Chronotropic response of isolated atria to acid base alterations. The effect of acid-base alterations on spontaneous rate was analysed using isolated atria exposed to cumulative degrees of acidosis produced either by adding HCl or by increasing PCO2 in the incubation medium. Frequncy vs. pH curves were made to assess chronotropic response to acid-base changes. Heart rate was increased in alkalosis and decreased when the pH of the medium was lowered. Both "respiratory" and "metabolic" alterations affected the contraction rate to the same extent. Decreasing pH from normal values seemed to decrease heart rate more than the enhancement produced by the same change in pH towards the alkalotic side. When frequency was plotted as a function of hydrogen ion activity (aH+) a more linear relationship was obtained, either with pure "metabolic" or with "respiratory" acid-base alterations. Increasing (aH+) from normal values seemed to decrease heart rate to the same extent (respiratory alterations) or even less (metabolic alterations) than the enhancement produced by the same change in (aH+) towards the alkalotic side. Neither the increase in rate produced by alkalosis nor the decrease induced by acidosis were prevented by blocking the neurotransmitters by atropine or propranolol.


[Use of a method of staining sections with methyl green-pyronine for histobacterioscopic purposes]. A method of staining of histological sections with methyl green-pyronine for histobacterioscopic detection of microflora in tissues is proposed. A bright red staining with pyronine of microbial agents and yeast-like fungi of the genus Candida due to a high content of RNA in them permits elective detection of their insignificant amounts in tissues even at low magnifications of the microscope.


Histological and histochemical changes of the skeletal muscle in human chronic Chagas' disease. Gastrocnemius muscle biopsies were performed in 7 subjects with chronic Chagas' disease. On clinical and laboratory grounds the selected patients were judged to be healthy, being the only abnormality found the presence of positive serum tests for Chagas' disease. Fibre type grouping of either type I or type II was observed in 5 of the 7 patients. Furthermore, in 2 of the 5 patients showing muscle fibre groupings, angular fibres reacting with NADH and non-specific sterase were also found. These observations strongly suggest denervation associated with reinervation. This picture often can be observed in the skeletal muscle of patients with well compensated denervatory conditions who did not show clinical evidences of denervation.


The embedding of ossicular chain specimens in plastic. The following objects were specifically designed for teaching material to the medical students. Dried human auditory ossicles were bleached in 30% hydrogen peroxide solution and then, embedded in a good quality plastic. The plastic mass was shaped into a ball 5 cm in diameter, and the surface of which was polished with a good finish. The merits of the objects may be listed as follows; 1) A permenant preservation of the objects is possible because they are embedded in heat- and chemical-resistant material. 2) The objects are handy for use in the lecture room and easily observed by the students because of the magnifying effect of the plastic covering.


Detection, purification and characterization of a human cancer-associated galactosyltransferase acceptor. A low-molecular-weight acceptor of galactosyltransferase activity was detected in sera and effusions of patients with extensive maligant disease. This substance was purified to homogeneity from both human serum and effusion by using sequential charcoal/Celite and DEAE-cellulose column chromatography. The purified acceptor was shown to act as substrate for both purified normal and cancer-associated human galactosyltransferase (EC 2.4.1.22) isoenzymes, but had a higher affinity for the cancer-associated isoenzyme (Km = 20 microM) than for the normal isoenzyme (Km = 500 microM). The substrate was found to be a glycopeptide with mol.wt. approx. 3600 determined by polyacrylamide-gel chromatography. Carbohyydate analysis demonstrated only the presence of glucosamine and mannose. Amino acid analysis revealed that the peptide moiety consisted of eight different amino acids, including two residues of asparagine and one residue of serine, but no threonine. These structural data suggest that the acceptor is a fraction of an asparagine-glucosamine type of glycoprotein.


Comparison of three immunoassays for immune complexes in rheumatoid arthritis. Three widely used radioassays that depend on different principles for the measurement of circulating immune complexes (CIC) in biologic fluids are the monoclonal rheumatoid factor (mRF), Raji cell, and C1q binding tests. A comparison of the ability of these methods to measure immune complex-like material in 71 sera and 30 synovial fluids of 91 patients with rheumatoid arthritis (RA) was carried out by a group working in adjacent laboratories in a single institution. The highest number of abnormal levels in the seropositive group was detected by the C1q binding assay (91%). Levels of CIC by the mRF and Raji cell tests were elevated in 81% and 76% of the patients, respectively. The closest correlation was between the Raji and mRF tests (r = 0.44 and P = 0.002) although one depends on complement fixation and one does not. Though significant correlations between the levels of CIC determined by the C1q test and either the mRF (P = 0.2) or Raji cell (P = 0.3) assay were not found in this group, 59% of the samples had elevated levels by all three tests. The frequency of CIC in the sera of patients with seronegative RA was much lower, with the C1q test again giving the highest number of abnormal results (29% versus 16% and 12% for the Raji and mRF tests). In view of the technical problems associated with these tests, particularly lack of a uniform reliable standard, it is likely that all three tests measure the same material in most RA sera and that some of the differences observed are related to inherent variability in the tests themselves rather than intrinsic differences among the CIC detected in these samples.


[The antiexudative and anti-edematous action of sympathomimetics]. The anti-exudative and anti-edematous effects and the dose-effect relationship of a-sympathomimetics, especially of l-phenylephrine-HCl (PE), have been demonstrated using as model the rat pad-carrageenan edema and the histamine liberator test, the dosage being administered cutaneously, orally, and intraperitoneally. The beta-receptor activating compound bamethane sulfate was not effective when used on the same models. Evidence of percutaneous penetration of PE was provided both on isolated skin and in viro. PE, when injected intravenously or subcutaneously, produced a rise of blood pressure in experimental animals. When PE was administered orally and cutaneously, it did not, however, alter the blood pressure. The anti-inflammatory and anti-exudative effects remianed unchanged.


[Therapy and prophylaxis of experimental staphylococcal nephritis of the rabbit with gamma-globulin and F(ab')2-fragments (author's transl)]. Prophylactic and therapeutic effects of immunoglobulins and their combination with ampicillin were studied in an experimental bacterial nephritis model after i.v. injection of Staphylococcus aureus into rabbits. Untreated human IgG and a preparation containing F(ab')2-pieces were investigated on therapeutic effects. Both antibody preparations prevented the lethal purulent inflammation of kidneys when injected immediately before the infection. The course of the pathological process after infection was positively influenced by both immunoglobulins. The globulin containing F(ab)2-fragments was more effective. Repeated applications showed also a more significant therapeutical effect than did single treatment. Ampicillin was less effective against this resistant staphylococcus strain, in combination with immunoglobulins it was fully effective. F(ab')2-pieces of the human IgG activate the alternate pathway of the complement system, too, and are able to penetrate into cells. The antibacterial effect may be caused by this behaviour.


BUdR as a tracer of the possible mutagenic activity of Pb++ in human lymphocyte cultures. The method of SCE was used as an indicator of the possible mutagenic activity of lead acetate in in vitro cultures of human lymphocytes. After determining the optimal concentration of BUdR, the cultures were treated with increasing doses of the metal. The negative results obtained with this method led us to examine other parameters, in particular the classical cytogenetic aberrations.


The interaction of thermal tolerance with drug cytotoxicity in vitro. The effect of preheating EMT6 cells in vitro on their response to cytotoxic agents of either 43 degrees C or 37 degrees C has been investigated. Preheating for 3 h at 40 degrees C produced measurable protection (thermal tolerance) to subsequent treatment for 1 h at 43 degrees C. This preheat treatment was further found to reduce cell killing by BLM and BCNU (drug tolerance) present during 1 h at 43 degrees C. In contrast, no such heat-induced drug tolerance was seen with ADR. An additional effect with ADR was the apparent elimination of heat-induced thermal tolerance at toxic drug doses. However, preheating under these conditions had no effect on the subsequent cytotoxicity of any of these drugs at 37 degrees C. Also, preheating for 1 h at 43 degrees C was found to sensitize cells to BLM and BCNU toxicity at 37 degrees C but to protect against ADR toxicity. The results are discussed in relation to known mechanisms of cell killing by heat and of thermal tolerance.


Cancer of the prostate: early diagnosis by zinc and hormone analysis? Zinc, testosterone and dihydrotestosterone concentrations have been measured in normal prostatic tissue and in specimens obtained from untreated patients with benign prostatic hyperplasia (BPH) and carcinoma of the prostate (CaP). The metal--androgen relationship was examined and related to the pathological condition of the patients. The evidence suggests that discriminant analysis combining the hormonal data into a single variable is a reliable test for distinguishing between BPH and CaP patients. We have observed that the high Zn values found in BPH specimens were always associated with a DTH:T ratio greater than 1. Androgen tissue ratios less than 1 were characteristic of all CaP specimens, and these were usually preceded by a reduction in prostatic Zn concentration. Since these patterns, particularly those associated with neoplasia, precede the clinical manifestations, they may be used as an index for predicting the onset of carcinoma in the prostate gland. They may also be of value in monitoring the progress of the disease.


Histamine release from platelets for assay of byssinogenic substances in cotton mill dust and related materials. Previous reports suggest that byssinosis, an asthma-like condition among textile workers, may be mediated in part by histamine liberated following inhalation of dust. A simple, sensitive, and reliable procedure using pig platelets which contain the unusually high concentration of 0.8-1.6 microgram histamine/10(9) cells has been devised for the assay of histamine-releasing factors in cotton mill dust and related materials, and has yielded results generally in accordance with earlier assays using chopped lung tissue. As little as 50--100 microgram of total extractable substances from cotton mill dust can be measured. The activity of the extract is associated with the non-dialysable high molecular weight portion. However, conditions of acid hydrolysis do not destroy the activity. Extracts of leaves from different varieties of plant are highly potent, which suggests that the factors responsible for byssinosis are widely distributed plant components, present in textile fibre plants and converted to a respirable form by handling processes. Ellagic acid and sodium metasilicate release histamine from pig platelets, and represent new classes of compounds with possible roles in the aetiology of byssinosis.


