[Different types of cells in tissue cultures of normal bovine iris (author's transl)].

Calf iris tissue cultured in TCM 199 was studied under the electron microscope at 5, 14, and 21 days. Melanocytes and clump cells (macrophages), and mast cells could be distinguished, as well as fibroblasts, round cells, and non striated muscle cells, Melanocytes lost their pigment granules temporarily, but not their ability to produce new ones again after a certain adaptation period. The morphology of clump and mast cells was only slightly altered. In incubation, fibroblasts and round cells were found to have spheroid surfaces. After a 3 week's culture, muscle cells showed signs of growth.


Splenic vein thrombosis: an unusual case of gastric bleeding.

Acute and/or recurrent gastrointestinal bleeding due to ruptured gastric varices from an isolated thrombosed splenic vein is a distinct entity. Incidence of this syndrome is probably less than 1%. Typical clinical features of this syndrome include evidence of splenic hypertension without liver disease and no demonstrable cause of gastrointestinal hemorrhage. Diagnosis can easily be missed unless the surgeon is familiar with this syndrome. Typical findings at the time of surgery are an enlarged spleen, varicose veins usually involving the upper third of the stomach, and pancreatic and peripancreatic inflammation. Portal vein and portal pressure will be normal. Meso-portography is a convenient and safe procedure and will lend support to suspicion when a retrograde nonfilling of the splenic vein is present. Splenectomy offers the expectation of a long-range cure. A representative case of a 39-year-old man is discussed. He had at least six episodes of gastric bleeding in less than 3 years. At a previous laparotomy, the cause of bleeding could not be determined. A splenectomy in December 1970 has been able to control the gastric bleeding since then.


T lymphocytes in children with allergic respiratory disease.

The role of T lymphocytes in atopic disease is of considerable importance because animal studies indicate that cells of this lymphoid series may influence reaginic antibody response. T lymphocyte subpopulations were studied in a group of 76 children with allergic respiratory disease. There was no statistical difference between atopic children with asthma and those with allergic rhinitis as compared with an age-matched control population of 20 non-atopic children in terms of levels of active T lymphocytes or total T lymphocytes. The results of this study do not support the concept of a T cell immunodeficiency in children with allergic respiratory disease.


Influence of barbiturate anesthetic induction on circulatory responses to morphine.

Circulatory responses to morphine (1 mg/kg, 10 mg/min) following a barbiturate anesthetic induction (thiamylal 4 mg/kg) were determined in 7 patients with coronary artery disease. Compared with awake measurements, the mean arterial pressure and cardiac index were decreased 16 and 42 percent, respectively (p less than 0.05) and systemic vascular resistance increased 35% (p less than 0.05) at the end of morphine infusion. The same dose of morphine, but without prior barbiturate, has been previously reported not to significantly alter these measurements. Therefore, the assumption that morphine is associated with minimal circulatory changes may be invalidated by the prior administration of thiamylal.


[Ocular fixation test in pendular vestibular stimulation. Hearing and balance laboratory (author's transl)].

The effects of ocular fixation on per-rotatory nystagmus were studied during the course of the pendular vestibular stimulation test. If the subject is normal, or the disorder is peripheral, the nystagmus is abolished. Examples of these different results are presented.


[Benign intracranial hypertension in childhood (author's transl)].

Eight patients with clinic manifestations of benign intracranial hypertension are reported. Every patient was studied to the point where all other causes were ruled out. Epidemiology, pathogenesis, clinical picture and complementary studies are analysed. Finally, authors examine management and prognosis of the syndrome.


[Congenital arachnoid cysts of the posterior fossa midline. On their anatomoradiological variability (author's transl)].

An anatomical and radiological study of seven cases with arachnoid cyst of the midline of the posterior fossa in which there was either necropsic or surgical anatomical diagnosis is reported. A great variation was observed regarding their location, size and extension, which covers all the zone existing between the foramen of Magendie and the quadrigeminal lamina, presenting two cases which rose to the medial fossa, one of them entering in a left lateral ventricle and implanting itself in its' walls. Associated to arachnoid cysts other malformations have been found, such as type I Chiari, partial agenesis of the vermis, cerebellar dismorphies and stenosis of the aqueduct. Neuroradiological examination is fundamental for diagnosis, existing important alterations both in plain skull X-Rays and pneumoencephalography, ventriculography and vertebral and carotid angiographies. There are no strict neuroradiological patterns in the study with air nor in arterial and or venous times of angiography which define assuredly, presence, of the arachnoid cyst, since each case shows different findings. However, the conjugated analysis of all explorations must give correct diagnosis in almost the totality of cases.


[Congenital tuberculosis (author's transl)].

The present case report concerns a term infant, born of a mother with an undiagnosed tuberculosis, which was proved by inguinal lymph node biopsy to have numerous acid-fast bacilli; sequence of events supports the thesis that infant had acquired infection transplacentally. Literature is reviewed and diagnosis, prophylaxis, chemotherapy of this infrequent disease with a grave prognosis is commented, emphasizing success of chemotherapy in this infant.


Sensitive, rapid, and specific bioassay for the determination of antilipogenic compounds.

A sensitive and rapid bioassay for the determination of the antilipogenic compounds cerulenin and CM-55 is described. The bioassay is based on the inhibitory effect of cerulenin and CM-55 on the in vivo luminescence of an aldehyde-requiring mutant of the marine bacterium Beneckea harveyi. A total quantity as low as 0.1 mug of cerulenin can be determined within 15 min with an error of +/-2%. The bioassay, as presented, is specific for compounds that are known to inhibit fatty acid biosynthesis and, as such, it might be used as a general screening method for the detection of antilipogenic compounds.


Pyruvate production and excretion by the luminous marine bacteria.

During aerobic growth on glucose, several species of luminous marine bacteria exhibited an imcomplete oxidative catabolism of substrate. Pyruvate, one of the products of glucose metabolism, was excreted into the medium during exponential growth and accounted for up to 50% of the substrate carbon metabolized. When glucose was depleted from the medium, the excreted pyruvate was promptly utilized, demonstrating that the cells are capable of pyruvate catabolism. Pyruvate excretion is not a general phenomenon of carbohydrate metabolism since it does not occur during the utilization of glycerol or maltose. When cells pregrown on glycerol were exposed to glucose, they began to excrete pyruvate, even if protein synthesis was blocked with chloramphenicol. Glucose thus appears to have an effect on the activity of preexisting catabolic enzymes.


Pathology of Ménière's disease as it relates to the sac and tack procedures.

The rationale for endolymphatic sac and tack operations is to prevent the further accumulation of endolymph, the former by draining endolymph into extralabyrinthine tissues and the latter by shunting endolymph into the perilymphatic space. While the basic concepts are reasonable enough, the probability of achieving these objectives seems remote. There appears to be adequate clinical evidence, however, to show that these procedures are of therapeutic value in selected cases. It is tempting to speculate that these surgical insults to the labyrinth, with the associated inflammatory and biochemical changes, alter the function of cells which control fluid physiology and thus improve the symptoms of Ménière's disease.


Current status of surgical decompression and drainage procedures upon the endolymphatic system.

At the present time, neither the etiologic factors nor the pathophysiologic mechanisms that lead to the development of endolymphatic hydrops or Ménière's disease are known; however, the anatomic-pathologic alterations in the labyrinth that are produced by the disease are well-documented. Undoubtedly, what is not known is responsible for the controversial status and questionable benefit that surgical decompression and drainage procedures have upon the endolymphatic system.


Evidence for a T-cell reaction in the cutaneous sarcoid granuloma.

In cutaneous sarcoidosis, mononuclear cells of granuloma have been freed and characterized by immunological tests. Of these cells, 84% of them were found to be T dependent by the E rosette test. A similar value (79%) was obtained in the stroma of delayed hypersensitivity reaction, whose immunological functions are well known. However, the physiological role of the thymus dependent pupulation within granulomas must be established.


Diagnosis of perioperative myocardial infarction after coronary artery bypass.

The diagnosis of perioperative myocardial infarction (POMI) in patients undergoing coronary artery bypass is uncertain because the criteria of infarction are unclear. Fifty patients who underwent coronary artery bypass were evaluated preoperatively and postoperatively with serial ECGs, creatine phosphokinase (CPK), isoenzyme determinations, and technetium pyrophosphate myocardial scans. Clinical evaluation correlated with exercise testing and postoperative angiography supported the diagnosis in questionable cases. Thirty-five patients (70%) had no evidence of POMI by any criteria, and 2 patients (4%) had unequivocal evidence of infarction by all criteria. Our studies indicate the complexities of diagnosing POMI. We believe that the serially recorded ECG is the most useful diagnostic technique. CPK isoenzyme determinations may be useful but are difficult to interpret in the operative setting. Preoperative cardiac scans are necessary so as to avoid a high incidence of false-positive scans postoperatively. In doubtful cases, postoperative coronary arteriography and left ventricular angiography may provide the most definitive information.


Management of sudden coronary death.

Twenty-three survivors of out-of-hospital sudden coronary death (SCD) have been followed subsequent to initial hospitalization, cardiac catheterization and coronary angiography, and ultimate coronary revascularization (11 patients) or medical treatment (12 patients). All were treated at the Milwaukee County Medical Center. History of previous myocardial infarction (10 patients) and predominance of triple coronary artery disease (20 patients) with associated ventricular dysfunction (21 patients) demonstrated advanced coronary disease in both groups. Selection for revascularization (mean, 3 grafts per patient) was not randomized, but was based on precarious coronary anatomy and was reinforced by post-SCD ventricular dysrhythmias and angina. During an average follow-up of 13 months, there were 2 perioperative surgical deaths (1 recurrent SCD) and 3 medical deaths (2 recurrent SCDs), giving a mortality rate of 22%. This is an improvement over reported post-SCD natural history and may support a policy of offering revascularization to all SCD patients who have precarious coronary anatomy.


Problem-oriented medical record: experience in 238 rehabilitation institutions.

Since Lawrence Weed introduced the Problem-Oriented Medical Record (POMR) nine years ago, several individual rehabilitation institutions have reported their experiences with POMR. As the author was interested in learning of the experiences of other rehabilitation institutions throughout the United States, questionnaires were sent to 253 such institutions. Replies were received from 238 (94%), 132 University-Affiliated Departments of Physical Medicine and Rehabilitation (UAD) and 106 Free-Standing Rehabilitation Centers (FSRC). Some experience with POMR was reported by 155 institutions (65.1%), including 94 (39.5%) which currently use POMR. A comparison between UAD and FSRC revealed statistically significant differences in experience with POMR, the use of POMR for medical and rehabilitation problems, the use of functional evaluation systems and POMR, and professional areas of responsibility. There were no statistically significant differences between UAD and FSRC in the use of POMR for medical audit, the need for formal workshops to orient staff to POMR concepts, the advantages of POMR, and the principal reason an institution was not using POMR. The implications of these findings for patient care, medical education, record keeping, and research are discussed in detail.


