Prenatal methylmercury poisoning. Clinical observations over five years. Thirty-two infants prenatally exposed to methylmercury and their mothers were examined over a five-year period after the Iraqi methylmercury epidemic. Severity of poisoning in mothers was related to the peak mercury concentration in their hair and in the infants to the maximum concentration in maternal hair during pregnancy. In nine cases of cerebral palsy, methylmercury exposure occurred only during the last trimester. All infants except three (two were orphaned soon after birth and one was bottle-fed) were exposed postnatally via suckling. Whereas the mother's symptoms usually improved, the damage to the fetal nervous system appears to be permanent. Milder cases previously not identified in other studies are reported. The syndrome consists of varying degrees of developmental retardation in addition to exaggerated tendon reflexes and the pathologic extensor plantar reflex (minimal brain damage syndrome).


Protected environment-prophylactic antibiotic program for malignant lymphoma. Randomized trial during chemotherapy to induce remission. Fifty-eight patients with malignant lymphoma were randomly allocated to receive three courses of chemotherapy to induce remission with CHOP-Bleo on the protected environment-prophylactic antibiotic (PEPA) program (30 patients) or as controls (28 patients). The complete remission rate for all patients was 74 per cent, for patients with diffuse histiocytic lymphoma 78 per cent and for patients with nodular poorly differentiated lymphocytic lymphoma 65 per cent. There were no significant differences in response rates and duration of responses between those on the PEPA program and control patients. The frequency of infection was significantly lower among the patients on the PEPA program, and dosage escalation of the chemotherapeutic agents was accomplished more often among these patients. Dosage escalation did not increase the complete remission rate, but it did reduce the relapse rate and signficantly reduced the fatality rate. The duration of remission and survival was significantly longer for those patients who received dosage escalation.


Interpreting elevated amniotic fluid alpha-fetoprotein levels in clinical practice: use of the predictive value positive concept. The amniotic fluid alpha-fetoprotein assay (AF-AFP) is the single most valuable method of detecting anencephaly--spina bifida (ASB) in utero. A raised AF-AFP, however, is a more reliable indicator of a fetus with ASB in the pregnancy at high risk for ASB. Most women undergoing fetal chromosomal analysis are at low risk for ASB. Consequently, gravid patients who have an AF-AFP incidental to chromosomal studies, and elect abortion because of a raised AF-AFP, will more frequently abort a normal fetus than the gravida who has previously had an infant with ASB. For the obstetrician utilizing this assay, the predictive value positive (PVP) concept, a guide for the interpretation of a positive result, is introduced and then applied to examples of patients at varying risk for ASB in their offspring.


[First experiences with an instrument for TV display of digitally encoded text information for the visually handicapped (author's transl)]. An instrument was developed which allows the display, in enlarged form--especially for visually handicapped persons--of text information as used in modern data-acquisition systems such as writing automats, screen editing devices, and television text display (in England, Ceefax). With the aid of electronic character generators, the text is displayed continually, moving from right to left in one line on the screen of a commercial TV set. Rate of movement, size, brightness, and color of the characters (and/or the background) can be selected arbitrarily. We performed a preliminary reading test with visually handicapped subjects from a rehabilitation center for blind persons. The resulting optimal parameters for text display and the achieved reading speed with the system are discussed.


[Demonstration of the juxtaglomerular cell granules with phosphotungstic acid hematoxylin (author's transl)]. Author recommends for staining of the JGC-granula after oxydation with periodic acid a solution of phosphotungstic acid hematoxylin ripened by kalium iodate, and for counterstain an orange G solution.


Equine markers genes. Polymorphism for group-specific component (Gc). Polymorphism of equine Gc protein was demonstrated by immunofixation electrophoresis with a goat anti-human Gc antibody. Three different phenotypes, F, FS and S, were found. Family data supported the genetic theory of two autosomal codominant alleles, GcF and GcS. Both alleles occurred in Standardbred, Thoroughbred and Arabian horses and in Shetland ponies. A frequency of 0.23 for GcS in the American Standardbred horse indicates the system should be useful for problems of identification and parentage.


