The von Willebrand syndrome.

The von Willebrand syndrome. Five patients with an original diagnosis of von Willebrand's disease are described because of their levels of factor VIII related protein, Ristocetin-induced platelet aggregation and/or family studies differed from the main group of patients with classical von Willebrand's disease. Two had normal levels of factor VIII related protein with reduced Ristocetin aggregation when this was tested in platelet rich plasma. In one, however, this was due to a plasma defect and in the other to a platelet abnormality. After cryoprecipitate infusion all abnormal tests were corrected in both these patients. The first patient, however, failed to show a secondary rise of factor VIII whereas the second showed a secondary rise of both factor VIII and of factor VIII related protein. The other three cases, who were all very severely affected, have been separated from the main group as none of their families was segregating for classical von Willebrand's disease. It is suggested that the term von Willebrand's disease should be confined to those patients who have reduced factor VIII related protein and Ristocetin aggregation, and that von Willebrand's syndrome should be used for the various sub-groups that are emerging.


Senile pseudoexfoliation in aphakic eyes. In three eyes which underwent cataract extraction no evidence of senile pseudoexfoliation was found either before, or immediately after, surgery in spite of repeated biomicroscopical examination under full mydriasis. Years later pseudoexfoliative material was found on the anterior hyaloid and on the pupillary border. In another case, wherein the lens was traumatically dislocated and had lain in the inferior vitreous chamber for 53 years, pseudoexfoliative material was present on the anterior hyaloid, but not in the vitreous. Collating these clinical observations with data from electron and light microscopy studies, the authors refute the assumption that lens epithelium is the source of pseudoexfoliative material.


Dematiaceous fungal keratitis. Clinical isolates and management. Clinical and laboratory features of 16 cases of keratitis that were caused by dematiaceous pigmented fungi are reported. Management, including the treatment of nine cases with Natamycin (Pimaricin), resulted in corneal healing in 14 cases, and therapeutic surgery in two cases.


The management of patients with the Kasabach-Merritt syndrome. The Kasabach-Merritt syndrome comprises consumption coagulation associated with giant haemangioma. A case is reported of disseminated intravascular coagulation complicating dental extraction in a 36-year-old male with this syndrome. The pre-operative management with anticoagulant therapy is discussed and the relevant literature reviewed.


Classification of lymphocytes in nasopharyngeal carcinoma (NPC) biopsies. Seven biopsies taken from nasopharyngeal carcinoma tissues were analyzed with regard to infiltrating B or T lymphocytes. The vast majority of these cells had the capacity to form SRBC rosettes (mean 87%) and were thus classified as T cells. Varying proportions of these were blast-transformed suggesting that they represented a host response against the tumor.


[Study of nucleoside phosphorescence with a phosphorescent microscope]. Phosphorescence of ribonucleosides and desoxyribonucleosides is studied. The phosphorescence spectra of polycrystalline powders of adenosine, guanosine, thymidine, uridine, cytidine, desoxyadenosine, desoxyguanosine and desoxycytidine are recorded at -175 degrees C as well as the decay curves at -175 degrees C. The phosphorescence spectrum of guanosine is also recorded at room temperature. The decay times of phosphorescence of adenosine and cytidine are determined at room temperature too. Obtained values of pi for nucleosides are shorter than those for nucleic acid bases.


Measurement of the number of bone marrow multipotential stem cells in AKR leukaemic mice. Using the anti-theta serum to eliminate leukaemic cells, it was possible to assess the number of colony-forming units in the bone marrow of AKR leukaemic mice. It was found that the number of CFUs decreased in leukaemic mice.


[Absorption spectra of nucleic acids and related compounds in the spectral region 120--280 nm]. The absorption spectra of thin films of nucleic acids, nucleosides, nucleotides, D-ribose, Na3PO4 in vacuum ultraviolet region are measured. In the spectral region 280--160 nm the absorption spectra consist of the bands of nucleic acid bases. In the range shorter than 160 nm the absorption is determined by phosphate and D-ribose groups. The methods of thin films preparation are discussed.


[Separation of antirubella IGM antibodies by affinity chromatography]. The separation of IgM globulins by affinity chromatography has been used for serodiagnosis of primary infections with rubella or measles virus. IgM antibody activity levels are similar to those obtained when the separation has been performed by ultracentrifugation. The results of 50 cases examined by the two separation methods are in agreement. However, affinity chromatography is more specific than gel filtration methods and requires a smaller amount of serum. The anti-human IgM globulins, covalently bound to activated Sepharose, maintain their binding capacity to IgM for a long period; and the same gel could be used more than 40 times. Due to its specificity, sensitivity and reliability, affinity chromatography is an efficient tool for routine laboratory diagnosis of a recent primary infection.


[Effect of anti-inflammatory preparations on the process of thermogenesis in blood erythrocytes]. The authors studied the influence of the main antiphlogistic agents (prednisolone, acetylsalicylic acid, rheopyrine, chloroquine and indomethacine) on the thermal effect of the peripheral blood erythrocytes in man. It was found that only chloroquine possessed a significant inhibitory effect. This fact pointed to the inhibition by chloroquine of energy production in the process of glycolysis, which could be regarded as one of the factors of the therapeutic action of this preparation.


Use of logic-based flow patterns in the investigation and management of surgical disorders. The principles used in the development of logic-based flow charts for use by computer have been applied to the decision-making processes encountered in surgical disorders. Such charts do not attempt to alter one's decisions, but rather present a more logical approach to a given problem, based on past experience. It has been shown that the use of flow charts may be of material benefit to the patient, as well as having other advantages.


Meckel's bleeding diverticulum diagnosed by mesenteric arteriography. The details of a young adult patient who presented with chronic gastro-intestinal blood loss as a complication of Meckel's diverticulum are reported. A preoperative diagnosis was made using superior mesenteric arteriography. The merits of superior mesenteric arteriography and other investigations currently available in the diagnosis of Meckel's diverticulum are discussed.


Comparison of fibreoptic endoscopy in acute upper gastrointestinal haemorrhage in Africans and Europeans. The results of endoscopy in acute upper gastrointestinal haemorrhage were compared in a group of 138 Africans and one of 84 Europeans. Contrary to widely held clinical opinion, the incidence of gastric and duodenal ulceration was similar in the two races. Peptic ulcers were the main source of bleeding in both groups and were surprisingly more common than varices in the Africans. Bleeding from varices, however, was far more common in the Africans than in the Europeans. Stomal ulcers were confined to Europeans. Gastric erosions, often attributed to herbal medicines, were more common in the Africans but the difference was not significant. The study was not designed to determine reduced mortality since the introduction of endoscopy, but management, especially in the Africans, was aided by early recognition of haemorrhage from oesophageal varices and acute gastric erosions.


Isoimmune haemolysis in pathogenesis of anaemia after cardiac surgery. A patient who had received multiple transfusions developed antiglobulin-positive haemolytic anaemia due to a delayed haemolytic transfusion reaction. Many cases of haemolytic anaemia after cardiac surgery could be explained on this basis.


Antitrichomonal antibody in the vaginal secretions of women infected with T. vaginalis. Vaginal secretions from 508 women were examined for evidence of infection by Trichomonas vaginalis, and for antibodies directed against this organism; 42 women (8-3 per cent.) were found to be infected. Secretions from 29 of these women were assayed and antibody apparently directed against T. vaginalis was found in 22 (76 per cent.) of them. Eight out of nineteen secretions (42 per cent.) from apparently uninfected women also contained antibody. Amongst the infected women, no correlation could be found between the presence or absence of antibody and the degree of inflammation, duration of symptoms, use of oral contraceptives, or additional sexually-transmitted disease; there was, however, a slight suggestion that low parasite counts in the vaginal secretions were associated with the presence of antitrichomonal antibody.


Effect of light stimulation on the synthesis and release of GABA in rat and frog retinae. The effect of light stimulation on the incorporation of label from [14C]glucose and [3H]acetate into aspartate, glutamate, glutamine and GABA, both in vivo and in vitro, was studied in dark-adapted frog and rat retinae. With [14C]glucose as the precursor, but not with [3H]acetate, flashes of light appeared to increase the in vivo syntheses of both GABA and glutamine at the expense of glutamate in frog retina, while attenuating the labelling of aspartate and glutamate in isolated frog tissues. Amino acid synthesis from these radioactive substrates in rat retina was not altered under these conditions. Also, the turnover rates of amino acids derived from [14C]-glucose in rat retina in vivo were closely similar in light and darkness. Isolated retinae from both species accumulated [14C]GABA from the incubation medium. In each case the subsequent release of this compound was augmented by electric field stimulation, high K+ and by a high concentration of cold GABA in the medium, but not by a flashing light. Similarly, application of a light stimulus was unable to evoke the liberation of [14C]GABA derived from [14C]glucose in either species.


The effect of fluoride alone or fluoride followed by calcium and vitamin D on disuse osteoporosis of the rat tail vertebrae. The therapeutic effect of fluoride alone or fluoride followed by calcium and vitamin D administration on the osteoporosis induced by immobilization of the rat tail vertebrae was investigated. Tail immobilization was carried out by musclectomy and tendectomy. After 60 days of immobilization, the vertebrae were examined for bone mass, breaking strength, ash percentage and Ca, P and F content. NaF or Ca and vitamin D as separate treatments improved the osteoporosis induced by immobilization. The effects of NaF treatment were more pronounced than those of Ca and vitamin D. NaF and NaF followed by Ca and vitamin D produced a similar degree of improvement in breaking strength. The fact that fluoride and calcium and vitamin D were administered separately and sequentially may explain the absence of a synergistic effect.


An efferent effect on a rhythmic response in the frog retina. Extracellular recordings made from class III ganglion cells demonstrated a rhythmic bursting firing pattern in response to the offset of a mesopic level stimulus of 15, 30, and 90-s duration. The average bursting frequency was found to be 12.8 +/- 0.3 bursts per second. The bursts of action potentials had a duration of 0.047 +/- 0.002 s and separated by silent periods of 0.028 +/- 0.006 s. When a reversible cold block was applied to the optic chiasma, the rhythmic bursting pattern was replaced with a firing pattern that appeared random, indicating that efferent fibers to the retina might regulate the rhythmic response.


