Radionuclide studies in Hodgkin's disease and lymphomas. A rational, multidisciplinary approach to Hodgkin's disease and the non-Hodgkin's lymphomas has been responsible for major advances in therapy. Invasive diagnostic procedures and exploratory laparotomy, with their associated complications, make nontraumatic radionuclide imaging most appealing in both the clinical staging of disease and in evaluating therapy. Gallium-67-citrate, the tumor scanning agent of the early 1970's, has demonstrated a marked affinity for Hodgkin's disease and the other lymphomas. False positives are few, with sensitivity greater than 70% throughout the spectrum of Hodgkin's disease and the histiocytic lymphomas. In addition to confirming sites of suspected neoplasm, this agent has proved useful in the detection of occult involvement. Moreover, resolution of abnormal gallium-67 concentrations on follow-up studies functions as a visual ancillary index of therapeutic response. The value of wholebody gallium-67 scintigraphy is further enhanced when used in conjunction with routine technetium brain, bone, liver, and spleen scans. While the diagnostic accuracy of gallium-67 studies has been limited in the abdomen due to bowel activity, our attempts to improve these results with the tumor-seeking radiopharmaceutical indium-111-Bleomycin were unrewarding and subsequently were discontinued. Finally, radionuclide lymphography has also been explored. Its diagnostic usefulness in detecting pelvic and abdominal lymph node involvement warrants further investigation.


Rh erythroblastosis fetalis 1975.

Rh erythroblastosis fetalis 1975. Great advances have been made in the management of Rh erythroblastosis fetalis in the past two decades. Perinatal mortality has been reduced from 16.4% to 3.2%. However, perinatal mortality can only be reduced to zero if Rh erythroblastosis can be eradicated by prevention of Rh isoimmunization. Although prevention of Rh immunization by Rh immune globulin prophylaxis is now a reality, it does not appear that Rh immunization will be completely prevented with a single postdelivery injection. Antenatal treatment plus screening by the Kleihauer technique for massive transplacental hemorrhage may be necessary before complete suppression can be achieved. Low protein Rh immune globulin, and ultimately column-produced, very low protein, highly purified Rh immune globulin for intravenous use may prove to be the safest, most economical, and effective material for Rh prevention and total eradication of Rh erythroblastosis fetalis.


Ultraphotomacrography of whole histological sections. A simple inexpensive method. A short and simple inexpensive method for black-and-white reproduction is described, utilising an accurate electronic time device, a well-designed condenser enlarger with a good lens and an intense point-source lighting system for even better results. For good colour reproduction, however, expensive photomacrographic equipment is still the only satisfactory method.


Three dimensional fine structure of cultured cells: possible implications for subcellular motility. To determine the three dimensional fine structure of whole motile cells, rat embryo cells, cultured on Formvar-coated cover-glasses, were glutaraldehyde/osmium-fixed, mounted on grids, dehydrated, critical point dried and examined by transmission electron microscopy using stereoscopic techniques. Three dimensional arrays of organelles occurred in a filament-rich cytoplasmic matrix. Here, besides microtubules and elongate filaments, inter-connected filaments formed a widespread fine-mesh space network which attached to the plasma membrane and closely surrounded all organelles. Negative staining revealed a similar newtork in unfixed cells. It is concluded that this network represents part of the force-generating mechanism for various subcellular movements.


Ultrastructure of neurosecretory cells in the pars intercerebralis of Rhodnius prolixus (Hemiptera). Six neuron types are distinguished in the pars intercerebralis of the starved fifth instar of Rhodnius prolixus. All neuron types contain electron dense secretory granules derived from Golgi complexes which are of characteristic size and morphology in each type. The neuron types are not thought to represent stages in a secretory cycle. The variety of neuron types described is related to that revealed by staining sections of the same cells with paraldehyde fuchsin. Active synthesis of neurosecretory granules continues throughout starvation and the lysosomal system appears to be involved in the continual degradation of secretory granules. Some of the variations in granule morphology observed may be a consequence of granule fusion and the importance of cytoplasmic events in the development of neurosecretory granules is discussed.


Differences in the graft-versus-host reactivity of cells migrating through nonlymphoid tissue or lymph nodes. Normal lymphocyte transfer reactions have been used to test the alloreactivity of lymphocytes which have migrated through lymph nodes (i.e., in efferent lymph) or nonlymphoid tissues (i.e., in afferent lymph) in sheep. Lymphocytes from afferent lymph are much less efficient at inducing a normal lymphocyte transfer lesion than lymphocytes from efferent lymph. Afferent cell preparations (which contain about 10% macrophages) do, however, cause a greater inflammatory response in the early part of the reaction, but this is at least partly nonspecific. A mixture of afferent and efferent cells produces a lesion which is simply the sum of the two expected reactions, suggesting that the afferent lymphocytes are not merely inhibited by a factor or cell in the preparation.


[Linear differentiation of the human Y-chromosome]. During the final 5.5th to 7th hr of the cycle in presence of BUdR, the human Y-chromosome was delayed in its mitotic condensation in the distal part of the long arm. This part was not homogeneous by the degree of the delay. The unevenly condensed Y-chromosome stained with quinacrine was brightly fluorescent in its stretched part. The fluorescence seemed to correlate with the degree of stretching. The distal part of the long arm was heavily labeled with H3-deoxycytidine thus demonstrating presence of late replicating DNA with GC-base pairs.


Dependence of resting urethral pressure on age and sex of healthy subjects and on technical details. The authors used the Harrison and Constable variation of the original method of Brown and Wickham. The maximal pressure level in the urethral pressure profile depends on technical details. Younger healthy individuals show a higher resistance than older ones. Prostate enlargement leads to an elongation of the posterior urethra. The physiological elasticity loss disposes older persons to stress incontinence.


Preperitoneal herniorrhaphy. Adjunct to prostatic surgery. The preperitoneal approach to inguinal hernia repair can easily be applied at the time of open prostatectomy. Our experience with this procedure is reported, and the surgical technique is described. A preperitoneal herniorrhaphy is easy to perform through the same incision as the prostatectomy, avoids opening new surgical spaces, adds  little to the operating time, and saves the patient another surgical procedure. Good results have been achieved in that we have had only 3 recurrences in 38 hernias reparied by the preperitoneal approach.


Treatment of clinical mastitis: two intramammary formulations compared. Two hundred cases of mild clinical mastitis were treated on two farms using two intramammary preparations in quick release bases. One preparation contained penicillin and streptomycin, the other contained lincomycin, neomycin and prednisolone. Fifty-eight per cent of cases were clinically and bacteriologically cured using the first preparation; and 61 per cent of cases, with a similar range of organisms, using the second. The main infections were Streptococcus uberis and coliform; all but three of the infections were sensitive in vitro to one or both of the antibiotics in the preparations. On the farm with 100 cows, 69 per cent of cases were cured, while only 52 per cent were cured on the farm with 300 cows. It is suggested that it is more difficult to detect and treat cases in the larger herd. A number of reservations about the interpretation of the results are discussed.


Clinical tolerance and catabolism of plasmin-treated gamma-globulin for intravenous application. Plasmin-treated gamma-globulin of placental origin was tested in clinical and laboratory studies and found to be suitable for intravenous use both for prophylactic and therapeutic purposes. Plasmin treatment of gamma-globulin (IgG) results in proteolytic cleavage of 60-70% of the molecules into Fab and Fc fragments whereas 30-40% of the molecules are plasmin resistant. The antibody spectrum of plasmin-treated gamma-globulin is similar to that of standard gamma-globulin. Catabolic properties of the plasmin-resistant portion of this preparation and of standard gamma-globulin are identical. Plasmin-treated gamma-globulin has no anticomplementary activity and its intravenous administration is well tolerated even by highly sensitive immunodeficient patients.


[Characteristics of the interaction of Klebsiella rhinoscleromatis DNA with the sera from patient with systemic lupus erythematos]. In blood sera of patients with systemic red lupus there were observed antibodies, which interacted in reaction of indirect hemagglutination and in complement fixation test with DNA of scleromic bacteria and did not react with the other structures of bacterial cell: with Buaven antigen, detergent and capsular polysaccharides. Reaction of indirect hemagglutination of scleromic DNA with the sera was inhibited by DNA preparations of animal origin. The serological activity of DNA from scleromic bacteria depended upon its secondary structure and did not correlate with polymeric state and nucleotide composition of the biopolymer.


Stimulation with thyrotrophin-releasing hormone (TRH) during antithyroid treatment. During antithyroid treatment a total of 88 TRH tests was performed in 56 clinical euthyroid patients. 56% had negative response to TRH (i.e. delta TSH smaller than 2 muU/ml) after being treated in average 12.6 months and no relation between the duration of treatment and the outcome of the TRH test was found. In the group with positive TRH tests (i.e. delta TSH greater than 2 muU/ml) the mean T4 value was slightly decreased (5.8 plus or minus SD 2.5 mug/100 ml) while the mean T3 value was normal (121 plus or minus SD 32 ng/100 ml). The group with negative TRH tests had quite normal serum T4 values (9.3 plus or minus SD 3.1 mug/100 ml) but in general high normal or elevated serum T3 values (175 plus or minus SD 31 ng/100 ml). Our results seem to indicate that serum T3 is of greater importance than serum T4 with regard to the outcome of the TRH test. The majority of the cases with negative TRH tests, however, had serum T3 and T4 values within normal range. In almost all patients with a negative TRH test a negative T3 suppression of 131I uptake in the thyroid gland was found while a positive TRH test was not correlated with suppressibility of 131I uptake.


Origin of the ATP formed during the light-dependent oxygen uptake catalyzed by Rhodospirillum rubrum chromatophores. The oxygen uptake which is observed when Rhodospirillum rubrum chromatophores are illuminated under air and in the presence of reduced 2, 6-dichlorophenolindophenol (DCIP), 2, 3, 5, 6-tetra-methyl-P-phenylenediamine (diaminodurene, DAD) or N, N'-tetramethyl-p-phenylenediamine (TMDP) depends on the electron-donor concentration according to the equation of Michaelis-Menten. The apparent Km for the donor is lowered by the electron-transfer inhibitor 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO) which causes therefore a stimulation of the rate of the reaction at non-saturating concentrations of the donors. In contrast, the ATP formation which takes place simultaneously to oxygen uptake does not show an enzyme-like dependence on donor concentration. Moreover it is inhibited by HQNO to a variable extent, depending on the particular donor present and on its concentration. Therefore it appears that the HQNO-sensitive phosphorylation is coupled to a cyclic flow which coexists and competes with the non-cyclic flow from donor to oxygen. In the presence of HQNO, substrates and uncouplers of ATP formation accelerate somewhat the rate of the oxygen uptake supported by reduced DCIP and DAD. Thus part of the HQNO-resistant phosphorylation seems to be associated with the non-cyclic flow from those tow donors to oxygen. The lack of stimulation by phosphorylation or by uncoupling of the TMPD-supported oxygen uptake does not permit a conclusion as to whether this reaction is coupled to ATP formation or not. Another part of the HQNO-resistant ATP formation is independent of the presence of oxygen and appears to be associated to cyclic flows which bypass the HQNO site. This type of phosphorylation is most important in the presence of TMPD.


