Visceral vessel erosion associated with pancreatitis. Case reports and a review of the literature. Visceral artery erosion is an uncommon but disasterous complication of pancreatitis. When gastrointestinal bleeding or severe intra-abdominal hemorrhage is associated with pancreatitis and the usual sources of bleeding are not detected by endoscopy or barium studies, erosion of a visceral vessel should be suspected. We present nine cases and an additional 44 cases from previous reports are reviewed. A palpable abdominal mass was present in 59% of the patients; however, a pseudocyst was present in 78%. Arteriography was performed in 15 patients and the source of bleeding was evident in 14. The splenic artery was the most common site of bleeding, although the other branches of the celiac axis and the middle colic artery have been involved. Successful treatment consisted of ligation of the bleeding vessel and, if present, drainage of the pseudocyst. Depending on the location of the vessel and the pseudocyst, major resections may be necessary. When bleeding and the pancreatic inflammatory process involved the colon, ligation of the bleeding site, drainage of the area, and colostomy was the most successful form of treatment.


'Downhill' esophageal varices. A rare cause of upper gastrointestinal bleeding. The submucosal venous network of the esophagus is part of the collateral system that develops following superior vena caval obstruction from any cause. The direction of flow in these thin-walled, valveless veins is "downhill," towards the azygous vein or to the inferior vena cava. Bleeding from upper esophageal varices is extremely rare. This case report describes a patient with massive bleeding from upper esophageal varices secondary to superior vena caval obstruction by a malignant thyroid tumor. Total thyroidectomy relieved the obstruction, with cessation of hemorrhage and subsequent disappearance of the varices.


Amebiasis causing massive gastrointestinal bleeding. Three patients had massive bleeding from proved amebic ulcers in the cecum. One had an accompanying amebic liver abscess. All were successfully treated by emergency right hemicolectomy.


Sensitive neutralization test for virus antibody. 1. Mumps antibody. A sensitive mumps virus plaque neutralization test has been developed based on the potentiation of virus-antibody complexes by heterologous anti-immunoglobulins (AIG). The enhanced neutralization test was approximately 100 times more sensitive than the conventional neutralization test or the hemagglutination-inhibition test. Using AIG against human immunoglobulin G (IgG) or human IgM permitted determination of the relative titers of the two classes of mumps antibody. The test does not require special equipment or expertise and can be readily introduced in virological laboratories.


[Changes in the cellular elements of the thymus gland in experimental destructive pulmonary tuberculosis]. Lymphoid elements of the thymus were studied by scanning electron microscopy in rabbits susceptible to tuberculosis. The complete Freund adjuvant causes in the thymus a 3-fold decrease in the number of cells with the properties of T-lymphocytes, and an appearance of a new type of cell elements (polygonal shape, with cytoplasmic outgrowths). Formation of a focus of tuberculosis and the development of destructive changes in the lungs occur under conditions of a progressive decrease in the number of T- and B-lymphocytes in the thymus. Delimitation of the pathological process in the lung is characterized by an increase in the number of T- and B-lymphocytes and considerable predominance of polygonally shaped cells with cytoplasmic outgrowths of different lengths and their subsequent replacement by a cell form transitory between T- and B-lymphocytes. A relatively favourable course of the process in the lungs is observed when the cell elements in the thymus differentiate towards B-lymphocytes.


Tomography of the vestibular aqueduct in ear disease. A controversy exists concerning whether or not roentgenographic narrowing or nonvisualization of the vestibular aqueduct is a specific sign for Meniere's disease. Of 190 ears that were evaluated, abnormal aqueducts were seen in 42.9% of ears with Meniere's disease, 45.4% of contralateral, noninvolved ears from patients with Meniere's disease, 41.3% of ears with diseases other than Meniere's disease, 30.4% of ears with no disease, and 51.6% of normal ears. Narrowing or nonvisualization of the vestibular aqueduct is a nonspecific roentgenographic sign that is seen in diseased, as well as in normal ears, and should not be used to make a diagnosis of Meniere's disease. Indications for polytomography in Meniere's disease include (1) preoperative evaluation of the aqueduct prior to endolymphatic shunt procedures and (2) exclusion of acoustic neurinomas or other organic causes of vertigo.


Repeated shunt surgery in a patient with portal hypertension. A 45-year-old male with chronic active hepatitis and portal hypertension had a mesocaval interposition graft performed because of repeated uncontrolled bleeding from oesophageal varices. Acute hepatocellular failure, manifested by Grade IV encephalopathy and severe coagulopathy, developed early in the post-operative course despite the absence of hepatic necrosis or other precipitating factors. Both encephalopathy and coagulopathy resolved rapidly following ligation of the shunt. Variceal bleeding recurred and nine months later an emergency distal lienorenal shunt was performed. Post-operatively the shunt was shown to be patent and there has been no encephalopathy or recurrence of variceal bleeding. It is concluded that (1) the splanchnic haemodynamic effects of a mesocaval interposition graft can result in severe hepatocellular failure and this can be reverted by shunt ligation and (2) the distal lienorenal shunt, while effectively reducing the risk of haemorrhage from varices, may be less likely to result in post-operative encephalopathy than more conventional forms of portal decompressive surgery.


[Action of various mitogens (phytohemagglutinin, cocanavalin A, pokeweed mitogen) on a purified human B- and T-lymphocyte population mediating spontaneous rosette formation]. The activity of various most widely used "polyclonal" mitogens, such as PHA, Con A and PWM on human peripheral blood lymphocyte subpopulations, after separation into the two main functional classes (B and T), was investigated. For separating the lymphocyte subclasses, the E-rosette formation technique has been followed, using the gradient centrifuged pellet and supernatant respectively. For the mitogens' activity evaluation, micro-culture system has been performed and 3H thymidine uptake has been assessed. The PHA has been shown to be much more active on the T lymphocyte subclass, though the mitogen cannot be kept very selective. Very much higher activity on the T subclass shows the Con A, which could be considered selective, if we hypothesize an at least partial receptor competition at lymphocyte membrane level between the mitogen and sheep erythrocytes. The PWM has been found very little active and not specific for either lymphocyte subclass.


A controlled study of ANA+ RF- arthritis. Thirty (7.5%) of 401 adult rheumatoid arthritis (RA) patients were antinuclear antibody positive (ANA+) and rheumatoid factor negative (RF-), and 15 of 16 patients who were followed for a year or longer remained so. Clinical, other laboratory, and radiographic parameters were compared among this group and 90 matched RA controls divided into ANA+RF+, ANA-RF+, and ANA-RF- groups. All groups were identical, except the ANA-RF- group, which had significantly fewer nodules and less destructive disease than the other three.


[Effect of cytotoxic Hodgkin's serum on LIF activity of T- and B-lymphocytes]. The cytotoxic activity of Hodgkin serum on some isolated T and B lymphocyte populations of 10 normal subjects, 10 patients with Hodgkin disease in the acute phase and 10 patients with Hodgkin disease but in clinical and biohumoral remission was studied. The effect of citotoxic serum variously affects the two different lymphocyte populations, particularly inhibiting LIF activity of T lymphocytes, while stimulating LIF activity of B lymphocytes. The possible causes of this phenomenon are set forth as well as their clinical implications.


Mixed connective tissue disease in siblings. Mixed connective tissue disease (MCTD) was diagnosed in a brother and sister, and 18 additional family members spanning three generations were studied to detect evidence of autoimmune disease. Symptoms or signs of MCTD without complete expression of the disease were found in 8 relatives of the original cases. Antibodies to ribonucleoprotein and high-titer antinuclear antibodies were found only in the affected siblings. Tests for rheumatoid factor were positive in 9 of 17 relatives of the patients; the titers ranged from 1:160 to 1:2560. The brother and sister with MCTD had an identical HLA genotype--11,12/2,12. The same genotype was inherited by 3 of their siblings, who had impressive rheumatic complaints. This report emphasizes the association between inflammatory connective tissue disease and a specific HLA type within a single kindred.


[Chronic persistent HBsAG positive hepatitis in children. I. Subpopulations of T and B lymphocytes]. T and B peripheral blood lymphocytes were studied in five children affected by chronic persistent hepatitis HBsAg-positive. The percentage of E rosette forming cells was found decreased while thymidine uptake after Phytohaemagglutinin stimulation reached values higher than that of normal controls. Null cells were found increased. In all the patients the percentage of IgG bearing lymphocytes was markedly decreased. Such alterations of T cells compartement are similar to those found by other authors in the aggressive form. The hypothesis of a derangement of T cell subpopulation regulatory activity is put forward.


Reevaluation of the ammonium sulfate assay for DNA antibody. The ammonium sulfate assay has recently come under criticism for alleged inability to detect low avidity antibody to DNA. The results of this study verify that a small percentage of antibody binding is not detected when this assay is performed under the conditions generally used. However, when the values obtained by the ammonium sulfate assay were compared to those obtained by other assays on sera of known avidity for DNA, it was concluded that ammonium sulfate does not selectively dissociate low avidity antibody from DNA. Conditions of incubation buffer and duration, pH, concentration of reactants, and molecular weight of the antigen are described for optimal detection of DNA antibody by the ammonium sulfate assay.


Humoral immune stimulation and antiepithelial antibodies in Yersinia infection. Results consistent with a general humoral immune stimulation were found when 127 sera from 89 patients with Yersinia enterocolitica infection were studied. Significantly increased gammaglobulin concentrations and elevated isohemagglutinin titers were seen in these sera as compared to sera from normal blood donors and patients with streptococcal infection. Antinuclear and anti-smooth muscle antibodies were demonstrated in both yersinia and streptococcal infection. The prevalence of non-organ specific antiepithelial antibodies reacting with gastrointestinal and thyroid epithelial cells was significantly increased in yersinia infection.


[Chronic persistent HBsAg positive hepatitis in children. II. Effect of treatment with levamisole]. In five children affected by HBsAg positive chronic persistent hepatitis a treatment with Levamisole (LMS) modified several immunological parameters which had been found altered before treatment. In particular the percentage of E rosette forming cells increased while that of EAC decreased; B lymphocytes with SmIgG and the responsiveness to phytohaemagglutinin, which were significantly decreased and increased respectively, got normal values. Since the persistence of HBsAg was unaffected by LMS treatment and a light increase of transaminase serum levels (GPT and GOT) was observed during treatment, doubts are expressed about the opportunity of using LMS in children affected by such a form of hepatitis.


