Reconstituted energy transfer from antenna pigment-protein to reaction centres isolated from Rhodopseudomonas sphaeroides.

Efficient energy transfer has been reconstituted between an antenna pigment-protein and reaction centres isolated from the photosynthetic membrane of Rhodopseudomonas sphaeroides. The reconstituted system has fluorescence induction kinetics and fluorescence yields similar to those obtained from antenna bacteriochlorophyll in chromatophores. The results indicated that closed reaction centres quench fluorescence from the antenna pigment-protein, although not as strongly as photochemically active reaction centres. The measurement of fluorescence yields from chromatophores of the reaction centreless mutant PM-8 and of the parent strain Ga confirmed these observations. The fluorescence yield from the reconstituted system was approximately the same whether the reaction centres had been closed by photo-oxidation of the bacteriochlorophyll electron donor or chemical reduction of the primary acceptor, indicating a similar lifetime for the excited singlet state in both states of the reaction centres.


Secondary electron transfer in reaction centers of Rhodopseudomonas sphaeroides. Out-of-phase periodicity of two for the formation of ubisemiquinone and fully reduced ubiquinone.

Electron transfer between purified reaction centers from Rhodopseudomonas sphaeroides and exogenous ubiquinone has been studied in the presence of electron donors by measurements of light-induced absorbance changes following a sequence of short actinic light flashes. Each odd flash promotes the formation of a molecule of ubisemiquinone; after each even flash the semiquinone disappears and a molecule of the fully reduced quinone appears. We interpret these results by means of a model where a specialized molecule of ubiquinone is reduced by the primary electron acceptor in a one-electron transfer reaction after each flash, and is reoxidized by a molecule of the ubiquinone pool in a two-electron transfer reaction every two flashes.


Formation of a thymine photoproduct in transforming DNA by near ultraviolet irradiation.

Irradiation at 334 and 365 nm of a highly purified preparation of thymine-labeled transforming DNA from Haemophilus influenzae produced a photo product containing label from thymine but different from the cyclobutane dimer. The photoproduct is soluble in water and in ethanol and Rf values in a number of solvents are presented. The photoproduct has properties similar in a number of respects to those of the spore photoproduct, 5-thyminyl-5,6-dihydrothymine. The near ultraviolet photoproduct is more likely to affect the oxygen independent inactivation of transforming DNA rather than its mutagenesis, as judged by the quantitative relationship between amount of photboproduct and inactivation and mutagenesis.


Thymus-dependent lymphocytes and delayed hypersensitivity in low birth weight infants.

The proportion of thymus-dependent (T) lymphocytes in the blood and the ability to mount cutaneous delayed hypersensitivity to a battery of antigens were evaluated in 50 low birth weight (LBW) infants when they were 3 months to 5 years old. There was a significant reduction in the number of T lymphocytes. Delayed hypersensitivity was impaired. The abnormalities were more pronounced in those with persistent growth retardation. It is suggested that continued postnatal depression of cell-mediated immunity in LBW infants may contribute to increased frequency of infections and immunopathological diseases.


Elastic and viscous properties of resting frog skeletal muscle.

The mechanical properties of the resting, whole semitendinosus muscle of the frog have been characterized as functions of both muscle length and temperature. Measurements were made of pseudorandom white noise (PRWN) displacements (less than 10 A/half-sarcomere) applied to the muscle and the force responses to these movements. Signal correlation techniques were then used to obtain the dynamic modulus function for the muscle in the frequency range 2.44-320 Hz. This function was represented by a series combination of a Voigt element and a time delay element for tension propagation along the muscle. A dynamic elastic modulus (E), coefficient of damping (B), and tension transmission velocity (V) were measured for resting muscle on the basis of this model. For each of these parameters, a marked variation with sarcomere length (s) was found. The mean values for E and B at LO (s=2.25 mum) were 1.84+/-0.24 X 10(5) N/m2 and 2.33+/-0.25 X 10(2) Ns/m2, respectively. Further, B demonstrated a negative temperature dependence, Q10=0.78 (P less than 0.05), in the range s=2.6-3.0 mum, while E was not significantly temperature dependent. The length-dependent variations of E and B are interpreted as deriving from both passive muscle elements and attached crossbridges. Velocity was calculated at a single displacing frequency for every experiment; the mean value at LO and all temperatures was v=11.7+/-0.6 m/s. Velocity was also calculated as a function of frequency within several experiments: the results indicate considerable variation of v with frequency.


Spin-lattice relaxation times for 13C in isotope-enriched glycine accumulated in frog muscle.

Spin-lattice relaxation times (T1's) of 13C-enriched glycine accumulated in frog muscles were determined at 1 degrees C by the inversion-recovery (180 degrees -tau-90 degree pulse sequence) method and compared with the values obtained in free solution. The value of T1 for the alpha-13C nucleus of glycine in the tissue was 50% of that obtained in free solution. The observed value for T1 in the tissue was not concentration-dependent, and no difference in chemical shift was observed between tissue and free solution. Quantification of the area under the glycine peak suggested that the observed signal represents at least 80% of the intracellular glycine. An average nuclear Overhauser enhancement of 2.83 for intracellular glycine indicates that the relaxation mechanism within the cell is predominantly dipolar, as in free solution. The value of T1 for the 13C' nucleus of glycine in the tissue was 67% of that in a solution of similar concentration. A quantitative analysis of the findings suggests that the observed difference in the value of T1 between tissue and free solution results from a difference in viscosity. The data provide no evidence either for special organization of intracellular water or for glycine binding. It is proposed that intracellular diffusion coefficients may be determined from measurements of 13C T1's of 13C-enriched intracellular solutes.


Systemic mastocytosis in a patient with polycythemia vera treated with radioactive phosphorus.

Systemic mastocytosis occurred as a fatal event in a patient with long-standing polycythemia vera. The patient had been treated over the course of 21 yr with radioactive phosphorus. Possible relationships between mastocytosis and polycythemia vera, and also between mastocytosis and treatment with ionizing radiation, are discussed. Histopathologic and electron microscopic findings are illustrated. Difficulties in establishing the diagnosis of mast cell disease in this setting are also described.


Stimulation by human placental conditioned medium of hemopoietic colony formation by human marrow cells.

Medium conditioned by human placental tissue was found to stimulate granulocytic and monocytic colony formation by human marrow cells in semisolid agar cultures. The colony-stimulating activity of unfractionated conditioned medium was equivalent to the activity of standard peripheral blood underlayers. Placentas were a reliable source of active material, and one placenta provided enough material to stimulate 5,000-10,000 cultures of normal or leukemic cells. The colony-stimulating factor in human placental conditioned medium (CSFHPCM) was concentrated and purified 1800-fold using ammonium sulfate precipitation, calcium phosphate gel absorption, DEAE-cellulose batch absorption, gel filtration on Sephadex G-150, and polyarcylamide gel gel electrophoresis. The active factor behaved on gel filtration as a macromolecule with an apparent molecular weight of 30,000 daltons. The active factor in placental conditioned medium was not dependent on the presence of adherent marrow cells with endogenous colony-stimulating activity.


Selective deficiency of IgG and IgA as a first stage defect in B cell differentiation.

A case of selective deficiency of IgG and IgA, in a 13-year old girl, is described. Immunologic investigations, showing an almost complete absence of IgG- and IgA-bearing lymphocytes and significant amounts of IgD and IgM positive cells, suggest the possibility of a block in the shift from IgM to IgG synthesis at the B lymphocyte level.


Pulmonary host defense: coordinated interaction of mechanical, cellular and humoral immune systems of the lung.

The respiratory system has numerous ways to protect lung parenchyma from implantation of bacteria and subsequent infection. Such "natural" defense utilizes the clearing mechanism in the nose, larynx and upper airways and the cellular and humoral immune factors in the lower respiratory tract so effectively that the host is largely unaware of its tireless surveillance. Normal lungs are kept sterile. In contrast, pneumonitis, which represents the fully developed acute inflammatory reaction, signals the ultimate response to virulent bacteria, but can be considered as general failure of host defense also--depending on your viewpont. In between these extremes are gradations of response, which are unnoticed, perhaps. Thus, the inflammatory response in the lung to bacteria requires initiation, modulation and eventually suppression. This report attempts to dissect individual components of this response and to examine cell products which may have regulatory functions.


Monoaminergic inputs to frog motoneurons: an anatomical study using fluorescence histochemical and silver degeneration techniques.

The monoaminergic input to spinal segment 10 of the frog, Rana pipiens, was investigated anatomically. Fluorescence histochemistry demonstrated a diffuse and overlapping distribution of catecholamine (CA) and indoleamine (IA) containing terminals within the spinal cord. Terminal fields containing high numbers of CA and IA varicosities included the marginal plexus and the anterior commissural region. IA terminals were also abundant in the dorsal portion of the dorsal horn, while CA terminals were found in high number in the ventral part of the dorsal horn. IA varicosities were regularly observed within the lateral motor nucleus, but CA varicosities were not. Throughout the medulla IA neurons were located exclusively within the raphe nuclei. From the level of the obex to the entry of cranial nerve X, CA neurons were scattered in the intermediolateral and intermediomedial reticular formation. In Fink-Heimer preparations of segment 10, raphe lesions produced a pattern of terminal degeneration that corresponded to the fluorescent histochemical picture of IA varicosities. All levels of the medullary raphe nuclei projected to the lateral motor column. Other spinal regions received a differential raphe input. It is concluded that motoneurons of segment 10 have a somal input from the medullary raphe nuclei that is indoleaminergic in nature. It is also probable that distal motoneuronal dendrites and interneurons are contacted by IA and CA terminals.


Adaptive distortions in the generator potential of semicircular canal sensory afferents.

The generator potential in sensory afferents of frog crista ampullaris was extracellularly recorded from the cut end of the posterior ampullary nerve by means of suction electrodes. A servocontrolled turntable allowed suitable rotatary stimulations. The analysis of the recorded generator potential revealed a different time course from that predicted on the basis of the pendulum model. Adaptation and undershoots in the responses to velocity ramps, steps and sinusoids, were mainly responsible for the deviations, which became very evident only when fairly high acceleration rates were applied. Both adaptation and undershoots were produced presumably by the activation of an electrogenic pump, probably located in nerv terminals contacting the hair cells. In fact, the time course of the generator potential became much more consistent with the predictions from the pendulum model under treatments capable of hindering the ion pump activity.


Stereotyped behaviour patterns and hyperactivity induced by amphetamine and apomorphine after discrete 6-hydroxydopamine lesions of extrapyramidal and mesolimbic nuclei.