Concealed his bundle extrasystoles in congenital heart disease. His bundle electrograms were recorded in two children with cardiac conduction disturbances. In the surface electrocardiograms of one patient His bundle extrasystoles mimicked atrial nonconducted bigeminal rhythms; in the other, they simulated second degree atrioventricular (A-V) block and conducted junctional extrasystoles. In both instances the conduction disturbance was a result of concealed conduction of the His bundle extrasystole into the A-V junction. We conclude that the surface electrocardiogram is suggestive but not definitive in diagnosing His bundle extrasystoles and that His bundle recordings are indispensable for correct diagnosis and treatment.


Localization of growth hormone-release-inhibiting hormone (somatostatin) in the rat brain. Utilizing an immunoperoxidase technique at the light microscope level, growth hormone-release-inhibiting hormone (somatostatin) was localized in the external zone of the median eminence, the subcommissural organ, the organum vasculosum of the lamina terminalis and the pineal gland. No positive reaction was detected in any other brain area.


The history and sociology of the scientific study of acupuncture. Although known to the medical community in some form since the 19th century, acupuncture is only recently receiving concerted research attention from the conventional medical and scientific community. The overall amount of scientific acupuncture research has doubled since the mid 1960's. Employing the approaches of the science, history, sociology and social relations of science, this article explores factors which explain why the present time is conducive to acupuncture research by the scientific and medical community. Perhaps this study of scientific acupuncture research will provide the basis for a later theory of scientific development.


Soil ameba infection. Specific indirect immunoenzymatic (peroxidase) staining of formalin-fixed paraffin sections. This report describes the use of the immunoenzymatic (peroxidase) method to identify the species and to stain distinctively the amebas in formalin-fixed paraffin-mounted sections. This permits the use of hematoxylin and eosin counterstaining. The method, now well developed by others for many purposes, is an alternative to immunofluorescence and seems to offer a number of advantages and a lesser number of disadvantages.


Histochemical localization of intraocular copper foreign bodies. Alizarin blue, rhodanine, and rubeanic acid stains cytochemically localized copper in rabbit eyes harboring intravitreal copper foreign bodies of varying copper composition. Local deposition presented in small amounts in areas adjacent to the foreign body (fibrous capsule, adjacent retina, and surrounding macrophages). The localized pattern was more frequent with the 60% copper alloy, when the foreign body was encapsulated or shiny, and when the aqueous copper concentration was normal. The generalized pattern of copper deposition revealed moderate to marked amounts of copper within the following multiple intraocular structures: (1) the posterior cortex of the lens, (2) the vitreous cavity, either in free-floating macrophages or in macrophages on the surface of the retina, (3) the retina, either within monocytic macrophages, Müller cells, or extracellularly in granular clumps, and (4) the fibrous capsule around the foreign body. Copper in several eyes was observed within macrophages that were clustered around the central retinal vessels within the optic nerve. The generalized pattern was frequent with the pure copper foreign body, when the copper foreign body was tarnished, and when copper concentration of the aqueous humor was elevated. There was a direct toxic effect of copper on the lens in the form of local opacification in the posterior cortex but no clinical or histochemical evidence of a Kayser-Fleisher ring or a sunflower cataract.


Sensory integrative function in a child guidance clinic population. Recent researchers in the area of mental health have recognized the influence of biological defects on psychosocial development and on the child's ability to acquire social and academic skills. This study was designed to determine whether or not senxory integrative disorders occurred in a child guidance population, to explore the nature of sensory integrative disorders found in such a group, and to ascertain any relationships between certain types of sensory integrative disorders and specific behavioral traits. The subjects were 87 children between 5 and 9 years of age, the total number referred to a child guidance clinic in southern California because of problems in behavior. All subjects (Ss) were administered the Southern California Sensory Integration Tests and were observed for neuromuscular deviations. The date were subjected to factor analyses. Results showed a high incidence of sensory integrative dysfunction in this population. There was considerable evidence of a syndrome known as postural and bilateral integration dysfunction as well as a strong tendency in this group to show various kinds of behavior problems.


A comparative study of predictive criteria in the predisposition of homicidal adolescents. The authors evaluated the criteria that are cited in the literature as predictive of homicidal predisposition. They applied three categories of criteria--clinical, developmental, and environmental factors--to a study group of 10 adolescents who had committed homicide, 10 who had threatened or attempted homicide, and 10 hospitalized controls. Their findings did not support the presence of a well-crystallized predisposition for homicidal behavior in this population, but they did show that the adolescents who committed homicide were psychotic-regressive and those who threatened or attempted homicide were organic-impulsive. The study strongly suggests the importance of environmental factors in reinforcing homicidal behavior.


Primary immunodeficiency diseases and malignancy. The incidence of malignant tumors in the primary immunodeficiency diseases is dramatically increased. Four patients with primary immunodeficiencies who developed fatal malignancies are reported. Lymphoreticular tumors and leukemia predominate in most conditions, but epithelial neoplasms are the most common tumors in selective Iga deficiency, and they comprise over one-fourth of malignancies in common variable immunodeficiency. With the exception of common variable immunodeficiency and the Wiskott-Aldrich syndrome, hyperplasia of lymphoid tissue usually does not occur. Lymph node enlargement in any of the other immunodeficiencies is therefore most likely secondary to malignancy. Benign gastrointestinal nodular lymphoid hyperplasia occurs frequently in common variable immunodeficiency and in some instances may be impossible to differentiate roentgenologically from lymphoma.


Proliferation of non-neuronal cells in spinal cords of irradiated, immature rats following transection of the sciatic nerve. Transection of a peripheral nerve not only elicits changes in the injured neurons but also results in an increase in non-neuronal cells, considered by most workers to be neuroglia, in the region of these neurons. Since studies in this laboratory have shown that the neuroglial population of spinal cords of immature rats can be reduced markedly by ionizing radiation, the present investigation was undertaken to determine if this reaction would occur in the irradiated spinal cord following transection of the sciatic nerve. In order to answer this question the sciatic nerve was sectioned unilaterally at 17 days of age (14 days post-irradiation). Sham-irradiated littermates served as controls. Light microscopic examination showed an increase in non-neuronal cells throughout the gray matter on the side of axotomy in spite of a decreased neuroglial population in the 2,000 R and 3,000 R groups. These cells were scattered in the neuropil or were adjacent to injured neuronal perikarya in the anterior horn. Qualitatively similar reactions occurred in the 500 R and 1,000 R groups and in shamirradiated controls. Whether the magnitude of response is the same in all groups is currently under investigation, as are questions dealing with the origins of the reactive cells.


Neuromuscular disease in secondary hyperparathyroidism. Neuromuscular function was evaluated in six patients with osteomalacia or secondary hyperparathyroidism, or both, as demonstrated by bone biopsy showing osteomalacia or increased immunoreactive parathyroid hormone, or both. Each patient had weakness, atrophy, and fatigability of proximal muscles, especially of the lower extremities. Most also showed involuntary fine movements of the tongue, hyperactive tendon reflexes with abnormal spread, and decreased vibration sensation, abnormalities similar to those observed in primary hyperparathyroidism. Every patient studies had evidence of neuropathic muscle disease, either on electromyography or muscle biopsy studies histochemically or both. Muscle biopsies showed no definite myopathic features. Treatment of the osteomalacia improved muscle strength. Patients with osteomalacia therefore have a treatable neuromuscular disease that is neuropathic in nature and resembles closely that found in primary hyperparathyroidism.


Giant keratoacanthoma of the nose.

Giant keratoacanthoma of the nose. In a case of giant keratoacanthoma of the nose, the lesion was extremely invasive, destroying the entire nose. Electrosurgical removal of the grossly abnormal tissue was followed by rapid recurrence on two occasions. Radiation therapy was not effective. The lesion healed after the use of bleomycin sulfate.


Ultrastructural demonstration of cystine in guinea pig back stratum corneum. A method for the ultrastructural demonstration of cystine in keratinized epidermal cells of guinea pig back hairy skin is described. This involves selective oxidation by peracetic acid and formation of an electron dense reaction product with uranyl acetate. Unkeratinized cells were unstained except for nuclei and keratohyalin. The thickened plasma membranes of transitional and keratinized cells were darkly stained. In transitional keratinized cells darkly stained membranes occurred initially in relation to desmosomes and then spread to intervening membrane areas. Dense material, probably matrix keratin containing cystine was found in the interiors of basal horny cells but not in the intermediate or superificial cells of the stratum corneum. It is suggested that the unstable matrix keratin is probably broken down by enzymatic hydrolysis above the basal zone of the stratum corneum.


Congenital insensitivity to noxious stimuli. Cerebral-evoked potentials were used to study a 25-year-old man, the older of two siblings with congenital insensitivity to all noxious stimuli, gross impairment of temperature perception, and anhidrosis. Electrical stimulation of tooth pulp consistently eliciting pain  and cerebral responses in normal subjects evoked neither cerebral potentials nor painful or other sensations in our patient. However, ordinarily painful electric shocks to the skin of his face evoked cerebral responses as well as sensations lacking disagreeable qualities. Those cerebral potentials elicited by electrical stimulation of the median nerve, clicks, and light flashes were within normal limits. These findings strongly suggest that a defect in transmission of noxious impulses presumably involving first order sensory neurons exists in our patient.


Basic fuchsin picric acid method to detect acute myocardial ischemia. An experimental study in swine. Application of the hematoxylin basic fuchsin-picric acid method to human postmortem myocardium specimens failed to show reproducible results.  Therefore, controlled animal studies utilizing the method were undertaken.  Myocardial ischemia was induced in swine by ligation of the anterior descending coronary artery for periods of 30 minutes to six hours.  Electron microscopy was used to monitor normal and ischemic myocardium.  Reproducible results were not obtained by staining successive slides of the same block at the same time or at different times, whether stained in the same reagent bath or after changing the reagent solutions.  Fresh, frozen, unfixed material also yielded nonreproducible results.  The method in its present form is unreliable for routine use in the diagnosis of early myocardial ischemia.