Changes in expression of albumin and alpha-fetoprotein genes during rat liver development and neoplasia. Albumin mRNA was isolated and purified from rat liver polysomes by a combination of immunoprecipitation of specific polysomes, poly(U)-Sepharose 4B chromatography, and fractionation of the resulting poly(A)-containing RNA on a sucrose gradient. alpha-Fetoprotein (AFP) mRNA was isolated from Morris hepatoma 7777 by a similar procedure. The purity of the mRNA preparations was determined by analytical gel electrophoresis under denaturing conditions, analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the polypeptides synthesized in a wheat germ cell-free system, and the kinetics of hybridization to cDNA transcribed from albumin mRNA and AFP mRNA. The albumin mRNA possessed a chain length of approximately 2265 nucleotides and the AFP mRNA possesed a length of approximately 2235 nucleotides when examined under stringent denaturing conditions on agarose gels containing 10 mM methylmercury hydroxide. Analysis of poly(A) content by a hybridization assay with [3H]poly(U) revealed the presence in albumin mRNA of a poly(A) region containing approximately 100 adenosine residues. The AFP mRNA preparation was found to contain an average poly(A) tract of approximately 190 bases. Thus, albumin mRNA appears to contain approximately 330 untranslated nucleotides, and AFP mRNA appears to contain a similar number (approximately 285) of noncoding, nonpoly(A) bases. The purified albumin and AFP mRNA's were used as templates for synthesis of full-length cDNA hybridization probes. Both of the probes selectively hybridized to their templates with kinetics expected for single RNA species the sizes of albumin and AFP mRNA. ROt analysis was used to quantitate albumin and AFP mRNA sequences during normal liver postnatal development and liver oncogenesis. The number of polysomal AFP mRNA molecules per liver was found to drastically decrease during the first weeks of postnatal life, concomitant with a decline in the AFP synthetic capacity of the livers and in the serum concentrations of AFP. During this period, the concentration of albumin mRNA molecules per cell in the liver remained at high, approximately constant levels. In Morris hepatoma 7777, the concentration of AFP-specifying sequences was at least 10(3)-fold higher than that found in normal adult liver, whereas the content of albumin nRNA was four- to five-fold lower. These changes in concentration of albumin and AFP mRNA sequences closely correlated with a parallel variation in the specific protein synthetic capacity of the tissues.


Effects of surface charge on the conductance of the gramicidin channel. The electric conductance of the cation-permeable gramicidin channel in negatively charged phosphatidylserine membranes has been studied. At low electrolyte concentrations the single-channel conductance is much larger in the negatively charged membrane than in a neutral membrane. This enhancement of conductance is in agreement with theoretical expectations, although a complete description of the salt concentration dependence of conductance was not possible. The results of these experiments may be compared with previous studies of a negatively charged gramicidin analog (O-pyromellityl gramicidin). It is found that the electrostatic effect on the conductance is much larger for a neutral channel embedded in a negatively charged lipid than for the negatively charged O-pyromellityl analog (with three charges at the channel mouth) embedded in a neutral lipid.


Dicarbocyanine fluorescent probes of membrane potential block lymphocyte capping, deplete cellular ATP and inhibit respiration of isolated mitochondria. 3,3'-Dipropylthiodicarbocyanine iodide, a widely used fluorescent probe of membrane potential, was found to inhibit anti-Ig antibody, induced capping of mouse lymphocytes. The dye also lowered the cell ATP content. Experiments with isolated mitochondria revealed that the probe had a potent inhibitory action at site I of the respiratory chain. This mitochondrial blockade helps to explain the ATP depletion and blockade of capping, and gives cause for caution in the use of this dye as a probe of cell membrane potential. Three related dicarbocyanine dyes had similar toxic effects, but two cyanine dyes with much longer alkyl side chains, which have been used as probes of membrane fluidity, did not.


Human glomerular basement membrane. Heterogeneity of antigenic determinants. Human glomerular basement membrane was solubilized by digestion with proteolytic enzymes and immunoreactive components were quantitated and characterized by using rabbit antibodies raised against the particulate membrane. A number of antigens were demonstrated but they did not separate on gel filtration. However, two antigenic components in a collagenase digest of the membrane could be separated and isolated by Sepharose 6B chromatography. Chemical characterization suggests that both fragments are noncollagenous glycopeptides (molecular weights approx. 1,000,000 and 60,000--200,000, respectively).


Human glomerular basement membrane. Antigenic determinants in human urine. Antisera against particulate human glomerular basement membrane prepared from cadaver kidneys were raised in rabbits. It was shown that both normal individuals and patients with glomerular and tubular diseases excrete in their urine several antigens reactive with these antibodies. One antigen crossreacted immunologically with an antigen from human glomerular basement membrane while several others did not. One of the urinary antigens and the antigen crossreacting with the basement membrane were separated from the others by ion exchange chromatography and gel filtration, respectively. The pattern of anttigen excretion differed depending on the underlying renal disease but the multitude of different antigens detected complicates the interpretation of the patterns of excretion in different diseases.


Molecular control of rabbit follicular testosterone production. Role of protein and RNA after stimulation with luteinizing hormone. The time and dose dependence of the relationship between uptake of labelled precursors into protein and RNA and production of testosterone by rabbit follicles was examined. Although testosterone production was stimulated by luteinizing hormone at concentrations between 0.1 and 10 microgram/ml, the uptake of [3H]leucine into protein was significant only when the concentration of luteinizing hormone was greater than 2.5 microgram/ml. Increased production of testosterone was observed within 15 min of stimulation with luteinizing hormone whereas uptake of [3H]leucine was only significant at 90 min. Puromycin (40 microgram/ml) and cycloheximide (10 microgram/ml) in the presence of luteinizing hormone inhibited the synthesis of both testosterone and protein. However, lower concentrations of puromycin (0.1, 1 and 10 microgram/ml) and cycloheximide (1 microgram/ml) had no effect on luteinizing hormone-induced testosterone production but significantly inhibited protein synthesis by 58, 37, 31 and 71%, respectively. Actinomycin D (20, 80 and 160 microgram/ml) alone and in combination with 5 microgram luteinizing hormone/ml severely inhibited uptake of [3H]uridine into RNA without affecting testosterone production. However, with 1 microgram actinomycin/ml, testosterone production was significantly (P less than 0.01) greater than in the presence of luteinizing hormone alone. These results cast doubt on the obligatory role of RNA and protein synthesis in rabbit ovarian follicular steroidogenesis.


Properties of a goat anti-L antibody: further evidence for heterogeneity of the L antigen. The preparation and properties of an antibody (anti-L) against low potassium type (LK) goat red cells raised in a high potassium type (HK) goat are described. This reagent stimulated active potassium transport, but showed only weak serological activity against low potassium type (LK) sheep and goat red cells. The results are discussed in relation to the hypothesis that anti-L antibody has two specificities--a sodium pump-stimulating activity (anti-Lp) and a serological activity (anti-L1y).


Hepatic uptake and degradation of trace doses of asialofetuin and asialoorosomucoid in the intact rat. Asialoorosomucoid and asialofetuin were prepared by using sialidase, which was removed chromatographically before the proteins were labelled with radioactive iodine. After intravenous administration of a small amount oa asialoglycoprotein (3--4 microgram/100 g body wt.) protein-bound and non-protein radioactivities in plasmas and livers of rats were determined at intervals over a period of 30 min. Transfer of either tracer protein from plasma to liver was almost complete in 5 min. Proteolysis of asialofetuin was evident very shortly thereafter, but degradation of asialoorosomucoid commenced after a significant delay and was initially slow relative to that of asialofetuin. Studies in vitro with crude hepatic lysosomal enzyme preparations indicated that asialoorosomucoid was less readily digested than asialofetuin, and that desialylation of orosomucoid or fetuin did not noticeably increase the susceptibility of these proteins to protease action. Proteolysis of asialofetuin was also demonstrable in liver homogenates in conditions under which albumin and asialotransferrin were stable. A generalized mathematical model was devised to represent the uptake and degradation of asialoglycoproteins by the liver. The theoretical assumptions that gave the best fits with experiment are outlined and discussed.


Intracellular localization and degradation of asialofetuin in isolated rat hepatocytes. Analysis by isopycnic and differential centrifuging of the intracellular distribution of radioactivity following uptake of 125I-labelled asialofetuin by isolated rat hepatocytes showed that during incubations up to 1 h, most of the radioactivity was associated with structures which had a subcellular distribution pattern different from both the lysosomes and the plasma membrane. The latter two organelles were followed by means of enzyme markers. Ca2+ is necessary for the binding of asialofetuin to the plasma membrane, and it was also possible to differentiate between asialofetuin bound to the plasma membrane and that contained in intracellular structures by removing Ca2+ from the medium (by EGTA). Such experiments showed that asialofetuin became rapidly internalized. Practically all the labelled protein was located intracellularly in cells that had been incubated with asialofetuin for more than 30 min. When incubations were carried out for more than 1 h a peak appeared in the radioactivity distribution in the same place as the peak of activity of lysosomal marker enzymes. However, degradation of asialofetuin takes place in the lysosomes and this starts before the labelled protein can be found in the lysosomal fractions. Our data suggest that the rate-determining step in the cellular handling of asialofetuin is the transport of endocytized protein from the endocytic vesicles to the lysosomes.


[Effect of long-term cytostatic therapy on the hematopoietic stem cells]. CFU-C and diffusion chamber studies were performed in patients with teratocarcinoma, who underwent long term chemotherapy. No significant decline of bone marrow CFU-C or diffusion chamber cell recovery was found during twelve months of cytotoxic treatment. In contrast to the results in these patients the CFU-C-content of the remission marrow in leukemic patients showed a significant decrease in relation to the duration of remission and chemotherapy.


Lead contamination in street dust in Hong Kong. Lead levels were generally found to be high in road-side dust samples collected from different sites throughout the Island of Hong Kong during winter 1977-78. Considerable differences in the concentration of lead amongst these sites were detected but such differences bore no direct relationship to the traffic volumes of the sites from which the samples were collected. Different districts within the city varied widely in the mean concentration of lead in their road-side dust samples.


Transurethral resection of large prostates. A total of 102 men (mean age 70) underwent transurethral resection of the prostate. The weight of the resected prostate ranged from 40 to 152 g. Two deaths occurred in patients aged 85 and 92 years, both of whom were generally unfit. The average postoperative stay in hospital was 8.3 days. The proportions of patients with incontinence, infection, and stricture were similar to those in other series of resections and open prostatectomies, suggesting that resection of the larger prostate is safe and an acceptable alternative to an open operation.


Double-staining procedure for the fluorescent treponemal antibody absorption (FTA-ABS) test. The fluorescent treponemal antibody absorption (FTA-ABS) double-staining procedure was reproducible, comparable to the conventional test, and easy to read. We recommend the use of the FTA-ABS double-staining procedure for microscopes with incident illumination, the 100 x/1.30 oil achromatic objective and the 6.3 x ocular to obtain optimal fluorescence, and the KP560 as a barrier filter to exclude rhodamine emission when fluorescein fluorescence is read. With this system, errors related to poor focusing or failure to visualise treponemes on all smears should be eliminated.