Cogan's microcystic dystrophy of the cornea: ultrastructure and photomicroscopy. Corneal biopsy specimens from 3 patients with Cogan's microcystic corneal dystrophy were examined by light and electron microscopy. Specimens were taken from corneas showing microcysts, geographic or map-like areas, and refractile striae. In all samples there is a bilaminate subepithelial layer of fibrogranular material, the friability of which is probably the basis for recurrent erosions in this disorder. Histochemical and ultrastructural findings provide further evidence that Cogan's dystrophy, the finger print/bleb dystrophy, and Meesmann's dystrophy should be regarded as separate entities.


The incorporation of reaction centres into membranes from a bacteriochlorophyll-less mutant of Rhodopseudomonas sphaeroides. Reaction centres purified from a blue-green mutant R-26 of Rhodopseudomonas sphaeroides can be incorporated into bacteriochlorophyll-less membranes purified from an aerobically-grown bacteriochlorophyll-less mutant 01 of R. sphaeroides. This can be accomplished by raising the temperature of the mixture or by addition of the detergent sodium cholate and its subsequent removal by dilution or dialysis. Optimum conditions for the reconstitution are at 4 degrees C in the presence of 1% cholate and soybean phospholipid (2 : 1, w/w, with membrane protein). Isopycnic sucrose density gradient centrifugation of such preparations shows that reaction centres and light-harvesting pigment-protein complex bind to the membranes. Reconstituted membranes exhibit light-induced steady-state cytochrome absorbance changes resembling those observed in chromatophores prepared from the photosynthetically-grown mutant R-26. The effect on these absorbance changes of varying reaction centre content in the membrane has been studied, and the time course of the interaction between 01 membrane cytochrome c2 and added reaction centre examined. Cytochrome b photoreduction and cytochrome c2 photo-oxidation were observed in the reconstituted preparation; each increased following the addition of antimycin A, suggesting that a cyclic light-driven system had been reconstituted.


A mathematical model of air and water caloric nystagmus. A mathematical model is proposed to explain the induction of nystagmic eye movements in response to thermal stimulation of the ear by air and water. Laplace-transformed equations are set up to describe heat flow in the meatus lumen to the ear-drum and heat transmission into meatus wall. Heat transport to the lateral semicircular canal, resulting in convective endolymph flow, and the induction of reflectory eye movements are included in the mathematical description. Input of the model is the time-course of temperature at the irrigating tip, output is the time-course of eye position (in correspondence to experimental nystagmogramms). The predicted nystagmus is in good agreement with experimental results, thus supporting our assumptions on the thermal effects of air and water irrigations.


[RNA-binding proteins of Rana temporaria frog oocytes. Isolation by methods of affinity chromatography and radioactive labelling in vitro]. A fraction of RNA-binding proteins from the maturating frog oocytes of Rana temporaria was isolated by the method of affinity chromatography on poly-U coupled to Sepharose. These proteins were labelled with high activity by the method of reductive methylation using 3H-sodium borohydride. It was shown that during the isolation and labelling there is no loss or inactivation of any RNA-binding protein fraction which is essential for informosome-like particles formation. The molecular masses of polypeptide chains in the isolated RNA-binding protein fraction were determined.


T and B lymphocytes during pregnancy.

T and B lymphocytes during pregnancy. Peripheral blood lymphocytes from 105 pregnant and 99 control women have been tested weekly for the nine months gestation period. Our results show no significant variations between age-matched controls and pregnant group lymphocytes in terms of their capacities to make spontaneous E-rosettes, active rosettes, EA-rosettes, to stain with anti-immunoglobulin antiserum, in cytotoxic anti-T serum assay and to dissociate into high (T) and low (B) electrophoretic mobility fractions. Concomitantly, the effect of pregnant serum on rosettes formation has been tested and found not to be different from the effect of pooled control sera.


[Induction of immune tolerance to thymus-dependent antigen (sheep erythrocytes) in the absence of T-cells]. The role of T cells in B cell tolerance induction to sheep red blood cells (SRBC) was studied in intact adult mice, in lethally irradiated mice injected with singeneic embryonic liver cells and thymocytes (TB-mice) and in animals functionally deprived of T cells--thymectomized, letally irradiated mice reconstituted with embryonic liver cells only (B-mice). Tolerance was obtained by treatment of mice with SRBC and cyclophosphamide (Cy). Cy-induced tolerance to SRBC was shown to be the result of the absence of specific T cells and partially of immunocompetent B cells. Suppression of immunoreactivity was observed not only in TB-mice but also in B-mice subjected to tolerogenic treatment. Splenocytes of tolerant TB-mice did not suppress the immune response of intact spleen cells to SRBC. The results obtained suggest the conclusion that B cells tolerance could be formed in absence of T cells.


[Immune response and tolerance to S. typhi Vi-antigen in T-lymphocyte depleted mice]. Comparison of immune response to Vi-antigen in thymetomized letally irradiated and reconstituted with fetal liver cells mice and in control animals revealed no difference between the two groups. The absence of enchancement of antibody formation in T cell depleted mice favours thymic-independent regulation of immune response to optimal dose of Vi-antigen. The induction of cyclophosphamide tolerance to Vi-antigen did not depend on the presence of T cells: tolerogenic treatment was equally effective in T cell depleted mice and in control animals. Therefore cyclophosphamide tolerance was not due to the activation of T suppressors but to direct elimination of immunocompetent clones of B cells.


[Fluorescent probe-indicator of differences between T- and B-lymphocytes in the blood]. A new physico-chemical marker for the human peripheral blood lymphocytes was worked out. The lymphocytes were vitally stained with the fluorescent probe 3-methoxybenzanthrone and measured by microfluorometry. The blood lymphocytes population was found to be heterogeneous; this population consists of the two main groups of cells differing by the intensity of their fluorescence. By means of immunological lymphocyte fractionation it was shown that one of these cell groups was represented by T-lymphocytes, and the other one--by B-lymphocytes.


B cell acute lymphoblastic leukemia (ALL) with a 14q+ chromosome abnormality. An adult patient with acute lymphoblastic leukemia associated with a 14q+ marker chromosome is presented. The abnormality resulted from a translocation of material from the long arm of chromosome 11. The leukemic cells were found to be B cells on the basis of surface immunoglobulins, lack of receptors for sheep erythrocytes, and a characteristically low level of adenosine deaminase activity. In other patients with ALL studied by us or reported by others in whom chromosome banding was done, a 14q+ chromosome was present in only one instance, also a case of B cell ALL. These two cases are the only examples of B cell ALL studied with chromosome banding reported to date. The frequent occurrence of a 14q+ chromosome in other malignant lymphoproliferative diseases of B cell origin suggests that a general association may exist between the 14q+ abnormality and B cell neoplasms. Cytogenetic analysis may therefore be useful in defining subtypes of ALL and in relating specific chromosomal abnormalities to lymphoproliferative disorders.


Size of cerebral ventricles in 66 psychiatric patients. The routine air ventriculograms of 66 psychiatric patients, aged from 22 to 73 years, taken during the psychosurgical operation of stereotactic subcaudate tractotomy, were studied. Ventricular size was unrelated to progressive ageing, but a minority of patients over 60 years had abnormally large ventricles, not invariably associated with cognitive impairment on testing. Enlargement was associated with a clinical diagnosis of schizoaffective illness but not with past ECT.


Return to work after coronary artery surgery for angina. The working habits of 115 consecutive patients who underwent coronary artery vein bypass grafting for angina were assessed. Only 25 patients worked up to the operation. Seven patients were housewives and seven retired. Sixty-eight patients had had to give up work because of their angina pectoris, and most of these had been off work for between six months and two years. After the operation 75 patients returned to full-time work, 59 within two to six months. These 75 patients included 23 of the 25 at work preoperatively and 47 of the 68 unable to work preoperatively. Most patients returned to their original occupation. We conclude that coronary artery surgery, as well as bringing symptomatic relief, increases the patient's ability to return to and maintain gainful employment.


Colonoscopy for unexplained rectal bleeding. Two hundred and thirty-nine patients underwent colonoscopy for unexplained rectal bleeding. Local anorectal conditions were excluded by digital and proctosigmoidoscopic examinations and results of barium studies were negative for all patients. A cause for bleeding was found in 95 patients. Thirty-nine had adenomatous polyps, 24 had unrecognised inflammatory bowel disease, and most importantly 23 (10% of series) had carcinomas. Forty patients had diverticular disease, but nine of them were found to have an adenomatous polyp and four a carcinoma. Colonoscopy can contribute positively to the investigation and treatment of unexplained rectal bleeding and may prevent unnecessary laparotomy.


Wavelength discrimination in the leopard frog: a reexamination. A reexamination of wavelength discrimination using a 2-choice paradigm and both chromatic-chromatic and chromatic-achromatic stimulus pairs has reaffirmed the positive tendency of Rana pipiens to approach short wavelengths. In addition, a strong avoidance of long wavelengths was observed and an interaction of these two opposite response tendencies was demonstrated. Thus, the percent choice frequency for a short-wavelength stimulus depends upon the stimulus with which it is paired. A positive phototaxis appears to influence response to some short-wavelength stimuli when they differ in brightness from an achromatic stimulus, but not with wavelengths above 471 nm.


Immunological basis of close-contact sensitization to osteosarcoma. Normal persons with a history of close contact with cancer patients often given positive responses in tests for specific tumor immunity. The present study was designed to establish the immunological relevance of such reactivity. Leukocyte adherence inhibition analysis was used to test responses of osteosarcoma (OS)-positive donors to extracts of OS. The test system proved highly specific in that positive leukocyte adherence inhibition responses were detected in 35 of 39 OS patients while only 6 to 7% of the controls were positive. Thus, the test system appears useful in the detection of OS. Two persons reactive presumably because of exposure to OS tissue gave booster-like leukocyte adherence inhibition responses on each reexposure. Reactivity was abrogated by treatment of their leukocytes with a goat anti-T-cell immunoglobulin G. Thus, the reactivity of cells from normal donors exposed to OS probably has an immunological basis, since the test system was highly specific, reflected anamnestic reactivity, and depended on lymphocytes (probably thymus dependent). These findings suggest that an OS-associated antigen can be transmitted horizontally, but the relation of immunity to risk remains obscure.