Bactericidal substance produced by Haemophilus influenzae b. During bacteriophage studies on Haemophilus influenzer, it was observed that encapsulated type b and unencapsulated Rb strains released a bactericidal substance acitve against types a, c, d, e, and f H. influenzae, non-typable H. influenzae strains, other Haemophilus species, and certain members of the Enterobacteriaceae. The bactericidal activity was assayed by a plaque test utilizing an Rd strain as an indicator lawn and was also demonstrated in mixed broth cultures of a producer strain and an indicator strain. Immediately lysis of sensitive bacteria by the factor was not evident. The factor is sensitive to trypsin but resistant to deoxyribonuclease, treatment with 2-mercaptoethanol, lipase, alpha-amylase, and heating in a 100 degrees C water bath for 20 min. The activity is not dependent upon increased Ca2+ or Mg2+ concentration as is necessary for HP1C1 and S2 phage propagation. The bactericidal factor is not pelleted by high-speed centrifugation at 150,000  X g for 6 h. Treatment with ultraviolet light or mitomycin C does not result in observable phage, phage-like particles, or increased bactericidal activity. T-HE BACTERICIDAL FACTOR IS NOT A TYPICAL SMALL MOLECULAR WEIGHT "COLICIN-LIKE" BACTERiocin in that it is not inducible, has a wider range of activity, and does not kill by "single-hit" kinetics. On preliminary characterization, it is a thermostable protein toxic to certain bacterial strains.


Changes in the mitogen response of lymphoid cells with progressive tumor growth. The response of spleen and lymph node cells from tumor-bearing animals to a variety of mitogens was measured. It was found that progressive tumor growth diminished the reactivity of spleen cells to the mitogen phytohemagglutinin. Lymph node cells from the same animal were not consistently so affected. The reactivity of spleen cells from tumor-bearing donors to concanavalin A was somewhat depressed, while stimulation by bone marrow-derived lymphocyte mitogens such as endotoxin and pokeweed mitogen was in some cases enhanced. Theta antigen expression in tumor bearer spleens was found to decline steadily after 7 days of tumor growth. Cytological analysis revealed that the normal structure of the spleens of tumor animals was infiltrated by myeloid elements. The changes described occurred regardless of whether the tumors were chemically or virally induced. Excision of tumors after long periods of growth resulted in prompt return of splenic phytohemagglutinin sensitivity. The data suggests that loss or incapacitation of parts of the normal thymus-derived lymphocyte components may occur in the spleens of animals with progressive neoplastic growth.


Serum levels of colony stimulating factor(s) and the growth of bone marrow cells in diffusion chambers. Fluctuations in the body fluids of long-ranged humoral substance(s) capable of stimulating the growth of bone marrow granulocytic and macrophage-like cells in diffusion chamber cultures in vivo, was observed after whole body irradiation of mice. The fluctuation pattern was similar to that of the in vitro colony stimulating factor(s) of the sera of irradiated mice which indicates a relation between in vivo and in vitro active factor(s).


The presence of a CSF enhancing activity in the serum of endotoxin-treated mice. Serum taken from mice a few hours after injection of endotoxin is a potent source of a stimulator of in vitro myelopoiesis. By means of dose-response studies, the biological activity of this material was compared to that of a colony stimulating factor (CSF) from pregnant mouse uteri. Postendotoxin serum appears to contain two different activities: a stimulating activity which may be identical to CSF and an activity which augments the action of CSF. The separate nature of the two activities is demonstrated by differences in the rate at which they are diluted out and by differences in the time at which they are maximally present after indotoxin administration. It is therefore concluded that the colony-stimulating properties of postendotoxin serum are not due solely to CSF present in the serum.


Ultrastructure of the stellate cell in the pars intermedia of the frog, Rana pipiens. Stellate cells in the pars intermedia of adult Rana pipiens black-and-white background adapted for 14 days were studied by electron microscopy. This cell type forms a capsule around the secretory cells at the surface of the gland; it also lines the perivascular spaces between the pars intermedia and pars nervosa. The cells are characterized by the lack of well-developed secretory organelles and by the presence of long procesees which extend into the intercellular spaces of the secretory cells and reach the surface of the gland and the perivascular spaces. Occasional desmosomes link the stellate cells to adjacent cells; bundles of microfilaments occur frequently. Microtubules run parallel to the long axis of the cell processes. In light of the possibility that the stellate cells might be involved in transfer of substances to and from the melanophore-stimulating hormone (MSH) secretory cells, they were studied for indications of membrane activity. The stellate cell plasma membranes abutting the pericapillary spaces showed intense micropinocytotic vesicle activity. Also at the gland surface the electron-lucent vesicles were common at the plasma membranes of the stellate cells. Along the plasma membranes of the stellate cell processes extending between the secretory cells were vesicles which were somewhat electron-dense. This membrane activity would suggest an active role for these cells in addition to a sustentacular role.


Potassium influx in the frog atrium during the cardiac cycle. A method for measuring inwardly directed transmembrane tracer flow during the cardiac cycle was developed and applied to a study of 42K influx in frog atrial trabeculae. A fine frog atrial fiber was suspended in a stream of nonisotopic perfusate into which a smaller tracer-containing bolus could be injected, subjecting the fiber to a brief, controlled exposure to the tracer at any desired point in the cardiac cycle. In an experiment, the tissue was exposed to a fixed number of radioactive pulses at a selected point in the cardiac cycle; a brief flush with nonradioactive perfusate removed ambient and extracellular label and an extended wash and removed the remaining intracellular tracer for radioassay. The same procedure was repeated at different points in the cycle, and the resulting tracer uptake at each point measured the relative influx of the particular ion. In this way, a characteristic and reproducible 42K influx profile was demonstrated which exhibited a marked drop below diastolic values during the first 500 msec or so of the action potential followed by a rise and an overshoot above resting values. The time of return to the resting level was somewhat uncertain but was tentatively placed in the vicinity of rapid repolarization. We suggest that the rise and the overshoot reflect the activity of the membrane Na+, K+-adenosinetriphosphatase.


The lymphocyte response in Hodgkin's disease. Fifty cases of Hodgkin's disease have been investigated using the lymphocyte transformation response to phytohaemagglutinin (PHA) as a mesure of their immunological reaction. The average response of all the cases was lower than that of the controls and lower than that of cases of carcinoma from all sites. Cases of Hodgkin's disease which had been treated previously with radiotherapy and/or chemotherapy had a markedly lower response than that of the untreated cases. There was no definite correlation between the stage of the disease and the lymphocyte response with PHA. Seven cases were also tested with pokeweek mitogen (PWM) and the response of the lymphocytes to PWM was found to parallel that of the PHA.


The twenty-four hour pneumoencephalogram: with particular reference to ventricular size a series of 150 patients and a review of the literature. A series of 150 pneumoencephalograms which included 24 h films was reviewed to evaluate previous contradictory reports concerning changes in ventricular size at 24 h and to establish whether it is useful to obtain these follow-up films. In normal patients the changes in the ventricular index one day following the pneumoencephalogram ranged between -4-1 to +6-4% of the initial index. Statistically significant dilatation of the frontal horns of the lateral ventricles was observed in both patients who had dementia and central atrophy, and in those with normal pressure hydrocephalus. This ventricular enlargement was associated with deterioration in neurological status in some cases. Since ventricular enlargement at 24 h occurs in patients both with normal pressure hydrocephalus and with cerebral atrophy with hydrocephalus, it cannot be used to differentiate between these conditions although the most marked changes have been seen in the former. Significant dilatation did not occur in patients without intracranial disease, with cortical atrophy without hydrocephalus, or in a miscellaneous group. In patients with a history of trauma, enlargement at 24 h occurred only when there was some other encephalographic abnormality. The changes in size of the ventricles were not related to the age of the patient, the volume of the injected air, or to the presnece of subdural air. There were other findings on the follow=up radiographs. Cortical atrophy was more clearly demonstrated than on the filling films. Dilatation of the temporal horns, of questionable diagnostic significance, was not infrequently observed 24 h after air injection. The mechanisms of these changes are discussed; no single theory provides a complete explanation.


Prevalance of malocclusion in young Finnish Skolt-Lapps. Two hundred Skolt Lapps aged 8 to 16 years were examined for malocclusion with the aid of intraoral radiography and dental casts. The frequency of loss of permanent teeth was high, particularly of the first molars. Also, many teeth were severely defective because of caries. Hypodontia was very common. The frequency of tipped and rotated teeth was high, probably as a consequence of the frequent loss of neighboring teeth. Space and occlusal anomalies were roughly as common as in urbanized Swedish and Danish materials. The need for orthodontic treatment, judged on a 5-grade scale by one examiner, was found to be greater than in Swedish materials.


The potential for an improved prognosis with chemotherapy in osteogenic sarcoma. Effective chemotherapeutic agents administered shortly after definitive primary treatment to patients with osteogenic sarcoma have altered the natural history of the disease. In contrast to past experience, pulmonary metastases fail to develop in the overwhelming majority of patients receiving treatment with chemotherapy. Patients with established pulmonary metastases have also been salvaged. The therapeutic strategy advocated for different categories of patients is outlined. Although chemotherapy involves additional hospitalization and inconvenience, this additional mode of treatment appears to be the only mechanism by which a substantial improvement in survival of osteogenic sarcoma can presently be achieved.


The comparative evaluation of methoxyflurane and diethyl ether action upon an active sodium transport across isolated frog skin preparation. The experiments were performed in vitro using the frog skin preparations and Ussing method. The effect of methoxyflurane was investigated in 18 sections and that of diethyl ether in 16 sections. Both the anaesthetics were found to affect the frog skin preparation in qualitatively similar way. They both demonstrated a two-phase character of action. Decrease in active sodium transport is preceeded by a transient increase in bioelectric indices of the membrane. The effect was reversible after removing of the drugs from the medium. In order to determine the mechanism of action of the two anaesthetics they were given in combination with ADH and insulin. It was found that the mechanism of action of both the drugs is not analogical to any of the hormones examined, at least, as far as the first phase of action is concerned.


Seasonal differences in drug- and temperature-dependent changes of the sodium transport across the isolated frog skin. In two series of experiments, one conducted in the spring (May-June) and the other in the fall (September-November), the influence of raising temperature of the medium and of vasopressin, insulin and ethacrynic acid on polarized sodium transport in the epithelium of frog skin in vitro was studied. Net sodium transport was determined electrometrically, and active and passive components of transport by the isotope method. Raising temperature markedly increased both components, as well as net flux and electric potential of the membrane in the spring months, but not in the fall. Regardless of the character and extent of the effect on transport, raising temperature of the medium had no qualitative or quantitative influence on the action of vasopressin, insulin or ethacrynic acid.