Changes in stereotyped sniffing, biting and hyperactivity induced by apomorphine and D-amphetamine in the rat were determined after bilateral 6-hydroxy-dopamine (6-OHDA) lesions (8-16 micron/4micron6) of the extrapyramidal caudate-putamen (CP) (anterior and centre), globus pallidus (GP) and substantia nigra (SN), the mesolimbic nucleus (ACB), tuberculum olfactorium (TUO) and central amygdaloid nucleus (ACE). Lesions were also induced in the medial forebrain bundle in the lateral hypothalamus (LH). The 6-OHDA lesions of the CP reduced amphetamine biting but not sniffing or hyperactivity. Centrally placed 6-OHDA failed to modify any response to apomorphine but anterior locations reduced apomorphine biting. Both lesion locations led to a 45-65% reduction in striatal dopamine (DA) content, but the anterior location also involved depletions of mesolimbic DA. 6-OHDA lesions of the GP reduced striatal DA by 62% but initially potentiated before reducing both apomorphine and amphetamine biting. These lesions also potentiated amphetamine hyperactivity but other parameters were unmodified. The LH and SN lesions reduced striatal and mesolimbic DA by 75-80% and potentiated apomorphine biting. The LH lesions reduced amphetamine biting and hyperactivity but the SN lesions initially potentiated these responses. 6-OHDA lesions of the ACB reduced the DA content of this nucleus by 72% but had little effect on the TUO: these lesions reduced the duration of amphetamine hyperactivity and potentiated apomorphine biting. In contrast, equally selective lesions of the TUO (80% DA depletion) enhanced the locomotor activity response to both apomorphine and amphetamine; apomorphine biting was also increased but other parameters were unmodified. Lesions of the ACE depleted amygdaloid DA by at least 80% and reduced or abolished apomorphine and amphetamine biting in the chronic stage. The results indicate that the sites for mediation of sterotyped sniffing, biting or hyperactivity are not the same for apomorphine and amphetamine, and that each behavioural state involves the functioning of more than one DA-containing area.


Behavioral effects of lesions in the A10 dopaminergic area of the rat.

Experiments have been carried out with 150 rats in order to study some psychophysiological functions of the mesencephalocortico limbic dopaminergic A10 group. Lesions in the A10 area were made by using 6-hydroxydopamine (6-OHDA) local injections; 2 small volumes of injections were used at the same concentration (2 mug/1 mul or 1 mug/0.5 mul). In a first experiment the effects of these two injections were tested on locomotor activity measured in a circular corridor, 10 and 30 days after surgery. Injections provoked hyperactivity, mainly during nocturnal basal activity periods, but not during initial exploratory activity periods. The larger the injection, the more important the hyperactivity was. The larger injections induced important food spillage evidence through the wire floor of the home cage and perturbation in a passive avoidance learning. There was no change in body weight or in amount of ingested food. In a second experiment, the effects of local injection of 6-OHDA in the other CA structures or bundles situated in or near the ventral tegmental area were tested. Injections in the substantia nigra compacta, in the noradrenergic ventral bundle, in the dorsal periventricular system-tegmental radiations did not provoke locomotor hyperactivity. In a third experiment, a possible role of the median raphe (MR) nucleus in the A10-lesion induced hyperactivity was tested: first, radiofrequency MR lesions were made and no durable significant hyperactivity was recorded; secondly, 6-OHDA (1 mug/0.5 mul) was injected into the A10 area and activity was measured 10 days later: these injections provoked significant hyperactivity during the nocturnal basal and the diurnal basal activity periods. It might be concluded that neither the neighboring CA fibers nor the MR were directly involved in the ventral tegmental -- 6-OHDA lesions syndrome. Anatomical controls by using the Fink-Heimer silver impregnating method have demonstrated, first, that the 6-OHDA injections did not destroy fibers other than catecholaminergic and secondly, that the degenerations are found in the forebrain and cortical limbic A10 projections. Hypotheses are made about a possible general inhibitory role of the A10 in behavior, in the sence of selective and attentive arousal processes, often impaired in some mental illnesses.


Herpes zoster conjunctival ulceration.

A 70 year old man acquired a herps zoster infection of the ophthalmic division of his right trigeminal nerve. During the course of the illness he developed the rare complication of a cojunctival ulcer. It is suggested that the ulceration may be the result of an ischemic ischemic vasculitis.


Effects of diphenylhydantoin and phenobarbital on voltage-clamped myelinated nerve.

Diphenylhydantoin (DPH) and phenobarbital (PB) have a selective action in blocking spontaneous activity in nerves made hyperexcitable by lowering the calcium concentration of the bathing medium (Rosenberg, P. and Bartels, E. 1967 J. Pharmacol. Exp. Ther. 155, 532-544.). To investigate this further, we examined the action of DPH and PB on voltage-clamped single myelinated nerves at two different calcium concentrations. In 1.8 mM calcium Ringer, DPH reduced the sodium permeability (PNa) without affecting the potassium conductance (GK) or the voltage-dependent time constants of sodium activation (taum) and inactivation (tauh), and potassium activation (taun). PB was similar to DPH except that in addition to reducing PNa, it shifted taum in the direction of depolarization. When the calcium concentration was lowered to 0.36 mM, the curves relating taum and taun to membrane potential were shifted in the direction of hyperpolarization, as expected. However, the addition of DPH or PB reduced or abolished these shifts. It is suggested that both DPH and PB stabilize hyperexcitable membranes by an action on the parameter m, and that this may contribute to their antiepileptic action.


Predictive value of myocardial perfusion imaging for aortocoronary bypass surgery.

Evaluation of patients for aotocoronary bypass (ACB) surgery is dependent on the findings at cardiac catheterization. Left ventricular angiography cannot distinguish between reversible and irreversible myocardial dysfunction. In 82 patients macroaggregates of human serum albumin, labelled with iodine-131, were injected before operation into the right coronary artery and technetium-99m labelled microspheres were injected into the left coronary artery. Two weeks after operation similar technetium-99m and iodine-131 injections were made into the native vessels and grafted vessels, respectively. Myocardial distribution of the 20 mu particles was visualized with a rectillinear scanner. Sixty-seven patients had normal myocardial perfusion scans preoperatively even though wall motion was abnormal in 34. There was substantial improvement in the left ventricular function of these 34 patients; the mean ejection fraction increased from 56 to 65%. Fifteen patients had abnormal myocardial perfusion scans; improvement in left ventricular function in this group occurred only if the bypass graft was made to an area with normal regional perfusion preoperatively. It is certain that an adequate capillary bed is present if the myocardial scan is normal, regardless of the degree of critical stenosis in either coronary artery. The patient with a completely occluded coronary artery and normal myocardial scan is a good candidate for ACB and will show improved left ventricular contraction if the ACB is patent,


Influence of left ventricular function and other parameters on early and late mortality following coronary bypass surgery.

Left ventricular performance, coronary anatomy and postoperative clinical parameters of 66 control patients (group 1) randomly selected from 797 survivors of coronary bypass surgery were compared with those of 45 patients who died within 30 days of operation (group II) and 53 patients who died late (average, 22+/-2 mo) (group III). Average preoperative left ventricular end-diastolic pressure, ejection fraction and mean circumferential fibre shortening rate were significantly better in group I  than in group III patients. These same parameters were not significantly different when group I  was compared to group II. Abnormal wall motion was significantly less frequent in patients from group I than in patients from group II and III. Triple-vessel disease was present preoperatively in 40 (61%) controls compared with 37 (82%) early deaths (P = 0.021) and 35 (66%) late deaths (NS). The number of grafts placed per patient was not significantly different in the three groups. Perioperative myocardial in farction (MI) and low cardiac output syndrome were the commonest causes of early death. Late complications such as MI and heart failure occurred in 4 (6%) v; 15 (32%) (P = 0.0006) and 4 (6%) v. 27 (57%) (Pless than 0.0001) group I v. group III patients, respectively. Surgical expertise, preoperative ventricular performance and triple-vessel disease are important determinants of early mortality following bypass surgery and preoperative left ventricular performance is one of the most important determinants of late mortality,


Myocardial infarction following coronary artery bypass: factors influencing its occurrence.

The authors have undertaken a prospective study on postoperative myocardial infarciton (PMI) following coronary artery bypass (CAB) grafting. PMI was diagnosed from electrocardiographic findings by the criteria of the American Heart Association. From Aug. 1, 1975 to Feb. 27, 1976, 198 patients (177 men, 21 women) underwent CAB. Their ages ranged from 31 to 71 years (mean, 49 yr). Of these patients, 18 (9%) sustained a PMI. A number of factors were analyzed to determine their influence on the occurrence rate of PMI. Preoperative factors wuch as the New York Heart Association classification, measurement of left ventricular function and evaluation of the severity of coronary artery disease were not helpful in predicting a predisposition to PMI. Endarterectomy was the only operative intervention associated with a higher inicidence of PMI (18%). Other factors such as the interval of cardiac anoxia and cardiopulmonary bypass and the number of vessels grafted did not seem to be associated with PMI. Low blood flow (less than 40 ml/min) through the graft could be correlated with the area of infarction in only 5 of 18 patients. Among the operated patients, postoperative ventricular arrhythmia was much more common in patients with PMI. The hospital mortality in this group was 11% (two patients).


Experience with reoperation following coronary bypass grafting.

Thirty-four of 1152 patients who had undergone aortocoronary bypass grafting underwent reoperation. There were no early or late deaths among these 34 patients. The average time between operations was 14 months. At the initial operation, 88 grafts and 10 internal mammary implants were placed; at reoperation, 56 grafts and 8 internal mammary implants were placed and a left ventricular aneurysm was plicated. Twenty patients underwent reoperation for graft stenosis or occlusion, five because of progression of coronary artery disease, seven because of combined progression disease and graft occlusion and two to correct technical errors. Improvement in the class of angina was 1.1 classes at an average follow-up of 9 months. The authors believe that reoperation for coronary bypass grafting can be performed safely with symptomatic improvement for the patient.


Genetic control of antinuclear antibodies in mice infected with Rauscher leukemia virus.

The incidence of antinuclear antibodies after Rauscher leukemia virus inoculation was found to be significantly higher in C57BL/6 than in BALB/c mice and still greater in their F1 hybrids. The relationships among antinuclear antibody incidence, erythroblastic disease, Rauscher leukemia virus production, and the H-2 genotypes were studied in the F1 generation and backcrosses using different virus inocula. The results observed suggest that (a) at least two genes are involved in the control of susceptibility to Rauscher leukemia virus-induced erythroblastosis, one of them probably being H-2 linked, and that (b) a non-H-2-linked gene seems to control, at the same time, induction of antinuclear antibodies, focus-forming virus production in the spleen, and susceptibility to the disease. It can be concluded that C-type viruses play an active role in antinuclear antibody induction.