Human lung tissue and anaphylaxis: the effects of histamine on the immunologic release of mediators. The IgE-mediated, antigen-induced release of histamine from human lung tissue causes profound changes in lung cyclic adenosine monophosphate and cyclic guanosine monophosphate. Exogenous histamine similarly induces increases in both cyclic nucleotides; pretreatment with H-1 antihistamines prevents the increase in cyclic guanosine monophosphate, whereas H-2 antihistamines prevent the increase in cyclic adenosine monophosphate. Anaphylaxis of human lung in vitro is unaffected by the presence of 1-100 micron histamine, H-1 antihistamines, H-2 antihistamines, or combinations of these agents despite the production of selective increases in total lung cyclic nucleotides. Futhermore, selective histamine agonists (2-methylhistamine [H-1 agonist] or dimaprit [H-2 agonist]) also fail to significantly influence the immunologic release of mediators. Histamine examined in the presence of ethylenediaminetetra-acetate was no more capable of modulating mediator release than when in the presence of calcium, in contrast to previous studies involving the human basophilic leukocyte. Therefore, the human lung mast cell is unresponsive to histamine with regard to modulating the antigen-induced, IgE-dependent, generation of mediators.


[Syncope and transitory amaurosis during mastocytosis]. In a 70-year old man who has been suffering for 20 years from mastocytosis, a syncope followed by a transitory amaurosis occured. Neurological complications of mastocytosis are exceptionnal. Syncope is the most frequent, secondary to a drop in arterial pressure due to an inappropriate discharge of histamin.


Factors affecting the fibrinolytic response to surgery. The stress of injury and surgical operation results in an initial increase in the spontaneous fibrinolytic activity of the blood which is followed by a period of reduced activity in the postinjury or postoperative period. This 'fibrinolytic shutdown' is particularly marked in patients with malignant disease and occurs irrespective of whether or not they develop a deep venous thrombosis. It also occurs in patients with benign disease and in these patients is greater, though only on the first postoperative day, in those who develop deep venous thrombosis. Venous occlusion studies suggest that this reduction in spontaneous fibrinolytic activity may be the results of a reduction in the fibrinolytic capacity of the vascular endothelium resulting either from a deficiency of the enzyme plasminogen activator or an inability to release the enzyme from the endothelium. Changes in antiplasmins, the inhibitors of the fibrinolytic system, also occur as a result of the stress of operation. Plasma levels of alpha2-macroglobulin fall while those of alpha1-antitrypsin rise. These changes occur irrespective of the presence of malignant or benign disease and do not appear to influence the development of deep venous thrombosis.


Thyrotoxicosis during pregnancy.

Thyrotoxicosis during pregnancy. The diagnosis of thyrotoxicosis during pregnancy is difficult on clinical grounds only. The determination of free thyroid hormones or free thyroid hormone indices is important and is possibly supplemented by the TRH stimulation test. Thyrotoxicosis during pregnancy should always be treated actively. The authors recommend subtotal thyroidectomy if there are indications for surgical therapy (nodular goitre or large diffuse goitre). Otherwise treatment with thyrostatic agents is used. After operation during pregnancy, substitution with thyroid hormones should be given throughout the pregnancy in order to avoid deterious effects of possible maternal hypothyroidism. Antithyroid treatment should be continued till after delivery. Beta-receptor blockers are used only as adjuncts but are not recommended as the sole therapy.


[Effect of S-adenosyl-L-methionine on the cerebral and peripheral metabolism of L-dopa]. The repartition of [3H]L-dopa administered i.p. does not change in rats treated with S-adenosyl-L-methionine (SAM). The effect was studied on the central nervous system and at the periphery. The biosynthesis of metabolites [3H]dopamine and [3H]3-O-methyldopamine are also unchanged. On the other hand, in the kidney an accumulation of [3H]norepinephrine +[3H]normetanephrine was observed, while in the brainstem + midbrain the synthesis of these metabolites was decreased after SAM injection. The relations between these biochemical results and the pharmacological antiparkinsonian effects are discussed.


[Kinetic properties of the fructose influx across the brush border of the rat jejunum. Effects of a diet rich in fructose]. The unidirectional influx (i.e. initial rate of uptake) of D-fructose across the brush border of rat jejunum is a saturable function of concentration, with a Kt of 125 mM, which implicates a carrier mechanism. This mechanism appears to be very specific for fructose in view of the lack of influx inhibition observed in the presence of large concentrations of the sugars or polyols, D-glucose, D-galactose, D-mannose, D-xylose, L-sorbose, D-tagatose, sorbitol or mannitol. D-Fructose uptake is inhibited by incubation, preceded by a 30-min preincubation in the same inhibitory conditions, in the absence of Na, or in the presence of metabolic poisons, NaF, 2,4-dinitrophenol, monoiodoacetate. Phloridzin (10-3 M), with or without preincubation, has no effect on uptake. D-Fructose influx is stimulated by fructose feeding, mainly because the augmentation of the number of active sites of transfer: Jmax is increased two-fold, Kt is more weakly affected.