Continuity between the ventricular and subarachnoid cerebrospinal fluid in an amphibian, Rana pipiens. Continuity between the ventricular and subarachnoid cerebrospinal fluid has been investigated in Rana pipiens. The structure of the posterior tela, a deficient membrane situated at the extreme caudal end of the roof of the fourth ventricle, has been studied using whole membrane mounts and by light microscopy of resin embedded tissue. The ependymal component consists of columnar and rounded cells which form a regular 'syncytium' enclosing round and oval fenestrations. Small fenestrations are covered on the subarachnoid side by elongated pial cells and thus do not give total continuity between the fourth ventricle and the subarachnoid space. Large fenestrations, on the other hand, are accompanied by equivalent pial fenestrations giving direct access between the fluid compartments. Towards the caudal end the fenestrations break up and the numbers of ependymal and pial cells decrease, the caudal end itself being characterised by a small remaining clump of ependyma and pia or of pia alone. Flow through the tela has been studied using fluorescein-labelled dextran placed in the intraventricular space. Infusion into the lateral ventricle and subsequent localisation by fluorescence microscopy shows the marker to be in the fourth ventricle, in the fenestrations of the posterior tela and in the subarachnoid space overlying the tela. Infusion of the marker followed by freezing and examination of the cut heads on a freezing microtome, shows fluorescence throughout the ventricular system, in the subarachnoid space adjacent to the posterior tela and also along the dorsal subarachnoid space of the spinal cord.


Human granulocyte specific nuclear antigen(s). I: Production of antisera and determination of specificity. Non-histone protein-DNA complexes isolated from human lung tissue were used to immunize New Zealand rabbits. Quantitative microcomplement fixation and immunocytochemistry revealed a specific chromatin antigen(s) in the human granulocyte nucleus. The study demonstrates the feasibility of employing antisera against chromosomal non-histone protein-DNA complexes for the immunochemical identification of cell types.


Nutritional anaemia--a major controllable public health problem. Nutritional anaemia, due chiefly to iron deficiency, is widely prevalent in many parts of the world. There is increasing evidence that even mild anaemia affects health and reduces productivity and that a high prevalence of anaemia has profound socioeconomic consequences. The pathogenesis of nutritional anaemia is now reasonably well understood. Measures avilable for combating it include: therapeutic supplementation for accessible population groups with a high prevalence of anaemia, such as pregnant women and schoolchildren; iron fortification of one or more widely consumed foodstuffs; management of those conditions, such as hookworm infestation, that increase requirements for haemopoietic nutrients; and education of the public, and of all categories of health personnel, regarding the importance of anaemia and the ways of controlling it. Experience has shown that there is no simple solution to the problem and in each area where iron deficiency anaemia is prevalent it will probably be necessary to develop and combine many or all of these measures. In each community it will be necessary to introduce these measures so that their effectiveness can first be studied in a pilot trial. When this has been successfully completed it should be followed by a field trial under realistic conditions, and only when this has proved successful should a regional or national programme be introduced. However, the problem is complex and it is only by sustained effort of all concerned that it will prove possible to develop adequate public health control of nutritional anaemia.


Role of veterinarians in modern food hygiene. Veterinary services and veterinary education and training must keep pace with the constantly changing patterns of agriculture and food processing. Changes in methods of animal production are associated with many problems of food processing and food quality. Veterinary supervision of the animal feed industry and of meat and distribution is essential. Quality testing of meat, milk, and eggs requires the introduction of suitable routine sampling systems, laboratory procedures, and complex evaluation procedures.Food hygiene problems have changed in recent years not only as a result of new methods of animal production, but also because of changes in food processing technology and in the presentation of food to the consumer, increased environmental pollution, increased international trade, and increased tourist travel.Food hygienists must adopt an active and progressive policy and change the scope of food control from a purely negative measure into a positive force working towards improved food quality and the avoidance of losses during production. A modern food hygiene programme should cover all stages of production, processing, and distribution of food and also other ingredients, additives and the water used for production and processing. Veterinarians should also be involved in the registration and licensing of enterprises and this should take into account the premises, the procedures to be used, new techniques in animal husbandry, machines and equipment, etc.In order to facilitate the microbiological analysis of foodstuffs, new mechanized or automated laboratory methods are required, and consideration must be given to adequate sampling techniques.


Statistical principles of monitoring and surveillance in public health. Monitoring and surveillance are seen as statistical procedures that will help health authorities to achieve better health services with the existing resources-monitoring being an integrated system of making observations on health and environmental factors and of scrutinizing, storing, and retrieving those data, and surveillance being a closely associated system for collating and interpreting the data.Monitoring should be an action oriented activity and may encompass a wide range of health activities, for example, communicable and noncommunicable diseases, environmental pollutants, and specific problems in health care delivery systems. Both monitoring and surveillance systems have to be related to control measures, and, since the available resources are usually limited, a scale of priorities has to be developed by the statistician in cooperation with the competent authorities.MONITORING MAY BE PERFORMED ON EITHER THE INDIVIDUAL OR AGGREGATE LEVEL AND SHOULD BE PLANNED TO TAKE INTO ACCOUNT THE COURSE OF THE DISEASE UNDER CONSIDERATION: it is concerned with monitoring stimuli and events. Collection of data, for example, on exposure to a pollutant, may be continuous and automatically recorded, or regular or irregular through population sampling or registries. The statistical requirements of monitoring and surveillance systems are discussed and a checklist of features to be considered is given in an annex.


The effect of undernutrition on immunological responses to BCG and TAB vaccines in mice maintained on a natural diet. A study was made on the effect of undernutrition on the immunological responses of mice to TAB and BCG vaccines. The mice were fed a diet comprising chapati and germinated grams; one group was fed ad libitum while another was given half the quantity consumed by the first group. The undernourished mice showed a poor immune response to BCG: only 23% showed a positive reaction to foot pad challenge with purified protein derivative, compared with 76% of the well-fed mice. The mean increase in thickness of the foot pad was also significantly smaller in the undernourished mice. Antibody titres against the H and O antigens of TAB were essentially similar in both groups of mice. The results suggest that undernutrition depresses cell-mediated immune responses but not humoral responses.


Evaluation of a finger prick blood collection method for the seroepidemiology of hepatitis B. A finger prick-swab method of blood specimen collection was qualitatively and quantitatively compared with the conventional venipuncture method for HBsAg and anti-HBs determinations by radioimmunoassay (RIA). The new method consisted of pricking the finger, collecting 0.1-0.2 ml of blood with a cotton-wool swab, and eluting the swab in 1 ml of 1% bovine albumin in saline containing 0.1% sodium azide. Using chimpanzees seropositive for HBsAg or anti-HBs, comparisons were made of RIA results of: (a) whole blood, haemolysed blood, serum, and plasma; (b) paired finger prick samples and serum; (c) dilutions of finger prick samples and serum; and (d) different volumes of blood on swabs. Field studies were carried out at two institutions where hepatitis B was hyperendemic to compare results from paired finger prick and serum specimens assayed by the RIA and haemagglutination techniques. The laboratory studies showed that swab RIA values for anti-HBs were significantly lower than serum values and that for HBsAg, swab values were significantly higher than serum values. In HBsAg tests, the field studies showed 100% agreement between the two methods; in anti-HBs tests, the finger prick method showed 85% agreement with positive sera. Because of the logistics of collecting and processing blood serum, the finger prick-swab technique may be a valuable aid in large-scale seroepidemiological surveys for hepatitis B.


A serological survey of Lassa fever in Liberia. A serological survey was undertaken at four Liberian hospitals in 1974 in which serum samples were taken from 104 health workers and 61 patients. Six persons had Lassa fever antibodies: four midwives and two students of midwifery. Of those persons who had lived in Loffa County, 4 of 22 midwives were seropositive whereas none of the other 39 residents were positive (P = 0.014).


Proposed rules for the designation of immunoglobulins of animal origin. The present article sets forth proposed rules for the designation of immunoglobulins of animal origin and their chains. It also recommends symbols to be used for the designation of classes and types, subclasses and subtypes, (sub) subclasses and (sub) subtypes, and allotypes. Recommended rules for the designation of newly discovered polypeptide chains are also included.


Viral haemorrhagic fevers of man.

Viral haemorrhagic fevers of man. This article reviews the current state of knowledge on the viral haemorrhagic fevers that infect man, namely smallpox, chikungunya fever, dengue fever, Rift Valley fever, yellow fever, Crimean haemorrhagic fever, Kyasanur Forest disease, Omsk haemorrhagic fever, Argentinian haemorrhagic fever (Junin virus), Bolivian haemorrhagic fever (Machupo virus), Lassa fever, haemorrhagic fever with renal syndrome, and Marburg and Ebola virus diseases.


The need for closer collaboration between the medical and veterinary professions. It is increasingly apparent that physicians and veterinarians share the same pool of scientific knowledge and that diseases of animals have many direct and indirect connexions with human health. Nowadays it is realized that, given the opportunity, the veterinarian can make substantial contributions to the medical services by (a) controlling zoonoses, (b) supervising the hygiene of food, especially food of animal origin, (c) assisting in the detection and prevention of environmental pollution, (d) facilitating exchange of research information on analagous problems in man and animals, and (e) ensuring a supply of healthy, standardized laboratory animals. Appropriate administrative machinery at government level is necessary to enable the veterinarian to develop and exercise his potential in this field and to ensure full and effective collaboration between the medical and veterinary professions. Conventional veterinary education provides an excellent background for public health work, but special training is also necessary, at both the undergraduate and postgraduate levels, for veterinarians who are to assume responsibilities in public health. A fuller partnership between these two health professions, which have so much in common, should be encouraged in various ways, for example by sharing some courses during university education, and by joint meetings to discuss problems of mutual concern.


Guiding principles and recommendations on labelling of clinical laboratory materials: a WHO memorandum. Comprehensive labelling forms a vital part of any steps to control the quality of commercially distributed diagnostic reagents. This Memorandum gives recommendations for such labelling. Specifications are given for the recommended minimum information to be given on the label attached to the immediate container of clinical laboratory materials, kits and kit components, reference materials including calibrators and control materials, and where applicable, of general laboratory materials. A package insert or brochure is generally required for clinical laboratory materials, kits, and reference materials, and specifications are also given for information to be given in such inserts or brochures. Definitions of terms used in this Memorandum are given in an Annex. It is hoped that WHO Member States will use these specifications through their adoption as national regulations.


Recent advances in rheumatic fever control and future prospect: a WHO memorandum. The public health importance of rheumatic fever is not directly related to its prevalence but is a complex function of many factors, such as incidence, prevalence, mortality, disability, and cost to the community, all relative to the importance of other diseases. Although the prevalence is low and still decreasing in developed countries, rheumatic fever is the commonest form of heart disease in many developing countries. Advances in the bacterial and serological characterization of group A streptococci are reviewed in this Memorandum, with emphasis on the nature of the structural elements of the cell wall. Microbiological methods play an important role in the diagnosis and control of rheumatic fever and up-to-date information is given concerning methods of isolation and grouping and of typing. Methods of assessing streptococcal activity in the host, in particular the different antibody responses found in relation to different pathological conditions, are discussed, as are the improvements required in laboratory services to enable them to carry out these duties adequately. Current concepts concerning the pathogenesis of rheumatic fever are reviewed together with differences in the epidemiology related to the different sites of infection and the problems of carriership. The Memorandum is also concerned with the methods of prevention and treatment, including prevention of recurrences, control of the disease in communities, and strategies for nationwide control.