Relationship of surface immunoglobulin-bearing cells, plasma cells, and tumor development in anaplastic carcinoma-bearing A/J mice.

Since the humoral immune response has been shown to be associated with immunological enhancement of tumor growth, the study of surface immunoglobulin-bearing cells and plasma cell antigen (PCA)-bearing cells during neoplastic development may provide new approaches to the study of tumor immunology. Peripheral blood was collected every other day from normal and carcinoma-bearing mice. Lymphocytes obtained by Ficoll-Hypaque density centrifugation were assayed for immunoglobulin-bearing cells and PCA-bearing cells using either fluorescein-conjugated goat anti-mouse immunoglobulin or rabbit anti-mouse plasma cell serum and fluorescein-conjugated goat anti-rabbit immunoglobulin. A marked increase in immunoglobulin-bearing cells from tumor-bearing mice was observed by Day 6 and peaked at Day 10. An increase in PCA-bearing cells followed the immunoglobulin-bearing cells increas by 2 to 4 days. The immunoglobulin-bearing cells declined by Day 12, whereas PCA-bearing cells remained elevated through Day 20. Using rabbit anti-mouse plasma cell serum as an immunosuppressive agent, a 4-day prolongation of the mean survival time was observed in rabbit anti-mouse plasma cell serum-treated tumor-bearing mice. This suggests that tumor growth in this model may be related to an active humoral immune response and that suppression of the plasma cell population may prove to be beneficial in the treatment of certain tumors.


Lymphoid subpopulation changes in regional lymph nodes in squamous head and neck cancer.

Lymph nodes from 10 normal patients and regional lymph nodes (RLN) from 19 patients with squamous cancer of the head and neck were evaluated as to their lymphoid subpopulations. In comparison to normal lymph nodes, RLN from cancer patients demonstrated a marked increase in the proportion of cells with membrane immunoglobulin, the receptor for the third component of complement, and the receptor for the Fc portion of immunoglobulin G. The increased Fc receptor cells were not Fc-bearing thymus-derived lymphocytes, inasmuch as they separated with the non-sheep erythrocyte-lymphocyte rosette-forming population. The overall thymus-derived lymphocyte percentage in RLN was proportionally decreased. A transition from the normal lymph node composition to the altered lymphocyte profile seen in RLN was demonstrated on moving from distal lymph nodes to RLN within the lymphatic drainage of a tumor. Lymph nodes involved with tumor also showed the pattern of bursa equivalent cell population increases.


Is early revascularization following acute coronary occlusion a safe and effective surgical procedure?

Coronary bypass surgery performed after more than 3 or 4 hours of coronary occlusion may results in serious derangements of cardiac metabolic, contractile, and electrical functions, and lead to extensive myocardial infarction, as well as to an unacceptable mortality. If the operation can be performed within the first 1 to 2 hours of acute coronary occlusion, hospital mortality rate is acceptably low. Although the average infarct size was usually reduced by reperfusion after 3 hours of experimental occlusion, in about 30 percent of cases there appeared to be no salvage of jeopardized myocardium.


Ciliary function of the frog oro-pharyngeal epithelium.

The palate epithelium of the frog was examined by scanning electron microscopy, light microscopy and high speed cine micrography. The cilia remain stationary for much of time in the end-of-effective stroke position. Each beat cycle begins with a forwardly-directed recovery stroke lasting about 60 ms, followed by an effective stroke towards the oesophagus lasting about 12 ms. Activity can often be correlated with the presence of mucus, which is carried as strands on the tips of the ciliary effective strokes whilst the recovery strokes move beneath the mucus. Coordination of ciliary activity was very variable; local antiplectic metachrony of the recovery strokes could almost always be seen, and on very active epithelia effective strokes were associated with approximately diaplectic waves (either to left or right), but any particular pattern of coordinated activity was transient and quickly transformed to another pattern. Beating and coordination of these short cilia were compared with those of cilia  propelling water.


DNA-synthesizing cells in human fetal thymus.

Fragments and suspensions of human fetal thymus were incubated in the presence of 3H-TdR to permit study of the distribution and morphology of DNA-synthesizing cells. Results of light and EM autoradiography showed that 1. although DNA-synthesizing cells were present in the medulla, the vast majority of these cells were localized in the thymic cortex, 2. cells with the typical EM appearance of small lymphocytes and lymphoid blast cells both synthesized DNA, and 3. cells in S-phase were predominantly 8 to 12 mum in size.


A Golgi study on the hypothalamus of amphibia. The neuronal typology.

The neuronal typology in the hypothalamus of the frog and the crested newt was studied by the Golgi technique. In the newt, piriform, multipolar or cerebrospinal fluid (CSF)-contacting neurons of relatively primitive type, according to the classification of Ramón-Moliner, are encountered in the preoptic area. Moreover, magnocellular neurons are impregnated. In the frog the preoptic area shows a more varied typology. The posterior hypothalami of the frog and the newt exhibit mainly bipolar CSF-contacting and piriform neurons. These latter are generally "tufted", but some bipolar of multipolar cells are encountered, especially in the frog. The simple anatomical organization of the amphibian hypothalamus corresponds well with the pattern of "generalized" integrative area where multimodal sensory inputs converge--including visceral information from cerebrospinal fluid by means of hypothalamic CSF-contacting sensors--to regulate the neuroendocrine outflow.


The topography of the caudal part of the paraventricular organ in Rana temporaria.

The results of a monoamine-fluorescence study of the hypothalamus of Rana temporaria show that the brain area corresponding with the nucleus infundibularis dorsalis (NID), as described in other species, does not differ, neither morphologically nor histochemically, from the paraventricular organ (PVO), with which it is anatomically continuous. It is concluded that a nucleus infundibularis dorsalis does not exist as a separate entity in this species.


Status of the grafts and the native coronary arteries proximal and distal to coronary anastomotic sites of aortocoronary bypass grafts.

The status of the native coronary arteries at necropsy in the vicinity of the coronary anastomoses of saphenous vein aortocoronary bypass grafts in 20 patients with severe coronary heart disease is presented. Of the 37 graft systems (graft plus coronary artery into which graft inserted) analyzed, the lumina of 44% of the native coronary arteries within the first 2 cm distal to the anastomoses were greater than 75% narrowed in cross-sectional area by atherosclerotic plaques, and the native coronary artery at the site of the anastomosis was greater than 50% narrowed in cross-sectional area already by atheroclerotic plaque in 25% of the graft systems. The mean coronary arterial size distal to the site of the coronary graft anastomosis, even after correction for heart weight, was greater in the 13 men than in the seven women. The residual luminal areas squared per gram of heart weight, however, were similar in both men and women. These results suggest that 1) relative coronary vessel size is greater in men than women; 2) the luminal area squared per gram myocardial mass (a relative estimation of flow) is the same in the two groups of patients; and 3) less atherosclerotic plaque is necessary in women then in men to produce similar limitation to coronary flow. Thus, vessel size alone cannot account for the higher reported frequency of unsuccessful aortocoronary bypass procedures in women.


Application of isoelectric focusing in alpha1-antitrypsin phenotyping.

alpha1-Antitrypsin, the major protease inhibitor in human serum, occurs in a considerable number of variant forms, some of which are associated with lung and liver diseases. The identification of these genetic variants, generally called Pi-types, by means of isoelectric focusing is described, as well as investigations concerning the practical application of isoelectric focusing as a routine procedure for typing the variants of alpha1-antitrypsin. Finally, isoelectric focusing is compared with the most widely used Pi-phenotyping technique, namely acid starch-gel electrophoresis followed by immunoelectrophoresis in antibody-containing agarose gel.


Lymphocyte surface markers in acute rheumatic fever and post-streptococcal acute glomerulonephritis.

Lymphocyte cell-surface markers were examined in forty children with acute rheumatic fever (ARF) and twelve with acute post-streptococal glomerulonephritis (AGN) and compared to thirty-six normal controls of similar age. Cell-surface-marker studies included surface Ig using fluorescein-labelled F(ab)2 anti-F(AB')2, IgG aggregate binding cells, and EAC rosettes. T cells were identified both as 'active' rosettes and total E-binding cells. Proportions and absolute numbers of cells bearing surface Ig and Fc receptors were elevated in subjects with AGN (Pless than0-01-0-5), whereas proportions of cells producing EAC rosettes were diminished. Patients with acute rheumatic carditis or chorea showed a substantial elevation in proportions and numbers of active T-cell rosettes (Pless than0-01). Streptococcal antigen binding cells capable of forming rosettes with autologous cells coated with group A streptococcal membranes were elevated in the acute phase of both rheumatic fever and acute glomerulonephritis(Pless than0-01). The majority of such cells were removed by passage over insolubilized Ig-anti-IgG columns and appeared to be B cells.


Ecotaxis: the principle and its application to the study of Hodgkin's disease.

A study of the function, characterization and distribution of T and B lymphocytes in five children with Hodgkin's disease is presented. The results, indicating that lymphocyte depletion in the peripheral blood does not necessarily reflect an overall lack of circulating lymphocytes, are presented to demonstrate that failure of ecotaxis (normal lymphocyte migration and distribution) can occur in man. The underlying reasons for such failure and their relevance to the pathogenesis of Hodgkin's disease are discussed.


The effect of adherent and phagocytic cells on human lymphocyte PHA responsiveness.

The effect of small numbers of adherent and phagocytic cells on the human peripheral blood lymphocyte response to PHA was examined by depleting these cells from lymphocyte preparations. Lymphocyte preparations obtained by centrifugation on Ficoll--Triosil, which contained on average 85% lymphocytes, responded well to PHA. Depletion of cells adhering to nylon fibre, giving a population containing on average 95% lymphocytes, resulted in a considerably reduced response. Depletion of cells that adhered to plastic or ingested iron powder to give populations containing on average 90% lymphocytes, also reduced the PHA response, but to a lesser extent. Reduction in PHA responsiveness correlated with increasing lymphocyte purity. The responsiveness of nylon-column-filtered cells could be restored by adding a small number of cells from a monocyte-rich population.


Human lymph node lymphocytes fail to effect lysis of antibody-coated target cells.

Human lymphocytes prepared from peripheral blood, lymph nodes, spleen and thymus were titrated for ability to mediate lysis of human target cells coated with rabbit anti target antibody. Lymphocytes from blood and spleen produced efficient lysis of targets in the presence of antibody. Lymph node cells and thymocytes were essentially non-reactive in this system. Lymph node preparations from non-cancer patients contained approximately 25% of non-T cells with receptors for Fc,C3 and/or Ig. Regional lymph nodes from patients with primary tumours contained 37-50% non-T cells by the same criteria. Failure of lymph node lymphocytes to effect lysis of antibody-coated targets did not therefore correlate with content of Fc or C3 bearing cells per se. The effector cell in antibody-dependent cytotoxicity in other systems has been shown to carry Fc and C3 receptors, but not surface Ig. This cell type appears to be absent or non-functional in human lymph nodes.