Diastatic perforation of the cecum without distal obstruction. Case report and review of the literature.

We present the first reported case to our knowledge of diastatic rupture of the normal cecum following cardiac surgery. All other reported cases of diastatic rupture of the cecum are reviewed. Nasotracheal intubation, hypoxemia, and enemas are thought to contribute to this complication. Prophylactic cecostomy for cecal diameters greater than 9 cm is recommended. Cecostomy and drainage are generally the treatment of choice should perforation occur.


Herpes zoster ophthalmicus with contralateral hemiplegia.

A 48-year-old man developed left hemiparesis nine weeks after herpes zoster skin lesions had appeared over the right forehead. Cerebral angiography showed bilateral changes consistent with cerebral arteritis. The patient's condition worsened after the angiographic procedure. Reports from the literature as well as the present case suggest that arteritis and ischemia best explain contralateral neurological symptoms that occur suddenly following herpes zoster ophthalmicus.


Vitrectomy technique for removal of retained lens material.

A posteriorly dislocated lens or non-encapsulated lens material in the vitreous cavity may be difficult to remove. The pars plana vitrectomy instrumentation and technique provide new methods to remove this material, including use of the cutting-aspiration system of the instrument with or without use of a second instrument to mechanically crush firm lens material. Also, the vitrectomy instrument can be used to mobilize a dislocated lens or lens nucleus and position it in the anterior chamber prior to removal through a limbus incision. These methods provide considerable flexibility and usually permit the surgeon to achieve the objectives of lens removal, vitrectomy, and treatment of any retinal complications under optimum control during one operation. We have used these methods to successfully treat ten consecutive cases.


Anterior uveitis in juvenile rheumatoid arthritis.

The ocular and systemic characteristics of 160 patients with anterior uveitis and seronegative juvenile rheumatoid arthritis are reviewed. Chronic uveitis occurred in 131 patients, 76% of whom were girls. Both eyes were involved in 70% of the cases. Band keratopathy occurred in 41% of the eyes, cataract in 42%, and secondary glaucoma in 19%. Only 11 patients had uveitis before the onset of arthritis. Notable correlations included a pauciarticular onset of arthritis in 95% of the patients, and positive tests for antinuclear antibody in 82%. Of 29 patients with acute anterior uveitis, 27 were boys. The inflammation responded well to therapy, and serious complications did not occur. At follow-up 21 patients had typical ankylosing spondylitis, and five had sacroiliitis. The incidence of positive results of tests for HLA-B27 antigen was 94%.


Paradoxical pupillary responses in congenital stationary night blindness.

Three unrelated boys, ages 2, 6, and 10 years, who have congenital stationary night blindness with myopia and a Schubert-Bornschein-type electroretinogram finding, were found to show a "paradoxical" pupillary constriction in darkness. When examining room lights are turned out, the patient's pupils briskly constrict and slowly dilate. Older night blind male relatives of these boys did not show this abnormal constriction to darkness.


Therapeutic options in Meniere's disease.

Emphasis on treatment in Meniere's disease has recently shifted from a concentration on the vertiginous component alone to include conservation or improvement of hearing as well. This presentation tabulates the current otological literature's reports of both the medical and surgical options available in the treatment of Meniere's disease. The authors conclude that these more recent reports deemphasize the importance of the vertiginous component, while emphasizing the importance of the hearing component in evaluation of the efficacy of any therapeutic modality. Furthermore, these reports indicate that endolymphatic sac decompression and drainage surgery alone offers a considerable chance for conservation or improvement in hearing, as well as relief of vertigo in early cases of reversible, fluctuating hearing deficits. In nonreversible or nonfluctuating hearing deficits with hearing worth conserving, vestibular neurectomy with excision of Scarpa's ganglion yields the best results.


Erythrocyte autoantibodies induced in mice immunized with rat antigens.

Antibody reacting against syngeneic mouse erythrocytes could not be elicited (by rat erythrocytes) in athymic nude mice. Rat antigen preparations from heart, muscle testes, brain erythrocyte ghosts and foetal material failed to elicit detectable autoantibody reactivity against mouse erythrocytes, even though all these preparations (other than brain and foetal) induced a reduction in half life of syngeneic murine erythrocytes in vivo. We suggest that an unstable rat erythrocyte antigen is responsible for the induction of the autoimmune reactions, which in turn depend on T-cells for their generation.


Analgesic abuse, renal papillary necrosis and concomitant drug intake.

In a retrospective survey of 145 patients coming to autopsy at a hospital in Sydney, 31 were found to have some form of renal papillary necrosis (RPN). Sixteen (52%) of those abused analgesic mixtures, there being a marked female preponderance (70%). A further two cases may have been linked with indomethacin and phenylbutazone. On the other hand, 15 (48%) of other analgesic abusers did not develop RPN and the differences between the two groups of abusers and a thrid group of non-abusers were studied in the hope of isolating a precipitating factor. Examination of concomitant drug intake revealed a positive correlation of RPN in abusers with the taking of diuretics and antihypertensive drugs. While these drugs may have been precribed to correct the sequelae of a silent renal lesion, an interactive component in its pathogenesis cannot be ruled out. Analgesic abuse but not RPN was linked with an increased consumption of barbiturates, psychotropic drugs, cardiac glycosides and antacids. Drug interactive effects may partly explain the sudden emergence of analgesic nephropathy 20 years ago.


Hereditary deafness in the cat. An electron microscopic study of the stria vascularis and Reissner's membrane.

The auditory pathway potentials have been recorded from a series of 35 cats, and the stria vascularis and Reissner's membrane examined with the electron microscope. The ages varied from the first postnatal day to at least 6 years. 30 of the animals had white coats, and 45 of the ears lacked potentials. The basal processes of the marginal cells fail to develop in pathological ears, and appear displaced towards the endolymphatic surface by a layer of grossly swollen, chromophobe cells. These features are already present at the 3 day stage, and persist until the third week. Thereafter, increasing electron density of all cell types is found, with progressive reduction in strial thickness. Swelling of the capillary endothelial cells and descent of Reissner's membrane are found in the first coil at 2 weeks. Obliteration of capillaries, and development of vacuolar cytoplasmic outgrowths from the vestibular membrane mesothelial cells occur at later stages. These findings are discussed and compared with the pathological changes produced by a variety of endogenous 2nd exogenous factors in both man and animals.


Pneumocystis carinii pneumonitis successfully treated with trimethoprim-sulphamethoxazole.

Pneumocystis carinii pneumonitis was diagnosed on lung biopsy in an immunosuppressed patient. Therapy with trimethoprim-sulphamethoxazole led to a rapid clinical recovery with no side-effects being observed.


Antisomatogyral illusion.

Following voluntary ambulatory turning (eight turns), perceived motion is reversed in direction relative to a) aftereffects of passive rotation and b) predictions from current concepts of immer ear function. Following active turning, feedback from reflexes elicited by vestibulospinal activity apparently dominates the perceptual experience. Practical consequences in medicine and aviation are discussed.


Discordant distribution of IgM and IgG antibodies to DNA and RNA in monozygotic twins with systemic lupus erythematosus.

Sera from six sets of twins (five monozygotic and one dyzygotic) in whom one or both has systemic lupus erythematosus (SLE) were evaluated for antibodies to DNA and RNA. Sera from three monozygotic twin sets were further studied to determine the distribution of 19S and 7S antibodies to DNA and RNA. The presence of significant binding of polyriboadenylic acid and of native DNA correlated with the presence of clinical SLE in this study. Sucrose density gradient fractionation studies of the sera revealed that the clinically normal twins had some binding of Poly A and DNA limited to the 19S region, whereas the twins with SLE generally had significant levels of 19S and 7S antibodies to DNA and/or Poly A. On the other hand, concordance for presence or absence of antibodies to doublestranded RNA was demonstrated within each twin set irrespective of concordance or discordance for clinical SLE. These results suggest that genetic factors may be important in determining which nucleic acids antigens become immunogenic, but genetic factors alone do not determine the immunoglobulin class distribution of antibodies to nucleic acids.


Limited anti-DNA antibody specificity in systemic lupus erythematosus.

Twenty-one sera from 11 patients with systemic lupus erythematosus were titrated with 125I-ss-calf thymus DNA to determine their maximum capacity to bind DNA. Only five sera were capable of binding greater than 90% of the input DNA. The remaining sixteen sera showed maximum DNA binding levels between 35% and 85%. The inability of these sera to bind all of the input DNA is shown to be dependent on variations in the molecular weight of the DNA antigen. The possibility that these differences in DNA binding reflect differences in antigenic specificity is discussed.


Quantitative determination of sodium-o-[(2,6-dichlorophenyl)-amino]phenylacetate (diclofenac) in human blood plasma or serum.

A rapid method for the estimation of sodium-(o-[(2,6-dichlorophenyl)-amino]-phenyl)-acetate (diclofenac-Na, Voltaren) used in the treatment of rheumatic diseases, in human blood plasma is described. The substance is extracted from acidified plasma with dichloromethane and acetylated after evaporation of the organic solvent, whereupon quantitation is performed by gas-liquid chromatography on a 10% OV-17 column, F.I. detection.


Case report: fatal gastrointestinal hemorrhage in mixed connective tissue disease.

We have presented the case of a young woman with MCTD who died of gastrointestinal bleeding due to fibrinoid necrosis of blood vessels of the small and large intestine. This appears to be the first recorded occurrence of such findings in MCTD.


[Cr51: its use in the detection of blood in stools in the neonatal period].

In order to determine the sensitivity of the Cr51 method in intestinal blood losses, 11 normal term newborn babies were studied with no intercurrences in the perinatal period. A blood sample was labeled for each patient with Cr51 and during 3 day periods; daily blood samples were drawn and stools collected. It was observed a high degree of erythrocytes labelling and minimal blood losses were detected showing high sensitivity of the method.


[Determination of intestinal blood losses in full-term infants].

Eleven normal full-term newborn babies with no intercurrence in perinatal period were studied to determine the volume of intestinal blood losses during breast feeding in the first week of life. Cr51 labelling method was used. The conclusions drawn are: 1. Normal newborn babies have blood intestinal losses ranging from zero to 0.104 ml/24 hours. 2. Intestinal blood losses determined in this study are negligible and allow the conclusion that human breast milk is not responsible for intestinal blood losses that occur during the first week of life.