Influenza in China in 1977: recurrence of influenzavirus A subtype H1N1. Preliminary results from epidemiological and laboratory studies on the new H1N1 influenza virus show that the 7-20 years age group suffered the highest morbidity; some adults over 20 years of age were also affected. The influenza epidemic caused by the H1N1 virus was characterized by slow spread, unevenness of attack rates, and the occurrence of many mild cases and inapparent infections. At least up to the end of 1977 there was concurrent persistence and spread of both H1N1 and H3N2 viruses. The H and N antigens of the new H1N1 virus, as well as its behaviour toward nonspecific inhibitors, were found to be closely similar to the old H1N1 virus prevalent during the first half of the 1950s. Most of the new H1N1 isolates in eggs were found to be temperature sensitive.


Influenza in the USSR in 1977: recurrence of influenzavirus A subtype H1N1. Early in November 1977, several outbreaks of influenza were reported in the far eastern region of the USSR. The epidemic spread rapidly throughout the country affecting mainly people under the age of 20 years. Most of the strains of virus isolated were found to be influenza A subtype H1N1. The serological characterization of the strains is described in this paper.


Comparison of influenza viruses isolated from man and from whales. Four isolates of influenza virus strains from Moscow and Habarovsk that caused outbreaks of influenza in November and December 1977 in several cities of the USSR were studied and their haemagglutinins and neuraminidases were compared with those of other human and animal influenza viruses including A/whale/Pacific Ocean/76. In H1 tests these isolates, designated A/USSR/77, reacted with immune serum against A/FM/1/47 (H1N1) to the homologous titre, and with antiserum against A/whale/PO/19/76 virus to 1/8 of the homologous titre. In neuraminidase inhibition tests all A/USSR/77 isolates showed the presence of human N1 type neuraminidase, more closely related to A/sw/New Jersey/76 (Hsw1N1) than to A/FM/1/47 (H1N1) virus. The haemagglutinin of A/whale/Pacific Ocean/19/76 virus occupies an intermediate position between H0 and H1, but its neuraminidase is close to Nav2. The virus from whales multiplies better at low (28 degrees C) and at high (40 degrees C) temperatures than do the viruses of human origin that were tested.


Epidemiology of influenza during 1957--75 in the Union of Soviet Socialist Republics and the German Democratic Republic. Analysis of clinical data collected by the epidemiological services in both countries showed that each shift or drift in the antigenic structure of the influenza virus resulted in epidemics in both countries. Depending on the degree of antigenic change, both shifts and drifts influenced the speed of epidemic spread and the time of occurrence and the intensity of the epidemics. However, the analyses did not reveal a direct relation between degree of antigenic variation and the attack rate or the severity of the epidemic. The mean attack rate in both countries was about 13% depending on the pathogenicity of the circulating virus. The severity of the epidemics varied in the two countries but the fourth epidemic after each shift was severe in both countries. The reasons for this and other features of the epidemiology are discussed.


Combined vaccination with BCG and toxoid antigens. Simultaneous vaccination with different antigens has been widely practised in recent years. A notable example is percutaneous smallpox vaccination together with the intradermal injection of BCG. In contrast, the potentially even more time-saving procedure of combined vaccination (i.e., with vaccines mixed prior to injection) has been tried on only a very limited, experimental scale. Combined vaccination with a mixture of BCG and toxoids has not been suggested before. Such a mixed vaccine, with diphtheria and tetanus toxoids, was used in experiments in vitro and in vivo.No deleterious effect of toxoids on BCG was found in terms of reduction in oxygen uptake, germination rate, or counts of viable particles. The dose-response relationships (delayed hypersensitivity and skin reactivity) for BCG with and without added toxoids were compared in guinea pigs and no differences were found. The antibody response in guinea pigs to toxoids mixed with BCG or with aluminium hydroxide was measured following both primary and booster immunization. The primary response to toxoids was lower with BCG than with aluminium hydroxide. In booster immunization, the response was identical for the two mixtures.It appears that not only is a mixture of toxoids and BCG innocuous (given intradermally) but also BCG may have an adjuvant effect on the production of antibodies to the toxoids. Further experimentation is needed, first in the guinea pig model and later in pilot trials in man, to establish suitable dose levels. Furthermore, since BCG may act as an adjuvant both in producing antibodies and in the cell-mediated response to the toxoids, it would be desirable to clarify the possible interaction of these two immune responses in protection against disease.


Further observations from St Lucia on control of Schistosoma mansoni transmission by provision of domestic water supplies. Individual households in five settlements were provided with piped water in a pilot scheme to investigate the effect on transmission of S. mansoni in St Lucia. Nearby comparison settlements, in the same valley, were provided with water through a public standpipe system. The incidence of S. mansoni infection among children decreased in the experimental area, leading to lower prevalence rates and lower intensity of infection in all age groups. Over the study period, indices of infection increased in the comparison settlements, but by the end of the period development was making those settlements less suitable for comparison purposes and some reduction in transmission was occurring.The changes in human infection rates were reflected in the results of studies with sentinel snails. In the experimental area, infection rates gradually fell owing to reduced water contact and consequently less contamination of the river and its banks, and possibly to the gradual reduction in contamination potential of the community with reduced prevalence and intensity of infection. It is suggested that a piped water supply be considered as a method of schistosomiasis control, but that the cost should not be debited only to the control of this disease since a clean water supply has other medical and social benefits.


The use of a continuous cell line for the isolation of influenza viruses. Cultures of MDCK cells exposed to trypsin were as efficient as cultures of rhesus monkey kidney cells for detecting influenza virus, both in dilutions of infected allantoic fluids and in nose and throat swabs. We suggest that the MDCK cell/trypsin system provides a satisfactory alternative to monkey kidney cultures for the isolation of influenza viruses from clinical specimens.


Relation of creatine kinase isoenzyme MB to postoperative electrocardiographic diagnosis in patients undergoing coronary-artery bypass surgery. We compared (a) the frequency of detection of isoenzyme MB of creatine kinase (CK; EC 2.7.3.2) in serum of patients undergoing coronary-artery bypass surgery, (b) the interval during uhich its activity was supranormal in serum, and (c) an index of the amount of CK released into blood ("CK-MB area") with postoperative electrocardiographic changes in 80 patients. The frequency of detection of CK-MB is a function of frequency of sampling during the early postoperative period. Because the duration of appearance and the calculated CK-MB area increased as the electrocardiogram became more specific for infarction (p less than 0.01), a twice-daily sampling schedule proved clinically relevant. Only 5.4% of patients had electrocardiographic evidence of infarction when CK-MB was absent by the second postoperative morning. When CK-MB was still detected at that time, 69.6% of patients had persistent new Q waves, consistent with infarction. In three patients who died postoperatively, significant myocardial necrosis was demonstrated. All three had had persistently increased values for CK-MB, related to electrocardiographic changes of infarction in one patient and ischemic changes in two. Evidently CK-MB is a more sensitive indicator of myocardial necrosis than the electrocardiogram and CK-MB area should be a useful criterion in evaluating methods of intra-operative myocardial protection.


Experimental and clinical immunopathology of middle ear transplantation. On the basis of experimental data the concept of immuno-tolerance in allograft-tympanoplasty is discussed. Immunological interference in human tympanic membrane grafts has been demonstrated by means of T-cell membrane fluorescence.


Studies of surface immunoglobulins on human B lymphocytes. III. Physiological variations of SIg+ cells in peripheral blood. The levels of B lymphocytes bearing surface immunoglobulins in human peripheral blood were measured by immunofluorescence after pre-treatment of the cells with a low pH shock or washing at 37 degrees C. It was found that the percentages of SIg-bearing cells in an individual varied throughout the day and showed a circadian rhythm with a peak at 12 midnight and a depression at 12 noon. In addition, the levels of the B lymphocytes in the peripheral blood were shown to vary annually, being lower in the winter than in the summer.


Control of cloning of normal human T lymphocytes by transferrin, albumin and different lectins. Normal human T-lymphocytes can be induced to form colonies with a high cloning efficiency by seeding the cells directly in agar with normal human plasma and the lectins concanavalin A, phytohaemagglutinin, or pokeweed mitogen. The requirement for human plasma can be substituted, to different degrees with different lectins, by adding transferrin and albumin to foetal calf serum. This provides a useful system for identifying specific deficiencies in the requirements for normal T-lymphocyte colony formation.


The effect of levamisole on E-rosette formation by trypsinized lymphocytes. Normal lymphocytes treated with trypsin lost their ability to form normal numbers of rosettes with sheep red blood cells. The recovery in E-rosette forming capacity of trypsinized cells was considerably augmented when the cells were incubated in the presence of a wide range of concentrations of the immunostimulatory drug, levamisole. Substantial recovery of rosette-forming ability was seen 2 hr after incubation of trypsinized cells with levamisole.


Theophylline modulation of E-rosette formation: an indicator of T-cell maturation. The binding of unsensitized sheep erythrocytes is a characteristic of human thymus dependent T-lymphocytes. We have investigated the effect of theophylline on E-rosette formation using cells from normal individuals, and patients with immunodeficiency or acute lymphoblastic leukaemia, and have attempted to correlate the influence of the drug on distinct T-lymphocyte subpopulations. Three subpopulations of E-rosetting T-lymphocytes can be delineated: theophylline-sensitive T-cells which lose the capacity to form E-rosettes following treatment; theophylline-resistant T-cells which are unaffected by the drug; and theophylline-dependent cells which acquire the ability to form E-rosettes following incubation with theophylline. The action of theophylline was shown to be dose-dependent, temperature-dependent and reversible. Reversibility or re-expression of the receptor for sheep red cells could be blocked by the addition of puromycin. In peripheral blood, E-rosetting T-lymphocytes were roughly divided into two equal populations, one sensitive, the other resistant. Thymocytes were shown to be entirely theophylline-resistant, whereas a small population of cells in peripheral blood and bone marrow were induced to become E-rosetting in the presence of theophylline. Induction by theophylline may be effective at a distinct stage of precursor T-cell differentiation.