Mouse red cell rosette formation by subpopulations of human lymphocytes.

In six separate experiments, 7-2% of purified peripheral blood lymphocytes from normal donors were shown to form rosettes with mouse erythrocytes. Normal T and B lymphocytes were separated according to their membrane properties, by E-rosette formation or fractionation on anti F (ab')2 column. The results obtained in both the separation procedures used were in good agreement: T lymphocytes were never found to form mouse red cell rosettes. On the contrary, enrichment of suspensions in B lymphocytes resulted in an increased percentage of mouse red-cell rosette-forming cells. Under the technical condition used, peroxidase-positive monocytes were not shown to form such rosettes. However, if all the mouse red cell rosette-forming cells were shown to be of B nature, not all the B lymphocytes form rosettes with mouse erythrocytes and it is not unlikely that this marker could be characteristic for a subset of Ig-bearing B lymphocytes.


C3-reacted sepharose: a preparative method for separating T and B lymphocytes.

Lymphocytes bearing complement receptors were separated from lymphocyte suspensions using C3-reacted Sepharose columns as solid phase immunoabsorbents. The retained CR+ lymphocytes were recovered by elution of the column with IgG fraction purified from a rabbit antiserum to human C3. This paper describes the method, its specificity and the yields of cells obtained.


Correlation of smooth-muscle and nuclear antibodies in normal subjects.

By means of the indirect immunofluorescent method the occurrence of smooth-muscle antibodies (SMA) was studied in 582 normal subjects of different ages, and the incidence of SMA was compared with that of anti-nuclear antibodies (ANA). SMA were detected in 32 (5-5%) and ANA in 28 (4-8%) of the normal subjects. The antibodies were of the IgG and the IgM class, while no IgA antibodies could be detected. IgM-SMA were demonstrated in 2-9%, IgM-ANA in 4-1% and IgG-ANA in 1-2% of the 582 subjects, and the incidence of these antibodies increased with increasing age (P = 0-012, P less than 0-001 and P = 0-003, respectively). IgG-SMA occurred in 3-6% and the prevalence of these antibodies was not age-related. No relationship between the incidence of antibodies and sex could be established. IgM-SMA and IgM-ANA were often found together in the same individuals (P = 0-002), which suggests that the same factors may elicit the formation of both antibodies.


Lymphocyte reactivity to mitogens in subjects with systemic lupus erythematosus, rheumatoid arthritis and scleroderma.

The mitogenic reactivity of lymphocytes from subjects with systemic lupus erythematosus, rheumatoid arthritis and scleroderma was studied. Cultures containing either unseparated or separated lymphocytes were stimulated with phytohaemagglutinin, Con A and pokeweed mitogen after inhibitory serum factors were eluted from the cell surface. Incorporation of [3H]thymidine in patient cultures was compared to that of normal controls. Greatly decreased reactivity was found in SLE to all three mitogens. Significantly decreased values to some mitogens was also observed in rheumatoid arthritis and scleroderma, but the defect was less severe. Cultures of study subjects contained significantly fewer small lymphocytes than normal controls and this finding explained at least in part the decreased mitogenic reactivity.


Quantitative measurements of T- and B-cell function in "variable" primary hypogammaglobulinaemia: evidence for a consistent B-cell defect.

Improved methods for separating B and T cells have made it possible to assess the functions of these cells. These techniques have been used to study the T dependence of B-cell immunoglobulin production in response to pokeweed mitogen. In the present study they have been used to reinvestigate the pathogenesis of "variable" primary hypogammaglobuliaemia. Twelve patients were studied by separating their B and T cells and culturing them with normal cells. The results show that none of the patients have normal B cells. In cultures containing high ratios of T cells to B cells we have found a marked reduction in B-cell immunoglobulin production both with T cells from normal donors and from patients. T cells from all but one of the patients helped immunoglobulin production and reduced T helper activity was only seen in patients whose B cells were very abnormal. The results suggest that a B-cell defect is the cause of the low serum immunoglobulin in all the patients we have studied.


Nuclear deposits of immunoglobulins in skin of patients with systemic lupus erythematosus.

Skin nuclear speckled IgG deposition was noted in seven patients. The patients' clinical courses satisfied at least four of the preliminary criteria of the American Rheumatism Association for the diagnosis of systemic lupus erthematosus (SLE). Examination of approximately 700 additional biopsies from patiets with SLE, connective tissue and various skin diseases as well as normal individuals, failed to demonstrate similar nuclear immunoglobulin deposition. Sera from these seven patients had higher titre, complement fixing anti-nuclear antibodies (ANA) of IgG class which produced a speckled nuclear fluorescent pattern. In addition, the sera of all seven patients demonstrated by gel double diffusion precipitating antibodies against nuclear ribonucleoprotein (RNP) or Sm antigens.


Enhancement of in vitro immunoglobulin synthesis of human lymphocytes by lysosomal enzymes from polymorphonuclear leucocytes.

The effect of human peripheral blood polymorphonuclear leucocyte (PMN) extracts and PMN granule lysates on in vitro immunoglobulin (Ig) synthesis by autologous peripheral blood mononuclear cells was studied. The mononuclear cells were cultured for 3 days with or without autologous plasma. Newly synthesized Ig in the culture supernatants was measured using 14C-labelled amino acids by an immune coprecipitation method. Upon addition of a PMN extract to plasma-free cultures Ig synthesis was stimulated, the mean stimulation index (SI) of cultures from thirteen individuals, including nine normals, three patients with rheumatoid arthritis and one with psoriatic arthritis being 1-8 +/- 0-2 in comparison with control cultures (P less than 0-05). By contrast, in 10% fresh autologous plasma, PMN extracts yielded a mean SI of 0-9 +/- 0-1 indicating inactivation of the active extracts by plasma inhibitors. In experiments using PMN granule lysates containing high concentrations of beta-glucuronidase and cultured in RPMI 1640, the mean stimulation index was 3-2 +/- 0-7. Stimulation of Ig synthesis was also produced by trypsin. Stimulation of Ig synthesis was also produced by trypsin. Stimulating factors in PMN extracts were inhibited by Trasylol, a protease inhibitor. These results indicate that trypsin and proteolytic lysosomal enzymes in PMN increase Ig synthesis of human peripheral blood mononuclear cells. They suggest a possible new role of PMN in the potentiation of immunoglobulin synthesis.


Quantitative studies of immunoglobulin deposition in the kidney, glomerular cell proliferation and glomerulosclerosis in NZB/NZW F1 hybrid mice.

Using the NZB and NZB/NZW F1 (B/W) hybrid mouse as a model for systemic lupus erythematosus, an effort has been made to quantitate: (1) immune complex deposition in the glomeruli by immunofluorescent staining of immunoglobulin, (2) glomerular cellular proliferation by radioautographic measurement of [3H]Tdr incorporation into the glomerular cells in vivo, and (3) glomerular scarring by PAS staining. The relationship between these changes and increasing age has been examined. By radioautography it was observed that dividing glomerular cells were labelled in vivo after injection of [3H]Tdr. This provided a reproducible measure of the proliferative process in the nephritis of B/W mice. In C57B1/6J and CBA/J mice, which have a low incidence of glomerular disease, little change in the amount of glomerular cell proliferation was observed with increasing age. The NZB strain of animals showed a somewhat increased level of proliferation but this did not increase with age. In striking contrast, glomerular cell proliferation in the B/W mice increased rapidly with age. The earliest change observed in the kidney was the deposition of immunofluorescent material in the mesangium and glomerular capillary basement membrane beginning between 3 and 5 months of age and reaching a peak at 9 months. Increase in glomerular cell proliferation began about 2 months after the onset of immune complex deposition but also reached a maximum at 7 months. Glomerular sclerosis was the last change to appear and continued after the other two parameters measured has begun to decline. These data suggest that the deposition of immune complexes in the glomerulus may be an important triggering mechanism for renal cell proliferation and glomerulosclerosis in the B/W mouse. The techniques described would provide a sensitive and reproducible quantitative method for analysing the differential effects of various types of treatment of immune complex nephritis in animals.


Influence of antigenic competition on the development of antibody-forming cell clones.

Isoelectric focusing in polyacrylamide gels was used to investigate the anti-sheep red blood cell antibody responses of mice subjected to antigenic competition. A reduction in the number and intensity of antibody bands was found, even in situations where the suppression of IgG antibody titres was minimal, while with large reductions in titre, antibody bands were rarely seen. It thus appeared that the output of individual B-cell clones was severely depressed during competition. It was concluded that inhibition of clonal expansion is an important feature of competition, and that this may reflect a normal regulatory activity which acts to limit cellular proliferation during immune responses. This conclusion was supported by observations on the level of DNA synthesis, following immunization with sheep red cells, in the spleens of normal and suppressed mice.


Leucovorin in combination chemotherapy of breast cancer.

Cyclophosphamide, 5-fluorouracil, methotrexate, and prednisone were administered for 165, 28-day cycles to 33 patients with metastatic breast cancer. Because of serious infections (fever larger than or equal to 101 degrees F. granulocytes less than 1,000/mm, 3 and hospitalization) and 1 drug death in the first 4 patients, oral calcium leucovorin, 20 to 30 mg/m2 orally, was given 2 days after methotrexate in subsequent treatment cycles. There were 26 cycles without calcium leucovorin and 6 serious infections; 139 cycles with calcium leucovorin and 5 serious infections (p = 0.002). Objective response was seen in 13 of 18 evaluable patients with no previous treatment and in only 2 of 11 patients with previous treatment (p = 0.0065). Equivalent doses were given to all subsets of patients. It is concluded that leucovorin added to the above combination of drugs can preserve antitumor activity while decreasing serious infections and that prior therapy significantly decreases the response rate to this combination.


Arteriocolic fistula: an unusual cause of gastrointestinal-tract bleeding.

An unusual case of acute gastrointestinal hemorrhage secondary to an iliac-artery aneurysm to a sigmoidal fistula is presented. Diagnosis and pathogenesis are discussed, and a plan for treatment is proposed.


Hemorrhoidectomy--how I do it: experience with the St. Mark's Hospital technique for emergency hemorrhoidectomy.