Treatment of severe coronary artery disease with quadruple and quintuple saphenous vein grafts. Review of 100 consecutive patients. In order to establish the anatomic criteria, the functional results, and the safety of complete myocardial revascularization for severe coronary artery disease, 100 consecutive patients who received four or five saphenous-vein grafts were analyzed. Ages ranged from 37 to 75 years (mean, 56 years). Men predominated by a ratio of 12:1. As an indication of the severity of multiple-vessel disease, 28 percent were in functional class 4, and left ventricular function was classified as good in 47 percent, as fair in 44 percent, and as poor in 8 percent. Coronary arterial scores ranged from 9 to 15 (average, 12.2). Fourteen patients had significant left main coronary arterial obstruction. All 100 patients had grafts to the left anterior descending coronary artery; 96 to the right coronary artery; 94 to the obtuse marginal branch of the circumflex; 78 to a diagonal branch of the left anterior descending; and 27 to the distal circumflex. Operative mortality was 5 percent. Nonfatal perioperative myocardial infarction occurred in 10 percent, and only one of these had low cardiac output. Follow-up from 5 to 23 months showed 95 percent of the patients to be improved, with 70 percent free of angina. Two late deaths occurred, for an overall mortality of 7 percent.


The experimental and clinical use of intravitreal antibiotics to treat bacterial and fungal endophthalmitis. The experimental data involving the use of 14 antibacterial and two antifungal agents have been used to establish a rationale for the management of human endophthalmitis. Twelve cases of endophthalmitis treated under our supervision have been described in detail. The eventual visual outcome is related to two factors: (1) time interval between diagnosis and institution of adequate therapy, and (2) the severity of the organism and its ability to produce exotoxins and proteolytic enzymes. Eleven of 12 cases were cured with the recommended dosages of gentamicin and dexamethasone (400 mcg and 360 mcg, respectively). Eight of the 12 patients retained useful vision of 20/200 or better.


[Fatal course of generalized BCG histiocytosis in congenital immune deficiency (author's transl)]. Severe combined X-linked immune deficiency ended fatally in a six-month-old male infant after generalized BCG infection. Damage in the cell-bound thymus-dependent immune system is decisive in the development of "BCG histiocytosis". As a result of absent immunocompetent sensitized T-lymphocytes there is insufficient information and stimulation of macrophages to kill the micro-organsims. The disorder of bactericidal properties of macrophages results in an overwhelming infection with BCG bacteria with a characteristic morphological picture.


Binding by thyroid hormones by nuclei of cells from bullfrog tadpole tail fins. The regression of the tadpole tail is under the direct control of the thyroid hormones and offers a unique system for the study of the action of these hormones. We have examined the binding of L-triiodothyronine (T3) and L-thyroxine (T4) in vitro using tail fin bricks which included epidermal and connective cells. The binding of 125I-labeled hormones was followed in both nuclear and extranuclear fractions. High affinity and limited capacity sites for T3 and T4 were observed only for the nuclear fraction. Scatchard plots gave similar apparent dissociation constants for both hormones, about 10(-10)M. The maximum number of binding sites per nucleus for T3 was about 1500 and for T4 about 800. There was no significant change in the chemical identity of [125I]T3 and [125I]T4 which was associated with binding in the nuclei. Nuclear binding of [125I]T4 was inhibited competitively for both unlabeled T3 and T4, but unlabeled T3 displaced [125I]T3 significantly more than unlabeled T4. Thus, both binding and competition data support the conclusion that tadpole tail nuclei had more T3 than T4 binding sites.


Binding of thyroxine and triiodothyronine by nuclei of isolated tadpole liver cells. The binding of L-triiodothyronine (T3) and L-thyroxine (T4) to cytoplasm and nuclei has been studied in isolated Rana catesbeiana tadpole liver cells. Nuclear binding for both thyroid hormones occurred more slowly at 4 C than at 25 C, but reached the same level as at 25 S. Scatchard analyses suggest high affinity, saturable binding sites for both hormones in the nuclear but not in the cytoplasmic fraction. Apparent equilibrium dissociation constants were 6.8 X 10(-10)M and 4.6 X 10(-10)M for T3 and T4, respectively. The maximum number of binding sites per nucleus for T3 was about 12,300 and for T4 about 2,300. Unlabeled T3 competed for the binding of [125I]T3 to nuclei more effectively than unlabeled T4. No difference in the competition of [125I]T4 binding with non-radioactive T3 or T4 was found. Chromatographic analysis of the bound nuclear radioactivity demonstrated no chemical modification for either hormone.


Transport of amino acids in membrane vesicles of Rhodopseudomonas spheroides energized by respiratory and cyclic electron flow. Active transport of amino acids in whole cells and membrane vesicles from the facultative photo-synthetic bacterium Rhodopseduomonas spheroides is coupled to electron flow in the respiratory chain and in the cyclic electron transfer system. In vesicles made from cells grown aerobically in the dark transport of amino acids is most effectively energized by the oxidation of NADH and to a lesser extent by ascorbate or succinate in the presence of N,N,N',N'-tetramethyl-1, 4-phenyldiamine dihydrochloride or by ascorbate + phenazine methosulphate via the respiratory chain with oxygen as terminal electron acceptor. In membrane vesicles from cells grown anaerobically in the light the energy for active transport of amino acids is supplied upon illumination by photo-oxidation of bacteriochlorophyll and subsequent electron flow through the cyclic electron transfer system. The initial rate of transport increases with the light intensity upto saturation levels. In both vesicle preparations, inhibitors of electron transfer flow inhibit amino acid uptake. In order to obtain light-driven amino acid transport the isolation of membrane vesicles needs to be performed in a medium with a redox potential between 0 and 100 mV. Illumination of vesicles made from cells grown anaerobically in the light results in the generation of a membrane potential as is indicated by the uptake of the lipophylic cation triphenyl-methylphosphonium.


Amplification of colony stimulating activity in human serum by interaction with CSA from other sources. An enhancing interaction of a human serum factor with other sources of colony stimulating activity (CSA) is described. Using a mouse colony forming cell assay, the enhancing effect of human serum was observed with suboptimal doses of colony stimulating material from monocytic cell feeder layers, supernatant from peripheral monocytic cells, and L cell supernatant. The enhancing factor is heat stable, is not dialyzable and is not extractable by ether. The enhancing phenomenon does not require the presence of viable cells in the feeder layers. The observation of enhancing interaction makes interpretation of all data from CSA stimulation experiments more complex than previously appreciated.


Colony stimulating factor and the regulation of granulopoiesis and macrophage production. The growth and differentiation of  colonies of granulocytes and/or macrophages in semisolid cultures of hemopoietic cells requires the presence of "colony stimulating" factors (CS factors). Colony stimulating factors are found in serum, urine and certain tissues, and in media "conditioned" by certain cells in culture. A procedure has been developed for purification of CS factor from human urine. The 100,000-fold purified product is active on mouse bone marrow cells at less than or equal to 10(-11) M but is only partially purified. Polydispersity of activity has made further purification difficult and may necessitate use of biologically selective techniques such as immunosorbent chromatography. Knowledge of the properties of human urinary CS factor has been derived from the in vitro assay of biological activity following various treatments. It appears to be a sialic acid-containing glycoprotein of molecular weight 45,000-60,000 daltons that migrates electrophoretically between alpha1 globulin and albumin. Highly purified CS factor preparations can be iodinated with five atoms of iodine per molecule of protein without loss of biological activity. Neutralizing antibody to human urinary CS factor neutralizes CS factor from other human and monkey sources. Higher concentrations of antiserum are required for neutralization of murine CS factor. Human urinary CS factor has similar properties to human and murine CS factors that stimulate the early appearance of macrophages in murine colonies. It is physically distinct from CS factors that stimulate the formation of pure granulocytic colonies by murine or human hemopoietic cells. Purified CS factor of the human urinary type is being used to investigate the nature of the interaction of CS factor with target cells and its physiological role in vivo.


[Responses of the units of the nerve of the frog semicircular canal to caloric stimuli of changing amplitude]. In curarized frogs, responses of isolated neural units of the lateral semicircular canal (LSC) to short local caloric stimuli of different amplitudes (0.5--12.0 degrees C over the canal temperature) inducing excitatory current of endolymph in the LSC, were studied. Estimation of maximal (peak) frequences (PF) in the responses showed the PFs in responses of 52% units (group A) to be either linearly or semilogarithmically related to peak temperature (PT). In 22% units (group B), PFs revealed saturation at PT8--12 degrees C. In 25% units (group C) PFs were related to PT first in direct and then--along with increase of PT--in reverse proportion. Duration of the group A responses is linearly for semilogarithmically related to PT. This was not observed in groups B and C. Two cases of correlation between spontaneous activity and dynamic characteristics of units from groups A and C were found. The results are in agreement with the literature references concerning data obtained in frogs using angular acceleration and stop-stimuli. The controlled caloric stimulus was concluded to be convenient for studying the dynamic characteristics of the semicircular canals afferents.


[The relationship between changes in the temporal and frequency characteristics of parasympathetic nerve activity and effects on the heart]. Changes of chronotropic effect on the frog heart on stimulation of part of extracardiac pathways, were studied. On the 10/sec stimulation during 1.5 sec, cardioinhibitory effect occurred. On 0.2 sec duration of the stimulation the positive chronotropic effect was observed. The heart rate increased on the 3/sec stimulation of the undegenerating extracardiac fibers by the 21st day after bilateral vagotomy; of the frequency of the stimulation was increased to 10/sec, the negative chronotropic effect occurred, and sympathectomy and the reserpine injection had no influence on the positive chrontropic effects. Both the negative and positive chronotropic effects could be prevented by atropine. The different chrontropic effects on the heart are, probably, due to activity of but a single functional type of fibers: the parasympathetic nervous fibers.


Hemodynamic evaluation of patients with intrahepatic portal hypertension. Relationship between bleeding varices and the portohepatic gradient. Between January 1970 and December 1974, 389 patients presenting an upper gastrointestinal hemorrhage had an emergency fiberoptic esophagogastroscopy at the time of bleeding. In 103 cases, esophageal and/or gastric varices were visualized and in 46 of these cases, bleeding originiated from varices. Hemodynamic data was available in 19 cases of intrahepatic portal hypertension who had bled from varices. Combined hepatic and umbilicoportal catherterization was perform in 17 cases whereas only hepatic vein catherterization was possible in 2. The portohepatic gradient (FPVP-FHVP or WHVP-FHVP) was utilized as an index of portal hypertension. In the 19 patients studied, the portohepatic gradient ranged from 12 to 24.5 mm Hg with a mean of 18.4 mm Hg; only one patient had a portohepatic gradient of less than 14.5 mm Hg. The present data strongly suggest that in intrahepatic portal hypertension, bleeding varices are observed only if significant portal hypertension exists. Hemodynamic studies may thus be helpful in evaluating bleeding patients with varices particularly when the origin of the bleeding cannot be determined at endoscopy. The finding of a portohepatic gradient of less than 12 mm Hg would strongly suggest that varices were not the source of bleeding and in these cases, shunt surgery should not be considered.


