- 9.5 nmol (mean +/- SD, n = 6). The MCR-rT3 compares well to that of previous studies employing tracer kinetic methods. The disposal rate of rT3 estimated in the present study is considerably lower than found in some previous studies. The discrepancy is due to differences in the measured levels of serum rT3 in normal subjects.


Effect of glomerulopressin on ovarian blood flow in dogs. The present study showed that glomerulopressin decreased ovarian blood flow in normal dogs and that this effect could be inhibited by treatment with indomethacin. In addition diabetic dogs had a high plasma level of glomerulopressin and a low ovarian blood flow that was shown to increase after treatment with indomethacin. This suggests that the low ovarian blood flow in diabetis might be due to a prostaglandin synthesizing effect of glomerulopressin.


Effects of calcium deprivation and orchidectomy on bone composition in the rat. The effects of calcium deprivation and of orchidectomy, separately and combined, on body growth and bone composition have been examined in mature male rats. Calcium deprivation had no significant effect on the rate of body growth but femoral bone weight, bone ash weight and total calcium and phosphorus contents of the femora were reduced while the water content increased. However, there were no significant changes in the ratio of bone weight to body weight, in the amounts of calcium or phosphorus per unit weight of bone, or in the ratio of calcium to phosphorus in the bone. Orchidectomy reduced the rate of body growth, the mean weight of the femora and the ratio of bone weight to body weight. There was a 60% decrease in bone water and a 30--35% reduction in the calcium and phosphorus content of the femora but the amounts of ash, calcium and phosphorus per unit weight of bone fell by only 6--10%. The combination of orchidectomy and calcium deprivation had no greater effect on body weight, bone weight or bone composition than had orchidectomy alone. It is concluded that calcium deprivation and orchidectomy both cause osteoporosis in mature rats but the effects of orchidectomy are more severe than those of calcium deprivation and there appears to be some demineralization of the bone remaining after orchidectomy.


Elevated hypothalamic norepinephrine content in mice with the hereditary obese-hyperglycemic syndrome. There is evidence that hypothalamic norepinephrine (NE) plays a role in the control of appetite in the rat. Using specific and sensitive radioenzymatic assays, we determined if there was a difference in the tissue (hypothalamus, cerebral cortex and kidney) concentration of NE or of dopamine (DA) in mice with the hereditary obese-hyperglycemic syndrome (ob/ob) and their normal weight littermates, both when they were in the rapid growth phase (2--3 months of age) and when they were mature (6--7 months of age). The concentration of NE was similar in the cerebral cortex of obese and normal mice and in the kidneys of obese and normal mice. The concentration of DA was similar in the hypothalamus of obese and normal mice. The concentration of DA was similar in the hypothalamus of obese and normal mice and in the cerebral cortex of obese and normal mice. These observations support the concept that alterations in hypothalamic NE may play a role in the obesity of ob/ob mice.


The fluorescence scanning of microgels stained for glycoproteins with fluorescein isothiocyanate-labelled concanavalin A. A technique is presented which allows one to label and quantitate glycoproteins. Small amounts of protein from biological samples (0.5-2.5 microgram for mixtures and less for individual proteins) are separated by sodium dodecyl sulfate gel electrophoresis on 1-30% polyacrylamide gradient microgels. The gels are stained with Co-omassie Brillant Blue R250 to evaluate relative migration or fixed in 2-propanol/acetic acid and stained with fluorescein isothiocyanate-labelled concanavalin A. The microgels are then scanned using a fluorescence microscope controlled by a computer, although simpler configurations are possible. Standards of known carbohydrate composition (e.g., glucose oxidase) are used for comparative purposes. Glycoproteins in the order of 5-30 ng protein (or 1-5 ng carbohydrate) can be detected without difficulty. This technique may prove valuable in evaluating glycoproteins when the available material is limited.


Separation of the gonadotropic activity of crude choriogonadotropin from the inhibitory effect on lymphocyte transformation. The effect of choriogonadotropin of different purities on the transformation of peripheral human lymphocytes was studied. Various crude hormone batches inhibited lymphocyte transformation in a dose-dependent manner, both in the mixed lymphocyte reaction and in the phytohemagglutinin-induced stimulation. The inhibitory activity, however, was found not to be correlated with the gonadotropic activity of the crude hormone batches (2660-4300 IU/mg). Choriogonadotropin (13 000 IU/mg), which was purified in 3 steps, showed no inhibitory effect except at high doses (greater than 5000 IU/ml final dilution). More detailed investigations provided evidence that in the first step of the choriogonadotropin purification procedure (batch adsorption of crude choriogonadotropin on SP-Sephadex C-50), the inhibitory activity can be enriched in a fraction (Fract. I) which displays a very low gonadotropic activity (less than 500 IU/mg). A further separation of Fract. I was achieved by isoelectric focusing as well as by chromatography on DEAE-Sephadex A-25. By these means, the inhibitory potency could be enriched more than 100-fold. The substances which display inhibition of DNA synthesis in lymphocytes were proven to act in a nontoxic way. A preliminary characterization of the strongly inhibiting substances which show a dose-dependent suppression of lymphocyte transformation by about 99%, showed that this effect is probably exerted via non-dialysable sialoglycoproteins. By a fourth purification step entailing a chromatography of purified choriogonadotropin (13 000 IU/mg) on SP-Sephadex C-50, a highly purified choriogonadotropin (14000 IU/mg) could be obtained which showed no inhibitory effect on lymphocyte transformation (in both mixed lymphocyte reaction and in phytohemagglutinin-induced stimulation) up to a dose of 43 000 IU/ml. The components which were removed from choriogonadotropin in this step seem to be immunologically identical with the strongly inhibiting substances isolated by isoelectric focusing. These investigations demonstrate that biologically active, highly purified choriogonadotropin is unable to inhibit lymphocyte transformation. The inhibitory activity of crude hormone can be enriched in choriogonadotropin-free fractions. Therefore, it is concluded that the inhibitory activity of crude hormone is not a property of choriogonadotropin itself.


ATP citrate (pro-3S)-lyase in the rat; two-step purification procedure, properties, organ distribution. A simple two-step procedure is described for the purification of rat liver ATP citrate lyase with an overall yield of upt to 70%. In binding experiments on Blue Sepharose CL-6B (an affinity gel for nucleotide-specific enzymes) it is observed that ATP citrate lyase recognizes ATP and CoA as well as ADP, 3'-AMP, and NADP, but not 5'-AMP, NAD, or adenosine. It is further demonstrated that ATP citrate lyase is activated by various anions including chloride, hydrogen carbonate, and acetate. The highest specific activity of the enzyme is found in adipose tissue of various origins. Activities are also present in the adrenals, in liver, and intestine, in brain, and in lung. No measurable enzyme activity is found in heart, skeletal muscle, and in the kidneys.


Preparation and characteristics of porcine growth hormone. A recently published method for preparation of porcine growth hormone resulted in a highly purified protein with good biological activity. However, after storing for some months the originally homogeneous hormone separated again into several fractions when rechromatographed on ion exchange columns. The biological activity, found in one of these fractions, was clearly diminished compared with the activity of a freshly prepared hormone. In the present paper a modified procedure is described for the isolation of a more stable porcine growth hormone. The influence of ions, involved in buffers of the same molarity and the same pH, upon ion exchange chromatography of porcine growth hormone is discussed. The purified hormone shows high biological activity in the tibia test and is free of activities of other pituitary hormones. The molecular weight is about 20 000; only phenylalanine is found as N-terminal as well as C-terminal amino acid; the amino acid composition resembles neither that of porcine growth hormone described in literature nor that of human growth hormone.


[The primary structure of a dimeric hemoglobin (erythrocruorin); component CTT VI of Chironomus thummi thummi, Diptera (author's transl)]. The complete amino acid sequence of 147 residues was determined automatically for a major dimeric component (CTT VI) of the insect larva Chironomus thummi thummi (Diptera). The molweight was found to be 32411. All tryptic, maleylated tryptic and cyanogen bromide peptides were isolated. The handling of some large fragments was facilitated by maleylation and subsequent ion exchange chromatography. Some details of the primary structure are discussed. The alignment of the amino acid sequence with that of human alpha-chains shows only 29 identical positions.


The myth of advocacy.

The myth of advocacy. In an era in which advocacy has become a buzzword, both psychiatry and the legal profession have climbed aboard the advocacy bandwagon. Yet the American Psychiatric Association's notion of advocacy--championing the medical needs of patients--is often in direct conflict with the lawyers' notion of advocacy--championing the legal rights of their clients. The author observes that psychiatry has proved to be a weak adversary for patients' legal advocates; the result has been a one-sided advocacy system that has advanced patients' rights at the expense of their needs. He believes that if the APA is to become an effective advocate for patients, it must hire lawyers and work with them to reverse the trend of turning rights into needs.


The creative arts therapies as "real therapies". Elements of a standard definition of psychotherapy are used to support the argument that the creative arts therapies should not be characterized as adjunctive therapies, or discredited as not being "real therapies." Two concepts widely acknowledged as important in the application of the creative arts therapies are discussed: first, that the nonverbal media employed by creative arts therapists tap emotional rather than cognitive processes and evoke responses more directly and immediately than traditional verbal therapies, and, second, that creative arts therapies are reality-based and provide a more immediate and real link to a patient's experience than something he can portray only verbally.


Strategic planning has market orientation. A key element in the strategic planning process is active involvement of the hospital's key personnel so that each has a commitment to the ultimate plan.


Accounting principles, reporting rules, and payment practices. Misconceptions concerning the distinction between accounting principles, reporting rules, and payment practices and how they interrelate can lessen the effectiveness of hospital financial managers in these areas/clarification and recommendations are offered.