Effect of different preparation procedures on the ultrastructure of gonococci. Culture samples of Neisseria gonorrhoeae and Escherichia coli were studied by transmission and electron microscopy to evaluate the effect of different preparation procedures on the pilation of these organisms. Unfixed as well as fixed bacteria showed only few, long, filamentous appendages when investigated in ultrathin sections, negatively stained specimens, or critical-point dried preparations. Snap-frozen specimens of E. coli showed many short and thin pili after being shadowed with carbon and platinum whereas those of N. gonorrhoeae showed only some type-C-like pili. Thus, the number and morphological appearance of pili appear to be greatly influenced by the preparation techniques used for study by electronmicroscopy. Conclusions as to the type and the infectivity of a bacterial strain can, therefore, not be based on purely morphological criteria.


Transmitter histochemistry of the rat olfactory bulb III. Autoradiographic localization of [3H]GABA. The distribution of [3H]gamma-aminobutyric acid (GABA) labeled elements in rat olfactory bulb was studied by light and electron microscopic autoradiography. [3H]GABA was strongly taken up into glial cells and pericytes in all layers of the bulb. The neuronal uptake of [3H]GABA was mainly seen in certain types of nerve terminals. About one-third of the granule dendritic terminals, some nerve endings of short axon cells, and certain nerve endings of extrabulbar origin showed a strong labeling. Labeling was seen in a small population of the periglomerular, short axon and granule cell bodies. Most cell bodies of these 3 types as well as the mitral cells did not, however, accumulate any appreciable amo9nt of [3H]GABA. The labeling pattern seen after injection of [3H]glycine and [3H]leucine was clearly different from the pattern seen after [3H]GABA injection. The labeling was more uniformly distributed over the components of the neuropil with a considerably higher activity over certain cell somata such as the mitral cells. The present results demonstrate that neuronal uptake and accumulation of [3H]GABA occur into populations of olfactory bulb cells and processes, which from neurophysiological and/or immunohistochemical studies are supposed to use GABA as a neurotransmitter.


Observations of HRP labeling following injection through a chronically implanted cannula--a method to avoid diffusion of HRP into injured fibers. One of the limitations of the horseradish peroxidase (HRP) tracer method is the diffusion of HRP into injured axons resulting in unintended labeling of neurons not terminating in the injection area. To overcome this limitation, an experiment was designed to inject the HRP through an implanted cannula after degeneration and healing had taken place. It was shown that implantation of a cannula into the internal capsule significantly decreased the number of labeled axons in the injection site, thus limiting the unintended labeling of neurons from that injection. When injections followed implantation of the cannula by 24 h or more, fibers damaged by the cannula had healed or degenerated sufficiently that intraaxonal diffusion of HRP into those injured fibers did not occur. A significant difference between control (without the cannula) and experimental (with the cannula) injections was observed. Extensive axonal and neuronal labeling following the control injections was seen at the injection site and caudate nucleus, and in the thalamus and parietal cortex, respectively. Experimental injections resulted in sparse axonal and neuronal labeling evident mostly with the larger injections of HRP.


The use of neurotoxins to characterize the rates and subcellular distributions of axonally transported dopamine-beta-hydroxylase, tyrosine hydroxylase and norepinephrine in the rat brain. The effect of 6-hydroxydopamine (6-OHDA), colchicine and cytochalasin B on the transport and subcellular distribution of proteins, tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and norepinephrine (NE) were studied in the noradrenergic neurons of the rat locus coeruleus (LC). Four waves of transported 3H-labeled proteins and glycoproteins, defined by previous studies, as well as hypothalamic levels of TH, DBH and NE, were examined after injection of each neurotoxin into the ascending dorsal noradrenergic bundle. Blockade of subcellular components of TH, DBH and NE was compared to their endogenous hypothalamic distributions. 6-Hydroxydopamine variably blocked transport of all 4 waves of 3H protein and bilateral injections decreased hypothalamic levels of TH, DBH and NE by 58.2, 56.9 and 52.2% of controls, respectively. Cytochalasin B blocked transport of protein waves I (72--192 mn/day) and III (13--20 mm/day) and decreased hypothalamic levels of TH to 60.1% of control after bilateral injections. Colchicine blocked transport of waves I, II (24--48 mm/day) and V (1.4--2.9 mm/day) and blocked [3H]NE transport, while decreasing hypothalamic levels of DBH and NE to 56.6 and 69.3% of control after bilateral injections. Colchicine and 6-OHDA, but not cytochalasin B, caused a backup of DBH immunofluorescence proximal to the injection site. DBH and NE appeared to be transported primarily in particulate form, while TH transport was predominantly soluble in distribution. None of the toxins differentially affected the transport of one particular subcellular component of TH, DBH or NE. Based on the differential blocking effects of these toxins, DBH and NE appeared to be associated with wave II, and TH with wave III, travelling at 24--48 mm/day and 13--20 mm/day respectively.


Axonal transport of nucleosides, nucleotides and 4S RNA in the neonatal rat visual system. The axonal migration of RNA, the nucleoside uridine and its nucleotide derivates (NS/NT) were compared in neonatal and young adult rat optic axons. Tritiated uridine was injected into right eyes of developing (1- or 4-day-old) and young adult (40-day-old) rats which were sacrificed at times after injection ranging from 6 h to 20 days. Right and left lateral geniculates were removed and assayed for trichloroacetic acid soluble (NS/NT) and RNA radioactivity. Left minus right geniculate (L-RLG) radioactivity was used as an index of axonally migrating radioactivity. Results showed that uridine and its phosphorylated derivatives were transported along both neonatal and young adult rat optic axons. Greater than 90% of right geniculate (blood-borne) TCA soluble radioactivity was metabolized to volatile substances (probably 3H2O) by three days after injection, leaving approximately 3% of the neonatal and approximately 10% of the adult activity as [3H]NS/NT. In left geniculate fractions (containing transported material) approximately 15% and 40% of total TCA soluble radioactivity was present as [3H]NS/NT in neonates and adults, respectively. Thus, axonal NS/NT appears to be relatively protected from degradation when compared with blood-borne NS/NT. The amount of L-RLG [3H]RNA in the neonates was 10 times higher than in young adults. Peaks of neonatal [3H]RNA occurred at 5 and 10 days after birth, whether injections were made at 1 or 4 days of age indicating that this [3H]RNA may be linked to developmental events. Gel electrophoretic analysis of neonatal geniculate RNA indicated that a small portion of the [3H]RNA in the first peak represented axonally transported 4S RNA. The remainder of the L-RLG [3H]RNA in the neonates was probably due to a rapid and efficient incorporation of axonally transported [3H]NS/NT into extraaxonal geniculate RNA. In contrast, little or no axonal RNA transport could be demonstrated in the young adults.


Kainic acid-induced neurotoxicity in the striatum: a histofluorescent study. Histochemical observation of catecholamine terminals in the striatum following kainic acid injections revealed a 'sphere of influence' which contained an increased intensity of dopamine fluorescence as well as non-specific tissue destruction at the injection site. The area of involvement was approximately 50% of the striatum and varied somewhat at any one dose range depending upon the site of injection. It is suggested that destruction of regulatory neuronal systems (cholinergic, GABAergic, peptidergic, etc.) results in a net increase of dopamine in the sphere of kainic acid influence.


Effects of 16, 16-dimethyl prostaglandin E2 on bile-induced histamine release and permeability alterations in canine oxyntic mucosa. Damage to the stomach results in excessive movement of hydrogen ion (H+) out of the lumen, and increased movement of sodium (Na+) and potassium (K+) into the lumen. Histamine liberation during damage probably adds to the destruction by increased capillary permeability and formation of edema. Previous reports have shown that the synthetic prostaglandin analogue 16,16-dimethyl prostaglandin E2 (Dm PGE2) protects dog gastric mucosa from aspirin- and ethanol-induced gastric mucosa damage. The effects of dm PGE2 on bile salt (sodium taurocholate) induced injury has not been investigated. Using the canine Heidenhain pouch, the present study examined the action of dm PGE2 on gastric mucosal damage induced by 5 mM sodium taurocholate in 100 mM HCl. Bile salt damaged the pouch mucosa as evidenced by an increased loss of H+, and increased net fluxes of both Na+ and K+. There was also an increase in the histamine content of the fluid irrigating the Heidenhain pouch. Intravenous injection of dm PGE2 in the doses 0.1 and 1.0 microgram/kg 1/2 h before administration of the sodium taurocholate in HCl significantly reduced the net loss of H+ and the gain of Na+, K+, and histamine. It is concluded the dm PGE2 effectively protects the canine gastric mucosa from the damaging effects of bile salt and that the mechanism of dm PGE2 protection of canine oxyntic mucosa may be mediated in part via inhibition of the gastric mucosal release of histamine.


Periostitis associated with myelofibrosis. Two patients with myelofibrosis developed fever, leg pain and periostitis. The first patient had myelofibrosis with myeloid metaplasia and was symptomatic for months before x-rays showed periosteal new bone formation in the lower extremities. He subsequently developed periostitis of both upper extremities. Radiation of the lower extremities resulted in significant pain relief. The second patient had a past history of polycythemia vera and experienced painful periostitis of the tibiae and fibulae. 99mTechnetium pyrophosphate bone scans showed increased uptake in the involved bones in both patients. Asymptomatic or painful periostitis may be related to the increased bone blood flow associated with myelofibrosis. Radiation can afford successful palliation in the severely symptomatic patient.


Cobalt-57 bleomycin for imaging head and neck tumors. Cobalt-57 bleomycin imaging was performed in 11 patients with a history of head and neck cancer. Clinical and scan findings concurred on the presence and extent of tumor in 9 patients (82%); tumor was present in 7 and absent in 2 of the 9. In 2 patients (18%) the scan demonstrated tumor in the neck but failed to show metastatic sites. Cobalt-57 bleomycin images were of good technical quality, with remarkably low background activity at 24 hours after administration. Cobalt-57 bleomycin imaging appears to be a promising technique for evaluating patients with head and neck tumors.