Eighty-five patients who had thrombosed hemorrhoids underwent emergency hemorrhoidectomy according to St. Mark's Hospital technique, with very good results. All specimens were found to have dilated blood vessels filled with thrombi of different sizes, with irregular, fibrotic or hyalinized vascular walls. Early complications included urinary retention and painful defecation in some patients. Late complications included only skin tags. No sepsis was found among our patients. Although operative bleeding can be tedious during the hemorrhoidectomy, it was a complication in the postoperative period of only one patient. Segmental, open hemorrhoidectomy performed according to the St. Mark's Hospital technique has been shown to be an ideal operation for the treatment of patients who have hemorrhoidal thrombosis, prolapse, edema, and bleeding.


Role of self-carriers in the immune response and tolerance. II. Parameters of tolerance induced by haptenated lymphoid cells.

The induction of tolerance to the 2,4,6-trinitrophenyl (TNP) hapten has been studied in a system utilizing haptenated syngeneic lymphoid cells. Specific depression of the direct and indirect plaque-forming cell (PFC) responses to TNP-protein conjugates was regularly achieved by pretreatment of Lewis rats with 1 X 10(7) trinitrophenylated spleen cells or TNP lymph node cells. Very low numbers of haptenated lymphocytes were active since tolerance could be induced in vivo with as few as 10(5)-10(6) TNP spleen cells (SC). SC exposed to as little as 10-100 mug of the reactive hapten, trinitrobenzene sulfonic acid, were effective tolerogens in this system. Tolerance induced with 10(7) haptenated spleen cells persisted for at least 4 weeks. Viable intact cells did not seem to be required in this system since the cell-free supernatants from TNP-SC cultured for 24 h were also active. In addition, significant depression of an ongoing immune response could also be imposed by haptenated isologous lymphoid cells. Moderate suppression of the "hapten-specific" delayed hypersensitivity response (measured by the accumulation of 51Cr-labeled bone marrow cells) was also achieved with TNP-SC. These results suggest that haptenated isologous lymphoid cells are potent tolerogenic conjugates capable of inducing tolerance affecting antibody formation and delayed hypersensitivity. However, since we and others have shown that TNP-SC do not adversely affect the generation of hapten-specific cytotoxic T cells, it is postulated that there is a differential recognition of, or response to, hapten-modified self-carriers by functionally distinct lymphocyte subpopulations.


Tolerance in differentiating B lymphocytes.

These experiments demonstrate that the response of B lymphocytes to particulate antigen in vivo can depend on their stage of differentiation: B lymphocytes differentiating from stem cells were rendered tolerant, while mature B lymphocytes were primed under the same conditions. Lethally irradiated mice of one allotype were repopulated with 13-15 day fetal liver cells from congenic mice bearing another allotype and the effect of antigen on the emergence of responsive B cells determined. B cells, descended from the fetal liver inoculum (identified by their allotype) produced an antibody response to antigen in the presence of additional T cells, 15 days after transfer. Such a response could be prevented by injecting the recipient with alum-precipitated antigen (but not with deaggregated antigen) shortly after irradiation and reconstitution. This unresponsive state was specific and independent of afferent suppressor mechanisms. Mature B cells, on the other hand, were shown to be primed by alum precipitated antigen in irradiated reconstituted hosts. It was concluded that a fundamental difference exists between the responsiveness of mature and differentiating B lymphocytes. Tolerance in differentiating B lymphocytes is discussed in relation to the acquisition of self-tolerance.


Effects of several 'membrane stabilizing' agents on frog neuromuscular junction.

The effect of several "membrane stabilizing" agents was examined on spontaneously occurring frog miniature end-plate potentials (MEPPs). No consistent parallel was observed between the ability of agents to act as "membrane stabilizers" and their ability to alter MEPP frequency, Vinblastine, chlorpromazine and haloperidol increased MEPP frequency; the concentration of haloperidol which increased MEPP frequency consistently reduced the magnitude of the muscle resting membrane potential. Strychnine, methoxyflurane and practolol decreased MEPP frequency. Similarly, there was no consistent pattern of drug effects on the mean quantal content m of the end-plate potential. Chlorpromazine did not change m, and it is known that vinblastine does not influence m. Strychnine, methoxyflurane, haloperidol and practolol brought about a decline in m.


Replacement of serum by hemolysate as growth promoter for murine leukemic and normal hemopoietic progenitor cells in culture.

The present study was undertaken to test whether lysates prepared from rat erythrocytes can replace serum as in vitro growth promoters for murine leukemic and normal hemopoietic progenitor cells (CFU-C). Normal bone marrow and three leukemic cell lines (P-1081, L-1210 and P-815) were used in all the experiments. The soft agar technique for cloning hemopoietic cells was used to quantitate cell proliferation. Addition of hemolysate to the agar medium at a final concentration of 4% promoted the growth of a maximal number of leukemic colonies, similar to the number of colonies obtained when 20% serum was added to the soft agar. As for normal CFU-C, addition of 10% hemolysate or 40% horse serum promoted the growth of comparable numbers of colonies. Rat hemolysate can therefore replace horse serum as a growth promoter of leukemic and normal CFU-C present in culture. It was also shown that rat hemolysate cannot substitute for the colony stimulating factor (CSF) needed for the cloning of CFU-C. However, addition of hemolysate, CSF and serum to the soft agar cultures promoted a potentiated rather than an additive growth of CFU-C.


An estimate of the extent of depletion of colony-stimulating factors in culture.

Between 30 and 40% of the CSF present in agar was depleted after culture for 1 week with 50,000 nucleated murine bone marrow cells. No statistically significant difference could be detected between this depletion and the extent of depletion of CSF when the amount present was halved. There was no evidence that toxic or inhibitory products reduced the colony count in the group to which stored CSF had been added.


Colony-stimulating factor (CSF) in oestrogen-stimulated spayed murine uteri.

A form of colony-stimulating factor (CSF) is known to be present in extracts of the uteri of normal mice. In this study CSF was not found in extracts of the uteri of untreated spayed mice but was present in uteri of these mice treated with oestradiol. CSF could be detected 9 h after oestradiol-administration. The occurrence of CSF in the uteri of spayed mice treated with oestradiol did not require DNA synthesis. The same amount was detected in uterine extracts whether or not DNA synthesis of the oestradiol-treated spayed mice had been prevented by hydroxyurea.


Vestibular units during decompensation.

Typical modifications of the unitary discharge of vestibular units have been recorded following the transection of the spinal cord of hemilabyrinthectomized and compensated guinea-pigs. These results support the concept that the spinal cord is essential in the compensation of the symptoms resulting from a lesion of one labyrinth.


Weightlessness and gravitational physiology.

Physiologic responses to the weightless environment have been documented in Skylab astronauts. Significant changes in cardiovascular responses, calcium metabolism, fluid distribution, and red blood cell mass are described. Although the data are limited and the sample size small, several hypotheses can be formulated. Future studies in a space laboratory setting offer the potential for understanding the effects of weightlessness on human physiology and the implications to medicine.


Studies on the product of antigenic recognition. III. Nature of PAR.

The product of antigenic recognition (PAR) is an antigen-antibody complex with granulotactic activity. The following results support his conclusion. PAR generated by cell-bound or shed T-cell RS or the recognizing structure of T alloantisera and alloantigen can be destroyed by heating at 56 degrees C for 30 min, because of inactivation of a heat-labile component supplied by alloantigen. Recognizing partners of these complexes are heat-stable. They form PAR anew when alloantigen is added or upon confrontation with anti-RS serum. They are also capable of inducing anti-RS sera. PAR preparations composed of these recognizing structures and antibodies to them are heat-inactivated at much higher temperatures, because both partners of the complex are relatively heat-stable. Heat-labile PAR complexes (recognizing structures and alloantigen) pass anti-mouse immunoglobulin immunosorbent, but are retained specifically by alloantiserum (via alloantigen of the complex) or by anti-RS serum (via RS of the complex) bound to the column.


Studies on the product of antigenic recognition. I. Formation of the product in recognition of alloantigens.

A product of antigenic recognition (PAR) was produced whenever receptors for alloantigens from T lymphocytes or a principle present in T-cell dependent alloantisera interacted with alloantigen. With two forms of the PAR assay (direct and indirect) the mechanisms underlying these interactions have been analyzed. For the interaction of T-lymphocyte receptors with alloantigen measured with direct PAR assays, the following conclusion emerged: upon confrontation with alloantigen, receptors (if not already present in secreted form) had first to be released from T-cell membranes. Shed T-cell receptors interacted with alloantigens by solubilizing them. Both processes could be prevented by fixing cells with formaldehyde. Release of T-cell receptors was temperature-dependent, solubilization of alloantigens was not. Because in mixed cell cultures receptors had first to be shed, this process was considerably slower and, in concordance with temperature dependence of receptor release, took place only at 37 degrees C. Titration of T lymphocytes with 'bound' receptors by the direct PAR test revealed that in the presence of excess alloantigen 10(2) T cells sufficed to give measurable responses. Supernates of parental strain lymphocytes containing numerous T-cell receptor specificities could be depleted of one of them. Alloantisera raised in presence of T helper cells ('T alloantisera') contained a principle capable of recognizing alloantigens, alloantisera incited in the absence of T helpers ('B alloantisera') did not. The recognizing principle appeared to be IgG. Like T-cell receptors, it was capable of solubilizing alloantigens form target cell membranes. B alloantisera lacked this capacity and their alloantigen-recognizing moiety was found to be monomeric IgM. The mode of interaction of this IgM with alloantigen most likely consisted in fixation to and shielding of antigen.


Studies on the product of antigenic recognition. II. Formation of the product in recognition of alloreceptors.

Cell-bound or shed T-lymphocyte receptors for alloantigens and the IgG-associated, alloantigen-recognizing principle in T alloantiserum reacted with antisera raised against those structures to form a product of antigenic recognition (PAR) in which the receptor or its equivalent took the role of 'antigen'. Formation of PAR was tested for numerous combinations of reactants and in all the process was completed within 1-2 h at 37 or 4 degrees C. Cell-bound receptors were found to be solubilized by all three forms of antisera, with the exception of an antiserum lacking anti-T-receptor activity. The data indicated that PAR might be a complex of antigen and antibody with granulotactic activity whose formation was specific.


Studies on the mechanism underlying the influence of alanine infusion on glucose dynamics in the dog.