On call is not enough: social workers in ED. A study indicates that ED staff would refer to hospital social workers the increasing numbers of patients who need their services if, and only if, they are on duty in the ED itself.


New CHA head charts assertive future course. Interview by Alan L. Appelbaum. John E. Curley Jr. is determined that the Catholic Health Association will be a strong advocate in the defense and expansion of the work of Catholic health care institutions.


It's a question of ethics.

It's a question of ethics. Ethical considerations must be taken into account whenever a merger of the closing of a facility or service is contemplated/Some guiding principles that can help in making decisions.


Sharing management skills.

Sharing management skills. The SHARE program brings administrators, state hospital association officers, and others together in an effort to strengthen association work.


Independent congressman makes waves. Interview by Karen Rittinger. Representative Gramm's opposition to the Administration's Hospital Cost Containment bill and the National Health Plan Act reflect his independence of Democratic Party politics.


Metabolic support: how to form a service. Organizing a metabolic and nutritional support service requires consideration of issues such as departmental affiliation, staffing, and budgeting.


How to evaluate a PR program.

How to evaluate a PR program. A missing element in public relations programs is evaluation. This article describes how to evaluate the effectiveness of a program by developing measurable goals and objectives.


Cost cap bill topped agenda in first session of 96th Congress. Congress took up amendments to Medicare and Medicaid programs, extension of health planning law, funding for nurse training, and other issues during its first session.


Public press highlights clinical innovations. After years of what seemed to be a single minded concentration on cost issues, the public press in 1979 showed interest in clinical innovations and advances in hospital care.


Creativity in planning: prerequisite for success and survival. For successful planning, an organization and its planners must recognize and accept inevitable changes; eliminate unnecessary self-imposed restrictions that limit options in seeking solutions to problems; and take risks to promote future development.


Many benefits possible with vertical system. Although legal and practical barriers still remain before vertical integration of health care facilities can reach full potential, this kind of multi-institutional arrangement deserves greater attention.


Regionalized health systems in five nations. As shown by the health care systems in five industrialized nations, the method of national medical care financing determines the extent of regionalization and the degree of hospital's autonomy.


What to do when organized labor comes calling. Many hospitals are finding that personnel management skills per se are not enough to successfully manage their employee relations functions.


Workshops enhance role of volunteers.

Workshops enhance role of volunteers. Educational workshops were developed to enrich volunteers' services to a large metropolitan hospital. Staff also benefit by acquiring useful input from the volunteers.


Staff training vital for risk management. To ensure an effective risk management program, a hospital's staff must be trained to deal with a variety of continuous and special risks.


Using financial ratio analysis to compare hospitals' performance. Financial ratio analysis allows a hospital to evaluate its own performance over time and to compare itself with other hospitals. Through reclassification procedures, potential distortions are reduced, and administrative decisions can be based on more reliable rations. Step 1 reclassifies financial statements for analysis, step 2 computes and explains ratios, and step 3 combines ratios into patterns for interpretations. Step 4 describes the 209-hospital sample, step 5 compares three individual hospitals' ratios to industry ratios, and step 6 discusses behavior of two hospitals' ratios and industry ratios over time.


Holy Cross Hospital, Chicago: cost variance analysis in a health care setting. By knowing and understanding basic cost accounting techniques, administrators are better equipped to keenly judge their own institution's health care expenses. Managers who implement cost variance analysis are less likely to be forced into a defensive position with either board members or the general public.


Antenatal diagnosis of a de novo reciprocal translocation 46,XX,t(3;7)(q21;q11). A de novo reciprocal translocation 46,XX,t(3;7)(q21;q11), detected at amniocentesis performed because of advanced maternal age, is presented. Both parents showed a normal karyotype. The girl was delivered and has shown no phenotypic abnormality in the first year after birth. Problems encountered with the prediction of the fetal outcome in balanced structural rearrangements are discussed.


Partial duplication of the long arm of chromosome 5: a case due to balanced paternal translocation and review of the literature. A partial duplication of the distal segment of the long arm of chromosome 5 (q31 leads to qter) was observed in an infant with congenital malformations and dysmorphic features. The phenotypically normal father had a balanced translocation between the long arm of chromosome 5 and the short arm of chromosome 9: 46,XY,t(5;9)(q31;p24). The clinical and cytogenetic data obtained from six patients with partial duplications of two different long arm segments of chromosome 5 suggest that partial duplication of the distal long arm of chromosome 5 is associated with microcephaly, hypertelorism, epicanthus, strabismus, large upper lip, low-set, dysplastic ears, in addition to growth and psychomotor retardation. Partial duplication of the proximal part of the long arm of chromosome 5, on the other hand, is associated mainly with musculoskeletal abnormalities including muscle hypotrophy and hypotonia, scoliosis, lordosis, pectus carinatum, cubitus valgus, and genu valgum, in addition to psychomotor retardation. The dysmorphic features in this latter group include a bulging forehead, short nose, thick upper lip, low-set protruding ears and tapering, thin fingers.


Ring chromosome 15 syndrome.

Ring chromosome 15 syndrome. Two new cases of ring chromosome 15 are reported. A review of the nine cases described in the literature shows that ring chromosomes 15 are associated with a rather uniform phenotype characterized by slight to moderate mental retardation, marked pre- and postnatal growth failure, triangular face, and short hands and feet.


Presumptive long arm deletion of chromosome 8: a new syndrome? This communication describes an infant with growth and psychomotor retardation and severe congenital malformations, who was found to have an interstitial deletion of the long arm of chromosome 8: 46,XY,del(8) (q13q22). Comparison with the only other previously reported patient with a deletion of a similar chromosomal segment suggested that deletion of the long arm of chromosome 8 may constitute a clinically recognizable syndrome.


Partial trisomy 4 resulting from a complex maternal rearrangement of chromosomes 2, 4, and 18 with interstitial translocation. A woman presented a complex chromosome rearrangement with translocation between chromosome 2 and 4 in addition to an insertion of the band 4q12 leads to q13 in the long arm of chromosome 18. The authors present a case study of the daughter who displayed the abnormal chromosome 18 and trisomy of band 4q12 leads to q13.


Partial trisomy 11q as the result of sporadic translocation. Partial trisomy 11q due to a sporadic translocation was found in a mentally retarded girl with multiple abnormalities. The proportion of sporadic translocations involved in the total incidence of partial trisomies is discussed.


Trisomy 6p22 leads to 6pter due to familial t(6;13)(p22;q34 or 33) translocation. A newborn with a 46,XY,der(13),t(6;13)(p22;q34 or 33)pat karyotype, trisomic for the 6p22 leads to 6pter segment and, perhaps, monosomic for the 13q telomere, is reported. The balanced translocation is familial and was also encountered in the sister and paternal grandmother. The infant's phenotype was similar to that described in seven previously reported cases of partial trisomy 6p and further supports a partial trisomy 6p syndrome as proposed by Breuning et al.


Absence of chromosome breakage in patients with retinoblastoma. Mixed lymphocyte cultures were employed to assess the degree of spontaneous chromosome fragility in patients with retinoblastoma. There was no difference between the patients and their controls. If chromosome instability plays a role in the inherited tumour, more sensitive methods need be employed to elucidate it.


Detection of rare major genes in lipid levels. A statistical test of polygenic inheritance (TPI) against the alternative of a rare major gene is presented. It is designed for a random sample of quantitative observations on index cases and siblings of those index cases (probands) selected on the basis of the observed measurements of these probands. The test focuses on an increase of the variance of siblings of probands over its value under polygenic inheritance, such an increase being expected in the presence of a major gene producing a shift of the quantitative observations. Certain data on lipids are then analyzed by this test. A major gene can tentatively be confirmed for triglycerides but not for cholesterol. In addition, the values of all index cases are subjected to an analysis of a mixture of normal distributions (NOCOMP computer program), resulting in a significant second component for triglycerides but not for cholesterol. For both TPI and NOCOMP, the exponent in a power transformation is estimated by maximum likelihood simultaneously with all other parameters, so that these analysis methods are robust against a wide range of skewness in the data, which is demonstrated by manipulation of the observations and their reanalysis.


Frequency of sister chromatid exchanges in a balanced reciprocal whole-arm translocation. The frequency of sister chromatid exchanges (SCEs) in the centromere of chromosomes involved in a whole-arm translocation t(1:19) was evaluated in altogether 911 metaphases of translocation carriers (n = 5) and of normal controls (n = 6). Comparison of the two groups reveals no significant differences in the SCE rate (x2 = 3.06, nf = 1). The question as to whether the possible increase of the SCE rate at the translocation point could be detected by light microscopy is discussed. Parameters included in the discussion are the ratio of the SCE frequency at the translocation point to the SCE frequency at any of the possible breakage points in the centromeric region and the number of possible breakage points in the centromeric region.


Condensation of all human chromosomes in phase G2 and early mitosis can be drastically inhibited by 33258-Hoechst treatment. Condensation of human chromosomes in phase G2 and early mitosis is inhibited by the fluorochrome 33258-Hoechst. This inhibitory effect is most apparent in primary diploid fibroblasts and lymphoblasts and least pronounced in peripheral blood lymphocytes. Condensation of the human Y chromosome, which contains a large heterochromatic region rich in A-T base pairs, is drastically inhibited by 33258-Hoechst treatment of fibroblasts and lymphoblasts. The difference in sensitivity of human chromosomes in different cell types to 33258-Hoechst probably reflects differences in the cell-membrane permeabilities to 33258-Hoechst.