Peripheral blood and intestinal lymphocyte sub-populations in Crohn's disease. Immunofluorescent technques were utilized to study the nature and distribution of cells forming the inflammatory infiltrate in Crohn's disease. Tissue from resected ileum (one patient) or colon (one patient) when compared with corresponding control tissues contained increased numbers of B and T lymphocytes. B-cells of IgG class predominated in the mucosal lamina propria, whereas T-cells were prominent in the deeper layers of inflamed bowel. Inflammatory cells bearing Fc receptors were also present in these tissues. Decreased peripheral blood T-cell numbers returned to normal after resection of the involved bowel in the two patients studied. Impaired cellular immunity in Crohn's disease may thus result from T-cell sequestration in or loss from involved segments of intestine.


A prospective comparative trial between early endoscopy and radiology in acute upper gastrointestinal haemorrhage. A prospective comparative trial is reported between the results of early endoscopy and barium meal examinations in 150 patients admitted with acute upper gastrointestinal haemorrhage. One hundred and thirty-eight patients underwent both investigations within 24 hours of admission. Emergency endoscopy accurately identified the source of haemorrhage in 86% of patients and emergency radiology identified the source in 51%. Misleading positive reports were given endoscopically in 3% and by barium meal in 8%. The identification of the source of haemorrhage was possible in 126 patients (91%) by the use of both methods. Emergency endoscopy is shown to be superior to emergency barium meal examination, but the examinations are complementary and both should be used for these patients.


Does alpha-1-antitrypsin P1 null phenotype exist? A second case of Pi null alpha-1-antitrypsin (AA) deficiency is described. In fact, the serum's subject contains 5 mug of AA per millilitre. With radiolabelled specific antibodies, it is possible to describe the Pi phenotype associated to this deficiency. The pattern which is obtained is like the ordinary Pi M, but 500 times lower than normal values. In contrast to a common deficient variant (ZZ or MZ), the subject tissues do not contain periodic acid-schiff positive inclusion bodies. The "normal" pattern obtained after antigen-antibody crossed electrophoresis, would be in favour of a deficient anomaly hereditarily transmitted.


Chromosomal banding of cultured T and B lymphocytes. Chromosomes of thymus-derived lymphocyte (T-cell) cell lines (CCRF-CEM, CCRF-H-SB-2, MOLT-3, MOLT-4, and RPMI 8402) and the corresponding lines (CCRF-SB and RPMI 8392) with thymus-independent lymphocyte (B-cell) characteristics, were examined with banding techniques. The pairs of cell lines (CCRF-H-SB-2 and CCRF-SB, and RPMI 8402 and RPMI 8392, were confirmed to be of the same origin, respectively, inasmuch as their individual specific variant bands were identical. The MOLT-3 and MOLT-4 cells had common marker chromosomes, indicating that the chromosomal changes in these cells probably occurred in vivo. All of the T-cells had structurally abnormal marker chromosomes, whereas the B-cells were chromsomally normal. The structural changes of the chromosomes suggest that the malignant nature of the present T-cells may be essential for such T-cells to be established in vitro in a standard culture medium.


Induction of PHA response in mouse bone marrow cells by thymic extracts as studied by changes in the structuredness of cytoplasmic matrix. The specificity and cross-species activity of thymic extracts and some non-specific agents on some alleged properties of T cells and their precursors in the bone marrow cells of normal, nude and thymectomized mice was studied in vitro. The phenomenon of changes in the structuredness of cytoplasmic matrix (SCM) was used to assess the maturation and responses of cells treated with the extracts to mitogens, i.e. phytohaemagglutinin and concanavalin A. We found that thymic extracts of calf and mouse origin induce functional 'maturation' which can be abrogated by cycloheximide, an inhibitor of protein synthesis. The nonspecific agents used (spleen extracts, PPD, dBcAMP and endotoxin) had no effect on bone marrow cells, as measured by changes in the SCM. The method of measuring SCM using the technique of fluorescence polarization is described.


Distribution of electrophoretic mobilities of mouse thymocyte subpopulations in the presence of tumour cells. Analysis of the electrophoretic mobility of mouse thymus cells has showed two main populations, with mean mobility values of 0-77 +/- 0-023 mum s-1 V-1cm and 0-99 +/- 0-015 mum s-1P1cm; these absolute values varied slightly from one strain to another. Implantation of tumour cells caused the relative proportions of these two populations to change dramatically within 48 hours, when an increase in the fast-moving 'immunocompetent' thymocytes was observed. The ratio of slow to fast cells changed from 9:1 in the normal BALB/c animal to 2:1 in the presence of the tumour cells and this 2:1 ratio persisted throughout the remainder of the animal's life. However, inoculation of histocompatible spleen cells from a normal individual evoked only a brief response in the host's thymus. This change in ratio of slow to fast cells in thethymus was interpreted as an increased production of immunocompetent cells in response to the presence of the tumour cells.


RRBC and EAC rosette formation after DNCB contact sensitization and cyclophosphamide treatment of guinea-pigs. Rabbit erythrocyte (RRBC) and EAC rosette formation, for detection of T and B lymphocytes respectively, was studied in thymus, lymph nodes and peripheral blood of guinea-pigs. Using two different methods for preparation of RRBC rosette-forming cells (RFC) it was found that, without an incubation period at 37 degrees, a smaller part of the pool of T cells is detected which is particularly sensitive to cyclophosphamide (cy). RRBC rosette formation was studied for 2 weeks after treatment with one high dose of cy and appeared to be minimal in the thymus after 1 week and maximal in the lymph nodes at about day 3. EAC rosette formation was completely blocked by cy treatment. Four days after contact sensitization with 2,4-dinitrochlorobenzene (DNCB) no percentual increase was found in RRBC-RFC, suggesting a parallel increase of B and T cells in stimulated lymph nodes. Cytological screening of the rosettes at day 4 after DNCB showed that of the RRBC-RFC approximately 17 per cent were blast cells, while no difference was detected in the number of blast cells participating in EAC rosette formation as compared to the unstimulated lymph nodes. In contrast to RRBC-RFC the percentage of EAC-RFC in draining lymph nodes is increased after contact sensitization. The mechanism of this increase is discussed.


Antigen-initiated B-lymphocyte differentiation. VII. Quantification of AFC progenitor levels in adoptive and culture responses to NIP-POL antigen. Quantitative studies on B cells require a direct assay for antibody-forming cell (AFC) progenitor function, in which the number of AFC produced bears a simple, linear arithmetic relationship to the number of progenitors present. This might be expected under conditions where helper T-cell and accessory cell requirements are by-passed, or provided in excess. This possibility has been tested using as antigen the hapten NIP (4-hydroxy-3-iodo-5-nitrophenylacetic acid) on the carrier POL (polymerized bacterial flagellin), in adoptive transfer of normal and nude mouse spleen cells to irradiated recipients, and in cell culture. Primary and secondary IgM responses to this antigen are "T cell-independent'. The secondard IgG response is T cell-dependent but this function can be provided by 'carrier-primed' irradiated recipients. However in no case did the cell dose response curve show a linear, arithmetic relationship between cells transferred or cultured, and AFC produced. If less than 10 X 10(6) cells were adoptively transferred or cultured, a sigmoid curve was obtained, approximately linear with a slope of around 1-6 on a log-log scale. In adoptive transfer, a plateau was then seen above 10 X 10(6) cells, followed by a second sharp rise beginning around 15 X 10(6) cells. Addition of irradiated spleen cells as 'fillers' to maintain cell numbers constant produced a linear (arithmetic scale) dose response curve for the primary IgM responses, both adoptive and in culture. Lipopolysaccharide injection of recipients also produced linear regions in the adoptive transfer system. These techniques provide more direct, quantitative assay systems for the primary IgM responses to this antigen. However, arithmetic linear cell dose response curves were still not obtained for the secondary IgG responses, using irradiated filler cells.


Clonal proliferation of PHA-stimulated human lymphocytes in soft agar culture. The purpose of this investigation was the induction of clonal proliferation of PHA-stimulated normal human lymphocytes using a two-layer soft agar technique. Essential conditions for colony formation include preceding sensitization of lymphocytes with PHA, and continuous presence of PHA in the soft agar culture. Two types of colonies developed: large colonies which appeared 3-4 days after seeding and comprised, after 5-6 days, 200-500 cells, and small colonies which were seen after 6-7 days of culture, resulting in production of 50-150 cells. Morphological study showed that all cells were blast-like and the mitotic index exceeded that in liquid medium by a factor of 50. Comparison between the number of colonies developing from cultured bone marrow and spleen cells with those from peripheral blood showed that, in proportion to the number of lymphocytes seeded, a larger number of colonies developed from bone marrow cells and a lower number of colonies developed from spleen  cells. The time required for sensitization of lymphocytes in liquid medium with PHA was found to be no less than 12 hours. The greatest number of colonies appeared when the optimal concentration of PHA was placed in the lower agar layer. A linear relation between the number of cells seeded and the number of resulting colonies was found. One out of 2 X 10(3) or 3 X 10(3) lymphocytes in peripheral blood has the potential to develop as colony. The rosette-forming ability and morphological identification of the cells suggest that the colonies are composed of T lymphocytes.


Local immune response in experimental pyelonephritis in the rabbit. I. Morphological and functional features of the lymphocytic infiltrate. The cellular activity of circulating lymphocytes and lymphocytes isolated from the infected kidney of animals with experimental haematogenous pyelonephritis was evaluated. The incorporation of [3H-methyl]thymidine into DNA by lymphocytes was studied with mitogens such as phytohaemagglutinin (PHA), pokeweek mitogen (PWM) and goat anti-rabbit IgG (GARIG). Lymphocytes from infected kidney had a high baseline DNA synthesis compared to circulating lymphocytes from days 5 to 27 of infection. Infected kidney lymphocytes failed to respond to PHA, PWM, or GARIG, whereas circulating lymphocytes did respond to these mitogens. Uropod-bearing lymphocytes, which were shown to be T lymphocytes, were present from days 5 to 77 of infection. B lymphocytes, as determined by surface immunofluorescent technique, were present by day 12, coincident with the onset of local synthesis of antibody. These studies reveal that in pyelonephritis, the cellular response goes through sequential changes and indicate a dynamic interrelationship between T and B lymphocytes at an infected site.