These experiments have been designed to study the influence of alanine infusion of glucose dynamics in the dog and to further elucidate the role of pancreatic hormones in the interaction of alanine with glucose homeostasis. The primed constant infusion of  glucose-2-t was used in order to quantitate the rates of glucose production by the liver (Ra) and glucose utilization (Rd). In a first group of experiments, the intravenous infusion of alanine at the rate of 2 mg./kg./min. produced a moderate enhancement of plasma insulin (IRI), while pancreatic glucagon (IRG) increased more consistently. This different pattern of IRI and IRG response caused the insulin/glucagon molar ratio to decline progressibely throughout the experiment. Both rates of glucose turnover increased significantly during alanine infusion. Since Ra rose more rapidly thanRd did initially, hyperglycemia developed. Later, glucose production slowly decreased and, in spite of the sustained hyperglucagonemia, reached levels very close to the baseline in the second part of the experiment. A significant direct correlation between Ra and IRG was found, while the changes in Ra correlated inversely with those in I/G molar ratio. In a second group of experiments, alanine was infused at the same dose together with 0.4 microng./kg./min. of cyclic somatostatin. In the first part of the infusion, IRG fell more than IRI did, so that I/G ratio increased. Later, IRI levels maintained at low values while IRG returned slowly to the baseline and consequently I/G ratio significantly decreased. Glucose production fell rapidly soon after the beginning of the infusion, and therefore hypoglycemia developed. Later, Ra increased progressively to levels above baseline and plasma glucose returned to the preinfusion levels. As in the the first group of experiments, a significant direct correlation between Ra and IRG and an inverse correlation between the changes in Ra and I/G ratio were observed. These experiments demonstrate that alanine infusion produces an acceleration of glucose turnover and that a clear interrelationship between the release of glucose by the liver and the mobilization of pancreatic hormones exists. Finally, the experiments with somatostatin indicate that hyperglucagonemia is one  of the mechanisms underlying the stimulatory effect of alanine on glucose production.


Lymphocyte cytotoxicity for isolated hepatocytes in alcoholic liver disease.

To determine whether an autoimmune reaction to liver-specific proteins occurs in alcoholic liver disease, the cytotoxic effect of lymphocytes on isolated hepatocytes was determined in 27 alcoholic patients. Cytotoxicity was demonstrated in 15 of 17 patients with a histological diagnosis of alcoholic hepatitis, but in none of 10 with other forms of alcoholic liver disease. The ability of a liver-specific membrane lipoprotein to block the reaction suggests that sensitization to this antigen is responsible for the cytotoxicity. The demonstration of cytotoxicity using T cell-depleted, but not B and K cell-depleted, lymphocytes suggests an antibody-dependent cell-mediated reaction. There was a close correlation between cytotoxicity and the presence of alcoholic hyalin, liver cell necrosis, and piecemeal necrosis, but not with other histological features, immunoglobulines, autoantibodies, or standard liver function tests. The persistence of this autoimmune response, induced in some way by alcohol or one of its metabolites, may be important in the progression of acute alcoholic hepatitis to chronic liver disease.


Traumatic hemobilia: a complication of percutaneous liver biopsy.

Two patients with hemobilia are presented. The first patient, with alcoholic liver disease, had a percutaneous liver biopsy. Subsequently he developed jaundice, with an enlarged tender gallbladder, biliary colic, and gastrointestinal bleeding. Hemobilia was demonstrated by superselective hepatic angiography and bleeding was stopped by intraarterial infusion of epinephrine and propranolol. The second patient, with primary biliary cirrhosis at an advanced stage, had a percutaneous liver biopsy followed by gastrointestinal bleeding, severe abdominal pain, and finally death. In both cases hemobilia was suggested by gastroduodenoscopy.


[The influence of BCG-vaccination on the tuberculosis in children (Hamburg 1950-1971) (author's transl)].

Between 1950 to 1971 6 758 cases of active tuberculosis were registered among children under 15 years of age in Hamburg. 33 of them died, mostly due to tuberculous meningitis. In spite of a general decrease in tuberculosis during that period a relative increase of cases of pleurisy, nodular tuberculosis and tuberculous infiltrations of the lung were seen. On the other hand, cases of tuberculous meningitis and miliary tuberculosis as well as cases of glandular tuberculosis decreased. Rarely BCG-vaccinated children suffered from miliary tuberculosis, tuberculous meningitis and pleurisy. A statistical comparison between BCG-vaccinated and unvaccinated children of the birth cohorts 1954 and 1963 revealed that unvaccinated children suffered significantly more often from tuberculosis than the vaccinated. The duration of the protection by BCG-vaccination has been calculated for 7 years. BCG-vaccination after school age of the tuberculin negative persons is still debatable.


Immunological regulation of spontaneous antibodies to DNA and RNA. II. Sequential switch from IgM to IgG in NZB/NZW F1 mice.

Serial serum samples from NZB/NZW F1 mice were studied by sucrose density gradient ultracentrifugation and filter radioimmunoassay for the immunoglobulin class of antibodies to double-stranded DNA and single-stranded RNA (Poly A). These antibodies occur spontaneously during the course of an autoimmune disease resembling systemic lupus erythematosus. Study of serum samples at 2-week intervals showed the sequential appearance of 19S followed by 7S anti-nucleic acid antibodies, an earlier commitment to 7S antibodies in female compared to male mice, and an independent and unrelated switching mechanism (19S-7S) for antibodies to DNA and Poly A. The more severe disease in female mice is correlated with an earlier appearance of 7S anti-DNA antibodies. This sequential and ordered appearance of antibodies to nucleic acids is discussed in relation to possible immunologic regulatory mechanisms.


Analysis of the induction phase of contact sensitivity by footpad transfer of regional lymph node cells. Macrophages and radioresistant T-lymphocytes induce immunity.

The skin of CBA mice was painted with the contact sensitizing agent 4-ethoxymethylene-2-phenyloxazolone (oxazolone). One day later the regional lymph node cells were injected into the footpads of normal recipients. The recipients were tested 6 days later for contact sensitivity by challenging the ears with oxazolone and measuring the increase of ear thickness at 24 h. T cells and macrophages in the regional lymph nodes each independently gave rise to contact sensitivity in the recipient following injection into the footpad. This activity of T cells and macrophages was found in lymph nodes taken 1, 3 and 4 days after painting the donors. The role of T cells in the injected population was shown by purifying T cells by nylon-wool filtration and rosetting with sheep red cells coated with antibody and complement (EAC rosetting) and by destroyed T cells with anti-0 serum and complement. The activity of purified T cells resisted 2000 rad in vitro. The activity of cells from T-deprived (B) mice showed that a second cell type was important in the footpad transfer. This cell behaved like a macrophage, and not like a B cell, on EAC rosetting in the presence or absence of divalent cations and on treatment with silica and carrageenan--agents which damage macrophages. Our working hypothesis is that the footpad transfer may be caused independently by macrophages or T cells with oxazolone (probably linked to major histocompatibility complex antigens) on their surface and that these cells act by collaborating with T cells in the recipient which give rise to the effector cells for contact sensitivity.


Cytotoxic T cells specific for influenza virus-infected target cells.

Conditions are described for the generation and detection of murine cytotoxic cells which lyse influenza virus-infected target cells. A number of criteria indicated that most and probably all of the activity was due to T cells. These criteria included the susceptibity of cytotoxic activity to treatment with anti-theta ascitic fluid plus complement and the enhancement of lysis when there was H2 compatibility between donors of effector and virus-infected target cells.


Thymus dependency of antigen-binding cells in the bone marrow and spleen of chicks.

The bone marrow and spleen of normal unimmunized chicks contain lymphoid cells which form rosettes with quail red blood cells (QRBC), but not with sheep red blood cells. The number of these antigen binding cells (QRBC-ABC) were restored to their original level 2-3 weeks after whole body X-irradiation following a temporary but drastic decrease in the number of QRBC-ABC. This recovery is not observed in chicks which have been thymectomized or thymectomized and bursectomized. However, QRBC-ABC numbers were restored in bursectomized chicks comparable with those levels in control chicks. Anti-thymic serum effectively blocked rosette formation and it seems that QRBC-ABC are a thymus-dependent population. Possibly, the thymus exerts an influence on the genesis of QRBC-ABC through a humoral mechanism.


The relation of immune depression and B-cell stimulation during the development of delayed hypersensitivity to soluble antigens.

A comparison has been made of the effects of cyclophosphamide (CY) pretreatment and i.v. injection of a high dose of antigen on delayed hypersensitivity induced by proteins in Freund's incomplete and complete adjuvants. Five antigens have been studied: obalbumin (OA), bovine serum albumin (BSA) bovine gammaglobulin (BGG), DNP 5-BGG and DNP 50-BGG. A spectrum of reactivity has been detected depending upon the ability of the antigen to stimulate B-cell as well as T-cell activity. Bovine serum albumin and BGG behave as relatively weak antigens in which the T-cell response, measured by delayed hypersensityity, is easily suppressed by i.v. antigen and the B-cell modulating system, revealed by CY sensitivity, is poorly stimulated. These proteins, injected i.v., are weak stimulators of antibody-dependent Arthus reactions. On the other hand, OA, DNP 5-BGG and DNP 50-BGG behave as strong antigens, resisting suppression of the T-cell response by soluble antigen and exhibiting (in Freund's incomplete adjuvant) a strongly developed CY sensitive, B-cell modulating system. Strong Arthus reactivity is readily demonstrated following i.v. administration of these antigens. A dissociation has been demonstrated between B-cell modulation of T-cell function and unresponsiveness induced by i.v. antigen. The failure to reverse the latter type of unresponsiveness by cyclophosphamide pretreatment suggests that separate mechanisms are involved in these systems.


Colony growth in vitro of mitogen-stimulated mouse B lymphocytes.

The ability of mouse B-mitogen-induced lymphocytes to grow and develop into colonies in a soft agar system was studied. Prerequisite conditions for the colony formation of mouse lymphocytes from inguinal lymph nodes of strains ICR C3H/eB and C3H were their suspension in a liquid medium and stimulation with polyclonal B-cell activators such as bacterial lipopolysaccharide (LPS), purified protein derivative (PPD) or dextran sulphate (DxS) prior to being seeded on a soft agar culture medium. After 3-5 days of culture, colonies of 50-350 cells or more per clone developed. A linear relationship was found between the number of cells seeded and the number of colonies growing. Of the cells seeded, only a limited population of the mitogen-stimulated lymphocytes has the potential to divide and to develop into colonies. The largest number of colonies was obtained by culturing lymph node cells of ICR mice and using LPS as mitogens. Two sublines of C3H were found to respond differently to LPS: C3H/HeJ mice were low responders while C3H/eB mice were high responders. Experiments with inbred, congenitally athymic nude (nu/nu) mice known to be deficient in T cells showed that LPS-stimulated lymphocytes were capable of forming colonies. The morphology of the colony cells, as well as the fact that they stain positively for cell-membrane immunoglobulins, suggest that the colonies developed from B lymphocytes.


Selective response of lymphocytes from Treponema pallidum-infected rabbits to mitogens and Treponema reiteri.