Pharmacokinetic studies with benoxaprofen in man: prediction of steady-state levels from single-dose data.

1 Plasma levels of benoxaprofen were measured in eight subjects 2-168 h after a single oral dose of 100 mg. Pharmacokinetic parameters were estimated by the NON-LIN computer programme using the two-compartment open model. Mean half-lives of absorption, distribution and elimination were respectively 0.4, 4.8 and 37.6 hours. Volumes of distribution were 6.8 and 3.2 litres for the central and peripheral compartments respectively. 2 Eleven subjects in groups of three or four were given 25 mg/day, 50 mg/day or 100 mg two times daily for 11 days. Their plasma levels were compared with those predicted from the above parameters, which were adjusted for individual body weights and elimination half-lives. Steady-state plasma levels were predicted in each case, and a resonable degree of accuracy (mean 91%) achieved. 3 There was no tendency for observed and predicted levels to diverge as the dose was increased, and there was no evidence of any change in the disposition of benoxaprofen on repeated dosing. 4 The pharmacokinetic parameters were used to predict steady state plasma levels for various dosage regimens.


Preliminary studies of absorption and excretion of benoxaprofen in man.

1 Benoxaprofen is a new acidic anti-inflammatory compound which was well absorbed after oral administration to man. 2 Single doses of 100, 200 and 400 mg produced mean peak concentrations in the plasma of 13.0, 33.5 and 45.3 microgram respectively, and the plasma half-life of the compound was between 30 and 35 hours. 3 Multiple dosing with 25 and 50 mg every 24 h achieved an equilibrium conentration in the plasma after 6-8 days, while dosing with 100 mg every 12 h enabled equilibrium to be reached in 3-6 days. Plasma concentrations between 35 and 45 microgram/ml were achieved by giving 100 mg doses every 12 hours. 4 Absorption of benoxaprofen was delayed when the drug was given with food, but the total amount absorbed remained the same. 5 The effect of milling the material to small particle size (19 micron) was to increase the rate of absorption compared to that of unmilled material (58 micron). 6 Benoxaprofen was well tolerated by healthy male subject in the doses given.


Proenzymic C1s associated with catalytic amounts of C1r. Study of the activation process.

1. Proenzymic C1s isolated from human plasma by euglobulin precipitation and DEAE-cellulose chromatography is associated with trace amounts of C1r (0.5--1% on a molar basis). Incubation for 2 h at 37 degrees C leads to the proteolytic activation of C1s. The proteolysis is characterized by the sigmoidal appearance of C1s esterase activity and of the typical heavy (57 000-dalton) and light (28 000-dalton) fragments of C1s on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. 2. The C1s activation process observed is markedly temperature and concentration dependent, and the rate of activation is decreased by calcium and high ionic strength (I = 0.9). Diisopropyl phosphorofluoridate, benzamidine, polyanethol sulfonate and pentosane polysulphate inhibit the activation, which is also sensitive to C1-inactivator and anti-C1r IgC. From the kinetic experiments and from the inhibition characteristics, the activation of C1s can be attributed to the presence of C1r, which appears to undergo activation and then to activate secondarily C1s.


Alpha-1-antitrypsin. Plasma survival studies in the rat of the normal and homozygote deficient forms.

Alpha-1-antitrypsin from normal individuals (Pi type MM) from those with an inherited deficiency of circulatory protein (Pi type ZZ) were labelled with 125I and plasma clearance rates measured in rats either prior to, or following treatment with neuraminidase to remove terminal sialic acid residues. In addition, these proteins and the derivatives were tested for their ability to bind to an hepatic binding protein obtained from rabbit liver membranes that has been shown to be responsible for the clearance of serum asialoglycoproteins. Finally, the two native forms of alpha-1-antitrypsin were treated with galactose oxidase followed by reduction with tritiated potassium borohydride and then analyzed for tritium incorporation in the neutral sugar fraction. The results indicate: (a) clearance from plasma for both forms of alpha-1-antitrypsin is dramatically enhanced upon the loss of terminal sialic acid residues to the liver membrane protein; (b) Z protein does not exhibit terminal galactosyl residues; (c) the low level of Z protein in plasma cannot be accounted for by a faster rate of clearance relative to M protein. The relevance of these findings to the alpha-1-antitrypsin deficiency state are discussed.


AKR T-cell acute lymphoblastic leukemia: a model for human T-cell leukemia.

In humans, the T cell form of acute lymphoblastic leukemia has proved to be a difficult therapeutic challenge; long-term remissions are unusual. Spontaneous T cell leukemia of AKR mice bears many similarities to the human disease and is proposed as an experimental model in which to test treatment strategies prior to clinical trial.


Studies of some inflammatory and immunological responses of different strains of rat.

Differences were observed in the severity of AA in four inbred strains of rats. The interindividual correlation of the primary and secondary lesions showed an inverse relationship between the gravity of the primary and secondary lesions in the LEW and LE strains. WAG strain appeared to be the lowest responder in the intensity of the primary and secondary oedema in AA and in carrageenan oedema. This is in agreement with the low vascular permeability of this strain after PGE2 injection. When the values of the immunological tests in the individual strains (LE, WAG, LEW) were compared it was found that there is a reciprocal inverse relationship between them and the intensity of the secondary lesion. It follows from our results that the induction of AA requires very precise conditions, indicating that they might depend both on a critical state of immune reactivity and on the action of the pharmacologic mediators.


Light diffraction studies of sarcomere dynamics in single skeletal muscle fibers.

A position-sensitive optical diffractometer has been used to examine the diffraction spectra produced by single skeletal muscle fibers during twitch and tetanic contraction. First-order diffraction lines were computer-analyzed for mean sarcomere length, line intensity, and percent dispersion in sarcomere length. Line intensity was observed to decrease rapidly by about 60 percent during a twitch, with an exponential recovery to resting intensity persisting well beyond cessation of sarcomere shortening; recovery was particularly prolonged at zero myofilament overlap. A number of single fibers at initial lengths from 2.5 to 3.5 MICRON EXHIBITED a splitting of the first-order line into two or more components during relaxation, with components merging back into a single peak by 200 ms after stimulation. This splitting reflects the asynchronous nature of myofibrillar relaxation within a single fiber. During tetanus, the dispersion decreased by more than 10 percent from onset to plateau, implying a gradual stabilization of sarcomeres.


[Electron microscopic study of the interaction between cytolytic T-lymphocytes and target cells].

Thymocytes stimulated in vitro in a mixed culture was sorbed by centrifugation on the surface of target cells for electron microscope study of cytology of immune T-lymphocytes and early cytolysis periods. A well developed Golgi apparatus was revealed in the cytoplasm of lymphocytes; there was also accumulation of tubular structures 50 to 60 nm in diameter which communicated with the cysterns of the granular endoplasmic reticulum, with "descended" vesiculi and plasmatic membrane of lymphocyte. This membrane formed numerous contacts with the membrane of target cells thus producing closed clefts. Taking into consideration these data and also current views on the intertransformation of membranes and intracellular transport a hypothetic scheme of the mechanism involved in the cytolysis of the target cell by immune T-lymphocyte was put forward.


[Ultrastructure and function of cellular elements of the thymus during formation of antitubercular immunity].

Electron microscope and histochemical study of the thymus was carried out on growing guinea pigs under normal conditions and during the immune responses in BCG vaccination. During the postvaccinal period, particularly at periods of from 3 to 7 days, there was a change in the cell elements ultrastructure of the hemato-encephalic barrier. The greatest shifts were noted in the reticular and endothelial cells. The data obtained pointed to the intensification of the morpho-functional activity of the mentioned cell elements of the thymus during the formation of antituberculosis immunity.


[Electrophoretic and immunologic characteristics of the Fc+ and Fc- fractions of the splenic cells of CBA mice].

Conditions have been established for the effective separation of Fc+ and Fc- cells, using sheep red blood cells monolayer, sensitized with the anti-SRBC antibodies. The method was modified by replacement of poly-L-lysine by protamine sulfate. The electrophoretic distribution profiles of Fc+ and Fc- mouse splenocytes were determined. Fc+ cells had mostly a slow electrophoretic mobility. The Fc- cells were more mobile. The Fc+ fraction contained mainly B-lymphocytes, and the Fc- fraction was enriched with T cells. There was an extensive overlap between the electrophoretic profiles of the Fc+ and Fc- splenocytes. After the splenocyte passage through the glass bead column the overlap became much less.


[Interaction and migration of T- and B-lymphocytes in immune responses during carcinogenesis induced by methylcholanthrene].

The (CBA X C57BL) F1 mice were injected intramuscularly with methylcholantrene (MCA) in a dose of 0.3 mg, and their T- and B-cells ability to cooperate in the immune response against sheep red blood cells, and also migration of these cells from the thymus and the bone marrow to the spleen were studied. The MCA immunosuppressive action proved to be associated with the inhibition of migration and cooperation of T- and B-lymphocytes in the immune response. A conclusion was drawn that the immunosuppressive effect developing during the carcinogenesis was complex and it was realized at various stages of immunogenesis.


[Properties and kinetics of a population of B-lymphocytes during rat ontogenesis].

Cells bearing surface immunoglobulins (Ig+-cells) detected by the indirect immunofluorescent method and cells forming rosettes (RFC) with sheep erythrocytes coated with antibody and complement (EAC rosettes) were found in the liver and the spleen on the 15th and the 20th days of prenatal life of rats. The percentage of Ig+-cells and RFC in the liver was high and remained unchanged and at about the same level during the whole postnatal life. The spleen and the bone marrow displayed an increase of the Ig+-cells and RFC increased throughout the 1st month of postnatal life with the maximum at the 30th day after birth; a sharp decrease occurred in old animals. In the thymus the Ig+-cells and the RFC were either absent or present in very small amounts only at some periods of study. Ig+-cells with "capping" were discovered in the spleen and bone marrow on the 5th--10th days of postnatal life; their count increased considerably in 30-day and adult rats. Such cells were absent in the lymphoid tissues of old 40-month rats.


[Inhibitors of trypsin-like proteolytic enzymes as means of preventing  the development of secondary necrosis in burn wounds].

The effect of proteolytic inhibitor contrical on the experimental burn wound healing was studied in rats using biochemical, histological and histochemical methods. In control untreated animals with flame burn of 20% of body surface wound healing was associated with development of secondary necrosis, marked inflammatory reaction, augmented activity of proteases and peptidases. The use of contrical prevented secondary necrosis; this effect was apparently related to reduction of tissue proteolytic enzymes activity.