Relationship between biological activities of polymers. I. Immunogenicity, C3 activation, mitogenicity for B cells and adjuvant properties. The effect of thirteen compounds on C3 activation, mitogenicity for B lymphocytes and on the immune response was tested. Only two of the compounds tested, polyinosinic acid and a high molecular weight polyethylenesulphate induced C3 conversion, while eleven out of the thirteen compounds (negative charged immunogenic or non-immunogenic polymers including the three C3 active compounds) stimulated thymidine incorporation in B lymphocytes. The non-charged (T-independent) immunogens dextran and polyvinylpyrrolidone neither activated C3 nor BP lymphocytes. With the exception of single-stranded polyribonucleic acids, the compounds stimulating DNA synthetis in B cells enhanced the response of the mice to a suboptimal dose of sheep red blood cells. In conclusion C3 activation, intrinsic (T-independent) immunogenicity and B-cell stimulation are not correlated properties. However, B-cell mitogenicity and adjuvanticity seem to be related.


Cell-mediated allograft responses in vitro. VI. Studies on macrophage-mediated cytotoxicity. Normal murine peritoneal macrophages were rendered cytotoxic against 51Cr-labelled allogeneic and syngeneic target cells by incubation with supernatant of selected cell cultures.'Active' culture supernatant was produced both by specifically sensitized cytotoxic T lymphocytes as well as by mitogen-stimulated T cells, but not by mitogen-stimulated B cells. The in vitro induced macrophage-mediated cytotoxicity was found to be non-specific in the sense that 51Cr-labelled target cells of different H-2 haplotype were lysed equally well.


A proposition on the distribution of antibody affinities, with implications for the mechanism of B-cell activation. For most antisera a linear relationship can be shown between log antigen concentration and the log concentration of antibody required to bind half the available antigen. This paper shows that the slope of this line, s, is related to the distribution of individual antibody clonotypes of different affinity. It is argued that the general form of the distribution approximates to exponential (rather than for example Gaussian) and that this indicates a requirement for some force to be exerted through the antigen-receptor bond in order to activate a B cell. An alternative parameter, A, which gives more weight to high affinity clonotypes, is offered in place of K0 as a measure of the avidity and biological effectiveness of an antiserum.


Quantitation of potential T-lymphocyte function in rats. A simple but dependable method for determining the functional capacity of thymus-derived lymphocytes is described. The in vitro culture conditions for determining the incorporation of tritiated thymidine after stimulation with phytohemagglutinin and other mitogens have been given in detail and the methodology confirmed by measuring the ablation of T-cell function in T cell depleted animals.


Increase in the proportion of cells with the C'3 receptor in BALB/c mice bearing mammary tumors. Transplantation of mammary tumors originally induced by dimethylbenzanthracene caused splenomegaly and an increase in the total population of spleen cells paralleling the increase in tumor size. At least part of the hypercellularity was due to in situ proliferation as evidenced by the increased number of blast forms and mitotic figures. The cells were characterized with respect to parameters generally associated with B-cells; surface immunoglobulins as detected by immunofluorescence and C'3 receptors as determined by rosette formation with sheep red blood cells (SRBC) coated with rabbit anti-SRBC and mouse complement. There was no change in the percentage of B-cells with surface immunoglobulin in tumor-bearing mice compared with the control animals. However, there was a profound change in the representation of cells bearing C'3 receptors. The percentage of the cells increased dramatically with tumor growth. Various possibilities regarding the nature and function of these cells are considered.


Evaluation of radiochemical protectors using urinary deoxycytidine levels. It is customary to use the LD50 response as a criterion for the evaluation of chemical radioprotection. The increased urinary excretion of deoxycytidine (CdR) in irradiated rats has been shown to be a linear dose--response relationship up to 200 R, thus making it possible to consider such a response as a criterion for chemical radioprotection evaluation at sub-lethal irradiation levels. Rats were subjected to 200 R X-irradiation with various chemical protectors and without chemical protectors, urine was collected 24 hours after irradiation, and the CdR levels in the urine analysed. A dose-reduction factor (DRF) was determined for 24-hour urinary excretions of CdR for chemically protected and non-protected rats. The DRF values are within 6 per cent of those reported by other investigators using LD50 as the response criterion for serotonin, cysteine, MEA and PAPP but differ for WR-2721 and WR-2823. Agreement using WR-2721 and WR-2823 compounds is only obtained when the toxicity effect of the compound is taken into consideration.


Effects of DTPA on the deposition of 65Zn, 60Co and 144Ce in pregnant rat and in foetoplacental unit. 65Zn, 60Co or 144Ce was injected intravenously into rats at various stages of pregnancy. Six hours later, DTPA chelate of non-radioactive Ca, Zn or Co was given intraperitoneally. Another group of pregnant rats received intravenous injection of 65ZnDTPA or 60CoDTPA. In both experiments, 24 hours after administration, the concentration of the radioactive material was determined in the dam's whole body as well as in foetuses, chorioallantoic placentae and yolk sacs. 65Zn and 60Co, which had been administered to the rats as DTPA complexes, were found not to pass from the dam's blood to the foetuses or to pass only in very minute amounts. This finding could be considered as an indirect indication that DTPA chelates do not penetrate the placental barrier. Nevertheless, the chelates injected intravenously into the dam early after an internal contamination, decrease the radionuclide content in the foetuses. No substantial effects of pregnancy on the effectiveness of the chelate in removing radionuclides from the body of rats were observed.


Lumphocytotoxins in leprosy.

Lumphocytotoxins in leprosy. Occurrence of cold lymphocytotoxins has been observed in 59 leprosy sera. In 46% of the patients, cold lymphocytotoxins were present whereas in helathy controls only 13% showed such antibodies. The highest incidence of alloantibodies was detected in lepromatous leprosy. Levels of autoantibodies, immunoglobulins and C3 were tested in parallel without finding any significnat correlations.


Radiation-induced decrease in influx rates of potassium ions into thymocytes in vitro in relation to decreased intracellular adenosine triphosphate concentrations. The relationship between decreases in the adenosine triphosphate content of irradiated thymocytes and observed changes in the influx constant values for potassium ions in the same cells have been investigated. The results suggest that radiation-reduced ATP concentrations may be a rate-limiting factor controlling the active transport of potassium, 3--4 hours after X-irradiation in the range 2--20 krad. However, over the first hour of incubation, post-exposure ATP levels have not decreased sufficiently to be a major factor in the observed decrease in active uptake of potassium ions. This evidence is supported by comparisons of the influence of 1 mM adenine on radiation-reduced ATP levels and influx constants over the first hour after irradiation.


Leprosy XII. Quantitative analysis of thymus-derived lymphocyte response to phytohemagglutinin in leprosy. The immune status of various leprosy patients was evaluated by using a micromethod to evaluate lymphocyte responses to phytohemagglutinin (PHA). In our study, whole blood was used and the degree of response to PHA stimulation was expressed in terms of unit volume of blood. A markedly decreased response to PHA stimulation was noted in patients with active lepromatous leprosy. Patients with active lepromatous  leprosy who have been proved drug (DDS) resistant showed less response than did those of drug sensitive patients with active lepromatous disease, while the patients with active lepromatous leprosy complicated by erythema nodosum leprosum (ENL) showed higher response than did those of patients with no complicated ENL. Comparing the results obtained to those obtained using other methods for T cell analysis indicates that these results reflect the number of T lymphocytes in the leprosy patient. Thus, this simple method is of value in assaying the presence and responses of T lymphocytes in the leprosy patient.


Alterations of liver DNA after x-irradiation. Reassociation and hybridization with nuclear RNA. When RNA is extracted either from irradiated or unirradiated rat livers, no difference in its ability to hybridize with DNA is detectable. In contrast, the ability of DNA extracted from irradiated animals to hybirdize with RNA is decreased, probably as a result of DNA fragmentation. Sedimentation of X-irradiated DNA on sucrose gradients separates a small (low molecular weight) from a large peak (high molecular weight). The hybridization capacity for RNA of the large molecular weight DNA is similar to that of unirradiated DNA, but that of the small molecular weight is DNA reduced. After irradiation of the denatured DNA in vitro, the reassociation of DNA is inhibited, which suggests that DNA strands have lost part of their complementariness.


Rdiotoxicity of intracellular 67Ga, 125I and 3H. Nuclear versus cytoplasmic radiation effects in murine L1210 leukaemia. L1210 leukaemia cells were labelled with various doses of 67Ga-citrate, 3H-thymidine, or 125I-iododeoxyuridine to evaluate the cytocidal effects of the intracellular decay of the three radionuclides. Based on radioisotope incorporation data, cellular dimensions, and intracellular radioisotope distributions (3H and 125I intranuclear, 67Ga cytoplasmic) the rates of deposition of cellular, nuclear, and cytoplasmic energy were calculated. In terms of energy absorption/cell, 67Ga (LD50: 2250 keV/hr; 69 rad/hr) was much less toxic than either 3H (LD50: 325 keV/hr; 10 rad/hr) of 125I (LD50:50 keV/hr; 1-5 rad/hr). In terms of energy absorption/nuclesu, 67Ga and 3H produced almost identical effects (LD50: 230 versus 255 keV/hr; 22-2 versus 24-6 rad/hr), but 125I remained much more toxic (LD50: 40 keV/hr; 3-9 rad/hr). These findings indicate that, although decay by electron capture in the cell nucleus (125I) is highly destructive, the same type of decay occurring in the cytoplasm (67Ga) is ineffective in killing L1210 cells. An analysis of the data suggests that the cytotoxic effects of the three radioisotopes result exclusively from nuclear damage. Cytoplasmic absorption of radiation energy appears to contribute little, if anything, to the lethal effects of ionizing radiations.


A comparative study of post-irradiation growth kinetics of spheroids and monolayers. Post-irradiation growth kinetics of gamma-irradiated spheroid and monolayer cells in exponential growth phase was investigated by means of dose-response curves based on cell counts after specified time intervals following irradiation. A mathematical model of cell-growth after irradiation was fitted to these curves. The model parameters (related to division delay and growth of non-surviving cellsy obtained from this analysis consistently indicate increasing resistance to sub-lethal damage of cells cultured as multicellular spheroids under conditions of increasing three-dimensional contact. In contrast, no indication of an increased radiation-resistance was found with cells cultured on a substratum under a variety of conditions.