On the presence of reverse transcriptase in myelo- and lymphoproliferative disorders. Buffy coats from 31 patients with a diagnosis of leukemia and 16 normal donors were tested for the presence of a viral-like reverse transcriptase. Eighty-five percent of fresh leukemic buffy coats were positive. Also tested were spleens from 16 patients with hematological disorders and 5 spleens from patients without history of hematological malignancy. The 5 normal spleens were negative. Also negative were 4 spleens from patients with Hairy cell leukemia. From the remaining 12 spleens 7 were positive. Reverse transcriptase measurements can be used to distinguish leukemic from normal buffy coats.


Detection of reverse transcriptase activity in human cells. Samples of three nonmalignant and seven leukemic human cells were examined for DNA polymerase activity that could be identified as RNA tumor virus reverse transcriptase. Experiments on virus-infected model animal cells provided the basis for cell fractionation procedures, and reconstituted systems of known virus, added to human cells, established a threshold of virus detection by enzyme assay at 1 to 10 particles/cell. DNA polymerase activity with some properties similar to a reverse transcriptase was detected in some of the human leukemic cells. However, parallel analyses of nonmalignant cells showed sufficient similarities to raise serious questions about the specificity of the criteria. Reverse transcriptase activity has been reported to be present in white blood cells from a proportion of cases of leukemia; however, it is concluded from the present study that the usual enzymatic criteria using synthetic template primers, which were used in most of the studies reported, are not sufficient to identify a DNA polymerase activity as viral reverse transcriptase.


Isolation of rat alpha1-fetoprotein messenger RNA from Morris hepatoma 7777. A double-antibody procedure has been developed for the isolation of alpha1-fetoprotein (AFP)-synthesizing polysomes from Morris hepatoma 7777. The polyadenylic acid-containing RNA, subsequently purified by differential sedimentation on sucrose gradient and oligodeoxythymidylic acid-cellulose chromatography, migrates as a single 21S component in polyacrylamide gel electrophoresis; in a cell-free translation system, it yields a peptide product immunoprecipitable by anti-rat AFP antiserum, but not by anti-rat albumin, and which migrates slightly faster than serum AFP on sodium dodecyl sulfate-urea-polyacrylamide gels. This messenger RNA fraction was used for the synthesis of a radioactive complementary DNA. In hybridization assays, the complementary DNA reassociated with its purified template at a Cr0t1/2 [product of RNA concentration (mol of nucleotides per liter) X half-time (sec)] of 1.5 X 10(-2). By constitute 3,2, and less than 0.01% of total polyadenylic acid-containing polysomal RNA of Morris hepatoma 7777, 10-day-old-rat liver, and adult rat liver, respectively. The high specificity of the polysome immunoprecipitation system, the electrophoretic homogeneity of the isolated messenger RNA fraction, its selective translation into AFP, and the specificity of the hybridization probe indicate that the procedure described yields a highly purified rat AJP messenger RNA.


A clinical investigation of total-body hyperthermia as cancer therapy. Seventy-seven patients have been treated with total-body hyperthermia to 42 degrees C alone or in combination with chemotherapy, radiation, or immunotherapy in a preliminary investigation of this modality. Most had advanced cancers progressing despite other antineoplastic therapy. An overall objective response rate of 43% was obtained, with an additional 15% achieving good subjective palliation. Results did not differ significantly for hyperthermia alone or combined with another modality, but the trend favored combination therapy. Mortality and morbidity were acceptable and not significantly increased by adding chemotherapy or radiation to the regimen. These results are preliminary and need confirmation by further controlled studies.


Potential for therapy of drugs and hyperthermia. The interaction of hyperthermia (41--45 degrees C) and chemotherapeutic agents frequently results in increased cytotoxicity over that predicted for an additive effect, although to date only a very limited number of drugs have been examined for such a possible interaction. At 42 degrees C, the upper limit of temperature useful for whole-body hyperthermia, the most promising agents of those examined to date appear to be the nitrosoureas and cis-platinum. Insufficient data exist for cyclophosphamide, whose long plasma half-life makes it an attractive candidate. Localized heating seems optimum at higher temperatures (43--45 degrees C). At these temperatures, not only those drugs effective at 42 degrees C but particularly bleomycin and possibly amphotericin B become candidates. No data exist in the literature on possible "thermic sensitizers," i.e., drugs which are noncytotoxic at 37 degrees C but which become effective at elevated temperatures. Two special cases are Adriamycin and actinomycin D. These drugs may be contraindicated for clinical use, since not only synergism but also protection by hyperthermia have been demonstrated, depending upon the time-sequence relationships of the heat and drug treatments.


Increase in immunogenicity with concomitant loss of tumorigenicity of respiratory tract carcinomas during in vitro culture. Cell lines were established in vitro from respiratory tract carcinomas induced in rats by carcinogenic, polycyclic hydrocarbons. Propagation of the carcinoma lines in vitro lead to a progressive decrease in tumorigenicity. Tumor transplantation studies in X-irradiated, immunosuppressed recipients and in immunologically reconstituted recipients suggested that the cells are rejected because of their immunogenicity, since a high incidence of tumors was observed in X-irradiated recipients but not in normal or X-irradiated, reconstituted recipients. When immunologically competent rats were immunized with cells from an in vitro tumor line, strong tumor transplantation resistance resulted. Similar immunization with the corresponding in vivo tumor line caused very little if any protection, and immunization with a non-cross-reacting sarcoma line grown in vitro did not produce immunological protection against carcinoma cell lines. A single in vivo passage of the in vitro-adapted tumor line in immunosuppressed recipients fully restored tumorigenicity. The increase in immunogenicity of carcinomas cultured in vitro appears to involve preexisting angigens indigenous to the carcinomas rather than new antigens acquired during tissue culture, such as antigens related to retroviruses, mycoplasmas, or heterologous serum.


Rapid decay of tumoricidal activity and loss of responsiveness to lymphokines in inflammatory macrophages. The ability of inflammatory tissue macrophages harvested on glass coverslips implanted in the s.c. tissue of C57BL/6 mice to kill tumor cells in vitro has been examined. Macrophage present on coverslips implanted for less than 4 days are devoid of spontaneous tumoricidal activity but can be rendered cytotoxic for syngeneic and allogeneic tumor cells in vitro by incubation in vitro with lymphokines release by mitogen-stimulated lymphocytes. Inflammatory macrophages on coverslips implanted for 4 to 7 days show significant spontaneous cytotoxicity for tumor cells in vitro, and their tumoricidal activity is further increased by additional incubation in vitro with lymphokines. With progression, the inflammatory macrophages harvested on coverslips implanted for longer than 7 days lack spontaneous cytotoxic activity and are also resistant to activation by lymphokines in vitro. These alterations in tumoricidal activity and responsiveness to lymphokines are accompanied by a marked reduction in the number of peroxidase-positive macrophages within the population, suggesting that maintenance of tumoricidal activity requires continuous influx of new peroxidase-positive macrophages from the circulation. Previously activated macrophages which have lost their tumoricidal activity and become refractory to reactivation by lymphokines in the extracellular environment can be reactivated by treatment in vitro with liposomes containing encapsulated lymphokines.


Pulmonary toxicity of antineoplastic drugs. Drug-induced pulmonary disease is an infrequent, but clinically significant, toxic manifestation of antineoplastic drug therapy. Pulmonary toxicity has been associated primarily with bleomycin therapy but also has been described with busulfan, cyclophosphamide, methotrexate, and the nitrosoureas. Cytotoxic drug-induced pulmonary disease is characterized by an insidious and nonspecific presentation. The common pathologic process of a fibrosing alveolitis follows a variety of toxic mechanisms associated with the chemotherapeutic agents. The importance of close clinical monitoring is emphasized by the necessity for early withdrawal of the drug in order to halt or reverse potentially lethal pulmonary disease. At the same time, the nonspecific nature of the lung disease and the multitude of potential insults to the lung in a cancer patient make an early aggressive evaluation mandatory. A lung biopsy is almost always indicated.


Improved therapeutic index of bleomycin when administered by continuous infusion in mice. The effects of three different dosage schedules on both therapeutic effect and pulmonary toxicity of bleomycin were studied in mice. Therapy was assessed by both survival and decreased tumor size in mice bearing Lewis lung carcinoma. Lung toxicity was estimated in nontumored mice as increases in lung collagen content by measuring lung hydroxyproline concentrations. In the first set of experiments, bleomycin injections twice daily (low-dose, high-frequency) produced a significant (34%) increase in lifespan over controls, whereas the same total dose given twice weekly did not result in increased survival. Both schedules produced pulmonary toxicity. Continuous sc infusion of bleomycin via an osmotic minipump was compared to these two schedules of intermittent injection. Identical total doses of drug were administered in all three schedules. Continuous infusion for 7 days produced marked inhibition of tumor growth (T/C = 16%), which was significantly better than twice weekly or ten-times weekly injection of the same total dose. Furthermore, at a total dose of 40 mg/kg of bleomycin, continuous infusion did not result in measurable pulmonary toxicity, whereas both schedules of bolus injection produced significant increases in lung collagen content. Thus, continuous infusion of bleomycin improved its therapeutic effect against Lewis lung carcinoma and also reduced its pulmonary toxicity.


Limitations of a portable infusion pump in ambulatory patients receiving continuous infusions of anticancer drugs. The delivery of continuous iv bleomycin and 5-fluorouracil to nine ambulatory cancer patients was studied using a novel disposable portable infusion pump, the Ar/Med Infusor. Although the chemotherapy was satisfactorily delivered, there were significant mechanical problems, including variable flow rates and substantial drug cartridge failure. Most therapy courses were finished within 10% of the projected termination time. Serious venous complications did not occur. While further exploration of various devices for outpatient continuous iv therapy is recommended, we believe that technical improvements are needed for the Ar/Med Infusor.


Successful re-treatment with cis-dichlorodiammineplatinum(II) after apparent allergic reactions. Allergic or anaphylactic-like reactions have been reported with cis-dichlorodiammineplatinum(II) (CDDP) administration. We have seen two reactions among 50 patients treated in a phase II trial of CDDP. Both patients were successfully rechallenged after diphenhydramine pretreatment, and therapy was continued. In vivo and in vitro studies performed in one patient included negative skin tests, negative histamine release assay, negative lymphocyte stimulation studies, and normal total serum IgE. We conclude that some apparent allergic reactions occurring with CDDP may have a non-allergic etiology, and treatment may be successfully continued in selected patients.