Cell-mediated immunity in mice vaccinated against malaria. Mice vaccinated with a formalin-fixed preparation of either Plasmodium berghei or P. yoelli exhibited delayed type hypersensitivity (DTH) to the homologous antigen. This manifested itself in increased delayed thickening of antigen-challenged pinnae of the vaccinated mice as compared to the non-vaccinated controls. DTH was also evident in the vaccinated mice using the homing of radio-labelled bone marrow cells (BMC) to the delayed lesion as a criterion of reactivity. When P. yoelii vaccinated mice were given a live infection P. yoelii, a marked migration of BMC into the spleen occurred, with a peak at 48 hr, and it is suggested that this was a systemic response of DTH. The splenic T-cells of P. yoelii-vaccinated animals transformed in vitro with a soluble extract of the homologous parasite. The potential function of cell-mediated mechanisms in immunity to malarial infections is discussed.


Long-term antiarrhythmic therapy with N-acetylprocainamide. The effects of long-term NAPA therapy were evaluated in 6 patients with chronic PVCs known to respond to this drug during a previous placebo-controlled, dose-ranging trial. Underlying cardiac status was evaluated every six months by switching each patient from NAPA to placebo. Placebo period PVC frequency after one year of NAPA therapy was reduced, compared to baseline placebo values. Mean PEP/LVET, measured while the patients received placebo, was elevated at the beginning of the study but was normal after one year of NAPA therapy. Comparison of NAPA and placebo period observations indicated a reduction in PEP/LVET when NAPA therapy was begun. This effect, however, could not be demonstrated one year later when mean placebo period PEP/LVET was normal. The apparent dependence of this effect on underlying status of left ventricular function suggests that the initial reduction in PEP/LVET represents an an indirect effect of NAPA rather than a direct inotropic action. NAPA therapy was well tolerated by the 6 patients and ANA titers became abnormal in only one, in marked contrast to reported experience with procainamide.


Chloroquine serum concentration and side effects: evidence for dose-dependent kinetics. Serum concentrations of chloroquine were determined fluorometrically in 100 rheumatoid patients who had been treated with 0.25 gm daily for at least 2 mo. The total dose varied between 3.7 and 400 gm. No patient received more than 75 gm annually. In 15% of the patients side effects were noted. There was a relationship between serum concentrations and side effects but not with the total dose administered. Chloroquine displayed dose-dependent kinetics, which may indicate that close monitoring of serum concentrations is an aid to the safe and rational use of the drug.


The double contrast barium meal in patients with acute upper gastrointestinal bleeding. One hundred and seven patients admitted to a small general hospital with acute upper gastrointestinal bleeding have been examined by the double contrast barium meal technique. The presumed bleeding site was identified in 75 cases (70%). Twenty-eight patients showed radiological evidence of recent or active bleeding. These were all examined within 24 h of the bleed and 18 (64%) continued to bleed or had a further bleed whereas of 79 patients who did not show radiological signs of recent or active bleeding only 10(13%) continued to bleed or had a further bleed. The radiological features of recent or active bleeding seen on the double contrast barium meal are: 1, Blood clot in an ulcer or adherent to a recently bleeding lesion. 2. An artery in the base of an ulcer. 3. Active bleeding during the course of the examination seen as a dynamic alteration or disturbance in the barium as it flows over the bleeding site.


Acute and subacute toxicology and safety evaluation of triphenyl tin hydroxide (Vancide KS). The acute oral LD50 in Sprague-Dawley rats was determined to be 171 mg/kg (100-295) for males and 268 mg/kg (205-344) for females. 2. A 1 ppm dietary supplement of Vancide KS for 90 days did not induce any abnormalities in weanling Sprague-Dawley rats. The parameters evaluated were serum glutamic-oxaloacetic transaminase (SGOT), hematocrit, differential white blood cell count, food consumption, and weight gain, along with histologic studies of the myocardium, spleen, liver, kidney, stomach, and small intestine. A 500 ppm diet was lethal. Weanling and older rats subjected to 1000 and 10,000 ppm diets died within 5 days. 3. Vancide KS induced no acute dermal toxicity, nor did it exhibit percutaneous absorption in New Zealand strain albino rabbits. 4. Vancide KS induced no chronic dermal toxicity in New Zealand strain albino rabbits. 5. Vancide KS was not shown to be teratogenic. It exhibited an antifertility action, especially in Sprague-Dawley rats dosed on day 1 through day 7 of timed-pregnancy. 6. Vancide KS was shown to be an eye irritant which induces corneal opacity. 7. Acute oral toxicity studies in Sprague-Dawley rats indicate that Vancide KS should be classified as a toxic substance as defined in the regulations under the Federal Hazardous Labeling Act. 8. The intravenous administration of 25 mg/kg of Vancide KS to New Zealand strain albino rabbits induced death preceded by topic convulsions. 9. Vancide KS did not induce skin sensitization in male adult guinea pigs.


Colonoscopy in the diagnosis of unexplained rectal bleeding. Colonoscopy, done to elucidate the cause of obscure rectal bleeding not diagnosable on the basis of previous proctoscopic and barium-enema examinations, resulted in a 17 per cent diagnostic yield in this series. Eighteen of 105 patients were found to have lesions: three had carcinomas, six had polyps, eight had colitis, and one had proctitis. When bleeding had been demonstrated by the physician, there was a higher diagnostic yield (22.5 per cent) than when bleeding was reported only by the patient (5.9 per cent). Descriptions of the bleeding by the patients were found to be somewhat unreliable. Sixteen of the 18 lesions revealed by the colonoscope were in the left colon. It is suggested that the basic work-up for undiagnosed rectal bleeding should include history, physical examination, sigmoidoscopy, and barium-enema study. When no diagnosis is found, subsequent work-up must be individualized. When bleeding has been demonstrated to originate from the colon, colonoscopic examination should be performed.


Endoscopy of the cerebellopontine angle. Endoscopy of the pontine angle ("cisternoscopy") is a decisive diagnostic procedure in clarifying pathological processes in this region. Using a retrolabyrinthal approach through the mastoid bone, the endoscopes are inserted in the pontine angle for direct inspection. In the present study the method and the diagnostic results are described.


Central and peripheral antialgesic action of aspirin-like drugs. The peripheral and central effects of some non-steroid anti-inflammatory drugs, aspirin, indomethacin, paracetamol and phenacetin were studied by comparing their intraplantar and intracerebroventricular effects on hyperalgesia induced by carrageenin injected into the rat paw. Hyperalgesia was measured by a modification of the Randall-Selitto test. The agents tested had antialgesic effects when given by any route. Their intraventricular administration enhanced the antialgesic effect of anti-inflammatory drugs administered into the paw. Previous treatment of one paw with carrageenin reduced the oedema caused by a second injection of carrageenin in the contralateral paw. In contrast, it had no effect on the intensity of hyperalgesia but shortened the time necessary for it to reach a plateau. Administration of a prostaglandin antagonist (SC-19220) in the cerebral ventricles, in the rat paw or in both sites, significantly inhibited the hyperalgesia evoked by carrageenin. The maximal hyperalgesic effect of intraplantar injections of prostaglandin E2 could be further enhanced by its cerebroventricular administration. It was suggested that carrageenin hyperalgesia has a peripheral and a central component and that the cyclo-oxygenase inhibitors used may exert an antialgesic effect by preventing the hyperalgesia induced by a peripheral and/or central release of prostaglandins.


Distribution of CSF (colony stimulating factor) in kidney of the mouse. The activity of the colony stimulating factor (SCF) was measured in kidney subcellular fractions in mice. The highest activity was noted in microsomes. From other fractions, the cytosol had large amounts of CSF. On the basis of literature data, and the findings presented we suggest that the kidney is at least one of the organs of CSF biosynthesis.


Benign neoplasmia and pseudoneoplasia of the small bowel in children. Fourteen cases of benign tumorous conditions in children are reported. Each new growth described represents a true neoplasm, one simulating a true neoplasm, or one simulating a true tumor, clinically and/or radiographically. The series contains 10 female and 4 male patients. Most patients were under 1 year of age when manifestations--most often, gastrointestinal hemorrhage or bowel obstruction-- of the condition became evident. Nine patients had roentgenographic findings which indicated the presence of an abnormality, but the specificity of the findings was low. Some patients had cutaneous or other somatic clinical stigmata found in known syndromes also containing gastrointestinal pathology. These stigmata and the awareness of the clinical and roentgenographic manifestations of small bowel tumors in children should lead to an early diagnosis and hopefully to the usually successful surgical therapy.


Highland and lowland populations of Lesotho. It has not been possible to demonstrate significant sero-genetic differences between lowland and highland Sotho populations; the differences which do exist may well be attributable to random genetic drift. The study shows that the Sotho have received an appreciable genetic contribution from the San they have absorbed but their sero-genetic profile remains eminently Negro. A low frequency of the PTC non-taster allele was found (t = 0.142 +/- 0.029) as was the overall frequency for colour blindness (cb = 0.013 +/- 0.009).


Human lymphoid cell lines as targets for DRw. Cultured human lymphoid cell lines (LCL) are useful as a source of target cells in several immunologic assays. More recently such cells have been used for the serological characterizations of the HLA-DR antigens. Typing of the same LCL in various laboratories during the VII Histocompatibility Workshop has given comparable results with a discordancy rate of less than 10%. This discordancy is likely to reflect the different sources of complement that can greatly alter the results of cytotoxic assays. The presence of naturally occurring antibody in rabbit complement to human cells can be avoided by: (a) absorbing with human cells at 0 degrees C; (b) dilution with human serum; (c) dilution with heat-inactivated rabbit serum; (d) repeated freeze-thawing of the complement; or (e) careful selection of complement by screening procedures. Comparison of the results of HLA-DR typing of LCL with peripheral B-cells of the same donor show good correlations. However, LCL will occasionally give extra reactions perhaps due to the expression of new antigens. LCL can be coated with F(ab')2 fragments from antihuman beta2-microglobulin antibodies that block reactions of HLA-A, -B and -C antibodies allowing for discrimination of anti-DRw activity.


Studies of hyperthymic mice. II. The influence of thymus grafts on cell flow through the peripheral T-cell pool. In normal CBA/H mice implanted under the kidney capsule with eight CBA/H.T6T6 neonatal thymus lobes it was observed that the percentage of marked thymus-graft derived T cells in the periphery, after building up to a peak, showed a biphasic exponential decline. The initial decline was very rapid and appeared to be due to loss of the thymus-graft derived cells from the system. The later decline was slower and was the same as that of an introduced cohort of lymph-node lymphocytes. The second rate of decline was, however, considerably more rapid than that of lymph-node cohort in non thymus-grafted mice. We conclude that in multiply thymus-grafted mice the flow of cells through the T-cell pool is more rapid than in normal mice and that in this sense the thymus can be thought to drive the lymphoid system.