Loss of adjuvanticity in rats for the hyperacute form of allergic encephalomyelitis and for reaginic antibody production in mice of a phenotypic variant of Bordetella pertussis. The adjuvanticity of a phenotypic (C-mode) variant of B. pertussis, known to be deficient in certain immunological and physiopathological properties, was compared to that of the normal (X-mode) strain. The X-mode vaccine was a potent adjuvant for induction of hyperacute experimental allergic encephalomyelitis to guinea-pig spinal cord in Lewis rats whereas C-mode vaccine was inactive. X-mode vaccine was also highly active in the induction of reaginic (both IgE and IgGl) antibodies to ovalbumin in mice while C-mode vaccine caused only a transitory increase in the IgE level. These data support the view that an adjuvant component of B. pertussis, which is probably identical with the histamine-sensitizing and leukocytosis promoting factor, is much diminished in C-mode cells while the lipopolysaccharide adjuvant remains unchanged.


A new approach to the thermodynamic study of ABO antibodies. Enthalpy change was determined for natural anti-A (B, O subjects) and anti-B (A1, A2, O subjects). Entropy change, free energy change and association constant were calculated according to the law of mass action and the Wurmser method. The concentrations of allohaemagglutinins were measured by the Wilkie and Becker method using an autoanalyser. 2-Mercaptoethanol was used to estimate the proportions of IgG and IgM and their respective contribution to the thermodynamic properties. The following results and conclusions were obtained. Individual enthalpy and entrophy changes are different for each subject so that only the average values of these thermodynamic parameters represent a characteristic of the phenotype. There is a correlation between enthalpy and entropy changes and the relative proportion of anti-A or anti-B IgG. There is heterogeneity of the values of association constant. Free energy change is about 10 kcal mol-1 for all anti-A and anti-B; this result confirms the low energy binding between antigen and antibody. All these results confirm the role of environment and red-cell phenotype in the synthesis of allohaemagglutinins.


Cell electrophoretic studies on mouse lymphocytes treated with concanavalin A: possible existence of two types of receptors on cell surface. Concanavalin A (Con A) induced biphasic changes in the electrophoretic mobility (EPM) of splenic lymphocytes of AKR mice. At low concentrations of Con A (2-12.5 mug/ml) there was a significant increase in there was a significant increase in the EPM which could be attributed to the redistribution and endocytosis of Con A receptors. At higher concentrations (greater than or equal to 15.0 mug/ml) a reduction in EPM below that of untreated cells was possibly due to the binding of excess Con A after redistribution. The biphasic profile of EPM was observed only in respect of Con A-treated T lymphocytes. The EPM of B lymphocytes, despite binding of Con A, remained unaltered at all concentrations of Con A. On the basis of these data, the existence of two types of Con A receptors on the cell surface is proposed and their status vis-a-vis the redistribution is discussed.


Time kinetics of the interaction of concanavalin A with splenic lymphocytes of normal AKR mice. Interaction of concanavalin A (Con A) with splenic lymphocytes of normal AKR mice led to concentration-dependent biphasic changes in electrophoretic mobility. Kinetics of this interaction suggested that at high concentration Con A interacted in two steps. The first step led to redistribution of Con A receptors and increased EPM. In the second step additional receptors to Con A became accessible and their interaction with Con A reduced surface charge density. The kinetic data thus provide additional evidence for the existence of two sets of qualitatively different receptors for Con A on AKR lymphocytes.


Viral protein sythesis by tissues from avian leukosis virus-infected chickens. II. Effect of passive neutralizing antibody in normal and agammaglobulinaemic chickens. Synthesis in vitro of avian leukosis virus (ALV) group (gs proteins, p27 and p12, by various tissues from chickens infected within a few days after hatching was studied by means of autoradiography of immunoelectrophoretic patterns. Viral protein was synthesized in all tissues of chicks examined between days 18 and 50 of age after which time liver, kidney, bursa, thymus, and spleen became negative. The lung and genital organs of the chicks, however, continued to synthesize viral protein up to 100 days of age, when the experiment was ended. Repeated injections of neutralizing chicken antibody to ALV (Ab) starting on day 26 or 37 caused gs protein to decrease in spleen, liver, and thymus within 5 days but not in bursa, lung, and genital organs. Agammaglobulinaemic (Agamma) chickens showed prolonged persistence of gs protein synthesis in the spleen and liver; this synthesis was abrogated by passive Ab. Liver from Agamma chickens, however, also became negative without Ab treatment. The relative roles of antibody and cellular immunity in influencing ALV replication during the initial phase of infection before lymphoma development are discussed.


Effect of Ascaris suum and other adjuvants on the potentiation of the IgE response in guinea-pigs. The potentiation effect of various adjuvants on the production of guinea-pig IgE was investigated using Freund's complete and incomplete adjuvant, the lipopolysaccharide of Escherichia coli and Salmonella typhosa, Bordetella pertussis, and the nematodes Nippostrongylus brasiliensis and Ascaris suum. While all the antigens had a variable effect on the potential of the IgG response, only infection with A. suum resulted in an enhanced IgE response to the antigen, egg albumin. Maximum potentiation occurred when primary immunization and nematode infection were accomplished simultaneously.


The role of Staphylococcus aureus cell-wall peptidoglycan, teichoic acid and protein A in the processes of complement activation and opsonization. The role of cell-wall peptidoglycan, teichoic acid and protein A in the processes of Staphylococcus aureus complement activation and opsonization was investigated. CH50 consumption studies reveal that, although all cell-surface fractions were capable of activating the classical C pathway, only peptidoglycan consumed C via the alternative pathway. Using a quantitative immunofluorescence assay, peptidoglycan was shown to bind C3 molecules via the classical as well as via the alternative C pathway and in the absence of IgG and IgA class antibodies. C activation via the classical and the alternative pathway could be distinguished by kinetic analysis. By comparing the rates of staphylococcal C consumption, C3 fixation and opsonization it was found that the CH50 consumption assay is a relatively insensitive method and may yield results that do not necessarily reflect the process of bacterial opsonization.


Isolation of serum amyloid P-component (protein SAP) in the mouse. Serum amyloid P-component (protein SAP) was associated from mouse serum and ascitic fluids by calcium-dependent affinity chromatography on Sepharose 4B (Pharmacia) followed by gel filtration on Ultrogel AcA44 (LKB) in the presence of EDTA. It was homogenous on gradient polyacrylamide gel and sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis, and induced a monospecific antiserum in the rabbit. The related protein, C-reactive protein (CRP) was present in only trace amounts not exceeding 2 mug/ml of serum even during acute phase reactions. It was isolated by calcium-dependent affinity chromatography on insolubilized pneumococcal C-polysaccharide. The availability of these isolated mouse proteins and their respective antisera will facilitate investigation of their function.


A quantitative method for measuring in vitro synthesis of IgA and IgG by human rectal mucosa: studies on normal controls and patients with hypogammaglobulinaemia. A quantitative technique has been developed for measurement of immunoglobulin production by human rectal mucosa in vitro. The technique overcomes the problem of serum proteins trapped in the tissue by parallel measurements of Ig and human serum albumin (HSA) over a 6 day period. IgG, IgA, IgM and HSA were measured in supernatant fluids using sensitive radioimmunoassays. The technique has demonstrated IgG and IgA synthesis by rectal mucosa in vitro. The conclusion that the observed IgA and IgG was synthesized in vitro was supported by the demonstration that the production could be increased by pokeweed mitogen and blocked by emetine. This culture system has been applied to measure Ig synthesis by the rectal mucosa of immunodeficient patients.


The genetic control of antibody affinity. Evidence from breeding studies with mice selectively bred for either high or low affinity antibody production. The genetic control of antibody affinity has been studied in mice selectively bred on the basis of the affinity of antibody they produce to protein antigens injected in saline. Two lines of mice have been obtained, one producing predominantly high and the other predominantly low affinity antibody. Breeding experiments have been performed with these two lines after ten generations of selection and the level and affinity of antibody to protein measured in parents, F1 hybrids and backcross offspring. The results indicate that antibody affinity is a genetically controlled parameter of the immune response and that this control is exerted independently of that controlling antibody levels. Furthermore, high and low affinity line mice have been typed for major histocompatibility complex antigens and the results show that the two lines are not significantly different. This therefore suggests that genes controlling antibody affinity are not linked to the major histocompatibility locus.


Reactivity of smooth-muscle antibodies with F- and G-actin. The reactivity of human smooth-muscle antibodies (SMA) with F- and G-actin was investigated by means of an enzyme-linked immunosorbent assay (ELISA). Polystyrene tubes were coated with F-actin, G-actin and tropomyosin. The optimal reactivity of SMA with F-actin was achieved with tubes coated with actin concentrations in the range of 10-100 mug/ml, while both lower and higher amounts yielded lower reactivity. The reactivity with G-actin was low and only a little higher than the reactivity with tropomyosin or uncoated tubes, but much lower than the reactivity with F-actin. The differences in reactivity of SMA with F- and G-actin could not be explained by differences in binding of the two forms of actin to the tubes, although 1.7-2.5 times more F-actin than G-actin bound to the polystyrene surface after coating with equivalent amounts of protein. SMA-negative sera did not react with F-actin coated tubes indicating that the binding of antibody was not due to non-immunological binding of immunoglobulins. Human SMA thus react better with F- than with G-actin.


Information carried by the DNA released by antigen-stimulated lymphocytes. Both antigen-stimulated and non-stimulated human blood lymphocytes release in vitro a DNA-containing complex which is not the product of dying or disintegrating cells. Lymphocytes obtained from different PPD or HBs positive or negative donors were incubated with one of these antigens and the DNA released in the culture medium was tested for its information content using, successively, two cell-free systems. The ability of the resulting protein product to bind specifically to the stimulating antigen was examined by immunoadsorption chromatography. Results show that DNA excreted by stimulated lymphocytes was transcribed into an RNA which coded for an antigen-binding protein, whereas DNA released by unstimulated lymphocytes did not. The protein produced in this system, using as template the DNA released after cell stimulation, bound specifically to PPD or HBs Sepharose 4B coated columns, depending on the stimulating antigen and on the cell response to this antigen. After elution from the column the protein sedimented at 19S in a linear sucrose gradient.