Radiation therapy in the management of cutaneous T-cell lymphomas. The lesions of mycosis fungoides (MF), a neoplasm of T lymphocytes, are extremely radiosensitive. The history of ionizing radiation in the treatment of MF is discussed in this paper. Low-energy X-rays have long been successful in the treatment of individual lesions and in the effective palliation of patients with this disease. The major breakthrough in the treatment of MF with ionizing radiation came with the development of the ability to treat the total skin by means of electrons which penetrate to depths of only 1--2 cm, thereby treating the epidermis and dermis while sparing more deeply situated tissues. The complications and results of this therapy are reviewed. The aggressive use of electron-beam therapy has resulted in many long-term remissions. It is important to use high initial doses of radiation and to treat patients when they are still in the early stages of disease. The potential use of other modalities of radiation, including total-lymphoid radiation with megavoltage photons, low-dose fractionated total-body radiation, and sequential hemibody radiation, are reviewed.


Combination chemotherapy for mycosis fungoides: a Southwest Oncology Group study. Between 1972 and 1977, the Southwest Oncology Group studied the following three chemotherapy programs for the treatment of patients with advanced forms of mycosis fungoides: (a) cyclophosphamide, adriamycin, vincristine, and prednisone (CHOP) (seven patients); (b) adriamycin, vincristine, and prednisone (HOP) (five patients); and (c) cyclophosphamide, vincristine, prednisone, and bleomycin (COP plus bleomycin) (12 patients). Among the 24 evaluable patients there was an overall objective response rate of 95% with seven (29%) achieving a complete remission. With the adriamycin-containing chemotherapy, five (42%) of 12 patients achieved a complete remission compared to two (17%) of 12 patients treated with COP plus bleomycin. The median duration of remission (partial plus complete) was longer with the COP plus bleomycin combination (median, 47 weeks) than with the adriamycin-containing combinations (median, 22 weeks; P = 0.03). The median survival for all 24 evaluable patients was 95 weeks and was similar regardless of remission-induction therapy. In summary, combination chemotherapy proved to be effective palliative therapy for advanced mycosis fungoides.


Effect of ventricular extrasystoles on idioventricular rhythm in patients with complete heart block. Idioventricular rate, QRS width, site of block and responses to premature ventricular stimulation were studied in 23 patients with chronic complete heart block in an attempt to distinguish patients liable to syncopal attacks. Seven patients were asymptomatic, five gave a history of presyncope and 11 of syncope. Although the mean idioventricular rate of the nine patients in the narrow QRS group (39.7 per min) was significantly faster than that of the 14 patients in the wide QRS group (35.3 per min; P less than 0.05), the rate did not distinguish symptomatic patients within either the narrow or the wide QRS groups. His bundle studies of the site of block and the effect of single and paired right ventricular stimulation upon idioventricular rhythm were also unhelpful. The response of the return cycle to increasingly premature ventricular extrasystoles, however, proved more complex than was anticipated.


Clinico-pathological and histochemical studies on hypertrophic duct epithelium in human non-endocrine pancreas cancer. The hypertrophic duct epithelium of the pancreas, including the pyloric gland metaplasia, mucous cell hypertrophy and ductal papillary hyperplasia were studied clinico-pathologically and histochemically to examine their precancerous character. A total of 180 surgical and autopsy specimens (90 pancreata with cancer and 90 pancreata without cancer) were analysed. The overall incidence of these three types of hypertrophic epithelium in the pancreas cancer was much higher than that in the pancreas without cancer. These hypertrophic lesions appeared most frequently in the interlobular duct. The histochemical study revealed the presence of a new type of glycoprotein in these hypertrophic duct epithelia, however, this substance was not detected in the cancer cells nor in the normal epithelium. This suggests that these hypertrophic lesions may not be the precursors of cancer but rather the coexistent lesions of pancreas cancer.


Horseradish peroxidase-labelled cells in cervical sympathetic and trigeminal nervous systems after injection of HRP into tooth pulp in monkeys and tree shrews (a preliminary report). The neurons that project to the tooth pulp were examined by the HRP-retrograde transport technique, using 4 monkeys (3 Japanese monkeys and a crab-eating monkey) and 6 tree shrews. The present investigation showed that a large number of sympathetic cervical ganglion neurons, small and medium in size, project to the molar teeth from the superior, middle and inferior ganglia and a small number of Gasserian gangilon neurons (small in size) to the molar teeth ipsilaterally.


The force-interval relationship of the left ventricle. We have carried out a quantitative analysis of the force-interval relationship of the human left ventricle and compared it to previous studies done in both intact subjects as well as isolated muscle. The characteristics of the force-interval relationship of the normal patient resembed those of normal isolated mammalian muscle (except when exposed to high levels of catecholamines). The relationship in group 2 (patients with increased left ventricular dimensions and normal pressure indices) resembled those obtained from isolated muscles from hypertrophied hearts. The relationship from group 3 (patients with increased left ventricular EDDs and depressed pressure indices, two of whom were in clinical heart failure) resumbled those induced in normal muscles exposed to high levels of catecholamines, and those obtained from experimentally induced heart failure. The force-interval relationships of the four patients who fell into the third group were strikingly different from the other groups. This suggests that the force-interval relationship may be useful to describe changes in the inotropic state of the patient's heart.


Dispersion of atrial refractoriness in patients with sinus node dysfunction. Abnormal atrial refractoriness was examined as a cause of atrial fibrillation/flutter (AFF) in patients with bradycardia. Refractory periods at three disparate right atrial sites were compared in 17 patients with sinus node dysfunction (SND) and 16 controls. Atrial pacing shortened refractory periods, but failed to decrease dispersion of refractoriness significantly. During sinus rhythm, duration and dispersion of refractoriness were greater in SND patients than in controls. These differences persisted with atrial pacing. For example, at the paced rate, dispersion of effective refractory periods in SND patients was greater than in controls (62.9 +/- 34 vs 36.6 +/- 21 msec, p less than 0.01). Six SND patients had AFF, but they did not have greater dispersion than other SND patients, or unusually short or long refractory periods. Thus, prolonged and nonuniform refractoriness were features of SND. Abnormal refractoriness in SND reflected atrial disease and persisted with pacing. These abnormalities were not unique to patients with AFF.


Specific IgE antibodies to platinum salts in sensitized workers. A RAST has been developed for the measurement of IgE antibodies specific to platinum chloride complexes in sensitized workers. Human serum albumin covalently linked to Sepharose beads by the cyanogen bromide method was reacted with ammonium tetrachloroplatinite (II) (NH4)2PtCl4. This conjugate was more suitable for the RAST, than conjugates of HSA and the platinum salt prepared in solution and then linked to the activated Sepharose, showing better sensitivity and giving lower levels of non-specific uptake of IgE from sera of non-exposed subjects with high total levels of IgE, e.g. allergic bronchopulmonary aspergillotics.


IgE mediated hypersensitivity to pancreatic extract (PE) in parents of cystic fibrosis (CF) children. On exposure to pancreatic extract (PE), four parents of cystic fibrosis (CF) children developed immediate hypersensitivity-like symptoms: i.e. rhinoconjunctivitis, asthma, and/or anaphylaxis. IgE to PE was demonstrated in the subjects by skin testing, leucocyte histamine release and radioallergosorbent test (RAST). No serum precipitating antibodies were found. Bronchial challenge caused an immediate asthmatic response. No delayed asthmatic response or hypersensitivity pneumonitis-like reaction occurred. The responsible antigen for PE induced asthma is unknown; trypsin failed to inhibit PE-RAST and is therefore not responsible in these subjects. Caution should be exercised in using PE for skin testing and bronchial challenge in subjects with suspected hypersensitivity to PE. Certain measures were found useful in preventing the occurrence of symptoms in the four subjects.


Passive transfer in man and the monkey of Type I allergy due to heat labile and heat stable antibody to complex salts of platinum. Passive transfer (P-K) tests have been performed in man and monkeys with sera from a group of six refinery workers sensitive to ammonium hexachloroplatinate and tetrachloroplatinite. Positive (P-K) prick test reactions were elicited in three human recipients and in the monkey to three of the test sera. The other three sera gave inconsistent results in man and negative results in monkeys and these differences appeared to correspond with the duration of exposure of the workers. The passive transfer tests did not correlate with the intensity of the prick test reactions in the donors or with the RAST; there was partial correlation with total levels of IgE. Antigen mediated histamine release from whole blood and washed leucocytes from two of the six subjects correlated with the RAST and P-K levels. These findings confirm the presence of specific IgE antibody to the platinum salts, and also of heat stable, short-term sensitising antibodies, presumably STS-IgG.


A radioimmune assay for human prostatic acid phosphatase-levels in prostatic disease. Prostatic acid phosphatase from human seminal fluid was purified to homogeneity. The enzyme was characterized as to its purity, molecular weight and amino acid composition. Analytical isoelectric focusing of purified enzyme on polyacrylamide gels resolved the enzyme activity into eleven discrete bands, apparently due to various amounts of sialic acid associated with the glycoprotein. Antisera raised against the purified enzyme produced only one precipitan arc on immunoelectrophoresis. A double antibody radioimmune assay was developed and used to evaluate serum prostatic acid phosphatase in 226 patients without prostatic disease, in 186 patients with benign prostatic hyperplasia and in 93 patients with prostatic carcinoma. No statistical difference was noted in serum prostatic acid phosphatase between patients with benign prostatic hyperplasia and in those without prostatic disease Serum prostatic acid phosphatase was elevated in 94% of the patients with metastatic prostatic carcinoma. Significant elevations were also found in carcinoma patients without metastases.


Microencapsulated antibodies in radioimmunoassay--I. Determination of digoxin. We describe the application of the microencapsulated-antibody technique to the radioimmunoassay of digoxin in serum. Droplets of emulsified rabbit antibody are microencapsulated in a semipermeable nylon membrane by an interfacial polymerization technique. The antibody microcapsules are incubated with 125I-labeled digoxin and unlabeled digoxin for 15 min at 37 degrees C, then free and bound digoxin are separated by centrifugation. Subtherapeutic, therapeutic, and toxic concentrations of digoxin in sera can be determined, with use of a standard curve prepared by use of known amounts of digoxin. With this technique we obtained an intra-laboratory correlation coefficient of 0.945 for 100 patients' sera and one of 0.940 for interlaboratory results for 21 sera (10 laboratories) when compared to a routine clinical laboratory radioimmunoassay for digoxin. Icterus, lipemia, hemoglobin, or disproteinemia had no effect on the analytical recovery of digoxin. The standard curve was linear to 6 microgram/L; the sensitivity was 0.25 microgram/L.