Effects of 5-azacytidine on nucleolar RNA and the preribosomal particles in Novikoff hepatoma cells. Examination of nucleolar RNA from cultured Novikoff hepatoma cells treated for 3 hr with 5 x 10-4 M 5-azacytidine shows that significant amounts of analog-substituted 45S RNA are processed to the 32S RNA species, but 28S RNA formation is completely inhibited. Under these conditions of analog treatment 37% of the cytidine residues in the 45S RNA is replaced by 5-azacytidine. During coelectrophoresis of nucleolar RNA from 5-azacytidine-treated and control cells, the analog-substituted 45S RNA and 32S RNA display reduced mobilities compared to the control 45S RNA and 32S RNA. Coelectrophoresis of analog-substituted and control RNA after formaldehyde denaturation shows no differences in electrophoretic mobility between the two RNA samples, suggesting that 5-azacytidine incorporation may alter the secondary structure of the 45S RNA and the 32S RNA. 5-Azacytidine at 5 x 10-4 M severely inhibits protein synthesis in Novikoff cells by 3 hr. After this length of treatment, however, CsCl buoyant density analysis reveals no difference in density of either the 80S or 55S preribosomal ribonucleoprotein particles when compared to normal particles. Also 5-azacytidine treatment does not appear to cause major changes in the polyacrylamide gel electrophoresis patterns of the proteins in the 80S and 55S preribosomal particles. These results together with previous findings suggest that 5-azacytidine's inhibition of rRNA processing is possibly related to its alteration of the structure of the ribosomal precursor RNAs and is not a consequence of a general inhibition of ribosomal protein formation.


The isolation and characterisation of a platelet-specific beta-globulin (beta-thromboglobulin) and the detection of antiurokinase and antiplasmin released from thrombin-aggregated washed human platelets. A protein fraction was isolated from the supernatant of thrombin-aggregated washed human platelets and was shown, by immunodiffusion techniques, to contain a platelet-specific beta-globulin (beta-thromboglobulin) as the major component. A molecular weight of 35 800 was determined for beta-thromboglobin from the measured sedimentation coefficient of3.0 S and Stokes radius of 2.85 nm. Beta-Thromboglobin was detected in the serum from whole blood and the supernatant of 48-h-old platelet-rich plasma and 28-day-old citrated whole blood, but not in platelet-poor plasma. The fraction containing beta-thromboglobulin was shown to possess an antiurokinase activity but was devoid of antiplasmin activity. A further fraction of approximate molecular weight 70 000 was also isolated which contained an antiplasmin but was devoid of antiurokinase activity.


On basic proteins in bovine peripheral nerve myelin. 1. Myeline proteins in bovine peripheral nerve migrated as two main band-(BF and BR protein) and one faint middle band (BM protein) on sodium dodecyls sulfate-polyacrylamide gel electrophoresis. The relative mobility of these two main bands differed from those of myelin proteins in the central nervous system. 2. The acid extract of the myelin fraction from bovine peripheral nerve was separated into one main peak and two minor peaks on a Sephadex G-75 column. The major component of the second minor peak was the BM protein; the major component of the main peak was the BF protein. The BR protein was not extractable by acid solution. 3. Molecular weights of the BF, the BM and the BR protein were determined as around 13 000, 20 000 and 28 000, respectively, by sodium dodecylsulfate-polyacrylamide gel electrophoresis. 4. The amino acid composition of the BF protein was quite different from the encephalitogenic protein and the Folch-Lees type proteolipid protein in the central nervous system. However the BM protein showed similar amino acid composition to the encephalitogenic protein. 5. The tryptic peptide maps of the BF protein and of the encephalitogenic protein were quite different. The results suggested that the amino acid sequences of these two proteins are different and that they contain no common tryptophan-containing peptide.


Acute leukemia with Burkitt's tumor cells: A study of six cases with special reference to lymphocyte surface markers. In six patients with acute leukemia (about 2% of the patients referred for acute lymphoblastic leukemia) the blast cells invading bone marrow and blood showed all the cytologic, cytochemical, and electron microscopy features of Burkitt's tumor cells. The presence of monoclonal surface immunoglobulins (their synthesis being proved by in vitro culture experiments), the binding of IgG aggregates, and the absence of rosette formation with sheep red cells documented the monoclonal B-cell origin of these blast cells which is in sharp contrast to the findings in common acute lymphoblastic leukemia. The course of the disease was usually rapidly fatal without chemotherapy-induced remission.


Canine cyclic neutropenia: Erythropoietin and platelet cycles after bone marrow transplantation. The marrow of a dog affected with cyclic neutropenia (CN) was transplanted into an unaffected supralethally irradiated litermate. Prompt engraftment occurred, and cyclic rises and falls in numbers of platelets, reticulocytes, and granulocytes were noted in the recipient soon after engraftment. Prior to transplantation and under hypoxic conditions, the donor had serum erythropoietin (ESF) peak levels at 11-12-day intervals. Following transplatation and under hypoxic conditions, cyclic peaks of ESF occurred in the transplanted dog.


Effects of erythropoietin on erythroid colony formation in uremic rabbit bone marrow cultures. Erythropoietin-responsive stem cell (ERC) kinetics in anephric uremic rabbits were studied in vitro using the growth of erythroid colonies in a methyl cellulose system in cultures with and without the addition of erythropoietin (ESF). Approximately 68 hr after bilateral nephrectomy, an increase in BUN and decreases in hematocrit and marrow erythroid cellularity were seen. However, the numbers of erythroid colonies formed in response to ESF on plates inoculated with 2 times 10-5 cells were greater in anephric rabbit marrows than in normal controls. In addition, the numbers of erythroid colonies produced by the uremic and normal marrows in the presence of ESF were increased in proportion to the number of precursors plated. These findings suggest that, in uremia, the concentration of ERC is increased and that the ERC are capable of responding normally to ESF. The increase in the number of erythroid colonies of uremia may be due to the undisturbed flow of uncommitted hematopoietic stem cells into the ERC compartment in the presence of a delay of differentiation of ERC into heme-synthesizing nucleated erythroid cells due to a lack of ESF.


Effects of testosterone and erythropoietin on erythroid colony formation in human bone marrow cultures. The effects of testosterone and erythropoietin (ESF) on erythroid colony formation in normal human  bone marrow cultures were studied in vitro using a methyl cellulose gel system.  Testosterone was found to produce a significant increase in erythroid colony formation at concentrations of 10-4-10-4M in vitro.  In this system, the numbers of erythroid colonies formed per plate increased in direct proportion to the increase in the number of erythroid precursors inoculated as well as to the increase in the dose of ESF in vitro.  In addition, a synergistic effect of a combination of testosterone and ESF on erythroid colony formation was seen when ESF was present at high concentrations.  These data suggest that a greater number of erythropoietin-responsive cells are available for ESF to differentiate into the nucleated erythroid cell line in the presence of testosterone, indicating that the effect of a combination of testosterone and ESF is greater in enhancing erythropoiesis than the additive effects of either agent alone.


Is 'second look surgery' justified in suspected recurrences of cancer of the stomach? A series of 23 patients with suspected recurrent disease following 'curative' resection for gastric carcinoma was submitted to further laparotomy (24 operations). Three patients were found to have hopeless dissemination, 10 had palliative procedures performed which relieved their dysphagia, vomiting and jaundice, 4 had recurrences resected and 1 had a second primary of the gastric stump excised. No less than 6 were found to have benign lesions. Although the reward as far as prolonged survival is small, we believe that the relief of symptoms and the occasional long term excellent result justifies judicious use of a 'second look' laparotomy for suspected recurrent disease.


Histological evidence for direct connections between the optic lobes of the cockroach Leucophaea maderae. Heretofore, descriptions of direct interconnections between insect optic lobes have been based on histological examinations of normal brains or on inference from electrophysiological or behavioral data. We present here what we believe to be the first demonstration of such monosynaptic connections by techniques of experimental neuroanatomy. Twenty-four to 39 h after extirpation of the left optic lobe, degenerating axons and axon terminals, as silvered by a modified Nauta technique, were abundant in the central portion of the medulla of the right optic lobe. The periphery of the medulla was free of argyrophilic debris as were the lobula and lamina. The distribution of neuronal somata with processes terminating in the the left optic lobe was established by retrograde axonal transport of horseradish peroxidase injected into the left lobe and by the development of distinctive perinuclear rings of RNA (a 'chromatolytic' reaction) by some cells within 1-2 weeks following amputation of the left lobe. Both techniques revealed distinct clusters of cells in the anteroventral and posterior regions of the right optic lobe, and in the medial portion of the right protocerebrum. The cells which interconnect the two optic lobes may be involved not only in the bilateral representation of visual information, but also in the coordination of optic lobe pacemakers which control a circadian rhythm of locomotory activity.


Tissue specificity of nuclear acidic proteins isolated from bovine brain and adrenal medulla. Nuclear acidic proteins from bovine brain and adrenal medulla demonstrated dissimilar electophoretic and chemical profiles. The amino acid analysis of the acidic nuclear protein fraction isolated from these tissues revealed some variation in the ratio of acidic to basic amino acids. The estimation of free carboxylic acid groups confirmed the more acidic nature of the brain proteins. In comparing the acrylamide gels either visually or optically, several electrophoretically specific bands were apparent. Although the total number of protein bands from each tissue was approximately the same, the adrenal medulla contained a larger proportion of the more positively charged proteins. These observations are interpreted to indicate that the nuclear acidic protein from brain and adrenal medulla may show functional variation.


Fungal fimbriae. I. Structure, origin, and synthesis. Fine hair-like appendages on the cell walls of the another smut Ustilago violacea are described. These hairs are termed fimbriae because of their close similarity to the fimbriae (pili) found on certain Gram-negative bacteria. Cells of U. violacea may carry more than 200 fimbriae varying in length from about 0.5 mum to over 10 mum, and having a diameter of about 60-70 A. Some fimbriae produce knobs similar to those found on bacterial sex fimbriae. Log-phase cells are the most densely fimbriated, while stationary phase cells are devoid of fimbriae. The cells can be defimbriated by sonication, high-speed agitation, or centrifugation through a 40% sucrose solution. The fimbriae can regenerate in these defimbriated cells in about 1 h. This regeneration is inhibited by both cycloheximide and rifampin, but not by chloramphenicol and therefore appears to depend on de novo protein synthesis on cytoplasmic ribosomes. Similar long fimbriae are found on U. maydis and Leucosporidium (Candida) scottii. Short fimbriae, about 0.5 mum long, were found on all the other species of yeast-like fungi examined (Rhodotorula, Saccharomyces, Schizosaccharomyces, Hansenula, Lipomyces, Nadsonia, and Torulopsis spp.).