[Effect of furosemide and ethacrynic acid on sodium transport and potassium secretion].

Ethacrinic acid (0.5 mg/ml), similarly to strophanthin K, increased sodium content and decreased potassium content in the tissue of the frog urinary bladder. Furocemide (0.1 mg/ml) failed to change the ionic content of this tissue. These data indicated a difference in the intracellular action of furosemide and ethacrynic acid.


[Resistance of the suppressor function of T-cells to agents possessing an antiproliferative action].

Administration of cyclophosphamide in a dose of 50 to 400 mg/kg to mice immunized with sheep red blood cells failed to decrease significantly the capacity of the splenic cells of these mice to suppress the primary immune response in transplantation to intact syngeneic recipients. Irradiation of the donors of immune splenic cells (ISC) in a dose of 900 r or treatment of ISC in vitro with mitomycin C failed to influence their suppressor activity. Supernatant obtained after the ultracentrifugation of ISC treated with ultrasound inhibited the primary immune response of intact mice. A conclusion was drawn that the suppressor effect of ISC was caused by the factor produced by T-cells. Active proliferation of these cells was not necessary for the realization of its action.


[Proliferation and cytotoxicity of thymus lymphocytes in vitro].

Proliferative and cytollytical activity of lymphocytes was compared in lymphocyte alloimmunization of the spleen and intact thymus. The count of live cells and DNA-synthesizing cells in the thymocyte monoculture was 10--15-fold, and in mixed thymus cell culture--about 5-fold lower than the corresponding amounts of spleen cells. The index of immune thymocyte stimulation was several times greater than that of the immune cells of the spleen. The cytotoxicity peak was observed on the 4th--5th day of stimulation when the cytolytic activity of the immune thymocytes approached the action of the immune cells of the spleen. Low DNA synthesis and a marked cytotoxic activity of immune thymocytes signified that stimulation of the thymus cells in vitro permitted to obtain cell population with a high content of cytolytic T-lymphocytes.


Recent advances in the cutaneous T cell lymphomas.

Classical mycosis fungoides and Sezary syndrome are part of a larger spectrum of cutaneous T cell lymphomas. Widespread infiltration of the skin, early invovlement of the T cell regions of lymphoid tissue and relative sparing of the bone marrow apparently result from the distinctive properties of the neoplastic lymphocytes. Clinically relevant suppression of normal T cell function has been noted in each patient in the leukemic phase and probably results from both dilution in the blood and lymphoid tissues of normal T cells and production of macrophage inhibitory factor(s) by the neoplastic cells. The abnormal T cells, which in these malignancies appear to be derived from normal "helper" T cells, which facilitate immunoglobulin production by normal B cells. Leukapheresis effectively reduces the tumor load and associated morbidity in selected patients but alone does not prevent the evolution to rapidly proliferating lymphomas. The basis of the remarkable affinity for the skin and identification of the primary site of proliferation and differentiation of the malignant T cells of these processes are important but, as yet, unresolved questions.


Morphological and functional differentiation and classification of cutaneous lymphomas.

It was the purpose of our studies to achieve reevaluation of the histo- and cytomorphology of cutaneous lymphomas by means of additional enzymecyto-chemical and funtional tests. Skin biopsy specimens from 99 patients with cutaneous lymphomas and pseudolymphomas were stained for HE, Giemsa, PAS, Gormori, several hydrolytic enzymes and peroxidase. In cell suspensions extracted from skin lesions B and T cell differentiation was performed using surface markers. In addition tissue cells were tested for PHA response in suspensions and for intracytoplasmatic Ig on smears. Based on these studies cutaneous lymphomas were filed, within the "Kiel" classification of low grade and high grade malignant lymphomas according to their histological, enzymecytochemical and immunological features. It got evident that B cell and T cell lymphomas of low grade malignancy in the skin both present distinct histological patterns, indicating areas B and T cell microenvironmental specificity.


Immunocytological characterization of the mycosis fungoides tumour cell.

The immunocytological identification of the mycosis fungoides cell was carried out on cells extracted from the tumorous nodules of a patient suffering from typical mycosis fungoides. Various techniques, such as E and IgM-EAC rosettes and examination for surface membrane immunoglobulins, were performed on the peripheral blood cells and the tumour cell. Membrane staining with a specific anti-T lymphocyte serum conjugated with peroxidase confirmed the thymodependent origin of the mycosis fungoides cell. However, the immunolabelling (with Fab peroxidase conjugate was found constantly negative.


Hypothalamic adrenaline synthesis after stimulation of the medial forebrain bundle.

1 The problem of whether locally released noradrenaline can be methylated to adrenaline in the hypothalamus has been investigated. 2 During stimulation of the medial forebrain bundle (MFB) the hypothalamic adrenaline content increased somewhat, but the increase was not statistically significant (13%, mean of 10 experiments). 3 After inhibition of the activity of monoamine oxidase and catechol-O-methyltransferase this increase was much larger (80%, mean of 9 experiments). 4 Adrenalectomy did not prevent the rise in hypothalamic adrenaline after stimulation of the MFB. These results suggest that noradrenaline released during activity of noradrenergic hypothalamic structures may be methylated to adrenaline in the hypothalamus.


Smoking and intermediate alpha1-antitrypsin deficiency and lung function in middle-aged men.

Lung function was evaluated in a representative population sample of 50-year-0ld men living in one Swedish city. Twenty-four smoking and 15 non-smoking men heterozygous for alpha1-antitrypsin deficiency--that is, with the protease-inhibitor (Pi1 phenotype MZ--were carefully matched for weight and smoking habit with Pi M controls. The pulmonary function of non-smoking Pi MZ subjects did not differ from that of non-smoking Pi M controls. In contrast, smoking heterozygotes showed a significant loss of elastic recoil, enlarged residual volumes, and increased closing capacity but no signs of obstructive ventilatory impairment. Most smoking Pi MZ individuals reported mild exertional dyspnoea.


Lysosomal properties during thyroxine-induced lateral motor column neurogenesis.

This investigation illustrates that thyroid hormones and not prolactin regulate lysosomal activity during hormone-stimulated lateral motor column (LMC) development in Rana pipiens larvae. Administering a single dose of DL-thyroxine to normal or hypophysectomized larvae induces a 4-8 fold increase in lysosomal acid hydrolase activity within 5 or 6 days. During this interval, the physical properties of lysosomes are dramatically altered with the granules becoming extremely labile to treatments known to disrupt membranes. Nevertheless, thyroxine does not directly influence lysosomal stability; it apparently induces the synthesis and packaging of enzymes through a mechanism requiring de movo production of RNA and protein. The subcellular distribution and physical characteristics of the enzymes are then modified during neuronal death. Such information suggests that changes in lysosomal activity which accompany LMC neurogenesis are indicative of neuronal death, and that these events are controlled by thyroid hormones.


Different behavioral responses to L-DOPA after anterolateral or posterolateral hypothalamic injections of 6-hydroxydopamine.

After bilateral microinjections of 6-hydroxydopamine (6-OHDA) into the anterolateral (AL) or posterolateral (PL) hypothalamus L-DOPA (1,3,4-dihydroxy-phenylalanine) produced running and rearing in AL6-OHDA rats and oral stereotypies in PL 6-OHDA rats. Since the same dose of L-DOPA had no behavioral effect in vehicle injected rats, the responses to L-DOPA of both AL and PL 6-OHDA rats are examples of behavioral supersensitivity. The different forms of behavioral supersensitivity correlated with different patterns of catecholamine (CA) denervation determined by fluorescent microscopy. The major regions of CA denervation in AL 6-OHDA rats were neocortex, hippocampus, limbic forebrain, anteromedial striatum and anterolateral hypothalamus. PL 6-OHDA had these same areas denervated and, in addition, had severe denervation of the entire striatum, parts of the amygdala and thalamus, and of the posterolateral hypothalamus. We conclude that the supersensitive behavioral response to a fixed dose of L-DOPA is determined by the pattern and/or extent of CA denervation.


The derivation and characterization of neuronal cell lines from rat and mouse brain.

This study shows that permanent cell lines can be established from rat and mouse brain by direct tissue culture methodology without the aid of exogenous chemical or viral transforming agents. These cells were derived from specific areas of the brain, such as the cerebellum and hippocampus, at chosen times during fetal and neonatal development. Success in establishing neuronal cell lines was dependent upon the use of selection pressures designed to keep the background of glial cells and fibroblasts at a minimum. These manipulations included care in the choice and processing of the original tissue, utilization of cytotoxic anti-glial sera, and continuous manual isolation of cells with neuronal morphology. Slow-growing nerve cells were thus allowed to adapt spontaneously to culture with a minimum of competition from faster-adapting cell types. Many of these cell lines are judged to be neuronal on the basis of their electrical excitability and their characteristic surface antigens. The cells respond positively in a sodium flux assay which has been shown to correlate well with the ability to generate an action potential, and also express one or more of three antigens previously found to be specific for nerve cells.


Relationship of a T-lymphocyte marker to phase of cell cycle and morphology of leukemic cells.

Spontaneous rosette formation with sheep erythrocytes (SRBC) was studied in the peripheral blood and bone marrow lymphoid cells from a patient whose leukemic cells appeared to be T-lymphocytes. Simultaneous morphological examination of the peripheral blood white cells indicated that they consisted of 21% lymphoblast; 26% prolymphocytes and 48% mature lymphocytes. The distribution of bone marrow cells within the cell cycle was determined by flow microfluorometry and 7 hours after treatment with vincristine consisted of 69% in G1, 21% in S, and 9% in mitosis. Since virtually all the cells both in marrow and blood formed rosettes with SRBC this implies that the expression of this T cell marker is independent both of the morphological appearance of these cells and their position within the cell cycle.


Immunologic characterization of chronic lymphocytic leukemia cells.

Immunologic characterization of the neoplastic cells in the circulation of patients with CLL suggests these cells show significant differences in membrane characteristics from normal B lymphocytes. Although the leukemic cells bear a homogenous membrane-associated immunoglobulin, they also react with an anti-human T cell serum. In all patients studied, 60-90% of the cells, were stained by this antiserum. This suggests that the leukemic cells share antigenic determinants with T lymphocytes. CLL cells, unlike normal B cells, showed a marked increase in mouse-complement receptors. No increase in receptors for guinea pig complement was observed in the leukemic cells. The population of SIg-bearing lymphocytes was significantly greater than that of complement-receptor bearing lymphocytes. The total number of E-rosetting cells was increased in all CLL patients. Mitogenic responses of the leukemic cells were depressed and delayed. These results suggest that neoplastic lymphocytes cannot be classified as T- or B-derived on the basis of criteria used to define normal lymphocytes.