The in vitro response of peripheral blood lymphocytes from rabbit infected with Treponema pallidum was examined using various mitogens and avirulent Treponema reiteri. For the first 4 weeks after treponemal infection, the response of lymphocytes from syphilitic rabbits to phytohemagglutinin and pokeweed mitogen was markedly reduced in comparison to uninfected controls. Lymphocytes from both groups of rabbits responded normally to class-specific immunoglobulin anti-sera (anti-immunoglobulin M and anti-immunoglobulin G) and T. reiteri.


Immunoprotective activity of ribosomes from Haemophilus influenzae.

Immunization with ribosomal preparations from Haemophilus influenzae type b elicited protective immunity in mice. Ribosomes from disrupted cells where isolated by differential centrifugation using sodium dodecyl sulfate. The washed ribosomes contained 25% protein and 75% ribonucleic acid and sedimented as a single peak on sucrose density gradient analysis with a sedimentation coefficient of 67S, using Escherichia coli ribosomes as a 70S marker. Immunodiffusion tests with antipolyribose phosphate serum showed that the ribosomes were free from capsular material. Mice immunized subcutaneously with ribosomes, with or without adjuvant, were challenged intraperitoneally with 100 to 1,000 mean lethal doses of H. influenzae type b suspended in gastric mucin. Significant protection was induced by ribosomes and was compared to that obtained after sublethal infection with live cells. The protection was greatly enhanced after incorporation of ribosomes into adjuvants. Maximum protection (90 to 95%) was observed at 1 to 2 weeks after immunization. Ribosomes from a nonencapsulated strain of H. influenzae were as immunogenic as those from the encapsulated strain, demonstrating that the capsular material is not responsible for immunogenicity of Haemophilus ribosomes.


Isolation of the capsular polysaccharide from culture supernatant of Haemophilus influenzae type b.

The capsular polysaccharide (CP) of Haemophilus influenzae type b is known to be spontaneously released from the cells in culture. The CP is precipitable from culture supernatant by the cationic detergent hexadecyltrimethylammonium. Most of the nucleic acid and some of the protein, but almost none of the endotoxin, in the supernatant are co-precipitated. Extraction of the precipitate with progressively stronger NaCl solutions separates nucleic acid and protein from the CP and also effects a molecular size fractionation. Residual endotoxin and protein can be reduced by extraction with cold phenol and ultracentrifugation. The resulting preparation has ribose, ribitol, and phosphate as principal components and contains less than 1% other sugars, protein, or nucleic acid; it elutes on Sepharose 2B as a symmetrical peak with Kav 0.51.


Enhancement of the allogeneic effect by irradiation of transferred cells.

Hot-pulse treatment of parental thymus or spleen cells increased their ability to simulate antibody synthesis in F1 hybrid recipient mice. A similar effect was obtained after treatment of the cells with low doses of X-irradiation. Likewise, irradiation of syngeneic unprimed cells made them stimulatory on antibody production. The production of IgG, IgM as well as IgE antibodies was enhanced after transfer of irradiated cells. The results suggest that irradiation preferentially inactivates suppressor cells, that are present in the thymus and the spleen.


Bovine reaginic antibody. II. Preparation of an anti-reaginic immunoglobulin.

A method of preparing an antiserum to bovine reaginic immunoglobulins is described. This procedure which relies on the ability of bovine reaginic immunoglobulin to attach to rat mast cells, results in an antiserum which upon absorption with rat and bovine materials did not react with rat serum or with bovine IgG1, IgG2, IgM and IgA. The antiserum did, however, react in a gel diffusion assay with PCA positive bovine globulin and a reaginic immunoglobulin-rich fraction of bovine serum. Further, anti-reaginic immunoglobulin activity was shown by binding to and degranulation of rat mast cells previously incubated with PCA positive bovine globulin. The antiserum prevented sensitization of calf skin in a PCA reaction while anti-human IgE did not.


Modified antibody response in rabbits treated with diphenylhydantoin.

Rabbits immunized subcutaneously with a mixture of normal human serum, Freund's complete adjuvant and a soluble diphenylhydantoin preparation, produced antibodies that differed qualitatively and quantitatively from the controls, as revealed by immunoelectrophoresis and passive haemagglutination. Antibodies in human IgG were greatly increased, whereas antibodies to human albumin were not influenced, and antibodies to some alpha- and beta-globulins were reduced.


Specificities in natural cell-mediated cytotoxicity by the cross-competition assay.

The cross-competition assay was applied to natural cell-mediated cytotoxicity in human, mouse and rat systems. The results showed that what appeared to be non-selective effects of natural cytotoxicity were in fact highly specific. Each effector suspension included natural effector cells (N cells) that recognized and reacted specifically with many different antigens on target cells, resulting in overall non-selectivity. The specificity of the reaction for each target cell was demonstrated by the selective inhibition of natural cytotoxicity achieved when competitor cells sharing antigens with the target cell were added. Sharing of common target antigens was demonstrated by selective cross-inhibition which was then used to investigate antigens on target cells.


Surface membrane changes in lymphocytes from patients with infectious mononucleosis.

Peripheral blood lymphocytes from 20 patients with acute infectious mononucleeosis (IM) were studied for cell aggregation and for cap formation by concanavalin A (Con A). The lymphocytes from these patients showed 5.2+/-1.5% cells with a Con-A-induced cap and a high degree of cell aggregation without Con A, compared to 27.7+/-3.2% caps and a low degree of cell aggregation with normal lymphocytes. The lymphocytes from IM patients were fractionated to enrich for T and B cells. There was a low frequency of cap formation in both T and B cells, but the high degree of celll aggregation without Con A only occurred with B cells. Studies with four patients in clinical remission from acute IM have shown that the frequency of Con-A-induced cap formation only returned to normal more than 3 months after the beginning of clinical remission and that even at 6 months the cells still showed a high degree of cell aggregation. The results indicate that a high degree of B-cell aggregation and a low percentage of B and T cells with a Con-A-induced cap were associated with acute IM and that the changes associated with a high degree of B-cell aggregation were by themselves not sufficient to cause the disease.


Evidence that MuLV-induced thymic lymphoma cells possess specific cell membrane binding sites for MuLV.

Cell-surface binding sites specific for thymotropic murine leukemia viruses were found in high concentrations on thymic lymphoma cell lines induced by this class of virus, but were detectable in much lower concentrations (if at all) in several non-T leukemias, plasmacytomas, and normal thymocytes or spleen cells. By specific comparison, Moloney leukemia virus (M-MuLV) binds to a lymphoma induced by M-MuLV, but not to a thymic lymphoma induced by Gross leukemia virus (G-MuLV); and G-MuLV binds to an AKR lymphoma but not to the M-MuLV-induced lymphoma. The material which binds to these T-lymphoma membrane sites is input virus, rather than a contaminant which copurifies with virus. Autoradiographic analysis demonstrates that a high proportion of T-lymphoma cells possess binding sites, whereas only a rare cell in the thymus binds murine leukemia virus to the same degree. We raise and discuss the hypothesis that each T lymphoma induced by thymotropic leukemia viruses may represent the clonal descendants of the few rate cells in the normal thymocyte population which also bind these viruses.


Survival of synchronized V79 cells treated with X-rays and cordycepin.

The survival of V79 Chinese hamster lung cells exposed to X-irradiation is reduced by co-treatment with cordycepin (3'-deoxyadenosine). This reduction is manifested principally by a decrease in the D0 of the X-ray survival curve from 199 rad in untreated cells to 106 rad in cordycepin-treated cells. Reduced survival is seen throughout the life-cycle when synchronized cell populations are exposed to both agents with cells in mid-S being especially sensitive.


Rescue of marker phenotypes mediated by somatic cell hybridization.

The effect of irradiation prior to virus-induced cell fusion on the frequency of hybrid production has been measured as a function of radiation dose. The Chinese hamster line wg3h (HGPRT-) was crossed with the TK- mutants; Chinese hamster A23 or mouse 3T34E, and hybrids were selected in HAT medium. Irradiation of one (marker rescue) or both (mutual rescue) partners before fusion yielded qualitatively different results. After X-irradiation marker rescue curves were of single-hit type, with D0 values about five-fold greater than the irradiated parent cell. Mutual rescue curves were of the multi-hit type, with zero-dose extrapolation value (n) greater than that of the more resistant partner, but no significant alteration in D0. Qualitatively similar results were obtained after U.V.-irradiation, but the probability of rescue per surviving parent cell was higher after U.V. than after X-rays. With both forms of radiation, reciprocal marker rescue curves were not significantly different. Control experiments showed that mutual rescue was not an artefact either of sensitization of parent cells due to TK- or HGPRT- mutations, or of the enhancement of recovery by feeder layers resulting from high-density mutant populations killed with graded radiation doses and HAT selection. Analysis of heterokaryon frequencies within 18 hours of fusion demonstrated that radiation doses up to four lethal hits, given to one or both parents of the cross wg3h x A23, did not increase heterokaryon formation.


The distribution of plutonium-214 in rodents.

Plutonium-214 citrate solution at pH 6-5 was injected intravenously or intra-peritoneally into hamsters and rats at a dose of 50 MBq kg-1 (1-35 mCi kg-1). The animals were killed 1 day or 1 week later, and tissues were removed for autoradiography and radiochemical analysis. Plutonium-241 was distributed in rats in the same way as plutonium-239, and is a suitable isotope for high-resolution tissue-section autoradiography. Plutonium deposits in cells consisted of a nuclear and a cytoplasmic component. In the hamster kidney cells, the amount associated with the nucleus was about 55 per cent of the total cellular plutonium at 24 hours after injection. Six days later, it was only about 30 per cent. Plutonium deposits were also characterized in hepatocytes, in the interstitial cells of the testes, in the cells of ovarian follicles, in chondrocytes and in bone cells, including osteoblasts and osteocytes. In bone there appeared to be both an extracellular and intracellular deposit. No evidence was found of substantial incorporation of plutonium into the mineral phase of bone.


Stage-sensitivity and dose-response study after gamma-irradiation of mouse primary spermatocytes.

The sensitivity pattern for X-ray-induced chromatid aberrations was analysed in primary spermatocytes irradiated at 1 to 11 days diakinesis-metaphase I of meiosis. Using a dose of 300 R (60 R/min) of gamma-rays zygotene (day 9) was found to be the most sensitive and leptotene (day 11) the most insensitive stage. The dose-response to gamma-ray-induced aberrations was evaluated in cells irradiated zygotene. Both the yields of rearrangement as well as the yields of fragments gave a best fit to a quadratic model.


Arterial wall damage by X-rays and fast neutrons.