An activity of hydrolyzing elastase substrate succinyltrialanine p-nitroanilide in human bile. Human hepatic bile contained an activity of hydrolyzing N-succinyl-L-alanyl-Lalanyl-L-alanine-p-nitroanilide, a substrate for elastase. The activity was associated with high molecular weight materials and was metal dependent. Neither elastase nor elastase-alpha 2-macroglobulin complex was implicated in this activity.


Development of antinuclear antibodies during acebutolol therapy. Eleven patients were studied while taking the beta blocker acebutolol for a period ranging from 12 to more than 24 wk. Control titers for fluorescent antinuclear antibody (ANA) were obtained in all. Serial testing was performed over the duration of the therapy and following its discontinuation. Additional immunologic testing was performed in most patients. The patients were observed closely for the development of clinical autoimmune disease. Using a sensitive assay, fluorescent ANA developed in 8 of 9 patients with negative values in the control period. In no patient has evidence of clinical autoimmune disease developed. In general, the titers of ANA tended to rise with increasing duration of therapy and decline after its discontinuation. Positive lupus erythematosus cell preparations were also observed in several patients. These data suggest that autoantibodies are frequently induced by acebutolol and, although no evidence of clinical autoimmune disease has been reported, immunologic surveillance is warranted.


Effects of DAPI on human leukocytes in vitro. DAPI (4'-6-diamidino-2-phenylindole), a fluorochrome specific for AT-rich DNA, was supplied for 24 h at various concentrations to human leukocytes in culture. This treatment caused the appearance on the chromosomes of specific areas lacking spiralization. In particular, the centromeric regions of chromosomes 1,9, and 16, a short region on the long arm of chromosomes 1 and 2, and the distal heterochromatic part of the long arm of the Y chromosome were despiralized. The despiralization pattern of DAPI is compared with those previously obtained with Hoechst 33258 and Distamycin A.


[Histamine concentration and diamine oxidase activity in the small intestine in superior mesenteric artery occlusion]. During intestinal ischemia in rabbits histamine concentration and diamine oxidase activity were altered in the intestinal wall and in the perfusate of mesenteric vessels. The results were interpreted as a histamine release and an increased catabolism of diamine oxidase. Thus, the combination of release of vasoactive histamine and partial elimination of a protective enzyme may contribute to the fatal outcome after mesenteric ischemia.


[Incidence of anaphylactoid reactions following infusion with colloid volume substitutes]. All available colloid volume substitutes carry the risk of anaphylactoid reactions with a total incidence of 0.033%. The frequency of life threatening reactions was 0.003% for plasma protein solutions, 0.006% for hydroxyethyl starch, 0.008% for dextran and 0.038% for gelatin solutions.


[Hormone concentrations in thyroid gland tissue and plasma in autonomous thyroid adenomas with and without thyrostatic pretreatment]. The thyroid hormone concentrations of T2, T3 (and the inactive R-T3) were determined in thyroid tissue of 20 patients with autonomous adenomas. High concentrations were found in scintigrafically decompensated adenomas without preoperative thyrostatic treatment. Decompensated adenomas after thyrostatic treatment, compersated adenomas and a group of 9 euthyroid goiters showed no difference in tissue-concentrations of T4 and T3. The amount of tissue-R-T3 seems to be lowered in autonomy. The plasma-concentration of T3, which was intraoperatively elevated in the venous effluent from decompensated adenomas without thyrostatic treatment, was significantly lower in the blood draining decompensated adenomas after thyrostatic treatment as well as compensated adenomas.


[Serologic breast cancer follow-up monitoring using pregnancy-associated alpha-2-glycoproteins]. The variations in serum level of the alpha 2-PAG (pregnancy associated glycoprotein) were studied during the treatment of 25 patients with breast cancer. A good correlation was found between alpha 2-PAG concentration and the course of the disease. Serum alpha 2-Pag levels rose prior to the clinical recognition of metastatic disease and decreased significantly on successful treatment. alpha 2-PAG appears to have potential as an indicator of the growth of micrometastases.


[Indications and results of percutaneous transhepatic bile-duct drainage]. Percutaneous transhepatic drainage of the bile duct (PTCD) is a method that has few complications and can successfully relieve an obstructive jaundice. As a palliative drainage in malignancies of the liver and the porta of the liver it reduces the jaundice and prevents the accompanying troublesome pruritus, thus prolonging life. It reduces postoperative lethality and complications by reducing the jaundice preoperatively. Suppurative cholangitis with bile duct obstruction is quickly relieved by administration of antibiotics into the drainage.


Management of postoperative constipation in anorectal surgery. Fifty patients who had undergone proctologic operations were given a combination of standardized senna concentrate and dioctyl sodium sulfosuccinate for prevention or treatment of postoperative constipation. Two Senokot S tablets were given on the evening of the first postoperative day. Patients who had bowel movements after receiving this dose and remained in the hospital received two tablets on each of the following two days. Otherwise, dosage was increased gradually to a maximum of four tablets twice daily until defecation occurred or until the four-day treatment period was over. All patients achieved bowel movements, and in no case was it necessary to administer an enema. No hard stools were passed on the day patients were discharged from the hospital. Overall results were excellent or good in 94 per cent of cases; these results were not affected by age. Ninety-two per cent had bowel movements after the first or second dose was administered. On the day of discharge from the hospital, fecal consistency was satisfactory in 88 per cent of cases and loose in 12 per cent. Except for a single instance of diarrhea after administration of one dose, which involved a patient with a history of loose stools, no side effect occurred during the trial. In view of these responses and the virtual absence of side effects in this study group, Senokot S tablets are judged to be of demonstrated value in the postoperative management of patients undergoing proctologic operations.


[Drugs and acute agranulocytosis: analysis of previous drug intake in 42 patients (author's transl)]. An exact account of previous drug intake was obtained in 42 patients with acute agranulocytosis. Because of the multiplicity of administered drugs, often in the form of multiple-drug preparations, it was possible in only a minority of cases to ascribe with any degree of certainty a causative role to any particular drug. But outstanding as possible causes were thyrostatic drugs and pyrazolone. But no conclusion could be drawn as to the incidence of agranulocytosis with regard to these drugs because of the absence of data on the frequency that they are prescribed and the unknown number of fatal cases of agranulocytosis. The results indicate that multiple drug treatment and in particular the unnecessary prescribing of multiple-drug preparations makes it difficult to ascertain side-effects of drugs. They once again point to the need of frequent checks of white cell count in the drug treatment of pyrexial diseases.


[Cancer of the colon: clinical and morphological aspects of local cryosurgery (author's transl)]. Using strict indications, local cryosurgery was performed in 86 patients with cancer of the colon. Survival time was longer both for patients with advanced tumour growth and distant metastases and those with potentially curable tumour but in too poor a general condition for operation. Other advantages of cryosurgery are the low complication and mortality rates, the possibility of out-patient treatment without anaesthesia, and thus a general reduction in cost. Indications for colostomy as final palliation can therefore be more narrowly defined than previously customary.


Local oxygen tension and its relationship to unit activity during penicillin interictal discharges in the bullfrog hippocampus. Simultaneous measurements of extracellular unit activity and pO2 were made with single polarographic microelectrodes in penicillin foci of the bullfrog hippocampus. Individual penicillin-induced ECoG interictal discharges were often associated with both a burst of extracellular unit activity and a simultaneous transient decrease in pO2. The bursts of unit activity lasted up to 1500 msec and were followed by longer periods of inhibition. Transient decreases in pO2 began within 150 msec of the onsets of the bursts but outlasted the bursts by many seconds. The durations of the pO2 transients ranged from 10 to 90 sec, depending upon the frequency of interictal discharges. When the frequency of interictal discharges increased, the pO2 transients summated such that pO2 approached 0 mm Hg. The magnitude of the pO2 decrease was related to both the baseline pO2 and the intensity of the associated burst of action potentials. The transient decrease in pO2 most likely represents increased local tissue O2 consumption. These measurements provide, for the first time, a means of assessing the relationship between local neuronal activity and local oxygen utilization in seizures.


Nerve fiber conduction-velocity distributions. I. Estimation based on the single-fiber and compound action potentials. A method is described for estimating the distribution of nerve-fiber conduction velocities in a nerve bundle. This method is based on a detailed general model of the nerve bundle compound action potential, which is characterized as a weighted sum of delayed single-fiber action potentials. The non-iterative estimation method is applied to two examples taken from existing literature, demonstrating the similarity of conduction velocity and fiber diameter distributions, sensitivity of the estimate to variations in important model parameters, and applicability to the differentiation of normal and abnormal nerve function.


Nerve fiber conduction-velocity distributions. II. Estimation based on two compound action potentials. A method is presented for estimating nerve fiber conduction velocity distributions from non-invasive measurements of the compound action potential at two distinct locations separated by a known distance along the nerve bundle. This method is based on a model of the compound action potential as a weighted sum of delayed single-fiber action potentials, but does not require explicit knowledge of the single-fiber action potential wave shapes in order to yield a unique estimate of the conduction velocity distribution. Illustrative examples are presented from normal and diseases nerves. This method appears to have clinical applications in the electrophysiological assessment of peripheral nerve function.


The estimation of conduction velocity in human skeletal muscle in situ with surface electrodes. In the present study new methods were developed for estimation of conduction velocity using surface electrodes. The subjects were 4 healthy male volunteers, aged 23-29 years. EMGs were recorded from m. biceps brachii by several surface electrodes and a fine-wire bipolar electrode during weak isometric contraction with the elbow positioned at 90 degrees. Surface electrodes, 8 mm in diameter, were spaced along the course of muscle fibers at 20 mm intervals, while the bipolar fine-wire electrode was inserted in the middle of the muscle. The EMGs recorded on the magnetic tape were analyzed through a mini-computer system (ATAC-2300). Triggering a signal averager by spike potentials of single motor unit recorded by the inserted electrode, the average contribution of the single motor unit to the surface EMG could be extracted. The conduction velocity in m. biceps brachii determined from the averaged waves of each surface electrode was 4.6 +/- 0.5 m/sec (mean +/- S.D.). The conduction velocity estimated from the cross-correlation analysis between the EMGs of two different surface electrodes agreed well with the value obtained by the averaging technique mentioned above.