Familial occurrence of colon and uterine carcinoma and of lymphoproliferative malignancies. II. Chromosomal and immunologic abnormalities.

Cytogenic and immunologic studies were performed on 20 members of a family who had an increased susceptibility to carcinomas of the colon, uterus and of lymphoproliferative malignancy. Chromosomal abnormalities such as small G and/or long submetacentric marker chromosome and other aberrations were observed in members who had cancer as well as some asymptomatic siblings. In addition, impairment of T cell function was noted in many of the members suggesting that defective cell-mediated immunity, which also may be genetically determined, played an important role in the expression of this disease.


Thallium-201 myocardial imaging before and after coronary revascularization: assessment of regional myocardial blood flow and graft patency.

Twenty patients underwent myocardial imaging with Thallium-201 (201Tl) before and subsequent to coronary artery bypass grafting. All patients had rest and maximal treadmill exercise imaging postoperatively. Eleven of the 20 patients had rest and exercise 201Tl images preoperatively; 9/20 only had rest imaging preoperatively. Postoperative regional exercise perfusion was improved in seven of the 11 patients who had preoperative exercise images, and was associated with regional graft patency in each case. Thirteen of 20 patients showed no new defect with postoperative exercise imaging. The remaining seven patients developed or had an increased defect with exercise. The patients with no new perfusion defects during postoperative study has 26/30 grafts patent (87%). Patients developing a new perfusion defect with exercise had fewer grafts patent (7 of 13 [54%]; P less than 0.03). In these seven cases, a new exercise-induced defect was associated with regional graft closure or residual nonoperated disease. Knowledge of the preoperative coronary anatomy allowed the distinction between ungrafted areas and regional graft dysfunction. We conclude that preoperative and postoperative 201Tl imaging may noninvasively predict graft closure and/or improved regional perfusion with patent grafts.


The contribution of alpha1-antitrypsin to the total antitrypsin activity of human serum. A study of two phenotype PI OO individuals.

Comparison of the levels of alpha1-AT, alpha2-M, inter alpha-AT, C1 inactivator and antiplasmin and global antitrypsin activity in a group of normal phenotype PI MM individuals, a group of normal individuals with phenotypes with intermediate alpha1 AT activities and alpha2-AT-deficient persons show that alpha1-AT contributes more than 90 percent of the total antitrypsin activity of normal plasma. AT III and fast reacting antiplasmin are shown to contribute to the remaining activity. It can be assumed that due to test conditions the antitrypsin activity of alpha2-M is not assessed. C1 inactivator and inter alpha1-AT do not contribute to a perceptible extent to the overall antitrypsin activity estimated according to the method of Eriksson (Eriksson, S. (1965) Acta Med. Scand. 177, 1).


Polymeric albumin in the urine of patients with nephrotic syndrome.

Dimer and polymer albumin was detected in the urine of a proportion of pantients with a nephrotic syndrome. Most of it was present as S-S bonded dimer and polymer; co-polymers, however, with IgG and alpha (1) anti-trypsin could be demonstrated. It is suggested that albumin polymerizes after it has passed the glomerular membrane. Albumin dimer was associated mainly with minimal change disease and early membranous glomerulopathy in patients, who in general responded well to therapy.


A bleeding syndrome in infants due to acquired prothrombin complex deficiency: a survey of 93 affected infants.

A bleeding syndrome due to severe prothrombin complex deficiency is reported in 93 infants. Most were breast fed (98 per cent), aged 2 weeks to 1 year and there were no serious preceding or associated diseases. Hemorrhagic diathesis, pallor and mild hepatomegaly were the major manifestations. The incidence of intracr anial bleeding was strikingly high (63 per cent) particularly with subdural and subarachnoid hemorrhage. Acute onset, short course and rapid clinical and laboratory improvement after vitamin K therapy were observed. Mortality rate was 35 per cent but has been reduced to 17 per cent since 1969. The location of bleeding, prompt diagnosis and early treatment are the major factors affecting prognosis. Severe prothrombin complex deficiency due to vitamin K deficiency accounted for the pathogenesis of bleeding. Possible causes of vitamin K deficiency were discussed but definite conclusions could not be drawn.


Metabolism of procainamide and p-aminobenzoic acid in patients with chronic liver disease.

Procainamide acetylation and hydrolysis, procainamide-derived p-amino-benzoic acid acetylation, and plasma hydrolysis of procaine were studied in normal volunteers and in 20 patients with chronic liver disease, Impairment of procainamide acetylation was evident in the patients, but no correlations were demonstrable between the degree of impairment and the severity of the disease. On the other hand, procainamide hydroylsis was diminished in liver disease, and as indicated by depression of serum albumin levels and plasma prothrombin activity this alteration did correlate with the degree of impairment of liver function. Procaine hydrolysis in plasma was also affected, the mean in vitro plasma half-life being prolonged in the patients with liver disease and correlating with the degree of hepatic impairment. A correlation of procainamide hydrolysis with procaine hydrolysis was also observed. Finally, acetylation of procainamide-derived p-aminobenzoic acid appeared to increase in patients with liver disease, the degree of acetylation increasing with decreasing procainamide hydrolysis capacity.


Concentrations of flurbiprofen in serum and synovial fluid from patients with active rheumatoid disease: some preliminary observations.

In 3 patients with active rheumatoid disease, concentrations of flurbiprofen of approximately 2 microgram per ml were achieved in synovial fluid 3 hours after a single oral dose of flurbiprofen (100 mg). The highest concentrations were seen between 3 and 9 hours after administration of the dose. The concentration of flurbiprofen in synovial fluid seemed to fall more slowly than in the circulation, but more extensive data would be needed to confirm this. In all 3 patients the drug was absorbed rapidly into the circulation, the highest serum concentrations of 7 microgram to 9 microgram per ml being seen in the first blood sample withdrawn 1.5 hours after administration of the dose. Serum concentrations fell with a mean apparent half-life of approximately 3 hours.


[Antireflux plasty. A new path in the management of bleeding esophageal varices].

Early results with a new operative method for the prevention of recurrent hemorrhage of esophageal varices are reported. The principle of this method is the protection of the esophaguscardia region against the corrosion of biliary reflux. The pathogenesis is based on clinical, endoscopic, and experimental findings. The term "Antirefluxplastik" (antirefluxplasty) is chosen for this type of operation. A segment of about 20 cm of the upper jejunum is excluded and retrocolically interposed between the resected stomach and the duodenum. The procedure is completed by a hemifundoplication and a proximal vagotomy. There were 25 patients treated because of recurrent bleeding of esophageal varices. A follow-up was done on 10 patients from 8 to 12 months postoperatively. Lower frequency of hemorrhage in comparison with the preoperative period is demonstrated. Bleeding recurred in only one patient. Endoscopically gastric erosions were proven the source of hemorrhage. These positive intermediate results encourage us to present the "Antirefluxplastik" as an alternative to conventional operations in the surgery of portal hypertension.


Rectal bleeding due to Oesophagostomum brumpti: report of a case.

A case of oesophagostomiasis of the large intestine in a black Ethiopian sailor is described. Diagnosis was made histologically by finding the parasite in nodules of the intestinal wall. A review of the literature is included.


The diagnosis of the site of upper gastrointestinal haemorrhage in patients with established portal hypertension.

Seventy-five endoscopies were performed in 60 patients known to have oesophageal varices presenting with further upper gastrointestinal haemorrhage during the previous 72 hours. A site of active bleeding was found at 50 of 75 examinations (67 percent) including all 21 patients endoscoped within three hours of the last haemorrhage. Variceal bleeding alone was seen in 40 of these 50 examinations (80%) and both varices and mucosal lesions were bleeding in 4 patients; mucosal lesions were an uncommon cause of haemorrhage and only 6 patients were bleeding from these alone. The adult GIF-D2 endoscope was used for 45 examinations. Variceal bleeding was restarted on three occasions and prolonged unconsciousness induced by intravenous diazepam in one patient. In contrast, the paediatric GIF-P endoscope (30 examinations) was easily tolerated with little or no sedation, did not impair diagnostic accuracy and was not associated with any complications.


Studies on the stimulation by Ca2+ and the inhibition by ADP of steroid 11beta-hydroxylation in adrenal mitochondria.

The Ca2+ stimulation and ADP inhibition of the steroid 11beta-hydroxylation enzyme system in adrenal mitochondria have been investigated. The Ca2+-stimulation is competitively inhibited by Sr2+. Nucleoside monophosphates were not inhibitory and among the nucleoside diphosphates and triphosphates only ADP, CDP and ATP were inhibitory with ADP being more inhibitory than the other nucleotides. None of the non-inhibitory nucleotides could reverse the ADP inhibition. ADP appears to inhibit the Ca2+-stimulation since the activity in the absence of Ca2+ was not inhibited by ADP. Ca2% and ADP did not affect the binding of deoxycorticosterone to the mitochondrial preparation. Ca2+ stimulated the reduction of cytochrome P-450 by NADPH and this was inhibited by ADP. These effects were independent of the presence of deoxycorticosterone. Neither Ca2+ nor ADP affected the reduction of adrenodoxin by NADPH. The inhibition by ADP with respect to Ca2+ or to deoxycorticosterone is non-competitive. The Ca2+ stimulation with respect to deoxycorticosterone is uncompetitive. Preincubation of the mitochondrial preparation with Ca2+ decreases the inhibition by ADP and preincubation with ADP increases its inhibitory effects. These data have been interpreted to indicate that Ca2+ and ADP influence the relationship between adrenodoxin and cytochrome P-450 so that a complex of increased activity in the reduction of cytochrome P-450 is formed in the presence of Ca2+ and a less active complex is formed in the presence of Ca2+ plus ADP.


Electron microscopic identification of postsynaptic dorsal root terminals: a possible substrate of dorsal root potentials in the frog spinal cord.

Dorsal root fibers were labeled with cobaltous chloride iontophoresis for electron microscopic investigations. In the base of the dorsal horn, where most of the coarser collaterals of dorsal root fibers terminate, many dorsal root terminals were found in postsynaptic relation to synapsing profiles. According to their morphological characteristics, three kinds of presynaptic terminals could be discerned in these complex synapses: axon terminals with spheric vesicles, axon terminals with flattered vesicles and presynaptic dendrites. These latter terminals contained relatively few flattened vesicles accumulated adjacent to a short synaptic articulation surface, and they were rich in cytoplasmic organelles. The functional significance of these structural specializations in the mediation of dorsal root potentials and recurrent inhibition is discussed.