Antibodies to the acetylcholine receptor in myasthenic dogs. Antibodies to the AChR and anti-striational antibodies have been detected in dogs suffering from myasthenia gravis. The detection of these antibodies adds to the known similarities between the human and canine disease. This animal model will facilitate investigation of agents involved in the induction of spontaneous myasthenia gravis.


Direct cytotoxicity against chicken erythrocytes in mice. III. Degrees of radiosensitivity and cross-reactivity of cytotoxicity, delayed hypersensitivity and antibody production. Exposure to 600 rad of X-irradiation 3 h after primary immunization with chicken erythrocytes (CRBC) abolished the production of antibodies and the generation of killer T cells, but scarcely affected the induction of delayed footpad reactions. Exposure to irradiation 3 h after secondary immunization reduced only slightly the generation of killer T cells and reduced slightly or substantially the production of antibodies. Delayed reactions persisted for long periods after elicitation in irradiated, boosted mice. Cross-reactivity between CRBC and quail erythrocytes (QRBC) was very weak with respect to the cytotoxicity, antibody and delayed reaction raised after primary immunization. Those raised after secondary immunization with the homologous antigen showed some degrees of cross-reactivity to another antigen. The booster with QRBC in CRBC-primed mice augmented the response to CRBC with respsect to the induction of cytotoxicity and delayed footpad reaction, but only weakly affected the response with respect to the production of antibody. Therefore, effector cells of cytotoxicity and delayed footpad reaction showed greater degrees of cross-reactivity than antibody-producing cells during an anamnestic response. The cross-reactive response raised after secondary immunization was radioresistant with respect to cytotoxicity and delayed footpad reaction but radiosensitive with respect to antibody production. Cytoxicity to immunizing antigen was inhibited by the unlabelled homologous antigen. Cytotoxicity to a cross-reacting antigen was inhibited by the same cross-reacting antigen and also by an immunizing antigen. There may exist heterogeneous populations of cytotoxic lymphocytes with different abilities to recognize antigens.


Species specificity in the binding of IgG to macrophages. The binding of human IgG1 and IgG3 and rabbit IgG to guinea-pig peritoneal macrophages was examined, and differences between the species, in terms of their binding mechanisms, were characterized. Rabbit IgG bound with high affinity (Kass = 3.11 +/- 0.45 x 10(6) M-1) to a finite number of receptor sites per cell (1.26 +/- 0.29 x 10(6)) and competitively inhibited the binding of guinea-pig IgG2. Heterogeneity in binding, with distinct high and low affinity components, was observed when human IgG3 was reacted with guinea-pig macrophages, while human IgG1 exhibited only low affinity binding. Neither human IgG subclass competed effectively with guinea-pig IgG2 for its cell receptor. Thus, rabbit IgG appeared to cross-react with a macrophage receptor for guinea-pig immunoglobulin, whereas the human IgG subclasses bound to macrophage membrane components that remained undefined.


Ontogeny of spontaneous antigen-binding cells in developing chick embryos. Bursal and splenic lymphocytes in developing chick embryos were examined for the number of antigen binding cells (ABC) which could form rosettes with red blood cells from sheep, guinea-pig, rabbit or dinitrophenylated sheep red blood cells (DNP-SRBC). The kinetics of the development of bursal ABC to rabbit red blood cells (RRBC) did not vary significantly with age of embryo examined, and its frequency was constantly high. The number of cells which bind to SRBC, guinea-pig red blood cells (GPRBC) or DNP-SRBC was low in the bursa and increased in an approximately linear fashion with the age of embryo. With the exception of RRBC-ABC, similar results were obtained with splenic ABC. Furthermore, the frequencies of ABC were higher in the spleen than in the bursa. These events suggest the antigen-independent generation of clonal diversity during the developmental stage of chick embryos.


Regulation of delayed-hypersensitivity response by the submandibular gland of male mice. This study was performed to determine whether the removal of submandibular gland (SG) of mice influences the delayed-hypersensitivity (DH) response. The removal of the SG increased the weights of thymus and spleen and enhanced DH response, only in males and not in females. The injection of crude extract of male mouse SG returned the enhanced DH response of SG-ectomized males to the normal level whereas the injections of crude extracts of female or castrated male mouse SG was not affected. These results suggested that SG of male mice contains an endocrine factor(s) to regulate DH response.


The effect of antiserum to eosinophils and susceptibility and acquired immunity of the guinea-pig to trichostronglyus colubriformis. There is much experimental evidence to suggest that eosinophils are important in resistance to parasitic infection. We tested this hypothesis by treating guinea-pigs with rabbit anti-eosinophil serum (AES) and determining the effect of treatment on susceptibility and acquired immunity to Trichostrongylus colubriformis. The treatment markedly reduced the number of eosinophils in peripheral blood and in the small intestine. The number of T. colubriformis present after initial infection and after a second infection was determined in animals treated with AES and in control animals. Administration of AES to guinea-pigs significantly increased the susceptibility of the animals to initial infection with T. colubriformis larvae; the number of worms recovered was nearly doubled. Similarly- administration of AES resulted in a significant diminution of acquired immunity to a secondary infection. These results are consistent with the view that eoxinophils are important in susceptibility and acquired immunity to infection with T. colubriformis in the guinea-pig. Because T. colubriformis infection is confined to the intestinal tract, our results also suggest that eosinophils may be involved in resistance to parasites at the level of the intestinal mucosa.


Complement activation by aluminium and zirconium compounds. The activation of complement by a number of metal compounds, previously found to induce chronic inflammation, was investigated. Results obtained were compared with complement activation by inulin and zymosan. It was shown that complement activation by these metal compounds did not necessarily involve either the classical or the alternative pathways. Some of these compounds were unable to activate complement in the absence of detectable plasminogen. The results obtained indicate a relationship between the ability of these compounds to induce chronic inflammation in the guinea-pig and to activate complement.


An immunogenetic basis for the tissue involvement in Behçet's syndrome. The multifocal involvement in Behçet's syndrome was grouped into a spectrum of four types, three of which appeared to have an immunogenetic basis. HLA-B5 was related to the ocular type of Behçet's syndrome (relative risk 7.3), HLA-B27 to the arthritic type (relative risk 12.1) and HLA-B12 to the muco-cutaneous type (relative risk 3.9). The concept that recurrent oral ulceration and Behçet's syndrome may belong to a disease spectrum is substantiated by the natural course of the disease. Furthermore, patients with recurrent oral ulcers share with the muco-cutaneous type of Behçet's syndrome a significantly increased frequency of HLA-B12 (relative risk 2.6). The HLA markers may also prove to be significant in the differential diagnosis and prognosis of a disease which may present under a confusing variety of clinical manifestations.


Vaccinia virus infection of the central nervous system in X-irradiated mice. The effect of X-irradiation on experimental vaccinia infection of BALB/c mice was studied. As compared with nonirradiated controls, the X-irradiated animals exhibited (i) a time lag in virus replication (delayed, but protracted replication); (ii) a delayed and repressed immune response: (iii) more severe acute cytocidal infection of leptomeninges, choroid plexus, ependyma, and vessels, with extensive damage to the brain-barrier system; and (iv) noncytocidal, latent infection of glial cells and neurons. Several animals developed acute or subacute demyelination disease, resembling experimental allergic encephalomyelitis or postinfectious encephalomyelitis.


Adherence of Actinomyces viscosus T14V and T14AV to hydroxyapatite surfaces in vitro and human teeth in vivo. Adsorption of Actinomyces viscosus strains T14V and T14AV to hydroxyapatite (HA) surfaces was studied, using an adsorption model based on the Langmuir adsorption isotherm. Data generally followed the adsorption model as judged by high correlation coefficients obtained for both strains to most of the treated surfaces studied. The number of binding sites for strains T14V and T14AV cells to human saliva-treated HA was similar to that for untreated HA. The affinity of strain T14V for saliva-treated HA was tenfold greater than the affinity of strain T14AV for that surface. To approximate the pellicle of the gingival crevice and margin and to determine whether adherence by strain T14V was to specific saliva or serum receptors, experimental pellicles were formed on HA by saliva/serum mixtures. The number of binding sites on the saliva/serum-treated HA remained the same as for the saliva-treated surface. Although the affinity of strain T14V cells for the saliva/serum HA surface remained generally the same as the affinity for the HA treated with saliva alone, the affinity of strain T14AV cells decreased further as the serum content increased. Strain T14V cell numbers adsorbed to serum-treated HA, and albumin-treated HA were less than those adsorbed to saliva-treated HA, indicating that the adherence by strain T14V was to specific saliva receptors. In vivo results from streptomycin-resistant mutants of both strains T14V and T14AV confirmed in vitro results using saliva-serum pellicles. Pretreatment of strain T14V with proteolytic enzymes and heat inhibited adherence to saliva-treated HA, suggesting that the adherence receptor(s) on the cell surface of strain T14V is protein in nature.


Demonstration of lanthionine as a natural constituent of the peptidoglycan of Fusobacterium nucleatum. Peptidoglycan was purified from the oral bacterium Fusobacterium nucleatum strain Fev 1, using boiling sodium dodecyl sulfate and pronase. The composition of this peptidoglycan was found to be similar to that of other gram-negative bacteria, except that it lacked diaminopimelic acid. Lanthionine, the monosulfur analog of diaminopimelic acid, was identified as the diaminodicarboxylic acid of this peptidoglycan. It is assumed that lanthionine replaced diaminopimelic acid. Thus, the peptidoglycan of F. nucleatum Fev 1 is one of the few known sources of naturally occurring lanthionine.