Radiation-induced atheromatosis has been studied for 200 kVp X-rays and 15 MeV neutrons. From the results of two earlier experiments, a r.b.e. less than 1 was expected for neutrons. (1) Irradiation of blood-vessels causes depolymerization of the mucopolysaccharides in the vessel wall, resulting in atheromatosis, and (2) in two other mucopolysaccharide systems a r.b.e. less than 1 is observed for fast neutrons. Irradiation of the carotid arteries of a total number of 120 hypercholesterolaemic rabbits, divided over several groups, with 500 and 1000 rad of X-rays or neutrons results, however, in atheromatous plaques which are more pronounced for neutrons than for X-rays at the 500 rad dose-level; for a dose of 1000 rad the effect of neutrons is less extensive than the effect of X-rays. These results lead to the assumption that the origin of the atheromatosis seems not only to be the mucopolysaccharide depolymerization, but that radiation induced damage at the cellular level must be taken into account.


O2 consumption of the amphibian cornea: relationship to Na+ and Cl- and the presence of drugs.

Thirty percent of the O2 consumption of the amphibian cornea was Na-dependent. Ouabain, in the presence but not the absence of this cation, inhibits O2 uptake by about 30 patient. In contrast, neither the presence of Cl- nor drugs which stimulate (adrenaline, theophylline) or inhibit (furosemide, thiocyanate) active transport of this anion had detectable effects on O2 consumption.


Small-angle light-scattering patterns of corneas of different species.

Light-scattering patterns of corneas of different species have been obtained. The different small-angle light-scattering (SALS) patterns were classified into four groups on the basis of the angular dependence of the intensity of light scattered in the I+ and I modes. At present, only two types of patterns can be explained on the basis of theoretical models. The need to develop a general model that can account for all four types of SALS patterns is discussed.


Polyene antibiotics. IX. An improved method for the preparation of methyl esters of polyene antibiotics.

An improved general method for the preparation of methyl esters of polyene antibiotics is discussed. Using this method methyl esters of pimaricin, nystatin, eurocidin, hamycin, hamycins A and B, aureofungin, partricins A and B, candimycin, candicidin, and amphotericin B have been prepared and their physical properties are reported. The biological activities of hamycins A and B and their methyl esters are also described.


Purification and properties of colony-stimulating factor from mouse lung-conditioned medium.

Colony-stimulating factor, which specifically stimulates mouse bone marrow cells to proliferate in vitro and generate colonies of granulocytes, or macrophages, or both, was purified 3500-fold from mouse lung-conditioned medium. Analysis by discontinuous polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate indicated that there was a single protein component. All of the colony-stimulating activity was coincident with the protein band. The molecular weight of colony-stimulating factor estimated by gel filtration was approximately 29,000 and by electrophoresis approximately 23,000. The specific activity of purified colony-stimulating factor from mouse lung-conditioned medium bound to concanavalin A-Sapharose, indicating that it is a glycoprotein. The small percentage of colony-stimulating factor in mouse lung-conditioned medium which did not bind to concanavalin A-Sepharose appeared to represent molecules which lacked the carbohydrate moieties required for binding to this lectin. It was necessary to include low concentrations (less than 0.01%, v/v) of polymers such as gelatin and polyethylene glycol, or nonionic detergents such as Triton X-100, in all of the buffers used throughout the purification scheme, otherwise colony-stimulating factor was lost from solution. At high concentrations (greater than 20 mug/ml) the factor stimulated the formation of granulocytic, macrophage, and mixed colonies from C57BL mouse bone marrow cells. As the concentration of purified colony-stimulating factor was decreased, the frequency of colonies containing granulocytes also decreased. At low concentrations of colony-stimulating factor (less than 70 pg/ml) only macrophage colonies were stimulated.


Quaternary structure of delta-aminolevulinic acid synthase from Rhodopseudomonas spheroides.

Delta-Aminolevulinic acid synthase (succinyl-CoA: glycine C-succinyltransferase (decarboxylating) EC 2.3.1.37) was purified from Rhodopseudomonas spheroides. The purity of the enzyme preparation was established by its behavior in disc electrophoresis in the presence and absence of sodium dodecyl sulfate and by analytical ultracentrifugation. The molecular weight of the enzyme as determined by sedimentation equilibrium was found to be about 80,300, a value similar to those obtained by gel filtration, polyacrylamide gel electrophoresis, and sucrose gradient centrifugation. The molecular weight of the enzyme, denatured with either sodium dodecyl sulfate or guanidine hydrochloride, was found to be about 45,000 and 41,000, respectively. The dimeric structure was supported by sedimentation in sucrose gradients. Further evidence for the dimetic nature of the enzyme was obtained by gel electrophoresis of the enzyme treated with dimethylsuberimidate and sodium dodecyl sulfate.


Effects of fibroblast growth factor and epidermal growth factor on the rate of growth of amniotic fluid-derived cells.

Primary cultures of cells derived from human and bovine amniotic fluid were cultivated in Dulbecco's modified Eagles medium with 20% fetal calf serum. Whereas increased concentrations of fetal calf or other types of serum bring about no further mitogenic response, the rate of growth of these cells was nevertheless sharply increased when 100 ng/ml of FGF was added to the culture media. The FGF induced mitogenic effect was statistically significant. EGF at 100 ng/ml had a less pronounced effect which, when analyzed statistically, was not significant. Amniotic fluid-derived cells are used for the prenatal detection of genetic disorders. The use of FGF to increase the rate of growth of these cells should reduce the time required between amniocentesis and diagnosis.


Modification of factor VIII complex properties in patients with liver disease.

Factor VIII procoagulant activity (VIII:C), factor VIII related antigen (VIIIR:AG), and the von Willebrand factor (VIII:WF) were measured in 19 patients with liver disease (8 cases of alcoholic cirrhosis and 11 with acute viral hepatitis). In both groups of patients the levels of VIII:C, VIIIR:AG, and VIIIR:WF were above normal or normal with mean values well above the normal range. VIIR:AG was also measured in the same patients by two-dimensional electrophoresis, and its biological properties were measured after purification by chromatography. By both methods the VIIIR: AG in the patients with liver disease differed from normal VIIIR:AG. In five of the patients with acute viral hepatitis, who were tested after they had recovered, the previously elevated levels of VIII:C, VIIIR:AG, and VIIIR:WF had dropped to normal limits and the qualitative abnormalities had disappeared.


Palindromic rheumatism: an allergic manifestation?

A case of palindromic rheumatism is presented in which the arthritic attacks usually were accompanied by an allergic vasculitis and dermatitis. Although this is not proof of the etiology, the constant association is suggestive of an allergic cause. A brief review of palindromic rheumatism in included.


Receptors for IgM-coated erythrocytes on chronic lymphatic leukemia cells.

Our experiments show that lymphocytes of CLL patients, having typical B cell characteristics, form rosettes with IgM-coated bovine erythrocytes. Of 18 investigated patients, 3 to 78% (mean 29%) of the isolated lymphocytes reacted with EA-IgM. With mixed rosette assays. EA-IgM bound to cells bearing receptors for IgG as well, but not receptor-bearing lymphocytes. Rosette formation could be completely blocked by addition of IgM at concentrations as low as 0.17 mg/ml. Ten milligrams per milliliter of aggregated human IgG had no effect on the rosette formation with EA-IgM but completely abolished the binding of EA-IgG. Adult human or rabbit serum blocked the EA-IgM binding, whereas cord blood serum and FCS had no effect. These inhibition data indicate that EA-IgM binding does not occur via a somewhat altered IgG-Fc receptor but reacts with different membrane structures. That EA-IgM receptor can be cleaved off with trypsin and can be reconstituted after overnight cultivation, also supports this viewpoint. In contrast to the situation in normal subjects, in CLL patients the IgM receptors are demonstrable before overnight cultivation and are found on cells with B cell characteristics.


T and B lymphocyte migration into syngeneic tumors.

The migration of splenic T and B lymphocytes into syngeneic tumors undergoing immunologic rejection was investigates. Spleen cells were obtained from normal BALC/c mice or BALB/c mice bearing tumors induced by murine sarcoma virus (MSV). Either whole spleen cells or immunoabsorbent purified T and B cells were radiolabeled with sodium chromate-51 and injected i.v. into normal or MSV inducted-tumor bearing syngeneic recipients. Twenty-four hours later the recipient mice were sacrificed and radioactivity was assessed for tumor, contralateral normal muscle, the lymph nodes draining the tumor and contralateral draining lymph nodes, peripheral lymph nodes, spleen, and liver. Both T and B lymphocytes from either normal or MSV tumor-bearing animals show greatly increased migration into the tumor when compared with normal muscle. Migration of T cells from both normal and MSV tumor bearers was 30 times that of migration to normal muscle. B cells from tumor-bearing mice, on the other hand, localized in the tumor itself only 50% as frequently as did B cells from normal animals. In addition, T cells from MSV tumor bearers were found in the highest proportion in the lymph node draining the tumor site. We conclude that T and B lymphocytes from either normal or tumor-bearing mice migrate to a syngeneic tumor undergoing immunologic rejection. In contrast, the migration of both T and B cells from tumor-bearing animals was decreased to the peripheral lymph nodes at the time of maximum tumor growth.


Antigen-initiated B lymphocyte differentiation. IX. Characterization of memory AFC progenitors by buoyant density and sedimentation velocity separation.

The characteristics of memory B cell antibody-forming cell (AFC) progenitors from long-term hapten-primed CBA mice were investigated by using sedimentation velocity and buoyant density separation to isolate physically distinct B cell sub-sets. The isolated fractions were assayed by the adoptive immune response to NIP-POL antigen, under conditions where neither T cells nor other accessory cells were limiting the IgM or IgG AFC responses. The results were compared to previous studies on the IgM AFC-progenitors of unprimed adult mice. Splenic IgM and IgG memory AFC-progenitor activity was largely found among the typical B cells of slow to medium sedimentation rate, in contrast to the fastre sedimenting IgM AFC-progenitor activity of unprimed animals. Splenic IgM and IgG memory AFC-progenitor activity was found among the medium to light density cells, and so resembled by this parameter the IgM AFC-progenitor activity in unprimed animals. Thoracic duct lymphocytes from hapten-primed mice also exhibited memory IgM and IgG AFC-progenitor activity in the slow-medium sedimentation range. However, in contrast to spleen, the IgM and IgG memory AFC-progenitor activity in lymph was found among very dense B cells. Two physically distinct sub-populations of memory B cells have thus been identified, namely: i) small, medium-light density, presumably tissue-resident B lymphocytes found in spleen; and ii) small, dense, presumably recirculating B lymphocytes found in lymph. Both physical forms include IgM and IgG progenitors. Both forms are distinct from the larger, medium-light density "virgin" AFC-progenitors in the spleen of unprimed adult mice.