Unilateral lesions of the olfactory tubercle modifying general arousal effects in the rat olfactory bulb. The centrifugal control exerted by different arousal states on the rat olfactory bulb was investigated. The olfactory tubercle was unilaterally coagulated with either 1 mA or 3 mA current. The vigilance state parameters and multiunit mitral cell activity were recorded in freely moving rats, stimulated either by their usual food odor or by isoamyl acetate, in a hungry or a satiated state. In each animal, a unilateral lesion affected resting activity and the relative proportion of positive (excitatory) and negative (inhibitory) responses in the same way in both olfactory bulbs; these effects were proportional to the extent of the lesion. In wakefulness, a nutritional modulation of the bulb responses for food odor existed in spite of a slight decrease in the general arousal level. In slow wave sleep (SWS), when compared to a control group, the rats with lesions showed an increase of neocortical desynchronization induced by olfactory stimulation, and a large decrease of mitral cell excitability. Inhibition of the olfactory input, which normally develops during SWS, could involve mainly mesencephalic neurons reaching the olfactory tubercle and the bulb via the ventral part of the medial forebrain bundle.


Electro-oculographic and electroencephalographic correlative study of optokinetic responses in brain lesions. Electro-oculograms of induced optokinetic responses (OKR) and EEG were recorded in 61 patients with either left or right hemisphere lesions. Of the 61 patients 55 showed focal EEG disturbances as follows: occipito-temporal (7 cases), parieto-temporal (10 cases), occipito-parieto-temporal (26 cases), temporal (9 cases) and frontal (3 cases). Symmetric OKR (21 cases) were recorded when no EEG changes were observed or when these were localized to left or right temporal and frontal electrodes and exceptionally when unilateral occipital and parietal regions were also involved. In 40 cases with unilateral hemispheric lesion a contralateral abnormal OKR was observed. Low frequency OKR with or without amplitude changes, especially of fast (saccadic) component, was mainly observed in parietal localization. In severely disturbed OKR, i.e. random jerks, the abnormal brain waves were mainly localized to the occipito-parieto-temporal region. In all cases in which no response was obtained the affected area was the occipito-parieto-temporal. These findings are discussed with reference to the regulating systems of slow (smooth) and fast (saccadic) eye movements triggered by visual stimuli.


Rapid eye movements in REM sleep--more evidence for a periodic organization. Density scores of horizontal eye movements during 70 REM sleep periods of normal young adults and combat neurotic patients were subjected to spectral analysis. The results showed that in 58% of all REM periods, spectral peaks occurred at either 0.1 c/min or 0.05 c/min. The percentages of spectral peaks at these frequencies were equal in both groups. Since the estimated variance at these frequencies was significantly higher than the estimated variance at the adjacent frequencies and the mean variance at the rest of the spectra, it was concluded that eye movements in REM sleep tend to cluster every 10-20 min. These results and findings on the neural control of REM ocular activity suggest that REMs in sleep are generated by at least 3 generators: a burst generator, an isolated eye movement generator, and a 10-20 min periodic generator.


Influence of diffuse brain stimulation (DBS) on human sleep. I. Sleep pattern changes. A 20 min period of diffuse brain stimulation (DBS), administered just before sleep onset in 16 human subjects, appeared to alter significantly the first sleep cycle of the succeeding sleep. The length of the first sleep cycle and the amount of REM sleep increased, whereas a shift from deep to more superficial NREM sleep occurred. This effect of DBS on sleep is discussed with respect to data from the literature on the effect of narcotic DBS and on that of local brain stimulation in animals.


Influence of diffuse brain stimulation (DBS) on human sleep. II. Sleep-induced periodic breathing with apnoea. The influence of diffuse brain stimulation (DBS), administered just before sleep onset, on the number of apnoeas and of sleep stage changes during the first part of the succeeding night sleep to the end of the first sleep cycle has been studied in 7 patients with sleep-induced periodic breathing with apnorea. When compared with the corresponding period of the baseline night, both phenomena are increased significantly. These results indicate that DBS influences slow periodic sleep phenomena. It is suggested that the target area for this influence might be mesencephalo-pontine reticular structures, which play a role in the generation of these slow periodic phenomena.


Short latency somatosensory potentials in humans. A sequence of high frequency potentials was averaged from the scalp of 10 normal human subjects during the first 25 msec following median nerve stimulation. There was a large positive component with a peak latency between 18.9 and 22.3 msec localized to the somatosensory area contralateral to stimulation. This was preceded by an early positive potential arising peripherally (peak latency 7.7-9.7) and at least 3 negative to positive deflections which appear to originate in multiple subcortical structures. When corrected for arm length, intersubject variability was less than 5% for all components. With further clarification, this method should allow one to study neural conduction along the entire somatosensory pathway.


Principal component analysis of ERP differences related to the meaning of an ambiguous word. Event-related potentials (ERPs) to the noun and verb meanings of/'led/in the single ambiguous phrase 'it was/'led/' were re-analyzed using principal component analysis (PCA). These data had previously been analyzed by SWDA and reported in this journal. PCA defined 3 meaning-related components, comprising 40.3% of the entire data variance. The N150 component was shown to be larger for the noun meaning than for the verb meaning; the P230 epoch differed in its anterior-posterior distribution according to meaning; and N370 for noun responses was relatively more negative at the right posterior lead and positive at the left anterior. All components taken together, the left anterior lead showed the greatest meaning-related difference. Previous analysis by SWDA had resulted in significant discriminant functions for left hemisphere ERPs, but this analysis did not yield a clear definition of the effects of meaning on specific ERP components or of the scalp distributions of meaning-related components. Thus, while the results of both analyses support the interpretation that the perceived meaning of words has a substantial effect on ERP wave forms, PCA appears to provide the clearest definition of the ERP component effects.


Projections on the cortical somatic I barrel subfield from ipsilateral vibrissae in adult rodents. In adult rats and mice, ipsilateral projections to SI posteromedial barrel subfield (PMBSF) are demonstrated for mystacial vibrissae (sinus hairs). Mechanical stimulation with angular deflections in the 2-5 degrees range, elicits potentials in limited areas of SI whose geometric centers coincide for two homologous (contra- and ipsilateral) vibrissae. The cortical domains for two adjacent vibrissae overlap one another slightly. Phase reversal of potentials and unit discharges are also present within the cortex. Ablation of the SI area contralateral to the SI area under study completely abolishes the ipsilaterally projected potentials. It is proposed that ipsilateral responses are mediated via the corpus callosum with an extra delay of 4-5 msec, thus giving each hemisphere the opportunity to compare vibrissal information originating from the two mystacial pads.


An optical scan system for encoding and tabulation of visually scored sleep data. A system for computer-assisted encoding, tabulation and analysis of visually scored sleep data is presented. The main features of the system are: (1) The use of computer-readable optical scan sheets for direct encoding of the sleep data. This eliminates the need for a separate transcription and/or key-punching operation. (2) The development of a visually scored data base, organized by NREM and REM periods, which contains all items of visually scored data indexed by time and/or page of occurrence in the sleep record. The advantages of this new system are: (a) the low cost per night of operation; (b) the facilitation of analysis of cycle phenomena and testing of new hypotheses that would usually involve retabulation of the data; and (c) the facilitation of analysis of real-time correlates of visually scored sleep stages.


A static charge sensitive bed. A new method for recording body movements during sleep. The principle of 'a static charge sensitive bed' method for recording body movements during sleep is described. We made records during 30 nights and the measured total number of movements per night (80-200) is in agreement with the findings of studies based on a combination of direct observation, EMG and videotape. The method is simple, inexpensive and very sensitive to all kinds of movement.


Release of growth hormone from purified somatotrophs: role of adenosine 3',5'-monophosphate and guanosine 3',5'-monophosphate. Studies were carried out to simultaneously measure cAMP and cGMP accumulation and GH release from acutely dispersed purified somatotrophs obtained from rat adenohypophyses. cAMP accumulation was dramatically increased by both prostaglandin E2 (10(-6) M) and 3-isobutyl-1-methylxanthine (a phosphodiesterase inhibitor, 0.5 mM) within 1 min of their addition, while there was a delay of 8--16 min before a significant increase in GH release was seen. SRIF (100, 10, or 1 ng/ml) completely blocked the stimulated release of GH. SRIF also consistently decreased the elevation of cAMP induced by the two secretagogues, but this decrease was small and not always significant. cGMP was unmeasurable (less than 0.02 fmol/1000 cells) in all of our experiments, while basal cAMP levels were about 1 fmol/1000 cells. We conclude that cAMP plays a role in the intracellular mechanisms governing GH release and that SRIF primarily acts subsequent to cAMP elevation, with a possible secondard or minor action on cAMP formation.


The use of Midodrin in the treatment of ejaculation disorders following retroperitoneal lymphadenectomy. The long-term treatment of retrograde ejaculation disorders following retroperitoneal lymphadenectomy with the alpha-sympathomimetic, Midodrin, administered orally, led to improvements in the intensity of orgasm and the degree of erection. Normal ejaculation was induced in 7 out of 12 patients and emission of spermatozoa into the posterior urethra was restored in 3 out of 12 patients by a single intravenous injection of 25--30 mg Midodrin.


Fine specificity of cytotoxic T lymphocytes: C57BL effector cells induced by autologous cells modified with hapten (4-hydroxy-3-nitro-phenyl)acetyl (NIP) are not heteroclitic. Cytotoxic T lymphocytes (CTL) were generated to syngeneic cells derivatized with serologically cross-reactive compounds (4-hydroxy-3-nitro-phenyl)acetyl (NP) and (4-hydroxy-5-iodo-3-nitro-phenyl)acetyl (NIP) in order to investigate the possible role of a heteroclytic T and B cell receptor marker (U. Krawinkel et al., Eur. J. Immunol. 1977. 7:566) in CTL-target cell interaction. The cytotoxic cells of both BALB/c and C57BL/10 (B 10) origin react specifically with target cells of the inducer type; no heteroclicity of the overall cytotoxic response can be observed by CTL of B 10 origin. The results suggest that in this system, the specificity of CTL is determined by a receptor with a variable region different from that of the isolated NP-specific antigen-binding T cell receptor material.


Antigen-specific suppressor cells in hapten-specific carrier-determined tolerance. Tolerance to the hapten 2,4-dinitrophenyl (DNP) induced by the injection of DNP coupled to isologous IgG (carrier-determined tolerance) is associated with a receptor blockade of antigen-binding lymphocytes. In the present study, hapten-specific suppressor cells were detected in the spleens of mice made tolerant by intravenous injection of 20 microgram DNP-IgG. When spleen cells from mice rendered tolerant to DNP were co-cultured with normal spleen cells in Marbrook cultures, the response to DNP-Ficoll was suppressed, while the response to sheep red blood cells was not altered. Depletion of T cells from these spleens restored the normal anti-DNP response. The suppressor cells were not detectable in the spleen lymphocyte population of mice in the early stages of tolerance but were present on day 7 after injection of tolerogen, and disappeared by day 14. Mice injected with larger doses of 1 mg or four weekly doses of 200 microgram DNP-IgG did not have detectable suppressor cells. Thus, it appears that a short-lived suppressor T cell is generated in carrier-determined tolerance. This cell most likely plays a minor role in the mechanism of carrier-determined tolerance and may be associated with the receptor blockade which is seen early in tolerance.