Influence of preformed antibody on experimental Streptococcus sanguis endocarditis. The influence of preformed, anti-whole organism antibody on the development of Streptococcus sanguis endocarditis was examined in both in vivo and in vitro systems. Antibody prevented, rather than potentiated, endocarditis in rabbits. The infectious dose in 30 control animals was 10(6.5) +/- 0.33 (mean +/- standard deviation); this increased to 10(7.71 +/- 0.05 in 36 immunized animals (P less than 0.01). No differences in bacterial clearance mechanisms were apparent between groups. Antibody also prevented the adherence of S. sanguis to the constituents of nonbacterial thrombotic endocarditis (fibrin and platelets) in vitro. When preincubated in high-titer antisera, adherence of S. sanguis was reduced compared with controls (adherence ratio mean +/- standard error of the mean, X 10(4): 174 +/- 5 versus 427 +/- 10, P less than 0.001). Preadsorption of immune sera with intact S. sanguis restored adherence to normal values, whereas preadsorption with dextran was partially effective. These studies demonstrate that preformed antibody had a protective role in vivo and suggest that a possible mechanism is blockade of adherence, a crucial early step in the pathogenesis of endocarditis.


Evaluation of Pseudomonas aeruginosa toxin A in experimental rat burn wound sepsis. The search for methods to achieve control of Pseudomonas aeruginosa infection continues with the introduction of aluminum-absorbed toxoid developed from P. aeruginosa exotoxin. This toxoid induces significant titers of neutralizing and precipitating antibodies for toxin A when given with appropriate adjuvants. These experiments show that immunization with aluminum phosphate-absorbed toxoid failed to protect burned rats infected with P. aeruginosa. These and previous experiments show that active immunization with live P. aeruginosa provides good strain-specific protection in the same model. No cross-protection was demonstrated between strains of P. aeruginosa in these experiments.


Modified polyriboinosinic-polyribocytidylic acid complex: sustained interferonemia and its physiological associates in humans. Fourteen patients with severe viral illnesses were given intravenous infusions of a modified interferon inducer, polyriboinosinic-polyribocytidylic acid-poly-L-lysine complexed with carboxymethylcellulose [poly)I:C.LC)], during a phase 1 clinical trial. The first eight patients received 0.15 to 0.30 mg of poly(I:C.LC) per kg of body weight daily for 5 consecutive days, and another received two courses separated by 1 week. A second group of five patients was given single intravenous infusions of 0.10 to 0.15 mg of poly(I:C.LC) per kg. Interferon was detectable in the serum 8 to 16 h after injection. Titers ranged from 15 to 800 U/ml and varied directly with the dose of poly(I:C.LC). Interferonemias persisted for 12 to 48 h. In patients receiving 5-day courses of poly(I:C.LC), lower levels of serum interferon (0 to 160 U/ml) occurred on days 2 through 5, characteristic of a hyporesponsive state. An exception was a 69-year-old patient with disseminated varicella zoster, multiple myeloma, and renal insufficiency whose serum contained 3,150 U of interferon per ml on day 3 of 0.3 mg of poly(I:C.LC) per kg. Fever (39 to 40.5 degrees C, rectally; 13 of the 14 patients) peaked 3 to 8 h after completion of infusions. Other toxic effects included lymphopenia (10 of the 14 patients), hypotensive episodes (7 of the 14 patients), and minor elevations of serum glutamicoxalacetic transaminase and lactic dehydrogenase.


A hamster-attenuated, temperature-sensitive mutant of Venezuelan encephalitis virus. Pathogenicities of 10 temperature-sensitive mutants of Venezuelan encephalitis virus were studied using the hamster model of human virulence. The parental strain and nine of the temperature-sensitive mutants produced lethal infections in hamsters. Strain ts 126 showed reduced hamster virulence. Deaths with the lethal mutants usually occurred 1 to 3 days later than with parental virus. Nine mutants produced lower levels of viremia than parental virus. Attenuation of ts 126 was related to restriction of viral growth in spleen and probably bone marrow and to absence of the usual pathological lesions in hemopoietic tissues and brain, but was functionally unrelated to temperature sensitivity since temperatures of both normal and infected hamsters remained within the permissive range of the mutant. Deaths did not correlate with titers of the 10 mutants in blood at permissive temperatures or with reversions of four temperature-sensitive mutants to non-temperature-sensitive virus in hamsters.


In vitro synthesis of humoral factors (immunoglobulins and complement) in lesional skin of leprosy patients. An in vitro culture technique was used to demonstrate the synthesis of immunoglobulins and complement in lesional skin of patients representing the entire clinico-histopathological spectrum of leprosy. The results indicate that immunoglobulin G is produced in different amounts in the various forms of leprosy. Classification of the patients according to the three main groups shows that a small amount of immunoglobulin G synthesis occurred in tuberculoid leprosy, a distinct amount occurred in borderline leprosy, and large amount occurred in lepromatous leprosy. Contrary to expectation, synthesis of C3 was found only in some of the cultures of these three forms of leprosy. The function of the locally synthesized immunoglobulin G and the findings concerning C3 synthesis are discussed.


Fractionation and properties of glucans produced by Streptococcus mutans. Water-insoluble (ISG) and water-soluble (SG) fractions of glucans produced by cell-free glucosyltransferase of Streptococcus mutans AHT (serotype g) were isolated by centrifugation at 20,000 x g for 15 min. No further resolution of slightly sonicated ISG was observed with gel filtrations on any Bio-Gel beads, including A-50m. Bio-Gel P-100 filtration subdivided SG into two fractions with higher and lower molecular weights (designated SG-A and SG-B, respectively). SG-A was further resolved into two subfractions, SG-A-I and SG-A-II, by 10 to 40% and 50 to 80% ethanol precipitation, respectively. Relative amounts of ISG, SG-A-I, SG-A-II, and SG-B were 66.3:9.4:4.4:19.9. The molecular sizes of these fractions were >1.5 x 10(7), >==1.5 x 10(7), <==5 x 10(6) (>1 x 10(5)), and <==1 x 10(4) daltons, and their alpha-1,3 glucosidic linkage contents were approximately 35, 35, 16, and 4% for fractions ISG, SG-A-I, SG-A-II, and SG-B, respectively. Both ISG and SG-A-I were resistant to hydrolysis by dextranase and possessed the ability to aggregate with concanavalin A and to agglutinate S. mutans cells. SG-A-II had extremely low dextranase susceptibility and significant agglutinating activities, whereas SG-B showed high dextranase sensitivity and neither aggregating nor agglutinating activity. These results indicate that SG of S. mutans AHT consists of three types of glucans with distinctly different molecular sizes and chemical structures and strongly suggest that the ISG and SG-A-I fractions are different physical states of an inherently identical glucan. Preliminary observations suggest that the glucans produced by other S. mutans strains of several serotypes may be similarly classified.


Cross-reactivity between antigens of Coccidioides immitis, Histoplasma capsulatum and Blastomyces dermatitidis in lymphocyte transformation assays. The cross-reactivity of four Coccidioides antigens, three Histoplasma antigens, and two Blastomyces antigens were determined in lymphocyte transformation assays of 11 coccidioidin-reactive, histoplasmin-nonreactive subjects (group I), 13 coccidioidin-nonreactive, histoplasmin-reactive persons (group II), and 13 subjects who were skin test negative to both antigens (group III). Mycelial and yeast (or spherule)-phase antigens of the three fungi were included. Significant cross-reactivity was obtained with both coccidioidins, spherulin, and the alkali-soluble, water-soluble cell wall antigen of C. immitis, to the extent that the responses of histoplasmin-reactive persons were not statistically different (P > 0.05) from those of coccidioidin-reactive persons. In contrast, optimal dilutions of Histoplasma mycelial and yeast-phase lysates effectively distinguished (P < 0.01) responses of histoplasmin- and coccidioidin-reactive persons. The alkali-soluble cell wall antigen of H. capsulatum showed extensive cross-reactivity at most concentrations and was markedly stimulatory to lymphocytes of skin test-negative persons. Blastomycin elicited significant cross-reactions in histoplasmin-sensitive subjects and to a lesser extent in coccidioidin-sensitive subjects. The alkali-soluble cell wall antigen cross-reacted in cultures of histoplasmin-reactive persons but not in those of coccidioidin-reactive persons. All antigens effectively distinguished (P < 0.001) homologous responses of skin test-positive persons (groups I and II) from those of skin test-negative persons (group III). The extensive cross-reactivity in lymphocyte transformation assays in the absence of cross-reactivity in skin tests suggests that these two immune responses may be mediated by different T lymphocyte populations, may be elicited by different antigenic components, or both.


Production of indirect hemolysin by Yersinia enterocolitica and its properties. Hemolysis was demonstrated on blood agar by adding lecithin from strains of biovar 1 Yersinia enterocolitica. Hemolytic activity was also observed in culture filtrates containing lecithin. No hemolytic activity was detected from any blood agar or culture filtrate without lecithin or after incubation at 37 degrees C. Crude indirect hemolysin was prepared by ammonium sulfate fractionation from culture filtrates, and two enzymes, phospholipase A and lipase, were fractionated by gel filtration. Hemolytic substances were recognized as lysolecithin and fatty acid when lecithin wad decomposed by phospholipase A and lipase, respectively.