A staging system for hepatocellular carcinoma: prognostic factors in Ugandan patients. A staging scheme for hepatocellular carcinoma was presented at an International Symposium on Liver Cancer in Kampala, Uganda in 1971. Historical, clinical, and laboratory aspects of that staging scheme were examined for prognostic significance in 72 untreated patients with this disease studied at the Uganda Cancer Institute. The median survival for the entire group was 1 month. The presence of a serum bilirubin concentration of greater than 2 mg/100 ml or weight loss greater than 25 percent of body weight were the poorest prognostic features. Other factors with prognostic significance were visible abdominal collateral circulation, ascites, tumor differentiation, and serum levels of alkaline phosphatase, SGOT, alpha fetoprotein, and proline hydroxylase. A modified staging scheme is presented which defines three prognostically different groups of Ugandan patients. It is hoped this staging scheme will serve as a stimulus for analysis of similar prognostic features in other populations of patients with hepatocellular carcinoma.


Carcinoembryonic antigen and alpha fetoprotein in malignant tumors of the female genital tract. Circulating carcinoembryonic antigen (CEA) and alpha fetoprotein (AFP) levels were measured by radioimmunoassay in 53 patients with carcinoma of the ovary, 16 patients with other malignant genital tumors, and 31 women with nonmalignant diseases of the genital tract. The serum CEA concentration was elevated (greater than 5 ng/ml) in 11 patients with ovarian cancer, 2 patients with endometrial cancer, 1 patient with carcinoma of the cervix, and 1 patient with a benign embryonal cystic teratoma. Elevated CEA levels were found only in patients with advanced malignant disease, while early stages were associated with normal CEA concentrations. AFP levels were normal in all but 1 patient. Both CEA and AFP levels were markedly raised in a case of advanced genital carcinoma arising probably from the ovary. Ascitic fluid of another patient with ovarian cancer contained a high concentration of CEA, giving an identical reaction in immunodiffusion with CEA from colon cancer. The present results indicate that while the increased expression of carcinofetal components takes place in some malignant tumors of the female genital tract, it is usually a late phenomenon.


Coagulative and fibrinolytic properties of ascitic fluid associated with ovarian tumors. Ascitic fluid samples from 19 patients with ovarian carcinoma, 3 with a benign ovarian tumor, and 5 with cirrhosis of the liver were examined for their content of coagulation factors and components of the fibrinolytic system. The concentration of trypsin inhibitors in the ascitic fluid was significantly higher in the presence of carcinoma. Large amounts of FDP were found in the ascitic fluid in all patients with malignant tumors, but not in the other two groups. Determination of FDP may therefore make it possible to differentiate between malignant and nonmalignant ascitic fluid.


Primary endodermal sinus (yolk sac) tumor of the liver. First reported case. A primary endodermal sinus (yolk sac) tumor of the liver occurring in an 18-month-old boy is described. Although several examples of extragonadal endodermal sinus tumors have occurred in other sites, this is believed to be the first reported instance of origin in the liver. An additional important feature was the detection of alpha-fetoprotein in a preoperative serum sample. Although no metastases were identified at the time of celiotomy, widespread metastases developed, and he died 6 1/2 months after an extended right hepatectomy was performed. Neither triple chemotherapy nor radiation therapy appeared to deter progressive spread of the neoplasm, although the metastases exhibited some radiosensitivity.


Studies on recovery from chemically induced damage in mammalian cells. The survival of plateau-phase or nondividing Chinese hamster ovary cells (in vitro) is reduced to a greater extent by treatments with nitrosourea compounds than are cells treated in the exponential phase of growth. The greatest decrease in the survival fraction occurred following treatments with 1-trans-(2-chloroethyl)-3-(4-methylcyclohexyl)-1-nitrosourea where approximately 128 times more cells were killed in plateau phase than in the dividing state (at the 10 mug/ml-for-1-hr dose). Only 5 times more cells were killed in plateau phase than in exponential growth when cells were treated with 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea. Cells treated with either nitrosourea compound failed to recover from potentially lethal damage and sublethal damage. The breakdown products of the nitrosourea compounds are known to inhibit DNA repair and may explain the failure of mammalian cells to recover from sublethal damage and potentially lethal damage induced by these chemicals. Both dividing and nondividing cells were able to recover from bleomycin-induced potentially lethal damage but not from sublethal damage. The recovery from bleomycin-induced potentially lethal damage by nondividing cells was twice as great as that exhibited by dividing cells; however, potentially lethal damage recovery was suffieiently high for cells in both growth states to conceal the true response to sublethal damage.


Elevated concentrations of serum alpha-fetoprotein in rats with chemically induced liver tumors. The study was undertaken to determine whether aflatoxin B1 (AFB1)-induced liver tumors in rats produced alpha1-fetoprotein (AFP) and whether the age of the animals would influence such as appearance, a finding suggested by data seen in man. Other liver carcinogens (N-hydroxy-N-2-fluorenylacetamide, N-2-fluorenylacetamide, and diethylnitrosamine) were tested for their ability to induce liver tumors producing AFP. The presence of AFP. The presence of AFP in the serum was determined by double diffusion in agarose and by comparison also by quantitative radioimmunoassay. Using double diffusion, AFP was detected in the majority of tumor-bearing rats that had received either N-2-fluorenylacetamide or N-hydroxy-N-2-fluorenylacetamide. Sera of diethylinitrosamine-treated rats with liver tumors were all positive, whereas sera of rats bearing AFB1-induced tumors were positive in only a few cases. However, all sera of tumor-bearing rats examined had elevated AFP levels by radioimmunoassay. Nonetheless, the average level of AFP in the sera of rats bearing AFB1-induced tumors was considerably lower, compared to the sera of rats with tumors caused by diethylnitrosamine, N-2-fluorenylacetamide, or N-hydroxy-N2-fluorenylacetamide. Rats started on AFB1 when 6 weeks old had more mixed liver tumors with neoplastic hepatocytes and bile ducts and higher AFP levels than did rats started at 26 weeks of age. However, the histological grade of differentiation of inducted tumors did not seem to influence the AFP level.


Immunofluorescent study on alpha-fetoprotein-producing cells in the early stage of 3'-methyl-4-dimethylaminoazobenzene carcinogenesis. The study was carried out to identify alpha-fetoprotein (AFP)-producing cells in the hepatic tissue by immunofluorescent antibody techniques during the early stage of 3'-methyl-4-dimethylaminoazobenzene ingestion. After 1 to 3 weeks, cells fluorescent to AFP were undetectable in cholangiolar cells ("oval cells") and also in degenerated megalocytic hepatocytes. After 4 to 7 weeks AFP appeared in rat sera, and "transitional cells" and small hepatocytes proliferated markedly in the periportal areas of hepatic lobules. AFP was exclusively detected in the majority of the transitional cells and a small portion of the small hepatocytes. Some fluorescent cells appeared in small groups, and others were randomly distributed in the periportal areas. The typical oval cells and the megalocytic hepatocytes were not fluorescent. When AFP in sera became undetectable, the regenerated hepatocytes matured considerably and were not brightly fluorescent. In the hepatic tissue, where AFP-producing cells were observed by fluorescent antibody technique, hematopoietic cells were frequently observed but they were not fluorescent.


Ultrastructural characterization by ruthenium red of the surface of the fat globule membrane of human and rat milk with data on carbohydrates of fractions of rat milk. The fat globules of the cream fractions of human and rat milk were stained with ruthenium red. Under the electron microscope, discrete granules and an amorphous coat of lesser density are seen at the surface of the milk fat globules. Since ruthenium red binds anionic groups selectively, it is probable that the granules contain the greatest concentration of these groups. The cream fraction of rat milk contains hexoses, hexosamines, methylpentoses and sialic acid. Methylpentoses and hexosamines are significantly enriched in the cream fraction. It is concluded that the finding of a surface coat in milk fat globules is in keeping with the Bargmann-Knoop model and suggests a distinct mechanism for carrying certain complex carbohydrates in milk. The role of the negative charges at the outer surface of the membrane coat is maintaining fat globules in suspension and in binding certain cations such as calcium is suggested.


The adenohypophysis of the flounder, Pleuronectes flesus, and the minnow, Phoxinus phoxinus. The pituitary gland of the flounder, Pleuronectes flesus, showed several unusual cytological features. Between the RPD and the PPD was a zone of cells that stained purple with Alcian blue--PAS--orange G. Many of these cells were apparently degenerating. In the PPD the strands and coils of presumptive STH cells showed a tremendous variation in both size and staining properties. In the PI there were two cell types, the PAS-positive one bordering the neurohypophysis. Around the periphery of the PI was a zone of chromophobic cells, and throughout the PI were numerous intracellular and extracellular acidophil spheres. No well defined ACTH cells were found in the RPD of the minnow (Phoxinus phoxinus). The Alcian blue--PAS--orange G technique distinguishes between blue TSH cells and purple GTH cells in the RPD and PPD. GTH cells from animals collected in the winter were vacuolated. The PI contained two cell types whose staining reactions and ultrastructure were extremely variable. Intra- and extra-cellular acidophil spheres were present.


The length of the fibroin gene in the Bombyx mori genome. Radioactive anti-messenger DNA (3H-cDNA) complementary to silk fibroin mRNA has been synthesized using reverse transcriptase. This 3H-cDNA has been found to be a specific and sensitive probe for the detection of fibroin genes in the genome of Brombyx mori. Actinomycin-CsCl gradients give a large separation of the high GC fibroin genes from the bulk DNA. This density shift of fibroin genes has been measured as a function of DNA molecular weight. The data support a model in which a single high GC fibroin gene of 11.6 times 10-6 daltons is surrounded by at least 6 times 10-7 daltons of low GC DNA (30-39 percent). This finding, along with saturation hybridization studies (Suzuki, Gage, and Brown, 1972), demonstrate that the fibroin gene is present in a single copy per haploid genome.