Heterogeneity of the B cell subpopulation operationally defined by (a) differentiation antigen(s) common to MOPC 104E and mature IgM plasma cells. An antiserum raised in rabbit against MOPC 104E myeloma cells was extensively absorbed by murine IgM, thymocytes and spleen cells. Using indirect immunofluorescence, the distribution of the corresponding mouse plasma cell antigen(s) (MPCA) was determined among immunized spleen cells. Only 1-3% of the cells were stained but this MPCA-bearing subpopulation included all IgM plasma cells, a sizable proportion of IgG plasma cells and about one third of the antigen binding cells identified following deliberate immunization. It is therefore proposed that MPCA is transiently expressed during the antigen-induced differentiation of virgin B lymphocytes into memory cells as it does during the maturation process into Ig-producing cells and thus reflects the ontogenic relationship of these two differentiation pathways.


The role of the thymus for maturation of transferred bursa cells into immunocompetent B cells in chickens treated with cyclophosphamide. Chickens injected with cyclophosphamide and X-ray irradiated in the newly hatched period were immunized with a mixture of sheep red blood cells, Brucella abortus and Salmonella pullorum at 4, 5 and 6 weeks of age, and were examined for serum antibody titres, serum immunoglobulin concentration and bursal and splenic structures at 7 weeks of age. The neonatal treatments suppressed completely or almost completely antibody responses, immunoglobulin production and formation of bursal follicles and splenic germinal centres. The transplantation of bursa cells into the chickens immunologically impaired by the treatments restored these functions and structures. In contrast, the transfer of bursa cells into chickens thymectomized, cyclophosphamide-treated and X-ray irradiated did not result in efficient restoration of the bursa-dependent immune system; 10-day-old bursa cells hardly restore the system, although 4-week-old bursa cells did so slightly. The chickens thymectomized, cyclophosphamide-treated, X-ray irradiated and repopulated with 10-day-old bursa cells were examined for the existence of functional B cells with the use of a syngeneic cell transfer system. The experiments verified that immunocompetent B cells had not developed in the chickens thus treated.


Why do protease inhibitors enhance leucocyte migration inhibition to the antigen PPD? Epsilon amino caproic acid (EACA) and Aprotinin significantly increase the inhibition to PPD in the leucocyte migration test (Burden, Stacey, Wood & Bell, 1978). This is not due to increased leucocyte inhibition factor (LIF) generation. The proteolytic inhibitors prevent the loss of activity of LIF due to prolonged incubation in the presence of serum. Both have a significant effect on the target cells (Polymorphonuclear leucocytes) used in the assay to exaggerate the migration inhibition to LIF. These actions can be explained by decreased proteolytic breakdown of LIF.


Titration and circulation of cells mediating delayed type hypersensitivity in normal and cyclophosphamide treated mice during response to sheep red blood cells. The delayed-type-hypersensitivity (DTH) reaction observed in mice primed i.v. with low doses of sheep red blood cells is not detectable after priming with large doses. Estimation of the number of DTH mediating cells (DTH-C) was performed by titration (D50 determination) using local adoptive transfer of these cells mixed with antigen into footpads of unprimed mice. In opposition to the absence of peripheral DTH-C, splenic DTH-C were more numberous after large doses than after low doses of antigen. The ability of cyclophosphamide to restore DTH responsiveness in mice primed with large doses appeared to be related to an increased cirulation of DTH-C. These cell circulated only in mice without any detectable B-cell response or in which this response was depressed. Thus a B-cell dependent splenic retention process of DTH-C is discussed as the origin of DTH unresponsiveness observed after priming with large doses of antigen.


Bone marrow suppressor B cells in vitro. Marrow cells from intact CBA and (CBA x C57B1)F1 mice added to a culture of syngeneic splenocytes at its initiation suppressed the generation of anti-SRBC antibody-forming cells. Removal of cells bearing surface immunoglobulins and/or MBLA led to a substantial reduction in the marrow suppressive activity. Marrow cells from mice pretreated with five doses of hydroxyurea did not suppress the immune response of syngeneic splenocytes to SRBC. While the total number of cells in the marrow of such animals declined 5-fold, the relative content of Ig-positive cells was somewhat increased. The proportion of blast cells dropped from 10.3 to 2.4%. No blast cells bearing surface immunoglobulins were observed. The proportion of cells incorporating [3H]-thymidine fell 40-fold. We conclude that suppression of the immune response to SRBC in vitro is mediated by immature precursors of the B lymphocyte series which are present in the bone marrow of intact mice.


Effects of transfer factor in patients with advanced cancer. Eighteen patients with advanced cancer were given subcutaneous injections of pooled dialyzable transfer factor (TFd) from normal donors for periods of from 9 days to 6.5 months. Minor tumor regression was observed in only two patients, an effect of no therapeutic significance. However, treatment with TFd was associated with at least a temporary increase in delayed hypersensitivity reactions in 12 of 17 patients tested, including four patients who became responsive to 2,4-dinitrochlorobenzene. In general, in vitro tests of immune function were not changed after treatment with TFd except for levels of C1q, and/or C3, which were increased in 6 of 10 patients tested. We conclude that TFd may augment delayed hypersensitivity in patients with advanced cancer, and that its effects are, at least in part, immunologically nonsepcific.


A sulfone beta-lactam compound which acts as a beta-lactamase inhibitor. CP-45,899 [3,3-dimethyl-7-oxo-4-thia-1-azabicyclo(3,2,0)heptane-2-carboxylic acid, 4,4-dioxide [2S-(2alpha,5alpha)]] has low intrinsic activity against most Gram-positive cocci, Enterobacteriaceae and Pseudomonas. It inhibits Neisseria at concentrations of 0.1 approximately 6.2 microgram/ml. The combination of CP-45,899 and ampicillin inhibited Staphylococcus aureus and Enterobacteriaceae resistant to ampicillin by virtue of beta-lactamase activity. Combination of CP-45,899 and cephalothin was synergistic less often, and CP-45,899 did not act synergistically with carbenicillin or ticarcillin against Pseudomonas resistant to these agents. CP-45,899 acted synergistically with ampicillin against Bacteroides. Synergy of CP-45,899 and ampicillin was demonstrated at varying concentrations suggesting that it may significantly enlarge the antibacterial activity of ampicillin against resistant bacteria.


Cell-free synthesis of rat parotid preamylase. Poly(A)-containing RNA from rat parotid gland directs the cell-free synthesis of several products in the reticulocyte lysate translation system including a very prominent 58,000-dalton polypeptide which is immunoreactive with anti-alpha-amylase. Purified alpha-amylase has a molecular weight estimated as 56,000 daltons. The 58,000-dalton, cell-free product and alpha-amylase share common peptides as determined by analysis of their limited proteolysis digests. The cross-reactivity and peptide homology suggest that the cell-free product may be a precursor of mature alpha-amylase. While the NH2 terminus of alpha-amylase is blocked, that of the 58,000-dalton product evidently is not, and automated sequence analysis has yielded its partial sequence as: Met-X-Phe-Phe-Leu-Leu-Leu-X-Leu-Ile-X-Leu-X-X-X-X-X-X-X-X-X-Phe-X-X-X-X-X-Ile-X-X-Leu-Phe. The highly hydrophobic nature of the NH2 terminus of the 58,000-dalton, cell-free product suggests that, like other secreted polypeptides, the extra piece may play a role in the transport and secretion of the mature alpha-amylase.


Intermembrane phospholipid transfer mediated by cell-free extracts of Rhodopseudomonas sphaeroides. The transfer of phospholipids between two membrane substrates catalyzed by a soluble protein fraction from Rhodopseudomonas sphaeroides has been demonstrated. The assay employs purified intracytoplasmic membrane (ICM) vesicles derived from cells of R. sphaeroides grown on [3H]acetate as the phospholipid donor substrate and phosphatidylcholine (70%)/phosphatidylethanolamine (30%) unilamellar liposomes containing [14C]triolein, a nontransferable marker, as the acceptor substrate for transferred phospholipids. Incubation of these two membrane substrates with a 40 to 80% (NH4)2SO4 protein fraction from R. sphaeroides results in the transfer of tritium-labeled ICM phospholipids to the acceptor membrane substrate. Upon completion of the incubation period, the donor ICM vesicles are quantitatively separated from the acceptor liposomes by precipitation with antibody prepared against whole, purified ICM vesicles. Phospholipid transfer is linear with respect to time and protein concentration, is inhibited by trypsin and heat, and shows an absolute dependence upon the presence of acceptor liposomes and the 40 to 80% (NH4)2SO4 protein fraction. Control experiments indicate that no fusion of the donor and acceptor membrane occurs during the incubation period and that, following prolonged incubation there is no detectable degradation of the labeled lipid components. Preliminary data on the phospholipid specificity of the transfer reaction is also presented.


Validity of an assumption underlying the confirmed bleeding day measurement in an experimental clinical trial. In a clinical trial involving gingival bleeding, one measurement proved far better than the others. It was assumed that this measurement would help to eliminate certain sources of error. This paper examines data from the trial which give support to the assumption.


Electro-oculography, dark adaptometry, and laser interferometry. This paper discusses the basic concepts of electrooculography (E.O.G.), dark adaptometry, and laser interferometry. Included is the clinical relevance of each procedure.


Visual evoked potential (VEP): basic concepts and clinical applications. The Visual Evoked Potential is the objective measurement of visual function monitored at the level of the occipital cortex with scalp electrodes. This article summarizes many of the recent clinical applications of the VEP. Included are basic concepts of the VEP and its clinical utilization for the objective assessment of refractive error, visual acuity, amblyopia, binocularity, demyelinating diseases, psychogenic disorders, pre-surgical prediction of post-surgical visual function, visual fields, color blindness and neurological development.


Ophthalmic ultrasound as a diagnostic tool. Ophthalmic ultrasound is a diagnostic technique which has a clinical utility in ocular distance measurements and in the evaluation of the structural integrity of eyes with translucent or opaque media. This article summarizes briefly the principles of ultrasonography, available instrumentation and clinical applications.