Wheat germ agglutinin blockage of chlamydial attachment sites: antagonism by N-acetyl-D-glucosamine. Addition of 2 to 10 micrograms of wheat germ agglutinin (WGA), a lectin from Triticum vulgaris specific for N-acetyl-D-glucosamine, per ml to suspensions of mouse fibroblasts (L cells) blocked the attachment of 14C-labeled Chlamydia psittaci 6BC to the L-cell surface. WGA and strain 6BC competed for similar sites on L cells, but once bound, one was not replaced by the other. N-Acetyl-D-glucosamine, but not other monosaccharides of related structure, antagonized the blocking action of WGA. Lectins with specificities other than that of WGA prevented chlamydial attachment only at much higher concentrations or not at all. Exposure of L cells to trypsin and to high multiplicities of strain 6BC decreased the amount of subsequently added 3H-labeled WGA that was bound by these cells. WGA also blocked the attachment of strain 6BC to other established cell lines of murine, simian, and human origin. A lymphogranuloma venereum strain (440L) of C. trachomatis was just as sensitive to the blocking action of WGA as was strain 6BC. It appears that the attachment of both C. psittaci and C. trachomatis to host cells of diverse origin involves an N-acetyl-D-glucosamine-containing entity that binds WGA with high affinity.


Appearance of rosette-forming macrophages in the lungs of influenza virus-infected mice. Approximately one fifth of the macrophages obtained from the lungs of mice infected 2 to 5 days with influenza A/HK virus were found to rosette well with either unmodified human, chicken, or guinea pig erythrocytes, but not with erythrocytes from hamsters, sheep, or mice. Rosette-forming macrophages were seldom seen in suspensions from uninfected mice (3+/-3%) or mice infected 24 h previously (3+/-3%). Rosette formation was not due to virus hemadsorption, as indicated by the failure of specific antiserum to influenza virus to block rosette formation; by the induction of comparable levels of rosette-forming macrophages in the lungs of mice infected with herpes simplex virus type 2, a nonhemadsorbing virus; and by the inhibition of rosette formation at 4 degrees C. Instead, rosette formation appeared to be directly related to macrophage elicitation or activation since nonstimulated macrophage populations such as peripheral blood monocytes, macrophages from uninfected lungs, or noninduced peritoneal macrophages were not observed to rosette to any significant extent. Furthermore, peritoneal macrophages induced with filter-sterilized normal horse serum rosetted at levels comparable to that observed with cells from infected lungs. These results indicate that hemadsorption alone can not be used as a criterion of virus infection of macrophages. However, rosette formation may serve to identify macrophage subpopulations which are active in host defense against viral infections.


Serum antibody prevents lethal murine influenza pneumonitis but not tracheitis. This paper reports studies showing the effects of serum antibody upon influenza infection at two different sites: the trachea and lung. Tracheal desquamation, pulmonary consolidation, death, and virus shedding were examined after infection of mice with a lethal A/Port Chalmers/1/73 (H3N2) influenza virus. Immune serum administered intraperitoneally before infection prevented death and pulmonary consolidation and also significantly lowered lung virus shedding as compared with controls receiving normal serum. However, this protection did not extend to the ciliated epithelium of the trachea because serum antibody did not prevent desquamation of the trachea or significantly decrease viral yield from the trachea. These results indicate that serum antibody is protective against severe pulmonary parenchymal disease but not for disease of the ciliated epithelium.


Human neutrophil migratory function: modulatory effect of interactions with opsonized particles. Human polymorphonuclear neutrophils preexposed to cytotaxin or to phagocytizable particles exhibited reduced spontaneous and chemotactic migratory responses. This influence of cytotaxin appears to be related to toxic effects of by-products of hexose monophosphate shunt stimulation. To determine whether a phagocytic stimulus may inhibit subsequent neutrophil migratory functions by the same mechanism, we assessed spontaneous and chemotactic migratory functions of neutrophils from individuals with chronic granulomatous disease exposed to antibody-opsonized sheep erythrocytes. Our results showed that phagocytosis of such particles did not alter these migratory responses of chronic granulomatous disease neutrophils and suggest that phagocytic stimulation of normal neutrophils may modulate migratory function by some mechanism dependent upon hexose monophosphate shunt stimulation.


Sites that bind polymerized albumin on hepatitis B surface antigen particles: detection by radioimmunoassay. Antibodies to polymerized human albumin (poly-HSA) could not be detected by using sensitive methods (enzyme-linked immunosorbent assay and radioimmunoprecipitation) in sera from chronic carriers of hepatitis B surface antigen (HBsAg) or in serial bleedings from one chimpanzee infected with type A hepatitis virus and one infected with non-A, non-B hepatitis virus. By a solid-phase radioimmunoassay, receptor sites for poly-HSA could be detected on HBsAg particles from sera containing either hepatitis B "e" antigen (HBeAg) or anti-HBe. Blocking experiments showed that monomeric HSA did not bind to this receptor. In general, the HBsAg particles from sera with HBeAg had more poly-HSA receptor sites or relatively more particles carrying this receptor compared with HBsAg from sera with anti-HBe. Microtiter plates coated with poly-HSA bound HBsAg from sera containing HBeAg with greater efficiency than did anti-HBs coupled to a solid phase (Ausria II beads), whereas with sera positive for anti-HBe, the two assays were equally sensitive. Decreased ability of HBsAg to bind to poly-HSA was seen in some sera which had been stored for a few years at 4 degrees C, whereas the binding to anti-HBs was unaffected. It is possible that polymers of albumin on the surface of hepatocytes could function as receptors for hepatitis B virus.


Colonization of the cementum surface of teeth by oral Gram-negative bacteria. By using in vitro assays, a group of related, filamentous gram-negative bacteria isolated from subgingival plaque deposits of patients with periodontal disease were found to colonize intact teeth. Tentatively identified as members of the genus Cytophaga, these isolates exhibited a preference for colonizing the cementum surface of the root. Examination of intact teeth after several weeks of colonization revealed that the root substructure had been extensively demineralized.


Thymosin restoration of cellular immunity to Blastomyces dermatitidis in T-cell-depleted mice. The immunopotentiating properties of thymosin in thymectomized, lethally irradiated, bone marrow-reconstituted mice (ThyXBM) were characterized, using footpad sensitivity to Blastomyces dermatitidis. Normal mice were shown to exhibit increasing delayed-type hypersensitivity responses to killed yeast cells of B. dermatitidis after injections of the organism on days 0 and 7, as measured by footpad swelling tests. The footpad response of normal thymosin-treated mice was similar to that of normal, non-thymosin-treated mice. ThyXBM mice were unable to elicit a footpad response when similarly injected and footpad tested with B. dermatitidis. Thymosin-treated ThyXBM mice responded to footpad testing at a level that was 62% greater than the response seen in non-thymosin-treated ThyXBM mice. This peak response occurred on day 12. The results indicated that thymosin was unable to enhance immune responses of normal intact  mice but could restore immunocompetence in a T-cell-depleted host, as measured by footpad sensitivity to B. dermatitidis.


Behavior of vaccine revertants of temperature-sensitive mutants of influenza virus in ferret tracheal organ culture. A live attenuated influenza vaccine candidate was not genetically stable when administered to some children who lacked antibody to surface proteins of the virus. To obtain additional biological information about these revertants, the vaccine strain, the wild-type parental strain, and isolates recovered from inoculated children during a vaccine trial were evaluated in ferret tracheal organ culture for effects on the ciliated epithelium and replication at both permissive and restrictive temperatures. The studies revealed that the vaccine strain destroyed cilia and replicated to high titer at its permissive temperature (33 degrees ) but caused minimal damage and replicated to very low titer at its restrictive temperature (37 degrees C). The wild-type parent destroyed cilia at both 33 and 37 degrees C. Isolates which were no longer temperature sensitive (ts(+)) destroyed cilia at both restrictive and permissive temperatures and grew to high titer. Isolates which retained the ts phenotype behaved as the vaccine strain in this system. The ts(+) virus recovered from volunteers behaved like the wild-type parent, which suggests that these viruses had not merely lost their ts phenotype, but had undergone reversion to wild type. Important information about the genetic stability of temperature-sensitive influenza vaccine strains recovered from volunteers can be obtained by evaluating them in ferret tracheal organ culture.


Baculovirus replication in a mosquito (dipteran) cell line. The baculovirus from the lepidopteran host Autographa californica (alfalfa looper) was shown to replicate in a dipteran cell line without the production of characteristic polyhedral inclusion bodies. The low level of replication could not be detected by 50% tissue culture infective dose titrations, but was apparent by [3H]thymidine labeling of the viral genome. Immunoprecipitation of the radioactive product confirmed baculovirus production.