[Changes in pacemaker cell activity during parasympathetic acceleration of the heart rate].

Acceleration of the heart rate can be elicited by stimulation of the extracardiac paraysmpathetic pathways (previous investigations). In this study, changes of the pacemaker APs were continually recorded with microelectrode in the sinus node cell. The data showed acceleration of the slope of slow diastolic depolarization (distolic prepotential) of the pace-maker AP during the parasympathetic positive chronotrct. The parasympathetic acceleration of the slow diastolic depolarization and of the heart rate was shown to be cholinergic by its nature.


[Assessment of the role of the sensory innervation of the intraocular muscles in habituation of vestibulo-oculomotor reactions].

In rabbits, participation of the extraocular muscles' sensitive innervation in extinguishing of the eyes nystagmus elicited by periodical rotations, was studied. Either limitation or complete abolishing of the above muscles motor activity were used, as well as the passive forced movements of the eyes. No considerable difference was revealed between the intact and the experimental animals. In both cases the course of nystagmus extinguishing was the same. On the other hand, repeated forced movements of the eye had no effect on the reflectory evoked nystagmus. The data obtained suggest that the proprioceptive afferents of the extraocular muscles take no part in the habituation of vestibular-oculomotor responses.


[A study on the incidence of post-partum hypopituitarism, (Sheehan's syndrome)].

The frequency of post-partum hypopituitarism (Sheehan's syndrome) is difficult to assess. It is the purpose of this paper to ascertain the incidence of post-partum hypopituitarism in Okayama Red Cross Hospital. Out of 19,302 women who delivered at the hospital during 10 years from 1961 to 1970, we selected 1.010 women who had a blood loss of 500 ml. or more at delivery. Firstly, after having sent the selected women a questionnaire by post, we investigated whether they had Sheehan's syndrome or not from the response of the 392 women who replied to the questionnaire. Secondly, 126 of the 392 women who came to our clinic when requested were examined concerning blood picture, fasting blood glucose, and the serum levels of cholesterol, thyroxine, cortisol, GH, TSH, LH and FSH. As a control, 24 women who had had a normal delivery were examined as described above. No advanced hypopituitarism or mild hypopituitarism was found in this follow-up study. Compared with Sheehan's series in England, it was found the incidence of post-partum hypopituitarism in Okayama was presumably less than in England. Suggestions for this discrepancy are as follows: (1) competent obstetric care and a rapid blood transfusion after gross post-partum hemorrhage in Okayama Red Cross Hospital. (2) an incomplete report of the incidence of post-partum hypopituitarism due to those who did not reply to the questionnaire. (3) the rarity of the frequency of Sheehan's syndrome in Japan.


Immunological studies in psoriasis. The quantitative evaluation of cell-mediated immunity in patients with psoriasis by experimental sensitization to 2,4-dinitrochlorobenzene.

Quantitative techniques of sensitization to 2,4-dinitrochlorobenzene (DNCB) was used to determine in psoriasis the intensity and frequency of allergic reactions to DNCB following primary challenge with 2,000 microgram allergen and secondary challenge with decreasing doses of DNCB. 56 patients with psoriasis and 23 healthy volunteers were examined. Frequency of positive reactions to DNCB was similar in both groups, since all normal controls were sensitized, whereas only 8 of 56 psoriasis cases failed to develop delayed hypersensitivity to DNCB. However, the intensity of acquired contact allergy was significantly diminished in psoriasis in comparison with controls. The patients with stationary skin lesions resembled the normal population in the intensity of reaction to DNCB. Decreased intensity of DNCB sensitization seemed to be related to the activity of the disease, but not correlated with the extent of the lesions. A relationship was found between reduced reactivity to DNCB and decrease in E rosette-forming lymphocytes. The data suggest that the impaired function of T lymphocytes in active psoriasis could be responsible for both, defective recognition of contact antigens, such as DNCB, and the alteration of secondary response to DNCB.


Drug and non-drug factors influencing adverse reaction to pyrazoles.

Some of the factors influencing the likelihood of the appearance of adverse drug reactions in patients are identified and discussed. Ways of quantifying some of the risks of adverse reactions in different patients are demonstrated. It is pointed out that adverse reaction data can give valuable guidance for both day-to-day patient management and for the initiation or guidance of research projects. In this latter connection, this study highlights the difference in time of onset between aplastic anaemia and agranulocytosis, suggesting different mechanisms of reaction. The majority of adverse reactions occur during the first three weeks of treatment and it is during this time that patients must be most carefully supervised. The old patients should be watched with particular care. It is concluded that age, sex, disease being treated, length of treatment and even the geographical location where the patient lives, can affect the time, type, frequency and outcome of an adverse drug reaction.


Butazolidin suppository medication in rheumatism; a clinical study in general practice.

One hundred and seventeen patients with various rheumatic conditions were admitted by eighteen general practitions to a multicentre open clinical trial of the acceptability, effectiveness and tolerability of Butazolidin Suppositories. Dosage was one or two suppositories (= 250 mg or 500 mg phenylbutazone) per day for up to eight weeks. Of the 117 patients entered onto the register for the study, 10 patients refused suppository medication and 8 had local conditions contra-indicating their use. Of the 99 patients starting the trial proper, half had excellent symptomatic relief and tolerance. Sixteen patients discontinued because of poor response. The remainder (34) whilst having significant improvement as shown by mean symptom scores, discontinued treatment. The majority who discontinued did so because of minor local discomfort or minor gastric symptoms, suggesting that acceptability of medication was at least as important as intolerance. No patient had a serious or persistent adverse effect. Comparison with previous studies suggests that gastro-intestinal tolerance to phenylbutazone is improved when it is administered by suppository.


Effects of anti-inflammatory drugs on serum chemistry.

1. Biochemical abnormalities are frequently observed when the sera of rheumatoid patients are analyzed. 2. The serum cholesterol, bilirubin, glucose and proteins are significantly related to the activity of the disease. 3. The possible effects of drug therapy are discussed and described. It seems that the way in which drugs modify a patient's serum chemistry may be used to assess the efficacy with which they control the inflammatory process.


Early steroid metabolism in Xenopus laevis, Rana temporaria and Triturus vulgaris embryos.

Embryos of Xenopus laevis, Rana temporaria and Triturus vulgaris exposed to radioactive pregnenolone have been found to convert it to progesterone. Incubations with radioactive progesterone showed that it was actively metabolized by oocytes and embryos. In Xenopus incubations progesterone was converted to 5alpha-pregnane-3,20-dione, 17alpha-hydroxy-4-pregnen-3-one, 4-androstene-3,17-dione and 17alpha-20alpha-dihydroxy-4-pregne-3-one, indicating 5alpha-reductase, 17alpha-hydroxylase, 19-20-desmolase and 20alpha-hydroxylase activities. In oocytes of Triturus and Rana no evidence of 19-20-desmolase was found. In Rana oocytes were also not evidence of 17alpha-hydroxylase activity. All identified activities except 20alpha-hydroxylase were common to embryos of all three species. It is suggested that the steroid enzyme activities present in the embryos are not solely derived from the oocytes but synthentized during early development. Possible meaning of this kind of metabolism during differentiation remains open.


[Immunological membrane markers in blasts and lymphocytes of patients with acute leukemia].

Acute lymphoblastic leukemia can be subdivided into groups with different prognoses and clinical courses by immunological cell-markers. Patients with blasts, expressing T-cell-markers in a high density (T-cell leukemia), had an increased incidence of meningeal relapses, tumor formation in abdomen or mediastinum. T-lymphoblastic lymphoma is the same disease only different in initial presentation. After combination chemotherapy over one year, lymphocytes populations with no immunological cell-markers are relatively increased. Regular monitoring of the immunological subgroups of lymphocytes seems to be unhelpful in early diagnosis of periods with increased risk of severe infections.


[New methods and findings in the measurement of intracranial pressure].

For measuring and continuous recording of intracranial pressure a small telemetric pressure transducer, called "Teleceptor" was developed and energized by inductive energy transmission. Technical problems with regard to in vitro calibration stability, zero-point drift and tissue reaction were examined and discussed. Clinically the "Teleceptor" has so far successfully been used in twentsix patients.


Alpha-1-antitrypsin phenotypes in a group of newborn infants in Somalia.

Pi phenotype determinations were performed on umbilical cord serum specimens of 347 infants born in Mogadishu (Somalia) hospitals. Phenotyping was performed by isoelectric focusing on polyacrylamide gel slabs. Sera containing alpha1AT in types other than MM were also studied by crossed antigen-antibody immunoelectrophoresis on agar. The most frequently occuring Pi allele proved to be PiM. With respect to the other African populations studied so far, there was a high prevalence of the PiS and PiZ alleles. In particular, three individuals were found with the ZZ phenotype, which had not yet been observed in Africa.


Classification of alpha1-antitrypsin (Pi) phenotypes by isoelectrofocusing. Distinction of six subtypes of the PiM phenotype.

Pi phenotypes have been determined by isoelectrofocusing in a sample of 538 healthy individuals from Southern Germany. Further subdivision of the common PiM phenotype is described. A procedure for the delineation of six common subtypes is presented. It is assumed that the six subtypes are determined by three alleles which are provisionally called PiMa, PiMb, and PiMc. Their frequencies in this sample were 0.75, 0.06, and 0.15, respectively.


New phenotypes of serum alpha1-antitrypsin in Japanese detected by gel slab isoelectric focusing.

The phenotype distribution and gene frequencies of serum alpha1-antitrypsin in 856 healthy blood donors in Tokyo were examined by gel slab isoelectric focusing (pH 4--6). The allele of the common subtype variant Pi M2 was present with a frequency of 0.1099 in Japanese. A study of 23 twin pairs and their parents was in agreement with the hypothesis of autosomal codominant inheritance of Pi M subtypes. Other rare variant alleles, Pi MF, Pi MS, Pi MN, Pi mv, Pi MX, Pi MZ were found in very low frequencies. The total concentration of serum alpha1-antitrypsin was compared among three different phenotypic groups (M1, M1M2, M2). Statistically significant quantitative differences were found among these three groups (P less than 0.01).


[Immune dysfunctions in aging (author's transl)].