Suppression and recovery of the immune response and CFU potential of mice during and after chronic low-dose-rate gamma-irradiation. Chronic exposure of mice to 60Co gamma-rays at rates of 1-6 R/hr and 6 R/hr leads to profound immunosuppression, as demonstrated with the Jerne haemolysis-in-agar technique. The immunosuppressive effect is dose-rate dependent. Thus, significant reduction of the number of splenic antibody plaque-forming cells (PFC) to sheep red blood cells is seen after exposure to 1005 R at 1-6 R/hr; at the higher exposure rate of 6 R/hr, PFC suppression is seen after a total dose on only 468 R. Although recovery of the PFC response is possible after accumulated doses of 1005, 1579, and 2138 R at 6 R/hr and 5256 R at 1-6 R/hr, normal immunological capacity had not returned 120 days after irradiation. In contrast the recovery of splenic colony-forming units (CFU), as an expression of the haematological potential of chronically irradiated mice, is found to be much more rapid than the repair of the immune system. Thus, 10 days after irradiation, at a time when little or no recovery in immune competence is apparent, the spleen of the chronically irradiatee animal has a CFU capacity about 50 times greater than has that of a normal, unirradiated mouse. The CFU's in the chronically irradiated spleens were also shown to be functional by virtue of their ability to protect lethally irradiated animals from haemopoietic death.


The effects of dose and field size on late radiation damage to the rat spinal cord. In groups of young adult rats, 1, 2, 4, 8 or 16 mm lengths of the cervical spinal cord were irradiated with 2000 or 4000 R of 60Co gamma-rays. All groups showed significant life-shortening, all but five animals (which died from intercurrent disease) showing signs attributable to radiation. The clinical signs, latent period, and histological changes observed were dependent on the dose and field size. The animals irradiated with the higher dose to 8 or 16 mm of the cord were the first to die, all had developed paralysis and were found to have extensive areas of necrosis in the white matter. When the field size was decreased using this dose (4000 R), or the dose reduced (all field sizes), no extensive necrosis was found and only a proportion of the animals developed paralysis; the others died suddenly. The interpretation of these results is discussed.


The modification of sodium excretion after abdominal irradiation. A dose-effect relationship study in the rat. Whole-body counting of radiosodium disappearance in the rat enabled us to evaluate the disturbance induced in sodium excretion after gamma abdominal irradiation at different doses (500, 750, 1000, 1250 and 1500 R). The observed modification is consistent with a sodium excretion loss depending on the dose. Two indices, the relative excretion rate (theta) and the recovery time (deltat), completely define the radio-induced modification and make feasible the quantitative assessment of its dependence on the dose. The development of the dose-response relationship established for each of these indices leads us to conclude that the reduction in the sodium-excretion rate, which can be related to a defect in sodium intake, is a linear function of the dose, while the time needed to recover the pre-irradiation sodium excretion rate increases exponentially with the dose.


Interaction of ethidium bromide with heparin. The reactions of hydrated electrons produced during pulse radiolysis have been utilized to investigate the binding of ethidium bromide to heparin. Complexes of ethidium bromide and heparin can be dissociated with salt. Divalent cations are more effective than monovalent cations in this respect. Pulse-radiolysis investigations at different temperatures indicate that the thermodynamic parameters governing the interaction of ethidium bromide with heparin are deltaH' = 11-6 kcal mole-1 and deltaS' = 42-6 cal deg-1 mole-1.


Pulse-radiolytic investigations of catechols and catecholamines. I. Adrenaline and adrenochrome. Adrenaline (epinephrine), adrenochrome and C4-substituted catechol model compounds were pulse-irradiated in aqueous neutral and alkaline solutions. Transient spectra are reported after oxidizing adrenaline and reducing adrenochrome. All species appearing during 20 msec interval after the pulse have been identified: the OH adduct with an absorption maximum at 300-310 nm, the semiquinone (at 245 nm), and adrenaline quinone (at 340 nm). The reaction of superoxide anions (O2-) with adrenaline was less efficient, compared with OH radicals. A novel oxidation product, derived from the semiquinone and O2-, has been identified as the 4-hydroxy-3,6-dioxo derivate. The pulse-radiolytic reduction of adrenochrome by hydrated electrons (eaq-) yielded the semiquinone of adrenochrome (absorbing at 470 nm), which subsequently decays by a second-order process. The dismutation products leuco-adrenochrome (absorbing at 300 nm, pH 9-8) and the adrenochrome tautomer (absorbing at 375 nm) are unstable, forming 5,6-dihydro-N-methyl indole and regenerating adrenochrome.


An investigation of chromosome damage in Vicia faba root tips after exposure to 1-5 MHz ultrasonic radiation. Roots of seedlings of Vicia faba were exposed to 1-5 MHz continuous-wave ultrasound at approximately 5 W cm-2 intensity, averaged over the dividing region of the meristem, thus producing considerable non-thermal cell damage and severe inhibition of root growth. No significant increase in the number of coventional chromosome aberrations was observed during the 26 hours after exposure, although a significant number of cells did show bridged, agglomerated or clumped chromosomes. It is concluded that after exposure to ultrasound, genetic damage in surviving cells is unlikely.


Storage of polyvinylpyrrolidone mimicking a congenital mucolipid storage disease in a patient with Munchausen's syndrome. A 24-year-old woman from the Sinai peninsula had, since the age of 15 years, repeatedly undergone surgery because of gastrointestinal bleeding. The bleeding was found to have been caused by self-inflicted injuries. Histiocytes in the patient's liver and lymph nodes (the spleen had been removed nine years previously) contained a peculiar material which was, in part, basophilic and had puzzling histochemical characteristics. The material is believed to be polyvinylpyrrolidone (PVP), which had been administered to the patient during one of her operations. The importance of recognizing this change, especially in countries where PVP is administered as a plasma expander, is stressed.


Specific effect of 5,6-dihydroxytryptamine on the monoamine fluorophore of the frog's gustatory cells. A specific formaldehyde-induced yellow fluorescence, suggesting the presence of serotonin-like monoamine has been demonstrated in the gustatory cells of the frog. The fungiform papillae of frogs were examined fluorescence-histochemically after intraperitoneal injection of 5,6-dihydroxytryptamine. The results indicated that the fluorophore of gustatory cells was affected selectively by the drug injection: the yellow fluorescence was transiently enhanced 3 hours after the drug injection, thereafter being reduced rapidly. The effect of 5,6-dihydroxytryptamine was long-lasting with the reduction of the yellow fluorophore persisting at least for the experimental duration of 14 days. A single injection of 6-hydroxydopamine induced a complete depletion of noradrenaline fluorescence from adrenergic nerve terminals, while the fluorescence of gustatory cells was not affected by a high dose of the drug. The present results with pharmacologic treatments further support the view that the gustatory cell of the frog contains a serotonin-like monoamine.


[Bleeding disorders and childhood ENT diseases (author's transl)]. The diagnosis and management of ENT patients with coincidental bleeding dyscrasias presents special difficulties, which include diagnosis of the disorder, arrest of the bleeding when unexpected, and the conservative and surgical care of affected children with ENT diseases. The dyscrasias most often found in childhood are haemophilia, von Willibrand's disease and ideopathic thrombocytopenic purpura. The clinical presentation and course of these disorders are described. Present day treatment is effective in arrest of the haemorrhage such that a fatality can almost always be avoided. The close cooperation between specialist paediatricians and haematology departments in the handling of these patients is stressed.


Vasocilators in the canine mesenteric circulation. Evaluation of a potential aid in the diagnosis of gastrointestinal bleeding. Radiodiagnostic potential of intra-arterially injected vasodialting agents was investigated by their effect on total and segmenal resistances (VR) of mesenteric vasculature, blood flow in superior mesenteric artery and its bleeding branch; heart rate and ventricular and systemic blood pressure. Dipyridamole, isoxsuprine, protricular and systemic blood pressure. Dipyridamole, isoxsuprine, prochlorperazine, lidocaine, meglumine diatrizoate and carbon dioxide were poor dilators. Phentolamine produced hypotension; glucagon and serpasil an extremely long dilation. A large and short vasodilation was produced with tolazoline and nylidrin, but both agents increased VR of the postcapillary segment and caused transient hypotension and arrhythmias, nylidrin's side effects were smaller. Oxygen produced large and long vasodilation and minimal systemic effects. It is concluded that oxygen or possibly nylidrin are suitable agents should an intermittently bleeding mesenteric artery be dilated for diagnostic purposes prior to angiography.


Immunoglobulin-containing cells in pig mammary gland. Peroxidase-labelled antisera to pig immunoglobulins A, G and M were used to study immunoglobulin-containing cell populations in pig mammary gland at different stages of gestation and lactation. Immunoglobulin-containing cells of all three classes were present several weeks before farrowing; IgA-containing cells were most numerous at all stages of lactation.


Glycerol dissimilation in Rhodopseudomonas sphaeroides. Rhodopseudomonas sphaeroides followed a diauxic growth curve when grown on a malate-glycerol medium, the first phase of growth being supported by malate and the second by glycerol. A soluble glycerokinase and a particulate, pyridine nucleotide-independent glycerophosphate dehydrogenase, were induced by the presence of glycerol in the medium, but neither was fully expressed nor functional until all malate had been consumed.


Interaction between ethanolamine ammonia-lyase and methylcobalamin. Half-site reactivity with an adenosylcobalamin-dependent enzyme. The adenosylcobalamin-dependent enzyme ethanolamine ammonia-lyase contains two active sites per molecule. The effects of methylcobalamin on the properties of this enzyme differ qualitatively depending on whether one or both of these sites is occupied by the cobamide. At 0.5 mol of methylcobalamin/mol of active sites, catalytic activity fell rapidly to approximately 30% of control levels, thereafter remaining constant for an hour. With the partially inhibited enzyme, Km values for ethanolamine and adenosylcobalamin were 5.5 muM and 1.6 muM, respectively, values that do not differ significantly from those of uninhibited enzyme. When the methylcobalamin per active site ratio was increased to 1, the decline in activity became progressive with time, eventually falling to levels much lower than seen at a cobamide per active site ratio of 0.5. Methylcobalamin also promotes the formation of a complex stable to gel filtration between ethanolamine and enzyme. Complex formation increased with increasing methylcobalamin per active site ratios up to a ratio of 0.7/1, at which point 0.5 mol of ethanol/mol of active sites was taken up. Ethanolamine uptake did not increase at higher methylcobalamin to active sites ratios. Methylcobalamin itself was taken up by enzyme, forming a complex containing 0.5 mol of methylcobalamin/mol of active sites that was stable to gel filtration. Measurement by the technique of Hummel and Dreyer ((1962) Biochim. Biophys. Acta 63, 530-532), however, showed one methylcobalamin binding site per active site. The formation of enzyme-ligand complexes stable to gel filtration was not affected by 5'-deoxyadenosine nor did 5'-deoxyadenosine by itself promote the formation of a stable complex between enzyme and ethanolamine. These observations were interpreted as evidence indicating half-site reactivity of ethanolamine ammonia-lyase with methylcobalamin. Comparison with previous results suggested that this half-site reactivity was an epiphenomenon not related to catalysis.