Live Victoria/75-ts-1[E] influenza A virus vaccines in adult volunteers: role of hemagglutinin immunity in protection against illness and infection caused by influenza A virus. To explore the relationship between neuraminidase immunity and the degree of attenuatíon of live influenza A virus vaccines, a comparative evaluation of three Victoria/75-ts-1[E] (Vic/75-ts-1[E]) recombinant viruses in serum hemagglutination-inhibiting-negative (titer, </=1:8) adult volunteers was performed. These three ts-1[E] viruses had a similar restriction of replication at 38 degrees C in vitro, and each possessed the two attenuating genes of the ts-1[E] donor strain (13). However, Vic/75-ts-1[E] recombinants 81 and 113 possessed both Vic/75 hemagglutinin (H3(75)) and Vic/75 neuraminidase (N2(75)), whereas Vic/75-ts-1[E] recombinant 67 had Vic/75 hemagglutinin but the N2(65) neuraminidase. Vic/75-ts-1[E] recombinant 67 was significantly more attenuated than Vic/75-ts-1[E] recombinants 81 and 113 in that fewer local and systemic signs and symptoms of illness were observed in those volunteers who received clone 67. These findings were consistent with our previous observations which suggested that the following two factors contribute to the attenuation of ts-1[E] vaccine strains in adults: (i) the attenuating effect of the two ts-1[E] genes and (ii) the neuraminidase immunity of the host. Vic/75-ts-1[E] recombinant clone 67 vaccinees developed an immunological response to the H3(75) hemagglutinin in the absence of a response to the N2(75) neuraminidase. To assess the role that anti-hemagglutinin immunity induced by an attenuated live virus vaccine plays in resistance to influenza A virus, vaccinees who received recombinant 67 were challenged with Vic/75 wild-type virus, and their responses were compared with those of Vic/75-ts-1[E] vaccinees who received recombinant 81 or 113. Each of the three groups of ts-1[E] vaccinees was significantly protected against illness induced by wild-type virus infection, although resistance was not complete. However, the clone 67 vaccinees were protected less against infection. The infection-permissive resistance induced by clone 67 resembled that previously described for inactivated neuraminidase-specific vaccines. These results suggested that a ts-1[E] recombinant that possessed the hemagglutinin of a new pandemic variant, the neuraminidase of the preceding subtype, and the two ts-1[E] ts genes would be satisfactorily attenuated for children and adults with neuraminidase immunity and could induce resistance to illness caused by the new pandemic wild-type influenza A virus.


Relationship of genotype of recombinants of influenza A/Hong Kong/68-ts-1[E]virus used as live virus vaccines to virulence in humans. Influenza A/Hong Kong/68-ts-1[E] virus is a temperature-sensitive mutant developed for use as a live virus vaccine (B. R. Murphy, E. G. Chalhub, S. R. Nusinoff, J. Kasel, and R. M. Chanock, J. Infect. Dis. 128:479--487, 1973). This virus and temperature-sensitive recombinants derived by mating it with A/Udorn/72, A/Georgia/74, or A/Victoria/75 wild-type virus have been administered to volunteers in clinical trials on the assumption that the ts-1[E] temperature-sensitive genetic lesions on a polymerase gene (P3) and on the nucleoprotein gene (NP) would determine a satisfactory and reproducible level of attentuation regardless of the genetic constitution of ts-1[E] recombinants at other loci (B. R. Murphy, D. D. Richman, S. B. Spring, and R. M. Chanock, Postgrad. Med. 52:381--388, 1976). In this paper, the parental origin of genes in the ts-1[E] recombinants was determined by using the technique of polyacrylamide gel electrophoresis of virion ribonucleic acid segments in the presence of a denaturing agent (urea). When tested in individuals who lacked immunity to hemagglutinin antigen, attenuation of the ts-1[E] recombinants appeared to correlate with inheritance of the ts-1[E] temperature-sensitive genes at the P3 and NP loci and with the level of preinfection neuraminidase immunity. There was no evidence that other genes from the ts-1[E] donor virus played a role in attenuation.


Arabinosylthymine: inhibitor of splenic lymphocyte macromolecular synthesis in vitro. The effect of arabinosylthymine on lymphocyte transformation was investigated. Arabinosylthymine was demonstrated not to be cytotoxic for hamster spleen lymphocytes but was found to inhibit the increase in deoxyribonucleic acid and protein synthesis usually observed as a result of mitogen stimulation. These findings suggest that, in addition to being a potent anti-herpesvirus inhibitor, arabinosylthymine is also an immunosuppressive agent.


Fatty acid composition of gliding bacteria: oral isolates of Capnocytophaga compared with Sporocytophaga. The extractable and bound lipids and cellular fatty acids of the gram-negative gliding bacteria, Capnocytophaga sputigena, C. gingivalis, and C. ochracea were compared to the non-host-related gliding bacterium Sporocytophaga myxococcoides. The extractable lipids represented between 17 and 28% of the cell dry weight, whereas only 2 to 4% of the lipids were in the bound fraction. The methyl esters of the cellular fatty acids were mainly aC15:0, which accounted for 69 to 73% of the total extractable fatty acids; S. myxococcoides had a similar distribution of branched-chain fatty acids; however, aC17:0 was the predominant fatty acid in this free-living gliding organism.


Reduction of contact sensitivity reactions to oxazolone in mite-infested mice. Oxazolone-sensitized mite-infested (SWR-M) and mite-free (SWR-J) mice were challenged with oxazolone on the skin of the neck and shoulder. The migration of radioactively labeled cells to the site of contact sensitivity reaction to oxazolone was significantly less in SWR-M than in SWR-J mice. Serum obtained from SWR-M mice suppressed the extravasation of cells into the skin site of SWR-J mice challenged with oxazolone. The decrease in cellular influx in SWR-M mice occurred in areas of mite infestation (skin of neck and shoulder) as well as in areas not infested with mites (the ears). SWR-M mice also gave evidence of enhanced vascular permeability. A possible role for histamine in the inhibition of contact sensitivity in mite-infested mice is discussed.


Phospholipid composition of gliding bacteria: oral isolates of Capnocytophaga compared with Sporocytophaga. The distribution of acetone-soluble (neutral glycolipid) and acetone-insoluble (phospholipid isoprenoids) lipids in oral isolates of gram-negative gliding bacteria of the genus Capnocytophaga was compared with those in a non-host-related gliding bacterium, Sporocytophaga myxococcoides. The acetone-soluble material accounted for 34 to 55% of the extracted lipids; the remainder was acetone-insoluble material. The major phospholipid was phosphatidylethanolamine (67%), with lesser amounts of lysophosphatidylethanolamine and several unidentified phosphate-containing compounds. Capnocytophaga also contained significant amounts of an ornithine-amino lipid.


Abortive replication of vaccinia virus in activated rabbit macrophages. During the course of infection of rabbits with vaccinia virus, macrophages obtained from the peritoneal cavity develop bactericidal activity and the replication of vaccinia virus becomes restricted in these cells. The abortive replication of vaccinia virus in the activated macrophages was characterized in the present study. The virus adsorbed to and was uncoated equally well in macrophages from both normal and infected rabbits. A burst of deoxyribonucleic acid synthesis of comparable magnitude took place 3 to 6 h after infection in both normal and activated macrophages. Although the production of viral antigens, as detected by immunodiffusion and immunofluorescence, was the same in both types of cells, very few virus particles were formed in activated as compared with normal macrophages. We conclude that a block in a late step of the virus replication cycle occurred in the activated macrophages.


Susceptibility of inbred mice to chronic central nervous system infection by Theiler's murine encephalomyelitis virus. The present study demonstrated that the clinicopathological expression of the late demyelinating disease due to chronic central nervous system infection by Theiler's mouse encephalomyelitis virus was dependent, at least in part, on the strain of mouse used as host. A range of involvement was observed, with late disease being most severe in the SJL strain, intermediate in the CBA and C3H/He strains, and least in C57BL/6 mice. The lack of clinical signs in seven other inbred strains of mice indicates that their response to chronic infection was similar to C57BL/6 mice. SJL, CBA, C3H/He, and C57BL/6 mice all generated similar levels of neutralizing antibody. A correlation between the severity of late disease and central nervous system virus content was not demonstrated, which indirectly suggests an immunopathological rather than a cytolytic mechanism of myelin injury during the late disease period. Finally, in addition to being more extensive, SJL demyelinating lesions contained a disproportionately large number of macrophages compared with those of similar lesions in CBA and C3H/He mice.


Multiplication of Leishmania in human macrophages in vitro. To facilitate in vitro studies of the immunology of human leishmaniasis, we developed a method of growing pathogenic Leishmania in human monocyte-derived macrophages. After 6 days of incubation, adherent mononuclear cells were infected with Leishmania donovani amastigotes obtained from infected hamster spleen cells or with L. tropica amastigotes obtained from infected BALB/c tissue mouse footpad. Forty-eight percent of the macrophages were initially infected, with a mean of 3.0 amastigotes per infected macrophage. After 6 days of incubation, 59% of macrophages were infected and contained 8.8 amastigotes per infected macrophage, representing 2.9-fold multiplication. Electron microscopy revealed the presence of dividing parasites within phagolysosomes. These observations indicate that Leishmania survive and multiply within human monocyte-derived macrophages despite fusion of secondary lysosomes with the parasitophorous vacuole.


Significance of abnormal rabbit ileal histology in the pathogenesis of diarrhea. In spite of several macroscopic criteria for predicting the presence of histological abnormalities in rabbit ileum, microscopic ileal abnormalities still can escape detection. The effect of histologically abnormal rabbit ileum was evaluated on basal intestinal absorption, on basal absorption, on basal adenylate cyclase activity, and on cholera toxin-induced secretion and cholera toxin-induced stimulation of adenylate cyclase activity. Compared to histologically normal rabbit ileum, the presence of histological abnormalities was associated with decreased basal intestinal water, Na, Cl, and glucose absorption, absent glucose-dependent water absorption, and elevated basal adenylate cyclase activities. However, histologically abnormal rabbit ileum responded to inoculation of purified cholera toxin with stimulation of intestinal water secretion and adenylate cyclase activity similar to that in histologically normal ileum. These data have implications concerning the design of experiments that attempt to study the pathogenesis of diarrheal diseases by correlating changes in ileal transport with changes in ileal mucosal adenylate cyclase activity. In spite of abnormal ileal histology, studies of intestinal secretory states which attempt to define the role of adenylate cyclase in secretory processes can be performed provided animals are used as their own controls. However, when groups of animals are compared, the presence of an histologically abnormal ileum can cause changes in basal and intestinal secretagogue-stimulated ileal water and electrolyte transport and in basal and intestinal secretagogue-stimulated mucosal adenylate cyclase activity which can lead to erroneous conclusions if the presence of the abnormal ileal histology is not considered.