Relationship between histocompatibility antigens, other surface determinants and the IgE receptor on rat mast cells. Rat alloantibodies recognizing classical transplantation antigens (CTA) or non-H-1 determinants were able to compete effectively with monomeric IgE or IgG-coated sheep erythrocytes for receptor sites on the rat mast cell surface. Inhibitory capacity, however, was entirely confined to anti-CTA antibodies of the IgG2a subclass, whereas IgG1 antibodies lacked this ability. Analogously, F(ab')2 fragments of anti-CTA antibody consistently failed to affect IgE binding, but exposure of cell-bound F(ab')2 to anti-rat IgG restored its inhibitory capacity. From these results it was concluded that receptor sites recognizing the Fc portion of the anti-CTA molecule are involved in the inhibition process. Based on a cytotoxicity assay and on comparative absorption studies on alloantisera, the existence and relative amount of CTA and I region-associated antigens on purified rat mast cells and lymph node cells were analyzed. Whereas the CTA concentration per unit surface area on both cell types was very similar, rat mast cells consistently lacked Ia antigens.


T cell-produced immunoglobulin binding factor (IBF) bears determinants coded for by the I region of the major histocompatibility complex and lacks allogeneic restriction. An immunoglobulin binding factor (IBF) produced by in vivo alloantigen-activated T cells suppresses in vitro antibody production to sheep red blood cells (SRBC) and probably represents the soluble form of the T cell Fc receptor. To study the relationship between IBF and the major histocompatibility complex (MHC), two types of experiments were undertaken. First, different IBF were prepared by injecting thymocytes from mice of H-2k, H-2b and H-2d haplotypes into irradiated, allogeneic recipient mice. Cells recovered in the spleens five days later were incubated for 2 h at 37 degrees C, and IBF was purified by affinity chromatography on insolubilized IgG. Purified IBF was then applied to immunosorbents prepared with anti-H-2 and anti-Ia anti sera. Acid eluates were then tested for their suppressive activity on in vitro antibody production to SRBC. Under these conditions, IBF was found to react with antisera raised against products of the whole MHC and of the I region of the MHC, but not with anti-H-2 D antisera. These data indicate that IBF bears, or is associated with Ia determinants. Second, IBF preparations from mice of H-2k, H-2b, H-2d haplotypes were tested for their capacity to suppress the in vitro antibody production to SRBC by spleen cells of a series of mouse strains. The data indicate that IBF is not genetically restricted in its function.


Induction and characterization of isogeneic anti-idiotypic antibodies to BALB/c myeloma S117: lack of reactivity with major idiotypic determinants. Isogeneic anti-idiotypic antibodies were induced by immunization of BALB/c mice with the BALB/c-derived myeloma protein S117 which binds N-acetylglucosamine-containing antigens including Group A streptococcal carbohydrate (A-CHO). Most BALB/c mice produced anti-S 117 idiotypic antibodies, as shown by various different immunization protocols. The antibodies of individual mice were of intermediate to high affinity (2.8 x 10(6) M-1 to 1.4 x 10(8) M-1). In isoelectric focusing, most individual antibodies were shown to consist of a small number of clonotypes, but each mouse produced its own unique set of clones so that the potential clonal repertoire of strain BALB/c is rather large. Most importantly, all isogeneic anti-idiotypic antibodies were directed against idiotypic determinants of S117 that are not shared with induced anti-A-CHO antibodies, whereas it has been shown previously that allogeneic and xenogeneic anti-S 117 idiotypic antibodies react with idiotypic determinants unique to S 117 as well as with those that are shared with anti-A-CHO antibodies. The data suggest that the expression of S 117 idiotypic determinants in strain BALB/c is not under antibody-mediated, anti-idiotypic feedback control.


Staining of granulopoietic agar colonies with a modified Papanicolaou technique. The aceto-orcein method of staining agar cultured granulopoietic marrow cells has not been entirely satisfactory. In staining whole colonies the original colony structure is lost and the colony cells have blurred cytoplasm. A slight modification of the Papanicolaou technique used for staining vaginal smears improved the quality of stained granulopoietic agar preparations. Smeared Papanicolaou stained colony cells could be compared with smeared ordinary Giemsa stained blood and marrow cells. The Papanicolaou method enabled staining of agar colonies without manipulation of the original colony structure. The preparations of transferred intact colonies had cells with distinct cytoplasm and clear cellular outlines and a background without disturbing masses of stained agar.


Histamine release in dogs by Emulphor EL620. A vehicle containing ethanol and Emulphor EL620 lowers blood pressure and increases heart rate in morphine-chloralose anesthetized dogs. These effects are associated with histamine release caused by Emulphor EL620.


Inhibition of fading in fluorescence microscopy of fixed cells. Following Hirschfeld et al., dithionite was added to mounting media to inhibit fluorescence fading. Excellent response is reported for fluorescein, acridine orange, 33258 Hoechst, acriflavine, berberine (and ethidium bromide), but not for quinacrine.


Association between pregnancy-associated alpha2-glycoprotein (alpha2-PAG) and mixed leucocyte reaction determinants on the leucocyte surface. alpha2-PAG is present on the surface on mononuclear blood leucocytes and can be demonstrated predominantly on B-lymphocytes and monocytes. Pretreatment of cells with antibody to alpha2-PAG leads to a marked reduction in Fc-rosette formation. Competitive blocking experiments with specific antisera reveal a particularly close association between alpha2-PAG and MLR (mixed leucocyte reaction) determinants on the cell surface. These findings suggest one mechanism whereby alpha2-PAG may modify cell-mediated immune responses.


Specific inhibition of formation of acid-fastness in mycobacteria by 3,3'-di-O-methylellagic acid. 3,3'-Di-O-methylellagic acid obtained from Euphorbia adenochlora selectively inhibited the formation of acid-fastness in mycobacteria without retardation of their growth. Gross reductions in contents of wax D, cord factor and free mycolic acids were found in the nonacid-fast bacilli compared with the normal ones.


A simple one-step procedure for staining the nucleolus organizer regions. A simple silver staining technique for routine use is described by which the nucleolus organizer regions of mammalian chromosomes, including those of mouse chromosomes, are stained selectively.


Alpha-fetoprotein in tumor-bearing mice assayed by particle agglutination inhibition. Serum alpha-fetoprotein (AFP) in hepatoma BW7756-bearing mice was measured by a new particle agglutination inhibition test employing AFP adsorbed to charcoal particles. The AFP levels and tumor weights showed nearly parallel increases to means of 2633 microgram/ml and 5.2 g, respectively, 28 days after implantation.


Colored filters for microphotography: a costless 'do-it-yourself' solution. Black and white microphotography makes use of several colored filters for an increased contrast. These filters are sometimes expensive and do not correspond exactly to the stain to be masked on the negative. Our specific colored filters contain a sufficient and adjusted amount of the dye used in the staining technic in an Agar medium.


Sequential quantitative studies on hyperplastic nodules in the liver of rats treated with carcinogenic chemicals. Sequential quantitative analyses were made of hyperplastic liver nodules induced by treating male Fischer rats first with diethylnitrosamine (DEN) and then with N-(2-fluorenyl)acetamide (2-FAA), alpha-isomer of 1,2-3,4,5,6-hexachlorocyclohexane (alpha-BHC), or phenobarbital. The test rats were injected ip with 200 mg/kg body weight of DEN, given basal diet containing 2-FAA, alpha-BHC, or phenobarbital from week 2 to 12, and subjected to partial hepatectomy at the end of week 3. From week 6, significantly higher percentage areas of hyperplastic nodules per unit area of liver were found in all experimental groups than in the control group. From week 8, significantly more hyperplastic nodules were found in all experimental groups than in the control group. Continuous administration of alpha-BHC or phenobarbital for 12 weeks with partial hepatectomy did not induce hyperplastic nodules. Continuous administration of 2-FAA did induce hyperplastic nodules, but the percentage areas and number of these nodules were significantly lower than in rats injected with DEN and then given 2-FAA orally, with partial hepatectomy.


Benign prostatic hypertrophy: When should you consider prostatectomy for your patient? Watching the way a patient voids is the easiest way to determine the presence of prostatic obstruction. Uroflowmetry can objectively measure the amount of obstruction. Clinical evaluation of prostatism should include an intravenous urogram, serum creatinine determination, urinalysis, urine culture, and cystoscopy. Suspicious areas of induration in the prostate should be biopsied. Surgery is not required for most men with prostatic hypertrophy. Indications for prostatectomy are severe symptoms of obstruction, significant renal or bladder decompensation, recurrent urinary tract infection, bladder calculi, and recurrent bleeding.


Angiocardiographic left ventricular volume determination in tricuspid atresia. Comparison of patients with and without palliative surgery. Group A (n = 10) had reduced pulmonary blood flow and no previous surgery, group B (n = 9) had decreased pulmonary blood flow despite a systemic-to-pulmonary artery shunt and those in group C (n = 10) had increased pulmonary blood flow 9 of whom had no previous surgery and 1 a large Waterston anastomosis. Left ventricular end-diastolic volume (LVEDV) and left ventricular systolic output (LVSO) were higher than normal in all 3 groups with an order of descending magnitude of group C (278 +/- 20% and 264 +/- 32%), group B (264 +/- 19% and 243 +/- 37%) and group A (189 +/- 14% and 190 +/- 13%). For the entire group A, left ventricular ejection fraction (LVEF) was normal (0.66 +/- 0.06 or 97 +/- 8%) with low systemic arterial oxygen saturation (SAO2) averaging 58%, but the LVEF of those infants less than 6 months with a mean SAO2 of 49% was lowered to 0.58 +/- 0.08 or 87 +/- 13% of normal. The ejection fraction was reduced to the greatest extent (0.50 +/- 0.04 or 81 +/- 6%) in group B patients who averaged 12.8 years of age and had undergone shunt procedures 10 months to 13.6 years, median 7.8 years previously. These findings indicate that a moderate degree of arterial desaturation appears to be better tolerated than a chronic volume overload in patients with tricuspid atresia.