Antigen-binding T cells: dose response and kinetic studies on the development of different subsets. We have described a number of the parameters involved in the in vitro induction of specific SRBC-binding T cells (T rosette-forming cells, T-RFC). Although T-RFC precursors pass through nylon, most of the induced cells do not; nor do detectable numbers of Ly 1+2, 3- cells bind antigen with sufficient stability to form rosettes. The ratio of Ly 2,3:Ly 1,2,3 T-RFC varies with time after immunization and with the dose of antigen used for stimulation. Relatively high or low doses of antigen selectively induce Ly 1,2,3 T-RFC. Ly 2,3 T-RFC, when they appear, follow Ly 1,2,3 T-RFC. Pretreatment of T cells with anti-Ly sera before RFC induction prevents formation by Ly2+ T-RFC. Since anti-Ly 1 treatment blocks RFC formation and since Ly 1,2,3, T-RFC always precede the appearance of Ly 2,3, T-RFC, our results suggest that some Ly 1+ cells (Ly 123 at least, but perhaps also Ly 1) may act as inducers, precursors, and/or amplifiers for Ly 2,3 RFC as they appear to do for Ly 2,3 suppressor and killer cells. Thus, our results confirm and extend the observed similarities between T-RFC and other Ly 2+ cells such as killer and suppressor cells as well as their differences from Ly 1+ helper cells.


Augmentation of mouse natural killer cell activity by interferon and interferon inducers. Interferon (IF), in addition to its anti-viral capacity, is increasingly being found to be a regulator of cell division, cell surface antigens, and cell function. To determine whether IF also plays a role in the regulation of natural killer (NK) cell activity in mice, the in vivo and in vitro effects of IF and IF inducers on NK activity were studied. We observed that pyran, lipopolysaccharide, and polyinosinicopolycytidylic acid (poly I:C) as well as crude and purified IF preparations significantly elevated splenic NK levels in normal mice within 3 to 24 hr of i.p. administration. Normal spleen cells treated with poly I:C or IF in vitro also had augmented NK activity. Poly I:C and IF were themselves not cytotoxic and their presence was not required during the lytic process, indicating that IF acts on lymphocytes to activate NK function. The addition of anti-IF in the incubation medium completely blocked the boosting of NK activity by poly I:C or IF. The characteristics of the effector cells activated by IF were consistent with those of NK cells rather than macrophages, since the boosted effector cells were not retained by a rayon column or removed by carbonyl iron. Moreover, they were resistant to treatment with anti-Thy 1.2 serum plus complement, which eliminated mature T cells.


The purification and quantitation of helper T cell-replacing factors secreted by murine spleen cells activated by concanavalin A. A microculture assay with low numbers of athymic (nude) spleen cells has been used to quantitate the helper T cell-replacing activity secreted by concanavalin A-treated spleen cells. This quantitation allows an estimate of the recovery of biologic activity during concentration from culture supernatants by salt precipitation, and purification by gel filtration, ion exchange chromatography, and isoelectric focusing. The T cell-replacing activity is found in protein of 30 to 40,000 daltons size as estimated by gel filtration, but shows heterogeneity in electric charge. The activity is active at concentrations of less than 10(-9) M, and is strictly antigen dependent in its mode of action.


The distribution of adenosine deaminase among lymphocyte populations in the rat. Adenosine deaminase (ADA) activity was determined in young rat lymphocyte populations. The ADA-specific activity (per 10(8) cells and per milligram protein) was 3- to 10-fold higher in thymocytes than in lymphocytes from thoracic duct, lymph node, spleen, and bone marrow. The high ADA activity in thymocytes appeared to be preferentially associated with cortical thymocytes. Enrichment or depletion of cortical thymocytes by density gradient centrifugation, cortisone treatment, or  selective lysis with anti-Thy-1 plus complement resulted in parallel increases or decreases in ADA levles. These results also suggested that medullary thymocytes have ADA levels similar to those of peripheral lymphocytes. "Immature" cortical thymocytes and thymocyte progenitors appeared to have low ADA activity; low enzyme levels were found in fetal thymus at 16 days of embryonic life, in the early phases of thymus regeneration, and in a "null" cell population isolated from bone marrow. This study demonstrates that ADA activity varies markedly during T lymphocyte differentiation and suggests that fundamental differences in nucleotide metabolism may exist in T cells at different stages of development.


Serologically defined subpopulations of human T lymphocytes. Several rabbit antisera to T cells obtained from various sources (thymus, peripheral blood, brain, T-derived leukemias) were studied with the aim to obtain reagents specific for a subset of T cells. Sera were first absorbed on human tissues and B cells; thereafter these T cell-specific sera were additionally absorbed with T cells of different origin and especially with leukemic T cells, which are likely to represent a clonal expnasion of a subset of T cells with potentially unique antigenic markers. Three antigenically distinct subpopulations of T cells were delineated. The relationship of these subsets with previously defined human T cell subpopulations (T subsets with a receptor for the Fc or IgG or IgM or with a receptor for a lectin from wheat germ agglutinin) was investigated.


Ontogeny of cell-mediated cytotoxicity: induction of CTL in early postnatal thymocytes. In this study we characterized the time when cytotoxic T lymphocytes (CTL) can be induced in the thymus and spleen from their immediate CTL precursors (CTL-P). In contrast to fetal or newborn thymus, the thymus of 1 to 2-day-old C57BL/6 mice contained cells that, after cultivation in vitro with allogeneic DBA/2 stimulating cells, exhibited high levels (as great or greater than that induced in adult thymocytes) of CTL activity as measured by the ability to lyse P815 (DBA/2) tumor target cells. However, CTL activity induced in spleen cells remained how during the first 5 days of life, increased sharply between 6 to 9 days, and reached adult levels at 11 to 20 days. Furthermore, early postnatal spleen cells did not suppress the adult splenic CTL response. These results suggest 1) that the full potential to generate CTL in response to an allogeneic stimulus commences in the thymus on the first day after birth and 2) a different temporal appearance of immediate CTL precursors in the thymus and spleen.


Immunochemical characterization of the Ly-8.2 murine lymphocyte alloantigen: possible relationship to actin. An immunochemical analysis of the Ly-8.2 antigen was performed. Reactivity of B10-radiolabeled T and B cell lysates with a C3H anti-AKR serum immunoprecipitates a major 43,000 m.w. protein detectable by one-dimensional gel electrophoresis. The protein can be radioiodinated, however, it does not appear to blind to lentil lectin. Two-dimensional gel electro phoresis and peptide-mapping studies indicate that this protein is structurally very similar to lymphocyte actin. The possible molecular relationship between Ly-8.2, other lymphocyte membrane antigens, and actin is discussed.


A receptor for IgA on human T lymphocytes. Receptors for IgA antibody-antigen complexes were demonstrated on 2 to 18% (mean 6.7%) of human peripheral blood T cells. The proportion of cells bearing detectable IgA receptors was low in freshly prepared T cells and increased in number after 18 to 24 hr of culture similar to the time course of appearance of the Tmu receptor. These T receptors were shown to be distinctly different from Fc-IgM and Fc-IgG receptors on T cells by blocking studies with purified IgA, IgG, and IgM.


Suppressor activity of splenocytes from mice treated with Ecteinascidia turbinata extract. Extracts of a marine tunicate, Ecteinascidia turbinata (Ete) were previously shown to be capable of suppressing humoral and cellular immune responses in vivo and in vitro. In the present work we have examined the mechanisms of suppression in Ete-treated DBA/2 and BALB/c mice. Treatment with Ete resulted in a significant splenomegaly accompanied by a diminished response to mitogenic stimulation, reaching coincident maxima of effect at days 4 to 6. Spleen size and blastogenic reaction returned to normal at 12 to 21 days. Cells from enlarged spleens inhibited blastogenic responses of normal splenocytes to Con A. Marker studies indicated that the suppressor activity was exerted by cells possessing T lymphocyte characteristics.


Helper cells activated by allogeneic H-2K or H-2D differences have a Ly phenotype distinct from those responsive to I differences. Helper activity for the primary in vitro response to sheep erythrocytes was induced by recognition of foreign major histocompatibility antigens. The Lyt phenotypes of helper activity induced by differences in the whole haplotype, K or D antigens, I region antigen, or by differences at the M1s locus were determined. All allohelper cells expressed Ly1. In a single spleen cell suspension helper activity generated in response to the whole haplotype, I region, or M1s antigens was derived from an Ly2-negative population, whereas helper activity generated to K or D alone was derived from a population of Ly2-positive cells. Mixtures of anti-Ly1 and anti-Ly2 treated populations were unable to generate helper activity in response to K or D differences. Such helper activity was therefore dependent on the presence of Ly12 cells. It was concluded that the Ly phenotype of the helper cells is not determined solely by the T cell function but is influenced by the region of the major histocompatibility complex that is recognized. Possible interpretations of these findings are discussed.


Macrophage factor controlling differentiation of B cells. Peritoneal macrophages of the mouse produce, in response to cell wall components of Gram-negative bacteria (lipopolysaccharide and lipoproteins), a factor that causes antigen-stimulated B cells of differentiate into antibody-producing cells. Unlike lipopolysaccharide, this factor is not mitogenic for B cells. Production of the macrophage factor does not depend on participation of T cells or other accessory cells since it is readily produced by several cloned macrophage cell lines as well as by peritoneal macrophages of athymic nude mice. The factor is active only in conjunction with antigen. T cells, although apparently not necessary, amplify its effect. The factor induces phenotypic differentiation of B cell precursors as selectively as thymopoietin induces differentiation of prothymocytes.


Primary in vitro cytotoxic response of F1 T lymphocytes against parental antigens. Spleen cells from adult female (AKR/J x BALB/(c)F1 mice can respond to mitomycin C-treated spleen from AKR/J mice and can generate effector CTL in a 5-day primary in vitro culture. The response is comparable in magnitude to the response to allogeneic H-2K or H-2D antigens. The response is T cell mediated and is directed to antigen(s) present only on the parental cells. The target cell must be homozygous at H-2Kk to be lysed and H-2Dk antigens do not serve as a target in this response. Spleen cells from (B10.BR x B10.D2) hybrids that have been stimulated with AKR/J lyse B10.Br as well as AKR/J target cells. Similar H-2k/d hybrid F1 anti-H-2k parent responses are seen in certain other strain combinations. A number of possible interpretations of these responses are discussed.


Characterization of in vitro T-lymphocyte colonies from spleens of nude mice. Cells residing in the spleens of nude mice that appear Thy-1 negative can give rise to Thy-1 positive colonies in an in vitro culture system. Cells from pooled colonies can provide an accessory cell required for the development of cytotoxic lymphocytes from their precursors in an in vitro mixed lymphocyte reaction. As few as 50 such colony cells produce detectable activity.