Full length and discrete partial reverse transcripts of globin and chorion mRNAs. Rabbit globin mRNA was copied by AMV reverse transcriptase in the presence of various concentrations of deoxyribonucleotides (dNTPs). The cDNAs were analyzed by electrophoresis under denaturing conditions in formamide-polyacrylamide gels. Discrete size products were detected, ranging from 65 to 650 nucleotides-that is, up to the full length of the mRNA template. Increasing the concentrations of all four dNTPs stimulated formation of full-length transcripts and made the incomplete copies less abundant. Hybridization and nuclease digestion experiments indicated that the full-size product is indeed a complete transcript of globin mRNA. Similar results were obtained with chorion mRNAs. The possible usefulness of the discrete partial transcripts is discussed.


Lack of sensing by demand pacemakers due to intraventricular conduction defects. A previously unrecognized form of failure of normally functioning noncompetitive R-inhibited or R-triggered pacemakers to sense is described in 30 patients. The failure was produced by the delayed arrival of ventricular depolarization due to intraventricular conduction disturbances at the site of pacemaker sensing electrodes. These patinets with late activation of the right ventricle exhibited right bundle branch block (RBBB) during conducted sinus beats or had ectopic beats arising from the left ventricle. In both these situations, several late occurring QRS complexes were not sensed by the pacemaker electrodes in the right ventricular apex (RVA) and thereby the R-inhibited pacemaker spike discharged into the initial portion of the QRS complex. Activation of the RVA was delayed up to 65 msec in sinus beats with RBBB and even longer in premature beats arising from the left ventricle. The depolarization front of such beats reached the myocardium around the right ventricular pacemaker electrodes late in the QRS and thereby were not sensed up to 65 msec after the QRS onset as seen on the surface electrocardiogram (ECG). It was found in this study that if the inscription of such QRS complexes began up to 65 msec before the next due pacemaker impulse, these complexes were not sensed, allowing discharge of normally functioning R-inhibited pacemakers within the QRS which mimicked pacemaker malfunction. Failure to sense due to this phenomenon was observed in patients with noncompetitive pacemakers, both R-inhibited and R-triggered; temporary and permanent; with unipolar as well as bipolar electrodes. Similarly, patients with left ventricular epicardial electrodes and left bundle branch block in sinus beats also exhibited this phenomenon. Complete evaluation proved that pacemaker function was normal in the entire patient group. Recognition that failure to sense was the result of intraventricular conduction disorder prevented the untimely replacement of the pulse generator. This phenomenon of nonsensing by noncompetitive pacemakers should be considered in the presence of wide QRS complexes before the pacemaker is concluded to be malfunctioning.


Observations on the staining of ceruloplasmin following disc-electrophoresis utilizing polyacrylamide gels. In our hands the dianisidine technique for the staining of ceruloplasmin in polyacrylamide gels following disc electrophoresis has proven unsatisfactory for three reasons. First, staining of ceruloplasmin could not be achieved with physiological amounts of the glycoglobulin present in serum. Second, even when relatively massive quantities of pure ceruloplasmin permitted effective staining there was loss of stain intensity with increasing electrophoresis time. Third, the necessity of utilizing dianisidine in alcoholic solution produces undesirable shrinkage and distortion of the gels. However, a modification of the p-phenylenediamine technique for the staining of ceruloplasmin in agar gels has been found to stain ceruloplasmin very effectively in polyacrylamide gels following disc electrophoresis.


Low molecular weight proteinuria in association with paroxysmal myoglobinuria. A patient with paroxysmal myoglobinuria presented with low molecular weight (LMW) proteinuria in association with an episode of exertional myoglobinuria. Since no signs of acute renal failure were present, the cause was probably competition between myoglobin and other LMW proteins for proximal tubular reabsorption. Agarose gel electrophoresis was found to be an excellent method for the investigation of myoglobinuria since this technique not only allowed the separation of myoglobin from hemoglobin but also myoglobin from metmyoglobin.


New developments in the laboratory diagnosis of gonorrhea. Various laboratory technologies have been developed to aid in diagnosing gonorrhea. Transport of viable specimens to the laboratory and the cultivation of N. gonorrhoeae on selective media are among the most effective diagnostic procedures in clinical microbiology. Several sensitive serologic procedures have been developed for detecting antibodies to N. gonorrhoeae.


Morphine binding to human plasma proteins. The interaction of (N-methyl-14C) morphine with purified human albumin and gamma globulin and with human plasma was studied by equilibrium dialysis. Morphine binds to albumin and, to a lesser extent, to gamma globulin, but probably does not bind to plasma lipoproteins. The per cent of drug bound to protein was dependent on protein concentration but independent of drug concentration in the therapeutic range of plasma morphine concentration. In this range, 34.0% to 37.5% of the drug is bound to human plasma, largely to the albumin fraction. This study is compared to an earlier one on the interaction of methadone with plasma proteins. It is likely that the lower lipid solubility of morphine compared to methadone is related to its decreased affinity for plasma proteins.


Double-blind study of the analgesic effect of indoprofen (K 4277). In a double-blind study, indoprofen was superior to placebo in decreasing pain in patients with primary and metastatic cancer and with neuralgia. A single oral dose of 200 mg was more active than a 100-mg dose. The preferences of patients proved to be a more sensitive parameter in this study than scores of pain intensity, pain relief, and other related measurements (SPID, TOTPAR, and Peak PID).


Indoprofen, a new analgesic and anti-inflammatory drug in cancer pain. Evaluation of the analgesic activity of indoprofen was carried out in patients with cancer pain under double-blind conditions and compared with aspirin and placebo. A randomized experimental design was followed. Single oral doses were given of the test drug (100 and 200 mg), aspirin (600 and 1,000 mg), and placebo. For measuring analgesia, 5-point pain intensity and pain relief semiquantitative scales were used. Potency ratio between drugs was calculated on SPID (sum of pain intensity differences) and TOTPAR (total pain relief)  and resulted in 10.3 by combination of estimates. In a group of only 24 patients, the data supported the following conclusions: indoprofen at 100 and 200 mg single doses is effective in relieving cancer pain; it displays a dose-related analgesic effect comparable to that of aspirin with only  one-tenth the dose.


Urologic complications after operations for anorectal cancer, with an evaluation of preoperative intravenous pyelography. Urologic complications arose in 23.7 per cent of 569 patients who underwent abdominoperineal or low anterior resection for anorectal cancer. No radomization of the two operations was attempted, low anterior resection being performed whenever resection 5 cm below the tumor was possible. Complications were more frequent after abdominoperineal resection and in men. Preoperative intravenous pyelography was performed in the cases of 541 of the patients, including 60 who underwent palliative colostomy. The pyelograms of 30 per cent of these patients were abnormal. The abnormalities were anatomic variations of the urinary tract in 25 per cent and urologic diseases in 75 per cent. None of the postoperative urologic complications was related to an abnormal preoperative pyelogram. No relation was found between the radicality of abdominoperineal and low anterior resection and the pyelographic signs of tumor involvement.


[Value of suppression test as a guide to prognosis of hyper-thyroidism treated with anti-thyroid drugs (author's transl)]. 23 out of 42 suppression tests in 39 hyperthyroid patients were positive. In these patients thyreostatic treatment was stopped. In the follow-up period of 1-4 years hyperthyroidism recurred in 4 cases. Eight patients out of another group of 20 patients in whom antithyroid treatment was stopped after an equally long period without previous suppression testing also had a recurrence of hyperthyroidism. Frequency of relapses after antithyroid treatment can thus be diminished considerably if treatment is stopped only after suppression of the thyroid has been demonstrated. Lack of suppression after 18 to 24 months of thyreostatic therapy is an indication for treatment with radioiodine or surgery.


The interaction of actinomycin C3 and actinomine with DNA. A small-angle x-ray scattering study. Small-angle X-ray scattering was applied to solutions of calf thymus DNA and calf thymus DNA complexed with various amounts of actinomycin C3 or actinomine in phosphate-saline buffer at pH 6.9 and I equals 0.2. From the measurements of DNA in the absence of dye, two cross-section radii of gyration of R-c equals 0.875 plus or minus 0.015 nm and R-c2 equals 0.81 plus or minus 0.02 nm, and a mass per unit length of M/l equals 1906 plus or minus 43 daltons/nm resulted. The investigation of DNA complexed with dye revealed a decrease of the cross-section radii of gyration as compared to those for the DNA in the absence of dye and a relatively low increase of the mass per unit length on binding of actinomycin and a slight decrease of M/l on binding of actinomine. The latter results are interpreted on the basis of a length increase of the DNA double helix by 0.47 plus or minus 0.03 nm per actinomycin molecule and by 0.355 plus or minus 0.03 nm per actinomine molecule bound. The results for R-c and R-c2 obtained for the various samples of complexed DNA were extrapolated to the limiting binding ratio where each dye molecule is associated with a minimum of six nucleotide pairs. According to this extrapolation, the cross-section radii of gyration of such a complex would amount to (R-c)b equals 0.805 plus or minus 0.015 nm and (R-c2)b equals 0.76 plus or minus 0.015 nm for the complex with actinomycin, and to (R-c)b equals 0.77 plus or minus 0.015 nm and (R-c2)b equals 0.75 plus or minus 0.01 nm for the actinomine complex. On the basis of a core and shell model for solvated DNA, these results can be understood as to indicate a decrease of the radial dimensions of both the core and the shell when the dye is bound. The experimental results are compared with theoretical data calculated from the atomic coordinates of the detailed intercalation model for the actinomycin - DNA complex as recently proposed by Sobell and Jain. The model proves to be consistent fairly well with our data on the length increase of the double helix, but it appears to be unable to explain the experimentally observed decrease of R-c2 on binding of dye.


Effects of stress and of autonomic blockers on gastric mucosal microcirculation in rats. Changes in gastric mucosal microcirculation in rats were studied by using the method of intra-aortic injection of India ink, followed by microdissection of the mucosa. Acute stress, induced by restraint and exposure to cold for 2 hr, caused marked and significant vasodilatation in the gastric mucosa. This vasodilatation was prevented by pretreatment with atropine or chlorpromazine, but not by alpha- or theta-adrenoceptor blocking agents. Phentolamine caused significant vasoconstriction in the gastric mucosa of non-stressed rats, but when animals were stressed phentolamine induced a greater vasodilatation than was obtained with stress alone. These observations provide added support for the hypothesis that stress induces vagal overactivity, probably of central origin. The resulting strong contractions of the gastric wall, and compression of the intramural vessels, are probably responsible for degeneration of the mucosal cells leading to the formation of stress-induced ulcers in the rat.