Nature of the fructan of Streptococcus mutans OMZ 176. The fructan of Streptococcus mutans OMZ 176 was shown to have a levan structure by comparing the chromatographic mobilities of the saccharides produced by partial acid hydrolysis of ghe fructan and known levan and inulin. This was confirmed by using concanavalin A as a lectin in a double-diffusion gel technique.


Chemotaxigenesis by cell surface components of Staphylococcus aureus. In an attempt to delineate the staphylococcal cell surface components of importance in chemotaxigenesis, we incubated intact Staphylococcus aureus H, crude cell walls, purified cell walls, peptidoglycan, teichoic acid, and cell membranes with human sera. The results reported indicate that both crude cell walls and purified cell walls, as well as peptidoglycan, were potent chemotaxigens. These particles led to the generation in normal human serum of a factor that was chemotactic for human polymorphonuclear leukocytes. Cell wall peptidoglycan and teichoic acid both appeared to play a role in chemotaxigenesis. Kinetic studies employing C2-deficient serum and immunoglobulin-deficient serum revealed that optimal chemotaxigenesis required the presence of an intact classical complement pathway, as well as antibody. Granulocyte aggregometry studies showed that significant levels of C5a were generated in normal serum and that this activated complement component appears to be a major chemotactic factor produced in serum upon interaction with staphylococcal cell wall components.


Effect of oral administration of glucosyltransferase antigens on experimental dental caries. The effect of oral administration of soluble antigen preparations containing glucosyltransferase on dental caries in hamsters was studied. Immunization was accomplished by feeding glucosyltransferase for 21 to 27 consecutive days. This immunization regimen resulted in the formation of salivary antibody, which was detected by functional inhibition of enzymatic activity and by a modified enzyme-linked immunosorbent assay. A serum response also occurred in two of the three experiments performed. After infection with cariogenic Streptococcus mutans strain 6715, glucosyltransferase-fed hamsters had significantly fewer S. mutans cells recoverable from molar surfaces on six of nine occasions, compared with buffer-fed control groups. Hamsters orally immunized with glucosyltransferase also always had lower mean caries scores and mean numbers of lesions than comparably infected sham-immunized groups. The results of this study suggest that significant protection from experimental dental caries can be accomplished by oral administration of soluble antigen preparations containing glucosyltransferase.


Kinetics and cell killing in dividing and nondividing leukemic cells in vitro and in vivo by natural splenic cytotoxic factor. Ultrafiltered splenic extracts (30,000-50,000 daltons) were specifically cytotoxic in crowded cultures of murine leukemic lymphoblasts but were only inhibitory toward normal lymphocytes or sparse cultures of leukemic cells. This expression of cytotoxicity was quantitated by (i) vital dye exclusion, (ii) cytochemical metods, and (iii) the increased survival time of mice injected with L-1210 leukemic cells incubated in vitro with splenic extract. Selective cytotoxicity of splenic extract was not due to medium depletion or to complement-mediated cytolysis and could not be replicated using similar extracts of other lymphoid and nonlymphoid tissues. The selective effect of splenic extract on leukemic cells was characterized by the progressive shedding of cytochemically demonstrable RNA and the corresponding inhibition of 3H-uridine and 3H-thymidine incorporation. The comparison of progressive and gradual expression of cytotoxicity by splenic extract with the S phase specific inhibitor cytosine arabinoside suggests that the former is cytotoxic against leukemic cells predominantly in the G1 phase of the cell cycle.


Cytochemical and ultrastructural alteration of leukemic cells by a naturally cytotoxic factor from spleen. Experiments were carried out to delineate the differences in cytocidal modality of natural splenic cytotoxic factor (30,000-50,000 daltons) and cyto-A against normal and human leukemic lymphocytes in vitro. It was found that the differences in the cytocidal action effected by cyto-A and splenic isolate were as follows: (1) the killing effect of cyto-A against leukemic cells was very rapid in contrast to the slowly acting spleen factor as evaluated by cytochemical and ultrastructural studies; (2) spleen factor was not cytotoxic against normal lymphocytes in contrast to the indiscriminatory cytotoxic cyto-A.


Ultrastructure of basophilic leukocytes and mast cells in normal and cutaneous basophil hypersensitivity-reacted guinea pig dermis. Basophilic leukocytes and mast cells in guinea pig dermis in normal and cutaneous basophil hypersensitivity (CBH) reaction were examined by light and electron microscopy. Basophils were rare in the normal dermis and predominantly revealed in the CBH-reacted skin. Some infiltrating basophils of the reacted displayed an immediate attachment to mast cells. Cytoplasmic continuities were partially seen between them. They were most plentiful at 48 h after phytohemagglutinin injection and decreased in number thereafter. The basophils contained three types of granule. The vast majority of granules were Type I granules, basophil-specific granules. Type II granules were less frequently encountered and resembled a mast cell granule in the fine structure. Type III granules were scant and small-cored vesicles.


Heterotoxicity of human serum. IV. Role of the alternative complement pathway and natural antibody in the lethal toxicity of human serum for mice. The toxic and often lethal reaction caused by intravascular injection of fresh human serum (FHS) was studied in mice. Decomplementation experiments indicated that the alternative complement pathway plays an essential role in the production of 'serum shock'. In vivo activation of human factor B and C3, following the administration of FHS into mice, was detected by immunoelectrophoresis. Absorption experiments suggested that natural antibodies were not required for toxicity, although they may amplify the lethal potential of normal sera by feedback mechanisms of C activation. Injections of FHS caused intravascular hemolysis of mouse erythrocytes and platelet aggregation in mice. The relationship of these observations to the mechanism(s) underlying lethality will be discussed in the following article.


Comparison of three hemodialysis procedures with unipuncture. Dialysis efficiency estimated by the decrease in urea, creatinine and phosphorus serum concentrations is higher with the double-lumen single-needle method, compared to the single-lumen needle unipuncture methods. This increased efficiency is related to a higher blood flow rate through the dialyser. Provided the superficial veins are not too small and/or tortuous, the double-lumen needle appears as a safe, simple and rewarding device and may be used in most patients on regular dialysis treatment.


Blood gas changes during carbon haemoperfusion. Treatment of experiment poisoning in dogs with haemoperfusion over activated carbon is reported. The blood gas concentrations were monitored and their changes during haemoperfusion considered with respect to their clinical significance.


The behavior of three different types of materials in vitro and in the dynamic physiological environment: review and analyses of critical parameters. A comparative analysis of smooth-surfaced synthetic materials, gels, especially those based on polyacrylamide, and polyester flocked surfaces indicate profound differences between these three types of materials when in contact with blood. Although many smooth-surface materials adsorb fibrinogen in vitro, several of these perform well in vivo without polymerization of fibrinogen to fibrin leading to clot-formation apparently because of enzymatic reactions in the physiological environment. Polyacrylamide hydrogels adsorb little plasma proteins, show negligible platelet adhesion and activation. In contrast, polyester flocked (fibril) surface deposit large quantites of fibrinogen that polymerizes to fibrin with entrapped formed blood elements (clot). The thickness of this layer is difficult to control and the build-up continues to a thickness up to approximately 400 to 600 micron depending on the species. The danger of clot-fragmentation and flock-detachment especially during flexing makes these rough surfaces inferior in blood-contacting application, in comparison to the best current synthetic smooth-surfaced biomaterials, hydrogels, and chemically modified natural tissue components.


Improvement of the quantitative determination of verapamil in human plasma. An improvement in the extraction procedure for the quantitative determination of verapamil in human plasma is described. The advantages are: 1. the time involved in the extraction procedure is only 1/3 of that in the original method allowing the extraction of 100 samples in a working day. 2. increased mean recovery from 67% to 85% over the range of 3--90 ng verapamil/ml plasma. 3. less expenditure a) use of cheap plastic tubes instead of stoppered glass tubes. b) 1/2 the amount of heptane required as compared to the original procedure. c) use of air instead of N2 for evaporation. The lower limit of detection is 3 ng/ml which is comparable to that for the original extraction procedure. Within-batch precision over the range of 9--90 ng/ml averages 5.8% and at the lower limit of detection 17.3%. Between-batch precision over the range of 9--90 ng/ml averages 8.5% and at the lower limit of detection 19.7%. No significant difference could be found in the quantitative determination of verapamil in 27 plasma samples from patients undergoing verapamil treatment using the original and modified extraction procedures. This improvement of the extraction procedures simplifies the determination of verapamil and the only additional material required is liquid nitrogen or a suitable solvent cooled with dry ice.


Interindividual differences of protein binding in man. Using equilibrium dialysis protein unbound fractions of phenytoin (390 patients) and phenobarbital (220 patients) revealed serum protein bindings of 8 to 40 per cent and 25 to 80 per cent, respectively. For the purpose of therapy service, this pronounced interindividual variability required in the case of a great number of patients the determination of the extent of protein binding so as to exactly evaluate therapeutical failures or occurrence of undesirable effects.


Evaluation of clinical efficacy and plasma levels of aprindine. The clinical efficacy of Aprindine, a new antiarrhythmic agent was tested in 37 patients with ventricular and supraventricular arrhythmias. The drug was shown to be effective - disappearance or prevention of relapse of the arrhythmia - in 80% of the cases with ventricular tachycardia, and in 67.7% of the cases with premature ventricular beats. No judgement was possible for supraventricular tachycardia due to the small number of patients. Effectivness of Aprindine was found to be related more closely to the administered dose expressed in mg/kg body weight than to plasma level in steady-state. Discontinuation of therapy was necessary in 5 cases due to the appearance of side effects (tremor and dizziness). Side effects were not found to be in relation with plasma levels or administered dose. In the period of observation (2 months) no effects on blood cells were noticed.