An in vitro study of vitamin D2 hydroxylases in the chick. An in vitro study of the liver 25-hydroxylation of vitamin D2 and the kidney 1- and 24-hydroxylations of 25-hydroxyvitamin D2 was undertaken in order to determine whether the discrimination against vitamin D2 seen in chicks in vivo is the result of a block of one or more of the steps in the activation of the vitamin D2 molecule. Vitamin D2 hydroxylation reactions in the chick are virtually identical with those observed with the vitamin D3 series. It is, therefore, concluded that the chick possesses the required enzymatic machinery to synthesize 1,25-dihydroxyvitamin D2 and that the discrimination must be because of some unknown metabolic reaction of the vitamin D2 compounds, to a defect in the transport of vitamin D2 metabolites, or to target organ discrimination.


Experimental correction of footdrop by electrical stimulation of the peroneal nerve. Footdrop was corrected in thirteen of sixteen patients with centrally paralyzed ankle dorsiflexor muscles who were treated by electrical stimulation of the peroneal nerve by means of surgically implanted electrodes. Four patients required surgical revision before a good result was obtained. After follow-ups of more than three years there was no evidence of any nerve damage caused by chronic electrical stimulation of the peroneal nerve.


Hemopoietic colony forming cells in regenerating mouse liver. Cells from regenerating mouse liver removed 2-25 days post 68% hepatic resection have been assayed for in vivo colony forming capacity in soft agar (CFU-C), proliferative capacity in liquid culture, and in vivo spleen colony forming capacity (CFU-S). These studies demonstrated low concentrations of CFU-C and CFU-S in normal and sham operated liver, with an appreciable increase of both in regenerating liver, reaching maximum values in tissue removed 5--7 days post hepatic resection. Colony formation in agar by regenerating liver cells occurred in the absence of exogenous colony stimulating factor. Separation of liver cells on the basis of adherent properties prior to culture indicated concentration of CFU-C in the non-adherent fraction, while cells producing colony stimulating factor were concentrated in the adherent fraction.  Foci of actively dividing cells of themacrophage and granulocyte series arose in liquid culture from preparations of sham operated and regenerating liver, although total cell formation was greater with regenerating liver. A small proportion of the colonies formed in agar from regenerating liver consisted of cords of epithelioid cells, which resembled hepatocytes and differed from the macrophages or granulocytes found in the majority of colonies, raising the possibility that regenerating hepatocytes form colonies in agar culture.


Separation of subpopulations of in vitro colony forming cells from mouse marrow by equilibrium density centrifugation. Equilibrium density centrifugation was used to characterise and separate subpopulations of mouse haemopoietic progenitor cells capable of producing colonies of granulocytes and macrophages in vitro. The material used to induce colony formation (CSF) was prepared from an extract of pregnant mouse uteri. This CSF preparation was found to be free of factors modifying the response. Under these culture conditions, in vitro colony forming cells (CFU-c) were found to be relatively homogeneous in their buoyant density. This homogeneity was independent of CSF concentration. A heterogeneous density profile of CFU-c was obtained when various cell fractions were cultured in the presence of CSF and rat blood lysate. The majority of the additional cells which responded to erythrocyte lysate were dense (modal density 1.080 g/cm3) compared to CFU-c which responded to CSF alone (modal density 1.074 g/cm3). It is concluded that in vitro colonies induced by CSF and in vitro colonies grown in the presence of CSF and erythrocyte lysate reflect two different populations of CFU-c.


Preparation and neutralization characteristics of an anti-CSF antibody. Nine of ten rabbits immunized with a partially purified L-cell CSF had demonstrable titers of anti-CSF activity. In vitro the antibody was markedly inhibitory to both post-endotoxin mouse sera and several mouse tissue extracts. CSF containing conditioned media prepared from a number of sources showed variable inhibition suggesting that murine CSF's may be characterized by marked antigenic differences. Human sources of CSF were also inhibited thus indicating a degree of cross-reactivity between murine and human factors. These studies may provide the initial steps toward definition of the role of CSF in vivo.


The value of moderate hypothermia during anoxic cardiac arrest for coronary artery surgery. Anoxic cardiac arrest, as opposed to induced ventricular fibrillation, greatly facilitates accurate distal anastomosis in aortocoronary bypass surgery. In order to diminish the anoxic insult, general and topical hypothermia may be used. In an attempt to establish the value of moderate hypothermia during anoxic cardiac arrest two groups of patients were compared. In group I coronary artery bypass procedures were performed under normothermic conditions with anoxic cardiac arrest. Patients in group II underwent similar procedures but under hypothermic conditions. General body hypothermia to an esophageal temperature of 30 degrees C and topical hypothermia with iced saline lavage were used. Using these techniques, the average intramyocardial temperature was 26 degrees C. Nonfatal cardiac complications did not occur more frequently in the hypothermic group. Operative mortality was decreased from 6.3% in the normothermic group to 1.5% in the hypothermic group. However, in group II, in addition to hypothermia, a second factor in the reduction of mortality was the completeness of the revascularization procedure: 58.5% of the patients had three or more bypass grafts in the hypothermic group. The mean anoxic arrest time was over 50 min for all patients--those who survived as well as those who died with postoperative low cardiac output or myocardial infarction. Therefore, anoxic arrest time should be kept as short as possible and certainly less than 50 min. Intermittent aortic occlusion and performance of the proximal anastomoses using a partial occluding clamp on the aorta are currently being used and, together with moderate hypothermia, provide a further reduction in postoperative myocardial complications.


Control of granulopoiesis in man.III. inhibition of colony formation by dense leukocytes. Cellular feeder layers, prepared from normal blood leukocytes, usually stimulate human marrow to form colonies. A significant increase in the stimulating activity of unseparated leukocyte feeder layers is brought about following the removal of dense leukocytes in a manner which avoids enrichment of any remaining cell type. Restoration of dense leukocytes to a dense leukocyte depleted leukocyte feeder layer results in the reduction of stimulating activity to that of an unseparated leukocyte feeder; however, addition of dense leukocytes to unseparated leukocyte feeder layers has no effect on the stimulatory activity, over the range of concentrations used in this study.


Emergency operations for left ventricular asynergy. Thirty-two patients were submitted to an emergency surgical procedure for left ventricular asynergy, following myocardial infarction. Indications and results are analyzed. All patients were operated upon with the aid of cardiopulmonary by-pass; an aneurysmectomy was performed in twenty-nine patients, a plication technique in three. These procedures were combined with mitral and/or aortic replacement in five patients, closure of ventricular septal defect in five, aorto-coronary saphenous vein by-pass in five. Early over all post-operative mortality was 75%: 80% in patients operated within three months from myocardial infarction, 67% in patients operated after three months from myocardial infarction.


Failure of tritiated thymidine incorporation into DNA to reflect the autoradiographically demonstrable calcium-induced increase in thymic lymphoblast DNA synthesis. Increasing the extracellular calcium concentration in thymic lymphocyte suspension from 0.6 to 1.8 mM stimulated the proliferation of the lymphoblast subpopulation as measured by increases in the proportion of cells autoradiographically labeled with 3H-TdR and in mitotic activity. However it was not possible to show this increased DNA synthesis by scintillometric measurement of the amount of 3H-TdR incorporated into extracted DNA. On the other hand, calcium did raise the incorporation of 14C-formate into the thymine residues of DNA, and increased the activity of isolated thymocyte thymidylate synthetase. In contrast to the mitogenic calcium ion, a thymidylate synthetase inhibitor, methotrexate, actually increased the incorporation of 3H-TdR into DNA. It is concluded that calcium increases the endogenous synthesis of thymidylate which in turn prevents the amount of incorporation of exogenous 3H-TdR from accurately reflecting the true level of DNA synthesis.


Colony-stimulating factor (CSF) in the uterus of the pregnant mouse. During the growth of the pregnant uterus of inbred Balb/c mice uterine colony-stimulating factor (CSF) content increased 800-fold. This corresponded to a 75-fold increase in CSF concentration. The size of the increase was the same in the uteri of inbred Balb/c mice and the uteri of F1 hybrid C57Bl/Balb/c mice mated with F1 males. It was about four times greater in these uteri than it was in the uteri of randomly mated TO mice. After unilateral Fallopian tubal ligation of TO mice the concentration of CSF was as great in the non-gravid uterine horns as it was in the gravid uterine horns. Physicochemical characterization of CSF from extracts of pregnant uterus showed that it differed from that found in foetal extracts and that found in the circulation. Its function in the pregnant uterus is discussed.


Identification of DNA-binding lymphocytes in patients with systemic lupus erythematosus. Antigen-ginding lymphocytes capable of binding native DNA (DNA-ABC) were identified in the peripheral blood of normal controls and patients with systemic lupus erythematosus (SLE) by autoradiography with 125I-nDNA. 12 patients with active SLE had 404 +/- 273 (mean +/- SD) DNA-ABC/105 lymphocytes, while 7 inactive SLE patients and 13 normals had 120 +/- 48 and 48 +/- 36, respectively. All three groups were significantly different from one another (p less than 0.01). No significant correlation was detected between the quantity of anti-native DNA (nDNA) antibody and number of DNA-ABC; however, most patients with large amounts of anti-nDNA antibody had both active disease and large numbers of DNA-ABC. Numbers of DNA-ABC and lymphocytes with surface immunoglobulin (Ig) did not change significantly after an 18-h incubation at 37degreeC. After depletion of B-lymphocytes by passage over bead columns coated with a complex of IgG and anti-IgG, the great majority of DNA-ABC were removed in both normal subjects and SLE patients. Labeling lymphocytes sequentially with 125I-nDNA, followed by an indirect fluorescence technique for identification of surface Ig, indicated that the great mahority of radiolabeled cells had surface Ig by fluorescence microscopy in four normals (average 93%) and five patients with active SLF (average 82%). The predominance of nDNA-sensitive B-lymphocytes in the peripheral blood of both normals and SLE patients is consistent with the concept that the induction of the anti-nDNA antibody response is due to the stimulation of preexisting nDNA-specific B lymphocytes by mechanisms other than those necessarily involving participation of nDNA-specific T lymphocytes.