Retardation of development including immunogenic expression of histocompatibility antigens in mice--by postnatal administration of antiandrogenic steroid. A specific antiandrogenic steroid cyproterone acetate was administered daily to mice of three different inbred strains starting from the day of birth until the age of 30 days. The total dose per mouse was 17.2 mg. This treatment resulted in developmental retardation which was manifested in a number of ways: at the age of 30 days, the weight of the body was well as spleen, testes and particularly thymus was significantly reduced; histologically, the normal proportion of the red and white pulp in the spleen was changes; spermatogenesis (but not oogenesis) was markedly retarded corresponding to the age of 12-15 days in normal males; also skin displayed a persisting immaturity as reflected by an abundance of mast cells. Minor signs of toxic changes were seen in the liver. Skin grafts from CA-pretreated donors had a subnormal immunogenicity; when transplanted across the MSA-barrier, they survived significantly longer than control grafts and about 23% took. Significantly prolonged survival was also observed with H-3 incompatible skin grafts from CA-pretreated donors, particularly from male donors. Across the barrier dicated, but did not reach a level of significance. The present study extends our previous observations concerning the androgen dependence of a normal immunogenic expression of H-antigens. The antiandrogenic effect of CA is comparable to the more complex effect of neonatal orchiectomy in terms of the subnormal immunogeneity of MSA-incompatible skin grafts from 30-day-old males which seems to be arrated at a stage typical for 1-2-day-old normal males.


Supportive effect of simultaneous immunization to other H-antigens on the female response to male-specific antigen in mice. The response of A-strain females to the male-specific antigen is very weak; both first- and second-set test skin grafts are rejected protractedly and about 50% of the grafts are retained in spite of generally present morphologic signs of cellylar immunity induced by these grafts. When the first-set skin graft involved H-incompatibilities additional to the MSA (such as H-2 only or H-2 plus multiple non-H-2) all the second-set grafts, which were only MSA-incompatible, underwent a markedly accelerated rejection. The possible mechanism of this adjuvant-like effect is discussed.


Effect of antibiotics on the mutagenic activity induced by chemicals. I. Chromosome aberrations during spermatogenesis in mice. Effect of antibiotics (rifampicin-RIF, puromycin-PU, chloramphenicol-CHF and cycloheximide-CYC) on the frequency of chromosomal abnormalities induced by TEPA [tris (1-aziridinyl) phosphine oxide] was studied by the cytogenetic analysis in mice during spermatogenesis. Males were given intraperitoneally a single dose of one of the following agents: TEPA 1 mg./kg. b.w., RIF 150 mg./kg. b.w., PU 100 mg./kg. b.w., CHF 500 mg./kg. b.w. and CYC 100 mg./kg. b.w., or a simultaneously administered TEPA and an antibiotic. Spermatocytes in the diakinesis/first metaphase stage and in spermatogonia. However, statistically insignificantly increased translocations in the group TEPA-CHF, univalents of X and Y chromosomes in TEPA-CHF, CYC and TEPA-CYC, and the increased frequency of metaphases with 19II + U and 20II + f were induced in the preleptotene stage. The decreased frequency of X and Y univalents in TEPA-RIF and TEPA-CHF was induced in spermatogonia. Results revealed a possible selective pressure against the cells carrying some abnormalities.


Infantile autism. I: A prospective study of the diagnosis. A prospective study was made of 200 consecutive children to evaluate the usefulness in the diagnosis of infantile autism of the behavioral scale reported by Clancy and coworkers in 1969. On this scale seven or more of 14 behavioral manifestations must be present before a diagnosis of autism can be made. Using this scale alone, 48 of the 200 children studied were "scale positive", i.e. could be considered autistic. However, further study of this group showed that only one child fulfilled the classical criteria of Kanner (1943) for a diagnosis of early infantile autism. Scale "positivity" was found to correlate with mental retardation and to be associated with other developmental defects, especially learning disorders and hearing loss.


[Myocardial infarction and diabetes (author's transl)]. 30 diabetic patients with myocardial infarction were compared with 30 non-diabetic myocardial infarction patients during their hospitalization in an Intensive Care Unit, to observe possible differences at the onset and throughout the course of the infarction syndrome. Myocardial infarction in diabetic cases has an oligosymptomatic onset, often painless, which evolves towards cardiocirculatory decompensation. The frequence of arrhythmias is higher, especially in serious or mortal arrhythmias. Possible pathogenetic reasons for this are briefly discussed.


Antigenic surface determinants of chicken lymphoid cells. II. Selective in vivo and in vitro activity of anti-bursa and anti-thymus sera. Appropriately absorbed turkey antisera to antigenic surface determinants of chicken bursa (ABS) or thymus cells (ATS) were assessed for their selective immunosuppressive activy in vitro and in vivo. The intraperitoneal injection of ABS or ATS into 2-3-week-old normal white Leghorn chickens entailed a significant depletion of B or T cells respectively from spleen and peripheral blood, while bursa and thymus themselves remained unaffected. The potential of this 'peripheral serological bursectomy and thymectomy' paralleled that found after the conventional surgical procedures with subsequent sublethal irradiation. The mean survival time of skin allografts from donors of genotype B4B4 onto B8B8 recipients was significantly prolonged by treatment with ATS (29 plus or minus 12 days) as compared to untreated (14 plus or minus 2 days), normal turkey serum (12 plus or minus 3 days) or ABS-injected (13 plus or minus 2 days) groups. This selective suppression of a T cell-dependent immune reaction by ATS was also confirmed in vitro by its inhibitory action on the graft-versus-host reactivity of adult peripheral blood lymphocytes in the chorioallantoic membrane assay, where normal turkey serum and ABS were again ineffective. Thus, ABS or ATS produced in avian species may serve not only to delineate B and T cells in vitro, but can also be used for selective manipulation of immune reactions of the chicken in vivo.


Carrier and hapten functions in immune deviation. I. Contrary effects of hapten and carrier on cellular and helper functions of T cells. Carrier and hapten functions have been studied in the immune deviation phenomenon. Delayed hypersensitivity to the carrier and anaphylaxis and Arthus hypersensitivities to the hapten and to the carrier were studied in guinea-pigs injected intravenously with large doses of carrier, homologous and heterologous hapten-carrier conjugates and subsequently immunized with the hapten-carrier conjugate in Freund's complete adjuvant. Pretreatment with DNP-BSA or with HGG were found to modify, in opposite directions, the hypersensitivity reactions induced by DNP-HGG in adjuvant. It is suggested that the hapten and carrier moieties of the antigen molecule might have antagonistic effects on the T cells responsible for cellular immunity as well as on T cells involved in helper functions for B cells.


Genetic control of sensitization to structurally unrelated antigens and its relationship to histocompatibility antigens in guinea-pigs. Five strains of guinea-pigs homozygous for GPL-A and 2/13 histocompatibility antigens were sequentially immunized with seven structurally unrelated antigens (TGAL, BPO-BGG, ASAN, BPO-PLL, phenetidine, horseradish peroxidase and DNP-dodecapeptide). Considerable differences in the immune response were observed, as measured by delayed skin reactivity and antigen-induced lymphocyte stimulation in vitro. The data presented show that when the progeny of various back-cross matings are analysed for their responder status, the responses to the antigens (TGAL, BPO-BGG, phenetidine and ASAN) are in most cases linked to the genes controlling the respective histocompatibility antigens. However, it appears that the responses to the various antigens in our system may be under polygenic control.


Studies of the inhibition of C56-initiated lysis (reactive lysis). IV. Antagonism of the inhibitory activity c567-INH by poly-L-lysine. The stable intermediate complex C56 can initiate the lysis (reactive lysis) of unsensitized erythrocytes (E) by the membrane attack machanism of complement. Certain serum constituents designated C567-INH inhibit reactive lysis by preventing the C567 complex, once formed, from attaching to a membrane surface. It is shown here that microgram quantities of poly-L-lysine (PLL), a synthetic polycation of molecular weight 180,000, can reverse the effests of C567-INH, and thereby potentiate formation of EC567 by erythrocytes, C56 and C7 in whole serum. Erythrocytes exposed to PLL in a preincubation step did not show either increased susceptibility to C567 or resistance to C567-INH, and reversal of C567-IHN by given amounts of PLL was not diminished as cell concentrations were greatly increased, indicating that the effect of PLL was predominantly directed against fluid phase rather than against erythrocyte membrane substrates. The effects of PLL and C567-INH were quantitatively reciprocal. Thus, PLL-induced potentiation of C56-induced lysis is a solute effect which seems to involve direct neutralization of naturally occurring serum inhibitors of the C567 trimolecular complex of complement. The use of PLL thus provides a suitable antagonist for C567-INH in reaction mixtures, and allows evaluation of the role of C567 and C567-INH in a variety of situations involving C-mediated lysis.


Biological properties of a canine distemper virus isolate associated with demyelinating encephalomyelitis. A canine distemper virus (CDV), DESIGNATED R252, originally recovered from a dog with demyelinating encephalomyelitis was shown to reproduce this disease in gnotobiotic dogs with a high incidence as compared to other CDV strains, which produced an acute fatal infection. In this investigation, R252 was propagated for the first time in Vero cells and compared to two known strains of CDV, Snyder-Hill (SH) and Onderstepoort (Ond). The results of this study revealed that intracellular R252 accumulated more slowly than either SH or Ond. There was extensive destruction of Vero monolayers infected with either R252 or SH. Each virus induced the formation of intracytoplasmic and intranuclear inclusions. Ond infection resulted in minimal cytopathic changes and intracytoplasmic inclusions. Immunofluorescence studies indicated that the spread of R252 infection within the monolayers was intermediate between the rapidly spreading SH and slowly spreading Ond. R252-infected cells developed characteristic immunofluorescent cytoplasmic inclusions. Initially, each stained homogeneously and later appeared as a non fluorescent body surrounded by a fluorescent ring. This characteristic pattern of fluorescence was observed only infrequently in thelate stage of SH infection and was absent in Ond-infected cultures. Reciprocal neutralization studies indicated that the three strains are of one serotype. These findings suggest that R252-CDV has biological properties which differ from two other strains of CDV and which may have bearing upon the in vivo capability of this virus to produce demyelinating encephalomyelitis.


Long-term Visna virus infection of sheep choroid plexus cells: initiation and preliminary characterization of the carrier cultures. Two sheep choroid plexus cell cultures were maintained and propagated for 413 days since being infected with strain K796 visna virus. The majority of the cells in these cultures contained visna virus-specific antigen on days 93 and 105 after infection. Reverse transcriptase-like activity similar to that present in visna virus preparations was obtained from these cultures when very little plaque-forming virus was being synthesized. The persistently infected cultures are resistant to the cytopathic effect which occurs in uninfected cultures upon exposure to visna virus. Persistently infected cells require more time than uninfected cells to become confluent. Less than 0.02 percent of the persistently infected sheep choroid plexus cells form macroscopic colonies within 14 days, whereas 20 to 30 percent of the cells from uninfected cultures form macroscopic colonies within this time.