The frequency of certain infectious diseases, autoimmune disorders and neoplasias reveals a correlation to age. Therefore, the potential pathogentic role of a series of humoral and cellular immune functions is discussed. The concentrations of immunoglobulins increase with age. Though the humoral immune response appears to be rather undisturbed, there is a definite and progressive reduction of delayed type hypersensitivity reactions in the elderly. Based upon own investigations the significance of cell-inherent factors on age-dependence of cell-mediated immunity is underlined, consisting of an age-correlated alteration of a T-cell subpopulation The increasing morbidity or mortality of infectious diseases, i.e. tuberculosis, salmonellosis and viral hepatitis in the older age-group, additionally, may be explained by a reduced cell-mediated immunity known to play an important role in immune defense mechanisms particulary in these diseases.


Haemophilus influenzae in otitis media and sinusitis: serotypes and susceptibilty to ampicillin and amoxycillin in vitro.

In order to obtain Haemophilus influenzae, other than Type b, with highly probable human pathogeniticy strains were collected that had been isolated in abundant numbers from middle ear or maxillary sinus secretions of 157 patients with otitis media or sinusitis. The distribution of serotypes was as follows: nontypable 33.8%, Type b 26.1% Type a 19.1%, Type c 7.6%, Type e 5.7%, Type f 5.1%, and Type d 2.5%. The type distribution did not depend on the source of the strains. All strains were susceptible to ampicillin and amoxycillin in vitro; the broth dilution minimum inhibitory concentrations (MIC) of both antibiotics varied only fourfold; typical MIC values were 0.125 microgram/ml and 0.25 microngram/ml, respectively. Susceptibility did not vary with the source or type of the strains nor with the presence or absence of concomitant penicillin therapy.


Pharmacokinetic studies of indoprofen in healthy volunteers and in patients.

The pharmacokinetics of indoprofen in healthy subjects after single oral and i.v. administrations is reviewed. During repeated administration of indoprofen to 6 normal subjects (200-mg tablet every 8 hours for 6 days) no variations in the disposition of the drug were found in comparison with single dose administration. In 6 inpatients, with rheumatoid arthritis, the pharmacokinetics of indoprofen was studied after single oral (tablet) and i.m. administration. As for oral doses, no difference in main kinetic parameters was detected between the patients and normal subjects except for a higher volume of distribution in the former population. The bioavailability of the drug given by i.m. injection was not significantly different from that observed after oral administration.


Uptake of 5-hydroxytryptamine by mast cells in vivo: a cytofluorometric study of mast cells and individual mast cell granules.

Uptake, distribution and turnover of 5-Hydroxytryptamine (5-HT) was studied by cytofluorometric analysis of whole mast cells and individual granules. Injection of 5-HT as well as 5-Hydroxytryptophan (5-HTP) intraperitoneally or subcutaneously resulted in a parallel uptake of 5-HT in cells and granules. Intraperitoneal injections of 5-HT in such small quantities that may be available under physiological conditions resulted in an increase in fluorescence intensity of the mast cells, indicating a very efficient uptake mechanism for 5-HT in vivo. Much larger doses of 5-HTP were required to obtain a corresponding uptake of 5-HT in the mast cells. The 5-HT was rather rapidly taken up in the granules and eliminated very slowly, at the same rate both from granules and mast cells. The low elimination rate confirms our previous findings that the turnover of 5-HT is much lower in mast cells than in other amine containing cell systems. The combination of an extremely efficient, rapid uptake of 5-HT with a slow elimination suggests a specific function for mast cells in the regulation of free amine concentrations in tissues.


Purification and properties of a NAD(P)H:flavin oxidoreductase from the luminous bacterium, Beneckea harveyi.

A NAD(P)H:flavin oxidoreductase, which produces FMNH2, one of the substrates for the luciferase reaction in bioluminescent bacteria, has been purified with the aid of affinity chromatography on epsilon-aminohexanoyl-FMN-Sepharose. The purified enzyme, isolated from Beneckea harveyi, had a specific activity of 89 mumol of NADH oxidized/min/mg of protein at 23 degrees in the presence of saturating FMN and NADH and appeared homogeneous by several criteria on polyacrylamide gel electrophoresis. A molecular weight of 24,000 was estimated both by gel filtration and and sodium dodecyl sulfate gel electrophoresis indicating that the enzyme is composed of a single polypeptide chain. Kinetic studies showed that the higher specificity of the enzyme for NADH than NADPH and for riboflavin and FMN than FAD was primarily due to variations in the Michaelis constants for the different substrates. Initial velocity studies with all pairs of substrates gave intersecting patterns supporting a sequential mechanism for the NAD(P)H:flavin oxidoreductase.


Structural study of circulating thymic factor: a peptide isolated from pig serum. I. Isolation and purification.

A circulating thymic factor (FTS) has been characterized by a bioassay based on its ability to render theta-negative rosette-forming cells theta-positive and azathioprine-sensitive. FTS was sequentially purified and finally isolated from 1000 liters of pig serum by ultrafiltration, gel filtration, and ion exchange chromatography. Its amino acid composition and apparent molecular weight estimated by Sephadex G-25 chromatography, indicate that FTS is a nonapeptide of composition lysine, aspartic acid (or asparagine), serine 2, glutamic acid (or glutamine) 2, glycine 2, and alanine.


Urogastrone levels in the urine of normal adult humans.

Urogastrone is a potent inhibitor of gastric acid secretion isolated from human urine and it is structurally related to and shares the properties of a potent mitogen-mouse epidermal growth factor. The amounts of urogastrone in 24 h normal human urine samples were measured using a specific and sensitive radioimmunoassay. Values varied from 120--1360 ng/kg/24 h but a significant difference (2P less than 0.005) was observed between urine samples from men (602 +/- 41 SE, n = 39) and women (782 +/- 44 SE, n = 33). A series of 24 h urine collections from male and female volunteers showed that wide variations could occur over a 4 week period in the individual output of urogastrone. Levels from women did not show variations that could be related to the stage of the menstrual cycle.


Relation between growth hormone levels and Von Willebrand factor activity.

The relationship between growth hormone and von Willebrand factor activity was studied in fasting plasma samples, nocturnal samples, and after intramuscular injection of growth hormone. A significant correlation was seen between the two levels in fasting samples. During sleep, peaks of growth hormone were associated with peaks of von Willebrand factor activity. A subject with isolated growth hormone deficiency had no peaks of either. Intramuscular injection of growth hormone produced a rise of von Willebrand factor activity in all subjects studied. These results indicate that there is a relationship between growth hormone and von Willebrand factor activity.


Beta-glucuronidase activity of lymph node imprints from malignant lymphomas and chronic lymphocytic leukaemia.

beta-Glucuronidase activity was semiquantitatively estimated in the cells of lymph node (LN) imprints from patients with Hodgkin's disease (HD), diffuse non-Hodgkin's lymphomas, chronic lymphocytic leukaemia (CLL), normal lymph nodes, and benign lymphadenopathies. In addition, in some of these cases beta-glucuronidase activity was semiquantitatively determined in peripheral blood smear lymphocytes. The beta-glucuronidase score (betaGS) was very low in the cells of the LN imprints from patients with diffuse non-Hodgkin's lymphomas. The LN lymphocytes of HD had a normal betaGS independently of the histological subtype of the disease, while in the LN imprint of CLL the enzyme activity was low, normal, or high. The betaGS of the lymphocytes in LN imprints of normal controls and HD were in general significantly lower compared with he lymphocytes of the peripheral blood smears in the same cases. The relation of our findings to the B and T cell origin of malignant lymphomas and chronic lymphocytic leukaemia is discussed.


Evaluation of a standardised assay for the measurement of antibodies to double-stranded (native) DNA.

A standardised commercially available radioimmunoassay kit for the detection of antibodies to native DNA (N-DNA) has been evaluated in clinical practice. This test system is shown to be a reliably reproducible method of detecting these antibodies. In addition, evaluation of the purity of the radiolabelled test antigen in this assay has shown it to be almost entirely double stranded (native) DNA with virtually no contamination with single stranded (denatured) DNA, and with few areas of single stranded breaks or ends in the duplex molecule. The inclusion of known standards and precise characterisation of the DNA has partially overcome variability in results and provides for interlaboratory standardisation which is lacking in the techniques used at present.


Demonstration of activation of B lymphocytes in New Zealand black mice at birth by an immunoradiometric assay for murine IgM.

We have developed a highly specific and sensitive "two-site" immunoradiometric assay for murine IgM in which antigen bound to immobilized antibody reacts with affinity-purified radiolabeled antibody. We utilized the sensitivity of this assay to study the rate of IgM secretion in short-term cultures by spleen cells from the autoimmune strains, New Zealand Black (NZB) and New Zealand Black by New Zealand White F1 hybrid (BW), and from normal (C57BL/6, DBA/2, NZW) mice. The temperature dependence of IgM secretion in short-term cultures, its pentameric structure, and the similar viability of NZB and normal strain spleen cells indicate that active IgM synthesis is being measured. We observed that the splenic B lymphocytes of NZB and BW mice, in contrast to normal strains, produce IgM in vitro at birth. By 6 to 8 weeks of age NZB and BW spleen cells produce 40 times more IgM than spleen cells from normal strains of mice. The IgM produced in vitro by splenic lymphocytes from NZB and BW mice is not absorbed by synegeneic or allogeneic thymocytes or erythrocytes.


The requirement of viable thymocytes for species-specific attachment to and release from macrophages.

The role of the thymocyte in its species-specific binding to macrophages has been explored. Although formalin treatment of macrophages resulted in loss of binding to thymocytes, formalin treatment of thymocytes did not have this effect. However, two differences between living and formalin-treated thymocytes were noted. Formalin-treated thymocytes bound to macrophages of any species whereas the binding of living thymocytes was species specific. Living thymocytes attained maximum binding in approximately 1 hr and then the fraction bound gradually diminished. Formalin-treated thymocytes remained bound to the macrophage and appeared to be phagocytized. Released thymocytes did not bind to fresh macrophages, but released macrophages bound to fresh thymocytes. The results suggest that the binding of thymocytes to macrophages results in maturation of thymocytes.


Spontaneous cytotoxicity of human peripheral blood mononuclear cells toward red blood cell targets. II. Time-dependent loss of suppressor cell activity.

In a previous paper we demonstrated that human peripheral blood mononuclear cells become strikingly cytotoxic toward a wide variety of red blood cell targets after 7 days of in vitro culture. The cell responsible for cytotoxicity does not rosette with SRBC and demonstrates both surface adherence and phagocytic properties. In this paper we wish to show that development of spontaneous cytotoxicity is due to a time-dependent loss of suppressor cell function. Fresh autologous lymphocytes, when added to cultured cells, abrogate the subsequent expression of spontaneous cytotoxicity toward RBC targets. The suppressor cell is radioresistant; requires 24 hr to suppress optimally; is inactivated by heating at 56 degrees C for 15 min, and is enriched in the non-T interface after SRBC rosette depletion over a discontinuous Ficoll-Hypaque gradient. Furthermore, the addition of a cell-free sonicate of fresh lymphocytes is capable of inhibiting spontaneous cytotoxicity toward RBC targets. However, if mononuclear cells are allowed to incubate in tissue culture medium for 7 days they are no longer suppressive after sonication. These data suggest that fresh mononuclear cells exert a potent negative regulatory influence on monocyte killing. Our culture conditions by removing this negative influence have produced a new model of spontaneous nonspecific killing by monocytes.