Thymus-dependent lymphocytes in human bone marrow. Human bone marrow is known to contain significant numbers of bursa-dependent lymphocytes. The presence of thymus-dependent (T) cells is controversial. Bone marrow cells obtained from healthy volunteers was fractionated by density centrifugation. A lymphocte-enriched subpopulation was shown to be reactive to alloantigens in mixed lymphocyte culture and to contain substantial numbers of T lymphocytes. The T lymphocytes were detected by cell surface markers (rosette formation with sheep RBC) and by response to the mitogens phytohemagglutinin and concanavalin A. Bone marrow T cells exhibited functional characteristics quantitatively different from peripheral blood T cells, suggesting that they may represent a particular subpopulation of T cells. The lymphocyte-enriched fraction additionally contained committed granulopoietic stem cells capable of colony formation in semisolid gel. The presence of T cells in human bone marrow is consistent with findings in other mammals and may explain the high incidence of graft versus host disease in bone marrow transplant recipients.


Autoradiographic studies of cerebellar histogenesis in the bullfrog tadpole during metamorphosis: the external granular layer. Spontaneously metamorphosing bullfrog tadpoles and those induced to metamorphose by injections of thyroid stimulating hormone (TSH) were given a single intraperitoneal injection of thymidine-3H (10 muCi/g body weight). The brains were dissected out at various periods 3 hours to 14 days later, and processed for autoradiography. At the 3-hour interval after thymidine-3H injection, ependymal cells were labelled, but not the external granular layer (EGL) cells. Furthermore, in all the metamorphosing tadpoles intense labelling was restricted to the ependyma of the marginal region of the cerebellar plate. At 48 hours, labelled cells were seen in the EGL of the marginal region. At the 4-day interval, most of the EGL was labelled, and labelled cells were also seen migrating from the EGL into the internal granular layer (IGL). By 14 days, several labelled cells were seen in the IGL. Although the sequence of cerebellar histogenesis in the frog is similar to the general pattern described in other vertebrate groups, the results indicate that the EGL of the frog cerebellum does not serve the function of a secondary proliferating zone.


Effects of cytotoxic immunosuppressants on tuberculin-sensitive lymphocytes in guinea pigs. The immunosuppressive activities of two phase-specific drugs, 6-mercaptopurine (6-MP) and methotrexate, and a cycle-specific agent, cyclophosphamide, were evaluated on the lymphocytic component of established tuberculin hypersensitivity in guinea pigs. In these animals, purified protein derivative (PPD)-sensitive lymphocytes are in an intermitotic phase of their proliferative cycle. Neither phase-specific drug significantly altered either the number or functional activities of these lymphocytes. By two in vitro criteria, PPD-induced lymphoproliferation and elaboration of migration inhibition factor (MIF), the responses of lymph node cells were equivalent to sensitized controls. In addition, these agents did not deplete pools of T lymphocytes, impair responses to phytohemagglutinin (PHA), nor inhibit cutaneous reactivity if employed before sensitization. In contrast, cyclophosphamide showed broader immunosuppressive effects including significant toxicities for intermitotic lymphocytes. This drug depleted pools of T cells and markedly impaired the in vitro proliferative responses of residual lymphocytes. The latter occurred with both PHA and PPD. Suppression of PHA reactivity was a dose-dependent phenomenon but was evident even with small quantities of this alkylating agent. The suppression of antigen-induced responses was independent of the proliferative status of target lymphocytes in vivo, after a single large dose, it persisted for more than 3 wk. In total, these results indicate that the effective use of cytotoxic drugs as immunosuppressants must include consideration of both the cycle specificities of the agent and the proliferative activities of the target lymphoid population.


Visual stimulation: effects on vestibular habituation. The influence of various forms of visual stimulation presented during the course of vestibular habituation to a caloric stimulus was studied. Eye movements which were either complementary or in opposition to the induced vestibular nystagmus were produced with an optokinetic drum. In addition, the effect of visual fixation during vestibular-response periods was studied. In all cases, the cats that received visual stimulation during the majority of the caloric trials habituated more slowly than did animals that received all the habituation trials in total darkness. These data conflict with previous reports of vestibular-visual interactions. Possible explanations for the discrepancy include species differences, distraction provided by the visual stimuli, and the transfer of learning from the dark to light conditions.


Aging, terminal decline, and terminal drop. Data from a 20-year longitudinal study of persons over 60 were analyzed by step-wise multiple regression to test for declines in function with age, for terminal decline (linear relationship to time before death), and for terminal drop (curvilinear relationship to time before death). There were no substanial terminal drop effects. All health measures had substantial declines with age, and the physician's physical function rating, as well as the individual's self-rating, showed additional terminal decline. Intelligence had a substantial decline with age and a small terminal decline. Most activities declined with age but had no significant terminal decline. Despite these declines in health, intelligence, and activities, there was little or no decline in satisfaction or adjustment.


Characteristics of the 'active rosette test'. I. Technical considerations of the test and comments. Using normal subjects, it was found that the 'Active Rosette Test' is both reproducible and consistent if performed exactly as described by Wybran et al. (1972). Variations in incubation time, centrifugal force while sedimenting rosettes, resuspension time after centrifugation, SRBC lymphocyte ratio, and increasing concentrations of fetal calf serum all produced marked deviations in test results. Rosette inhibition studies using a specific anti-T cell antisera showed inhibition of 'active' and total rosettes at the same level, suggesting that there are no marked variations in surface T antigens among respective T cells. Unless differences in functional activities can be demonstrated between the 'active' rosette-forming cells and the other T cells, the test will be of an empiric nature.


Action of staphylococcal epidermolytic toxin on mouse skin: an electron microscopic study. The ultrastructure of the skin of 3-day-old mice challenged with small doses of highly purified staphylococcal epidermolytic toxin was examined at various time intervals. Up to 130 min few changes were evident, but at this time wide gaps developed between cells in the horizontal planes of the stratum granulosum, and "bubbles" normally present in the intracellular spece were no longer apparent. Splitting of the desmosomes occurred after the development of distended intercellular spaces. After 20 hr, that is, in the "healing" phase, the appearance suggested that normal maturation of keratinocytes was altered. Also at this time a degree of cell separation was still apparent in the outermost actively maturing layer of the stratum granulosum. The proteinase inhibitor Trasylol was tested for its effect on the toxin when administered up to 45 min after challenge.


Second ear stapedectomy--a continued controversy. This paper presents the results of an investigation into the effects of stapedectomy on vestibular function. We detected a disturbingly high incidence of vestibular abnormality in the results of caloric tests at three months postoperatively. Surprisingly, this was not usually accompanied by concurrent vestibular symptoms and there was no evidence of cochlear dysfunction in most cases. Although the caloric response at 12 months postoperatively showed an improvement in most instances, the responses rarely became symmetrical. We believe that a long-term alteration in vestibular response is not uncommon following stapedectomy. Although most patients will compensate quite quickly after unilateral stapedectomy we suspect that this occurs much less readily after bilateral stapedectomy and not at all should bilateral impairment of vestibular function occur. On the basis of this evidence, we submit that bilateral stapedectomy is justified only when vestibular function can be shown to be normal prior to the second ear operation.


Reversible hearing loss in tumours of the cerebello-pontine angle. A survey of the literature concerning reversible hearing loss in tumours of the cerebello-pontine angle is presented together with two additional cases. After removal of one meningioma and one epidermoid tumour, respectively, the abolished acoustico-vestibular function returned in both cases. The pre-requisites for the restoration of hearing are discussed.


The Fc receptor on thymus-derived lymphocytes: II. Mitogen responsiveness of T lymphocytes bearing the Fc receptor. The responsiveness of purified Fc- and Fc+ T lymphocytes, isolated from normal spleen cell populations by cell sorting on the fluorescence activated cell sorter, has been examined. Although both Fc- and Fc+ T cells responded to phytohemagglutinin, the response to concanavalin A (Con A) was found to be a characteristic of the Fc+ T lymphocyte. The poor responsiveness of the Fc- T cells to Con A was shown not to be due to a requirement of either different concentrations of Con A or for adherent cells. The addition of Fc+ T cells to the Fc- T cells in a ratio of 1:3 resulted in a mitotic response not significantly different from that observed with the purified Fc+ T cells alone and up to 15-fold greater than that of Fc- T cells alone. It is suggested that the Fc T cells can be recruited into mitosis as a result of Con A stimulation of the Fc+ T cells.


In vitro cell-mediated immune responses to the male specific(H-Y) antigen in mice. C57BL/10 female mice were primed to the male specific antigen H-Y, either by grafting with syngeneic male tail skin or by i.p. injection of syngeneic male spleen cells. Primed female spleen cells, either unseparated or filtered through nylon wool to remove most of the B lymphocytes, were then cultured for 5 days in vitro with irradiated syngeneic male spleen cells and assayed against 51Cr-labeled target cells. Both unseparated and nylon wool filtered female cells displayed significant cytotoxic activity restricted to male target cells. Pretreatment of sensitized female cells with antitheta serum and complement just before assay abolished cytotoxic responses. We were unable to demonstrate cell-mediated cytotoxic responses into two nonresponding strains, CBA and B10.A, which fail to reject male isografts. The cytotoxic activity of C57BL/10 female cells was restricted to male target cells histocompatible with C57BL/10 over at least a portion of the major (H-2) histocompatibility complex. We conclude that secondary in vitro cytotoxic responses against the H-Y antigen are mediated by cytotoxic T lymphocytes, and that the H-Y target cell antigen may be specified by the H-2 complex.


Properties of antigen-specific suppressive T-cell factor in the regulation of antibody response of the mouse. I. In vivo activity and immunochemical characterization. An antigen-specific suppressive T-cell factor was extracted from physically disrupted thymocytes and spleen cells of mice that had been immunized with soluble protein antigens. The factor, when inoculated into syngeneic normal mice, could induce a significant suppression of IgG antibody response against a hapten coupled to the carrier protein by which the donor of the suppressor factor was immunized. The suppressor factor was found only effective in suppressing the antibody response of syngeneic or H-2 histocompatible recipients. The suppressive T-cell factor was removed by absorption with immunoadsorbent composed of the relevant antigen, but not with any of those of anti-immunoglobulin antibodies. The factor was successfully removed by alloantibodies with specificity for the K end (H-2K, I-A and I-B) of the H-2 complex of the donor strain, but not by those for the D end (I-C, SsSlp, and H-2D). The activity was removed by absorption with a heterologous antithymocyte serum. The mol wt of the suppression T-cell factor was between 35,000 and 60,000 as determined by Sephadex G-200 gel filtration. The suppressive T-cell factor was found to be a heat-liable protein.