Virion and soluble antigens of japanese encephalitis virus. Japanese encephalitis virions contain a 58 X 10-3-molecular-weight envelope glycoprotein antigen that can be solubilized with sodium lauryl sulfate and separated from other virion structural polypeptides and viral ribonucleic acid by gel filtration chromatography. The 58 X 10-3-molecular-weight envelope protein is the major antigen responsible for cross-reactivity of the virion in complement fixation tests with other closely related arboviruses. A naturally occurring soluble complement-fixing antigen is found in Japanese encephalitis mouse brain preparations after removal of particulate antigens. After partial purification by gel filtration and isoelectric focusing, the 53 X 10-3-molecular weight soluble complement-fixing antigen is more type specific than the Japanese encephalitis envelope antigen in complement fixation tests. Further, the Japanese encephalitis soluble complement-fixing antigen is stable to treatment with sodium lauryl sulfate and 2-mercaptoethanol, whereas virion complement-fixing antigens are unstable after this treatment.


Increased influenza pneumonia mortality of mice adoptively immunized with node and spleen cells sensitized by inactivated but not live virus. Syngeneic mice adoptively immunized intravenously with 25 million washed node and spleen cells from donors vaccinated subcutaneously with formolized influenza A PR8 had a higher mortality with influenza pneumonia after challenge with homologous virus than occurred in recipients of similar cells from unsensitized donors, and this increased mortality was prevented by treatment of the sensitized cells with antithymocyte serum. Mice adoptively immunized with cells from donors vaccinated with formolized influenza A PR8 also had a higher mortality than recipients of unsensitized cells after challenge with heterologous influenza B Lee. Mice who received PR8-sensitized cells and survived challenge with influenza B Lee developed antibody only to the challenge virus, and serum antibody titers to the challenge virus in surviving recipients of sensitized cells were similar to those of recipients of unsensitized cells in all studies. Influenza mortality of recipients of antibody-containing mouse serum after homologous virus challenge was similar to that of recipients of antibody-free mouse serum in this model. Washed node and spleen cells from donor mice who had survived respiratory infection or received subcutaneous vaccination with live influenza A PR8 and those from donor mice given typhoid vaccine subcutaneously all failed to alter mortality from that observed in recipients of unsensitized cells after challenge with influenza A PR8. These results suggest that subcutaneous vaccination with inactivated influenza establishes a reactivity of  the cell-mediated immunologic system which can increase the severity of influenza infection of the respiratory tract under certain conditions, and that sensitization by live influenza fails to produce this effect.


Ontogeny of lymphoid cell surface determinants in the chicken. Five antigenic lymphoid cell surface determinants (LCSD) were detected in hatched chickens using specific antisera. These LCSD were: thymus-specific surface determinants, bursa-specific non-immunoglobulin determinants, IgM-specific determants, IgG-specific determinants, and IgA-specific determinants (ASD). Viable cell suspension of embryonic yolk sac, bursa, thymus and spleen were tested by means of indirect or direct immunofluorescent staining procedures for the presence and frequency of LCSD during maturation. Experiments performed with liver cells. brain cells and red blood cells of embryos confirmed the specificities of the antisera used for determinants present on cells of lymphoid tissues. The results showed LCSD to occur on yolk sac cells on the 5th to 7th embryonic day (ED). This suggests the presence of a stem cell pre-committed for the lymphoid cell line already in the yolk sac. Furthermore, findings are reported indicating the presence of distinct lympoid stem cell populations or maturation stages in the yolk sac, which may be responsible for either populating the thymus or the bursa. The finding of ASD-bearing cells early in ontogenesis of the lymphoid system suggests the presence of two specificities in anti-chicken IgA sera, one of which may be directed against an antigenic site on a rudimentary immunoglobulin molecule, which becomes lost or hidden in later maturation. Studies on the bursa and the thymus show that covering, hiding, or loss of antigenic determinants plays an important role in lymphoid cell differentiation. Furthermore, the spleen is reached by B-determined stem cells as early as the bursa, but these stem cells seem not to proliferate in the former to any considerable extent until hatching. Finally, the sequence of the appearance of immunoglobulin classes as proposed by other authors is confirmed with reservaitons concerning IgA, and it is suggested that immunoglobulins are detectable earlier on cell surfaces than intracytoplasmatically.


Unique serum requirement for histamine release from human basophils. Basophil histamine release in a patient with an IgE-mediated seminal plasma allergy had a requirement for serum. Washed leukocytes, in the absence of serum, released little histamine on challenge with seminal plasma antigen. The addition of serum markedly enhanced the release. However, serum had only a mild effect on ragweed antigen induced histamine release from the same cells of this individual. Serum from normal donors was equally effective as autologous serum. Heating the serum and treating it with mercaptoethanol did not destroy this activity. The serum effect was unique in that another patient with seminal plasma allergy did not demonstrate this phenomenon. It is postulated that the effect of the serum is to stabilize the antibody-antigen step at the basophil surface.


Role of the chondrocytes in the breakdown of pig articular cartilage induced by complement-sufficient antiserum to pig erythrocytes. Previous work showed that in organ culture pig articular cartilage is not affected by complement-sufficient antiserum to pig erythrocytes (AS plus C') unless it is associated with soft connective tissue. In the presence of synovial tissue the matrix becomes depleted of proteoglycan and may finally disintegrate. Immunoglobulin fails to enter normal cartilage matrix, but penetrates depleted matrix and reacts with the surfaces of the chondrocytes. The present experiments were made to see whether the breakdown of partially depleted cartilage matrix would progress in AS plus C' after removal of the synovial explant. Affronted explants of articular cartilage and synovial tissue were cultivated in AS plus C' for 10 days (primary cultures). The synovial tissue was then removed and the isolated cartilage maintained for a further period either in AS plus or C' or in one of a variety of control media (SECONDARY CULTURES). The behavior of the isolated cartilage during the secondary culture period depended on the degree of breakdown attained at the end of the primary period. If only proteoglycan but not collagen had been seriously depleted, degradation of the matrix did not progress in AS plus C' and slight regeneration of metachromatic materal sometimes took place; regneration of matrix was greater after transfer to control medium. If collagen as well as proteoglycan had been destrobyed, the chondrocytes assumed a fibroblastic form, their surfaces no longer reacted with IgG antibody, and in secondary culture they failed to regenerate new matrix.


Immunochemical investigation on human ceruloplasmin. Partial explanation of the "heterogeneity". Human ceruloplasmin from fresh serum has been purified by chromatography on hydroxyapatite and Con A-Sepharose. Quantitative immunoelectrophoretic analysis of fresh serum, stored serum and fractions from the different purification steps for human ceruloplasmin has been carried out. A combination of the latter, advanced technique with amino acid analysis, molecular weight determination by size chromatography, urea treatment, staining for oxidase activity and enzymatic proteolysis, has revealed that: 1) human cerulplasmin is a heterogeneous mixture of two glycoproteins (x) differing only in their carbohydrate content and 2) the protein part contains at least one very labile peptide bond which upon enzymatic hydrolysis gives rise to peptides with molecular weights of 93,000 (y) and 24,000 (z) dalton, respectively. The two glycoproteins are immunochemically identical. The y peptide is immunochemically partially identical, and the z peptide immunochemically non-identical, with the parent molecule. The y and z peptides are non-identical. On the basis of these observations a simplified two-dimensional model of human ceruloplasmin is proposed.


Nonspecific inflammatory lesions of the prostate. Spectrum and patterns. Nonspecific inflammatory lesions of the prostate are described and illustrated. The spectrum of inflammation in this group of lesions caused by exposure of prostatic secretions to, or their escape into, the stroma has not been fully recognized. Inflammation ranges from acute, through chronic lymphoplasmacytic to granulomatous. The pathogenesis is primarily a defect in the acinar or ductal epithelium caused by prostatic secretion. The difference between these lesions and infective lesions is emphasized. The term ectasia prostatitis is suggested for this group of inflammatory lesions.


Accumulation of arsenate, phosphate, and aspartate by Sreptococcus faecalis. Uptake of arsenate and phosphate by Streptococcus faecalis 9790 is strictly dependent on concurrent energy metabolism and essentially unidirectional. targinine supports uptake only in presence of glycerol or related substances; glycerol is not directly involved in transport but depletes the cellular orthophosphate pool and thus relieves feedback inhibition of transport. Uptake of phosphate and arsenate is stimulated by K+ and by other permeant cations. The results suggest that electroneutrality is preserved by compensatory movement of either H+ or OH minus. Ionophores and N,N'-dicyclohexylcarbodiimide, which prevent establishment of a proton motive force, block the accumulation of thiomethylgalactoside and of threonine but not that of arsenate or phosphate. We conclude that arsenate accumulation requires adenosine 5'-triphosphate but is not driven by the proton-motive force. However, conditions and reagents that lower the cytoplasmic pH do inhibit accumulation of arsenate and phosphate, suggesting that uptake depends on the capacity of the cells to maintain a neutral or alkaline cytoplasm. We therefore propose that phosphate accumulation is an electroneutral exchange for OH driven by adenosine 5'-triphosphate or by a metabolite thereof. Accumulation of aspartate and glutamate also requires adenosine 5'-triphosphate but not the proton-motive force and may involve a similar mechanism.


Ultrastructure and adhesion properties of Ruminococcus albus. Morphological studies have shown that cells of the anaerobic rumen bacterium Ruminococcus albus have electron-translucent granules of reserve carbohydrate in their cytoplasm, and that they have a polysaccharide "coat" layer external to their gram-negative cell wall. This coat layer, which stains specifically with ruthenium red, forms a compact mat of fibers adjacent to the cell, and fibrous elements also project as much as 0.6 mum from the cells. These radial fibers are clearly visualized by freeze-etching, and can be seen to extend throughout the extensive intercullular space in centrifuged pellets of these bacteria. Cells of R. albus adhere to cellulose fibers added to the culture medium, and the coat material is seen to mediate this adhesion in addition to its function in the general protection of these cells.


Electron microscopy studies of the limiting layers of the rickettsia Coxiella burneti. Surface layers of Coxiella burneti studied at a high resoulution reveal a plasma membrane and an outer surface membrane 6 to 7 nm thick, and a thin, moderately electron-dense intermediate layer associated with the inner surface of the outer membrane of many cells. This layer appears to be unaffected by lysozyme treatment. Ruthenium red staining was used to delineate a layer of filamentous material external to the outer membrane; this fuzzy layer has a mean thickness of 20 nm and is not often seen on the surface of cells prepared by conventional means. Both antigenic phase I and II cells show a ruthenium red-binding surface layer. It is suggested that this fuzzy layer may be, among other possibilities, a highly branched mucopolysaccharide.