Evanescent delayed-type hypersensitivity: mediation by effector cells with a short life span.

Four days after i.v. immunization of mice with optimal low doses of heterologous erythrocytes (2 x 10(5) RBC), strong delayed-type hypersensitivity (DTH) responses can be elicited in the footpad. At later intervals after immunization, DTH responsiveness is progressively diminished and replaced by 4-hr antibody-dependent reactions. These evanescent T cell-mediated DTH responses, which are progressively replaced by antibody-dependent reactions, resemble Jones-Mote type delayed hypersensitivity responses of humans and guinea pigs. Since higher doses of immunizing antigen activate suppressor mechanisms that inhibit DTH responses, we examined the possibility that the evanescence of DTH in mice immunized with an optimal low dose of antigen might also be due to suppression. Using techniques that could clearly demonstrate the suppression produced by high antigen doses, we failed to find evidence for either humoral or cellular suppression in optimally immunized mice with declining of DTH responses. Thus, it appears that the evanescence of produced by optimal low dose immunization with RBC may be due to an intrinsic short life span of the effector cells rather than to the activation of an identifiable shut-off mechanism.


Cellular events in tolerance. VI. Neonatal vs adult B cell tolerance: differences in antigen-binding cell patterns and lipopolysaccharide stimulation.

The numbers and fate of antigen-binding cells (ABC) in neonatal and adult mice rendered tolerant to fluorescein (FL)-labeled heterologous gamma-globulins were studied. Similar numbers of FL-ABC were observed 1 day after tolerogen in both adult and neonatal mouse spleens: by 7 days after tolerization, no FL-ABC were observed in either case. Reinjection with FL-tolerogen at 7 days led to the detection of normal numbers of ABC in adult mice but significantly reduced numbers in neonates. This suggests that neonatal ABC either have been deleted or have failed to resynthesize surface receptors. Two weeks after tolerance induction, spleen cells from these tolerant mice were cultured with Escherichia coli lipopolysaccharide (LPS), a polyclonal B cell mitogen, or with specific antigen. Tolerant adult spleen cells made an equivalent anti-FL response to that of the uninjected controls when stimulated with LPS, but were unresponsive to specific antigenic triggering. In contrast, spleen cells from neonatally tolerized mice were unresponsive to either specific or nonspecific (LPS) stimulation. Thus, these neonatally tolerized spleen cells lose sensitivity to polyclonal-stimulating agents (along with their receptors), or more simply, are deleted.


Generation of T helper cells in vitro. III. Helper cell culture-derived factors are related to alloantigens coded for the I region of the H-2 major histocompatibility complex.

Thymocyte-macrophage cultures primed with carrier protein release an alloantigen-related factor that enhances the anti-hapten plaque-forming cell response of hapten-primed spleen cells in vitro. Use of immunoadsorbant columns made with a variety of alloantisera indicates that the relevant antigens in this system are coded for the the H-2 major histocompatibility locus in mice. The data indicate that in H-2d and H-2k strains the important genetic regions are in the I region between the K region and the I-C subregion and suggest, based on current understanding of Ia specificities, that the I-A subregion codes for these antigens.


A splenic requirement for the generation of suppressor T cells.

Tolerance to contact sensitization with DNFB may be induced by DNBSO3. This specific unresponsiveness may occur via one or both of two mechanisms--production of suppressor T cells or clone inhibition. We investigated the role of the spleen in this unresponsiveness. Splenectomized mice may be tolerized by i.v. injection of DNBSO3, but they are incapable of serving as donors of lymph node cells for transfer of tolerance to normal recipients. Kinetic studies indicated that the spleen must be present at least three days after tolerization in order to permit development of a significant number of suppressor cells in the peripheral lymph nodes. We interpret these results to indicate that 1) clone inhibition does not require the spleen, 2) the generation of suppressor T cells is dependent on the presence of the spleen, and 3) it is likely that tolerogens in this system induce suppressor cells in the spleen and some of these cells or their products leave the spleen to reach the peripheral lymph nodes.


Mechanism for induction of anti-DNA antibodies by bacterial lipopolysaccharides in mice; II. Correlation between anti-DNA induction and polyclonal antibody formation by various polyclonal B lymphocyte activators.

The capacity of various polyclonal B lymphocyte activators (PBA) to induce, in mice, the formation of anti-DNA antibodies was compared with their ability to mediate the release of DNA in circulating blood and to stimulate polyclonal antibody synthesis in vivo. Anti-DNA antibodies or polyclonal antibody synthesis were induced in mice after the injection of at least 10 microgram lipopolysaccaride (LPS) from Salmonella typhimurium, 1 mg dextran sulfate (DS), or 2 mg purified protein derivative of tubercle bacteria RT32 (PPD). Smaller quantities of LPS (0.1 microgram) or DS (500 microgram) were sufficient to cause the release of DNA in circulating blood, whereas PPD was not able to provoke such a release at any concentration used. The association of anti-DNA antibodies with polyclonal antibody synthesis in mice injected with various PBA contrasts with the lack of correlation between the formation of anti-DNA antibodies and the release of measurable amounts of DNA in circulating blood. These results strongly suggest that the induction of anti-DNA antibodies by PBA is a consequence of the polyclonal B lymphocyte activation.


The effects of neuraminidase and galactose oxidase on murine lymphocytes. I. Evidence for the differential delivery of signal(s) leading to cell proliferation and the differentiation of cytotoxic T cells.

The sequential treatment of normal C57BL/6 mouse spleen cell populations with neuraminidase (NA) and galactose oxidase (GO) resulted in cell proliferation, but not in the differentiation of cytotoxic T cells. In contrast, C57BL/6 spleen cells derived from animals primed 5 to 8 months earlier with alloantigen (P815 mastocytoma cells of the DBA/2 strain) both proliferated and demonstrated T cell-mediated cytotoxicity after NAGO stimulation. T cells differentiating into cytotoxic cells after NAGO treatment demonstrated properties similar to alloantigen-specific 'memory' T cells. These were: 1) cytotoxicity developed only from 'primed' cell populations, 2) cytotoxicity developed within 24 hr after NAGO treatment, 3) DNA synthesis was not required for the differentiation of cytotoxic cells during the first 24 hr of culture but both DNA synthesis and cell proliferation were required for the cytotoxicity developing after 24 hr, and 4) all cytotoxicity induced by NAGO showed specificity for the priming alloantigen. It was found, furthermore, that cytotoxicity could be induced at much lower GO concentrations than needed for increased DNA synthesis. We interpret this finding as an indication that NAGO can differentially deliver two 'signals' to T lymphocytes: one leading to cell proliferation, the other causing the differentiation of memory T cells into cytotoxic effectors.


Inhibition of antibody-dependent cellular cytotoxicity by autologous lymph node cells.

A population of lymph node cells that lack the usual T, B, or K cell markers was found to inhibit autologous spleen cells from mediating antibody-dependent cellular cytotoxicity (ADCC) to antibody-coated chicken erythrocytes. Inhibitor cells were not susceptible to treatment with anti-Thy 1.2 or anti-Ig and C; they did not adhere to Sephadex G-10, to nylon wool, or to monolayers of sheep erythrocytes (E) or erythrocytes plus 7S antibody (EA). After a brief (4-min) exposure to 45 degrees C, the ability to inhibit was lost whereas other cellular responses remained intact. ADCC mediated by nonadherent splenic effector cells (presumptive K cells) was highly susceptible to inhibition. Possible mechanisms for and implications of lymphocyte-mediated inhibition of ADCC are discussed.


A screening assay for rapid and quantitative measurement of immunoglobulins and anti-immunoglobulins in unknown sera.

A radioimmunoassay which can measure serum levels of antibody and antigen when both species are immuloglobulins (Igs) is described. The technique is based on competition between insolubilized Ig (bound to Sepharose) and serum Ig for radiolabeled Ig. For measurement of anti-Ig antibodies, labeled Ig antigen is added to a mixture of antibody-Sepharose and unknown or standard antibody solution. For measurement of antigen, labeled, purified antibody is added to a mixture of antigen-Sepharose and unknown or standard antigen solution. Although both assays can detect either Ig antigen or Ig antibody in a given unknown, the antibody assay requires 10-20 times as much antigen as antibody to achieve the same degree of inhibition, and the antigen assay is 100 times more sensitive for antigen than for antibody. By titrating the same unknown in both assays, one can determine whether inhibitory activity is due to antigen or antibody. The routine assay readily detects 1 microgram/ml of antibody or of antigen. A modified assay was also developed which detects levels of antibody as low as 5-10 ng/ml. The technique is simple and allows rapid screening of hundreds of serum samples in a short period of time. The assay can also be modified for measurement of anti-idiotype antibodies.


A simple and rapid method to distinguish between rosettes and non-specific aggregation of erythrocytes in the rosette assay for human T-lymphocytes.

A method is described for rapid and simple visualization of lymphocytes in the rosette assay for human T-lymphocytes. A drop of acridine orange (pH 7.4) is added to the suspension of rosetted cells and allowed to incubate for 1 min on ice. Nucleic acids are then visualized in a fluorescence microscope by excitation at 430-500 micrometer. Simultaneous semi-illumination with light microscopy allows enumeration of total lymphocytes and the number of lymphocytes rosetted with sheep red blood cells. The central lymphocyte is clearly distinguishable even when it is crowded with red cells, and this makes the differentiation between rosettes and non-specific aggregation of cells easy.


Polystyrene balls as the solid-phase of a double-antibody radioimmunoassay for human serum albumin.

Polystyrene balls have been incorporated as the solid-phase of a model double-antibody radioimmunoassay for human serum albumin. Purified IgG from the secondary antiserum is adsorbed on the 6.4 mm diameter balls. The solid-phase secondary antibody is then used to separate primary antibody bound iodinated antigen from unbound antigen. The secondary antibody coated polystyrene balls are easily prepared and manipulated; several hundred sample dilutions can readily be processed in a single assay. Assay background values of 1.5% or less are consistently obtained without extensive or special washing procedures.