Early stages of human marrow lymphocyte differentiation: induction in vitro by thymopoietin and ubiquitin. To study early stages of human lymphocyte differentiation, bone marrow cells were physically separated according to their density and size by gradient centrifugation and then velocity sedimentation. The isolated cell fractions were incubated with putative inducing agents and then assayed for their expression of an array of surface differentiation markers. The inducing agents used were two polypeptides, thymopoietin (Tp) and ubiquitin (Ub), and the cyclic nucleotide, dibutyril cyclic 3'5' adenosine monophosphate (cAMP). Tp, Ub, and cAMP each induced the ability to form sheep erythrocyte rosettes by small lymphocytes, which may thus represent T cell precursors. Ub and Tp induced rosette formation with mouse erythrocytes on lymphocytes of more heterogenous size, which may be "early" B cell precursors. Ub alone could induce surface IgM expression on small lymphocytes, which might be "late" B cell precursors. Both Tp and Ub induced Fc receptors on small lymphocytes. Complement receptors could not be induced on marrow lymphocytes by Tp, Ub, or cAMP. A number of lymphocyte precursors can thus be identified by their physical characteristics and their ability to respond to particular soluble factors with the expression of new differentiation markers.


Inhibition of human killer cells by autologous lymphocytes. Human antibody-dependent cellular cytotoxicity (ADCC) mediated by K cells is shown in this study to be inhibited by autologous lymphocytes. Inhibitor activity resides in a population of lymphocytes lacking Fc receptors, i.e., depletion of Fc receptor-bearing lymphocytes on immobolized enriches for inhibition. A T cell-enriched population does not inhibit. The effect is not steric inhibition since addition of large numbers of sheep or chicken erythrocytes does not decrease ADCC. Spontaneous cytotoxicity mediated by NK cells in the absence of added antibody is not inhibited by the FcR-depleted population, indicating that K and NK cells differ from each other in this respect.


Purification and biochemical characterization of human lymphocyte mitogenic factor (LMF). Supernatants of human T lymphocytes stimulated by TT antigen release two factors that induce mitogenesis in autologous and allogeneic B lymphocytes. These factors are precipitated by 60% ammonium sulfate and 50% ethanol, and are both destroyed by heating to 70 degrees C for 5 min. By equilibrium ultracentrifugation there was a peak of mitogenic activity in the fraction with a specific gravity of 1.3147 corresponding to a partial specific volume of 0.761. After ultrafiltration through an Amicon XM50 membrane, the concentrate was chromatographed on a Sephadex G-200 column. Mitogenic activity was found only in the post-albumin fraction. When the post-albumin fraction was run on an isoelectrofocusing column, two distinct mitogenic factors were identified. The major peak of mitogenic activity (LMF) had a pI of 6.68 +/- 0.05 and the minor peak (MMF) had a pI OF 7.27 +/- 0.05. Amino acid analysis of LMF identified it as a protein and PAGE showed that LMF probably was a tetramer with a m.w. of 80,000.


Immunodeficiency with thymoma: failure to induce Ig production in immunodeficient lymphocytes cocultured with normal T cells. Blood mononuclear cells of two individuals having immunodeficiency with thymoma (ID-THY) were cocultured with normal mononuclear cells or treated mononuclear cell fractions in an attempt to correct an imbalance of regulatory cells postulated to be responsible for the failure of pokeweed mitogen-induced Ig synthesis in vitro. Treatment included abrogation of suppressor cell activity by irradiation or incubation with prednisilone in vitro. T cell help was provided by cocultivating lymphocytes of related and unrelated persons, and in some cases autologous treated cells. Ig secretion failed to be induced by any experimental maneuver suggesting that the primary problem in the above ID-THY cells was related to defective or deficient B cells rather than an imbalance of T regulatory cells. Prednisilone treatment in vitro decreased suppressor cell activity in allogeneic cocultures of two ID-THY persons (S1 and S2) but not of an individual (S3) with variable immunodeficiency suggesting heterogeneity of suppressor cells.


The B lymphocyte-deprived mouse as a tool in immunobiology. The purpose of this article is to demonstrate that mice subjected to continuous injections with anti-IgM antibodies, initiated within one day of birth, or in adulthood, lose all their B lymphocyte functions, but retain T lymphocyte reactivity. The B lymphocyte status of these mice was analyzed by histology, radioautography, serum immunoglobulin levels, the antibody response and reactivity to LPS in vitro. T lymphocyte functions were evaluated in vivo by tumor allograft rejection, contact sensitization, and in vitro by reactivity to mitogens and allogeneic cells. On the basis of the results described, it is suggested that the anti-IgM-treated mouse may be considered as a balid and useful experimental model of the "T mouse".


Non-selective lymphocyte isolation from human blood by nylon wool filtration and density centrifugation. A new method was employed to isolate lymphocytes from human peripheral blood. Continuous flow filtration through a nylon wool filter, at a flow rate of 1.4 ml/min, produced a lymphocyte yield of 90.5% and a purity of 96% without any shift in the B-T cell ratio. Ficoll-Isopaque with a specific gravity of 1.085 g/ml instead of 1.077 g/ml could be used to remove the erythrocytes. An overall recovery, including defibrination, filtration, Ficoll-Isopaque centrifugation and washing step, of 74.5% was achieved.


Characterization of lipopolysaccharide of Haemophilus influenzae. Lipopolysaccharide from strains of Haemophilus influenzae was extracted and isolated by the hot phenol-water procedure. The preparations were relatively insoluble in water but could be solubilized with surface-active agents. The preparations contained carbohydrate (30%), fatty acid (29%), and phosphate (4.7%); protein content was less than 1%. Thin-layer chromatography, gas-liquid chromatography, and colorimetric assays detected glucose, galactose, glucosamine, heptose, and a 2-keto-3-deoxy-octonate-like molecule (less than 1%). Neither methylpentose nor dideoxyhexose was detected. The lipid portion was composed of fatty acids common to lipopolysaccharide of Salmonella. The preparations provoked positive dermal Shwartzman reactions and biphasic febrile responses in rabbits, responses typical of endotoxic activity. The 50% lethal dose for mice was decreased from 16.5 microgram/g to 0.015 microgram/g by concomitant administration of actinomycin D. The preparations were shown to be polyclonal activators of bone marrow-derived (B) cells. Limulus lysate gelation was seen with 8.0 ng of lipopolysaccharide. Preliminary hemagglutination data suggested at least three different antigenic factors associated with the lipopolysaccharide of H. influenzae type b. The H. influenzae lipopolysaccharide appeared biologically similar to that of enterobacteria but chemically different.


Palmerston North mice, a new animal model of systemic lupus erythematosus. This report describes a previously unrecognized animal model of SLE, the PN mouse. Although outbred PN mice were studied originally as models of polyarteritis nodosa, their inbred descendants have autoimmune disease which closely resembles SLE. In the current study, positive indirect immunofluorescence tests for ANA appeared when the mice were 5 months old, and 80% of mice were ANA-positive at 10 months of age. Anti-DNA were detected in sera from newborn mice and from 53% of mice under 2 months of age. Seventy-six percent of PN mice developed anti-DNA at the age of 10 months. Glomerular deposits of IgG, IgM, IgA, and complement appeared at 2 to 4 weeks of age, and examination of renal tissue by electron microscopy showed basement membrane thickening and dense intramembranous deposits. Neoplasms arose in 14% of PN mice. Female mice died earlier than male mice, and the most common causes of death were glomerulonephritis and arteritis. It was concluded that the serologic and histologic characteristics of disease in PN mice resembled SLE.


Vaccinia-specific cytotoxic T-cell responses in the context of H-2 antigens not encountered in thymus may reflect aberrant recognition of a virus-H-2 complex. BALB/c (H-2Kd-Dd) spleen and lymph node populations were specifically depleted of alloreactive potential by filtration through H-2 different, irradiated recipients. These negatively selected T cells were then stimulated with vaccinia virus in mice expressing the foreign H-2 determinants encountered previously in the filter environment. Strong virus-immune cytotoxic T-cell responses were seen in the context of H-2Kk and H-2Ks, but not 2H-2Kb. The T cells generated were not cross-reactive for the H-2Kk and H-2Kd alleles, and responsiveness was independent of concurrent presence of effector populations operating at H-2D. These findings are consisent with the idea that recognition is mediated via a complex receptor, part of which is specific for virus and part for self H-2. The capacity to interact with allogeneic, virus-infected cells may then reflect aberrant recognition of a virus-H-2-antigen complex by this single, large binding site. For instance, the T cell which would normally recognize H-2Kd-virus x, or H-2Dd-minor histocompatibility antigen Z, may now show specificity for H-2Kk-vaccinia virus. Implications for both the selective role of the thymus and for mechanisms of tolerance are discussed.


Regulatory functions of hapten-reactive helper and suppressor T lymphocytes. III. Amplification of a generation of tumor-specific killer T-lymphocyte activities by suppressor T-cell-depleted hapten-reactive T lymphocytes. 2,4.6-trinitrophenyl (TNP)-reactive T-cell activities were raised in mice by immunization with TNP-isologous mouse gamma globulin. After establishing that TNP-reactive T lymphocytes can serve as amplifier cells for induction of killer T lymphocytes in allogeneic system, we explored the possibility of this hapten-reactive T-cell system to amplify tumor-specific killer T-lymphocyte activity in the syngeneic system. We utlized relatively weak immunogenic syngeneic plasmacytoma X5563 in C3H/He mice. Analysis of the TNP-reactive T-cell activities revealed that such T lymphocytes express the biological functions of both major subtypes of regulatory T cells, namely suppressors and helpers, and that TNP-reactive suppressor and helper T lymphocytes, respectively, differ in their relative susceptibility to specific inactivation by TNP conjugates of the nonimmunogenic D-amino acid copolymer, D-glutamic acid, and D-lysine (D-GL). By taking advantage of the relative susceptibility-difference to TNP-D-GL, selective inactivation of TNP-reactive suppressor T cells was induced by appropriate treatment with TNP-D-GL, and the generation of TNP-reactive helper T-cell activity was amplified. The supplement of augmented TNP-reactive helper T-cell activity to the system at the immunization with syngeneic X5563 with TNP-haptenation, resulted in a striking augmentation of induction of tumor-specific killer T-lymphocyte activity, and a considerable number of hosts survived after the challenge with lethal dose of viable tumor cells. Thus, appropriate manipulations designed to induce potent hapten-reactive helper T-lymphocytes provided the potential for a very effective mode of immunoprophylaxis against tumor.