Release of extracellular enzymes from Bacillus amyloliquefaciens. Washed-cell suspensions of Bacillus amyloliquefaciens secrete significant amounts of the extracellular enzymes alpha-amylase and protease for about 15 min in the almost complete absence of protein synthesis. This apparently represents release of preformed enzyme en route to secretion. The release was independent of energy but was affected by temperature. Pulse-labeling experiments showed that newly synthesized enzyme molecules are either immediately released into the external medium or equilibrate with the preformed enzyme prior to eventual secretion. The results are compatible with a model of secretion whereby enzyme molecules emerging from the cell membrane become temporarily restricted by the cell wall so that a small pool of active enzyme accumulates in this region.


Mammary tumor virus induction by glucocorticoids. Characterization of specific transcriptional regulation. Dexamethasone (1,4-pregnadiene-9-fluor-16alpha-methyl-11beta,17alpha,21-triol-3,20-dione), a potent synthetic glucocorticoid, stimulates mouse mammary tumor virus expression 10- to 20-fold in tissue culture cells. This hormone effect was observed at concentrations as low as 1 times 10-10 M and was maximal at 10-7 to 10-8 M. The time course of induction indicated that detectable increases in extracellular viral DNA polymerase were first noted 18 to 24 hours following the addition of dexamethasone, and cells produced the highest polymerase levels at the time monolayers approached confluence. Steroid responsiveness was associated with specific increases in type B murine mammary tumor virus structural polypeptide (gp52(sl) expression and murine mammary tumor virus RNA that quantitatively paralleled the increase in extracellular virus production as measured by electron microscopy and supernatant RNA-dependent DNA polymerase activity. Another virally transformed murine cell line, KA 31, did not contain detectable levels of murine mammary tumor virus gp52(sl) or RNA before or after dexamethasone stimulation; thus induction was noted only in murine cells with pre-existing murine mammary tumor virus expression. No increase in basal levels of type C murine leukemia viral proteins or RNA was detected in dexamethasone-treated mammary cell lines which were producing increased levels of murine mammary tumor virus. Therefore, increases in murine mammary tumor virus gene products are specific for murine mammary tumor virus DNA sequences under these conditions.


A cleavage-associated neoantigenic marker for a gamma chain site in the NH2-terminal aspect of the fibrinogen molecule. The E fragment, derived from the NH2-terminal aspect of fibrinogen by plasmin cleavage (fg-E), possesses two generically distinct sets of antigenic expressions. The major set of antigens is expressed by the parent molecule as indicated by the capacity of a major subpopulation of antibodies present in antiserum to fg-E and reactive with fg-E to: (a) react with fibrinogen, and (b) be specifically absorbed by fibrinogen but appears following proteolysis with plasmin. These cleavage associated neoantigens (fg-E-neo) specifically react with a minor subpopulation of antibodies present in antiserum to fg-E.E fragments isolated after varying exposures to plasmin all expressed fg-E-neo, but early E fragments exhibited quantitatively less neoantigenic expression than more extensively degraded E fragments. The entire fg-E-neo expression is recovered on a single isolated constituent chain of the E fragment, and immunochemical analysis with antiserum to the isolated constituent chain-bearing fg-E-neo identifies it as a derivative of the gamma chain constituent, exhibits marked stability to physicochemical denaturation and enzymatic degradation. These properties suggest that the neoantigen may be associated with a specific amino acid sequence which is exposed by the cleavage process. The identification and localization of fg-E-neo provides a specific molecular marker site for the characterization of structural and conformational changes associated with catabolism and function of fibrinogen.


Conformational and structural modulation of the NH2-terminal regions of fibrinogen and fibrin associated with plasmin cleavage. Conformational and structural modulations of the NH2-terminal region of fibrinogen and fibrin associated with plasmin cleavage have been examined utilizing specific antibody probes. The E region derived from the NH2-terminal aspects of fibrinogen undergoes complex structural and conformational changes throughout the cleavage process as indicated by differences in the quantitative and qualitative expression of antigenic determinants by the E region of each isolated cleavage fragment. When the range of antigenic determinants recognized by the antibody probe is limited to a specific molecular marker on the gamma chain within the E region, fg-E-neo, evidence for a systematic and progressive modulation of this site during plasmin cleavage is observed. Fg-E-neo undergoes progressive exposure as the cleavage of fibrinogen proceeds from X to Y to D:E complex. Separation of the D:E complex into its constituent, D and E fragments, is associated with further exposure of fg-E-neo determinants. The sequential cleavage of fibrin by plasmin also leads to progressive exposure of the fg-E-neo site; however, comparison of corresponding fragments derived from fibrinogen and fibrin reveals significant differences in the character of fg-E-neo expression. Immunochemical differences between fibrin and fibrinogen E fragments are not abolished by further exposure of the fragments to plasmin, are apparently not due to the presence or absence of fibrinopeptides, and are maintained following denaturation and renaturation of the fragments. These results suggest that the differential expression of fg-E-neo by the E fragments may be primarily dependent upon differences in amino acid compositions of the fragments.


Biosynthesis of yeast mannan. Isolation of Kluyveromyces lactis mannan mutants and a study of the incorporation of N-acetyl-D-glucosamine into the polysaccharide side chains. One side chain in the cell wall mannan of the yeast Kluyveromyces lactis has the structure (see article). (Raschke, W. C., and Ballou, C. E. (1972) Biochemistry 11, 3807). This (Man)4GNAc unit (the N-acetyl-D-glucosamine derivative of mannotetroase) and the (Man)4 side chain, aMan(1 yields 3)aMan(1 yields 2)aMan(1 yields 2)Man, are the principle immunochemical determinants on the cell surface. Two classes of mutants were obtained which lack the N-acetyl-D-glucosamine-containing determinant. The mannan of one class, designated mmnl, lacks both the (Man)4GNAc and (Man)4 side chains. Apparently, it has a defective alpha-1 yields 3-mannosyltransferase and the (Man)4 unit must be formed to serve as the acceptor before the alpha-1 yields 2-N-acetyl-glucosamine transferase can act. The other mutant class, mnn2, lacks only the (Man)4GNAc determinant and must be defective in adding N-acetylglucosamine to the mannotetrasose side chains. Two members of this class were obtained, one which still showed a wild type N-acetylglucosamine transferase activity in cell-free extracts and the other lacking it. They are allelic or tightly linked, and were designated mnn2-1 mnn2-2. Protoplast particles from the wild type cells catalyzed a Mn2+-dependent transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to the mannotetraose side chain of endogenous acceptors. Exogenous mannotetraose also served as an acceptor in a Mn2+-dependent reaction and yielded (Man)4GNAc. Related oligosaccharides with terminal alpha (1 yields 3)mannosyl units were also good acceptors. The product from the reaction with alphaMan(1 yields 3)Man had the N-acetylglucosamine attached to the mannose unit at the reducing end, which supports the conclusion that the cell-free glycosyltransferase activity is identical with that involved in mannan synthesis. The reaction was inhibited by uridine diphosphate. Protoplast particles from the mmnl mutants showed wild type N-acetylglucosamine transferase activity with exogenous acceptor, but they had no endogenous activity because the endogenous mannan lacked acceptor side chains. Particles from the mnn2-1 mutant failed to catalyze N-acetylglucosamine transfer. In contrast, particles from the mnn2-2 mutant were indistinguishable from wild type cells in their transferase activity. Some event accompanying cell breakage and assay of the mnn2-2 mutant allowed expression of a latent alpha-1 yields 2-N-acetylglucosamine transferase with kinetic properties similar to those of the wild type enzyme.


Osteogenic sarcoma. A study of one hundred and thirty cases. One hundred and thirty patients with osteogenic sarcoma were studied clinically, roentgenographically, and pathologically. Prognosis by each of ten factors was analyzed with the Wilcoxon test. The test yielded p smaller than 0.05 in a comparison between the survival curves of patients fifteen years old or younger and that of patients over fifteen, but other comparisons did not yield p smaller than 0.05. The actual five-year survival rate was 25.5 per cent (twenty-six of 102 patients who had radical surgery). Our therapeutic regimen for osteogenic sarcoma at the present time consists of regional perfusion, amputation, bronchial artery infusion, and systemic administration of anticancer agents.


Immunological characterization of hypoxanthine-guanine phosphoribosyl transferase mutants of mouse L cells: evidence for mutations at different loci in the HGPRT gene. A large collection (105) of mouse L cell mutants lacking hypoxanthine-guanine phosphoribosyl transferase activity (HGPRT; E. C. 2.4.2.8) were analyzed for the presence of serologically cross reacting material (CRM). Antibody directed against highly purified mouse liver HGPRT was used for detecting DRM activity by two methods: (1) the standard precipitation-inhibition assay; and (2) a radioimmune-precipitation assay. The latter assay proved to have far greater sensitivity for the detection of altered forms of HGPRT. Approximately 40% of the HGPRT- cell lines contain CRM activity (i.e., were CRM+). This indicates that a minimum of 40% of the HGPRT- clones arose as a result of mutations in the HGPRT structural gene. The CRM+ cell lines were shown to contain different levels of CRM activity. Measurements of the heat sensitivity of CRM in the different HGPRT- cell lines showed a broad spectrum of CRM heat inactivation kinetics. These latter two observations provide strong evidence that the mutations giving rise to the HGPRT-CRM+ phenotype occurred at different sites in the HGPRT structural gene.


Control of transcription of the globin gene. This report provides a more rigorous proof of previous findings that the RNA transcribed in vitro from the chromatins of different organs shows different sequence specificities. Here the particular case of the globin gene is considered for a comparison of embryonic mouse liver chromatin and mouse brain chromatin using the reverse transcriptase cDNA copy of globin 9S m RNA as a definitive probe. It can be shown that globin sequences are transcribed in vitro from embryonic liver chromatin and not brain chromatin. This specificity in liver chromatin can be reconstituted after dissociation of the structural elements of the chromatin. It can be shown that the non-histone protein fraction of liver chromatin can confer specificity for the transcription of globin sequences from brain chromatin which otherwise lacks this ability. Preliminary results are described with the Friend cell system, in which haemoglobin synthesis can be induced in vitro in the presence of dimethylsulphoxide.