[Effect of dietary alpha-linolenic acid on the conversion of linoleic and gamma-linolenic acids (1-14C) into arachidonates in rats in vivo]. The effects of alpha-linolenic acid (9-12-15 octadecadienoic) upon the conversion in vivo of [1-14C] linoleic acid and of [1-14C] gamma-linolenic acid into arachidonate have been studied in adult rats. The two tracers have been administered by stomach tubing and the amounts of [14C]-radioactivity incorporated into arachidonate in the liver, kidneys and whole rat have been measured 48 h later. Three experiments have been carried out on rats fed on alpha-linolenic acid containing diets prior to the radioactive tubing. In these diets, alpha-linolenic acid was brought either as ethyl ester or in the form of Primor oil (erucic acid free rapeseed oil). In all of them, the ratio alpha-linolenic acid: linoleic acid did not exceed 0.45. Control animals were fed, in the same conditions, ethyl oleate or peanut oil respectively. Comparing the alpha-linolenic acid fed-rats to the control animals, we were able to observe the following results: (1) The exogenous supplies of alpha-linolenic acid used in the diets have not brought about any significant alteration in the amounts (weights) of arachidonic acid present in the liver, kidneys and whole animal. (2) Using [1-14C] linoleic acid as a precursor, the amounts of [14C]-radioactivity incorporated into arachidonate in the same organs as well as in the whole rat have been significantly lowered by dietary alpha-linolenate. (3) alpha-Linolenate, on the contrary, had no significant effect upon the amounts of radioactivity incorporated into hepatic, renal and whole body arachidonate following the administration of [1-14C] gamma-linolenic acid. These results lead to the conclusion that alpha-linolenic acid, when present in the diet of rats at a limited, phyisological level, partly inhibits the desaturation of linoleic acid in vivo but does not affect the subsequent reactions in the biosynthesis of arachidonic acid.


Non-electrolyte permeability through black lipid membranes with different surface charge. The permeability coefficients of some non-electrolytes has been measured across black lipid membranes of different composition. Large discrepancies between permeability coefficients and oil-water partition coefficients has been observed. The discrepancy level seems to be related to the degree of organization of the membrane.


Changes in muscle, liver, and plasma free fatty acid levels in the pigeon on acute exposure to cold. Adult pigeons were subjected to acute cold exposure (-25 degrees C; 30 min). Fully-plumed birds, showed a pronounced increase in the level of FFA in the blood, but not in the liver or muscle. Partially-defeathered (dorsum and pectoral regions) birds, likewise, showed a marked increase in plasma FFA level but failed to indicate any change in FFA levels in the liver or the muscle. It is concluded that even if the mobilized FFA may have supported calorigenic processes in the normothermic cold-exposed pigeon, lipid reserves are unlikely to have served as a significant source of energy for thermogenic functions in the hypothermic (defeathered) bird.


Neuromuscular blocking agents and spontaneous sympathetic activity. The action of neuromuscular blocking agents on the spontaneous sympathetic activity has been quantitated. "On line" spectrum analysis has been applied to the action potential of pre- and post ganglionic nerves of the coeliac plexus. The activity, the frequency spectrum and their changes after the injection of clinical and high doses of decamethonium, D-tubocurarine, succinylcholine, gallamine and pancuronium are determined.


[A study in twins of the urinary excretion of vanilmandelic acid, 17-keto-steroids, 17-hydroxycorticosteroids, and creatinine]. The 24-hour urinary excretion rates of creatinine, vanilmandelic acid (VMA), 17-hydroxycorticosteroids, 17-ketosteroids were analysed in 30 female twin pairs. For each of these variables, a comparative study based on the breakdown of total variance into interpair and intrapair variance was undertaken, first on identical (MZ) and fraternal (DZ) twins and then on twins who live together or separately. The comparison of test results shows a strong likeness in the MZ group and in the living together group with respect to the urinary excretion of 17-OH. The variability of VMA excretion can be explained by genetic factors while the variability of 17-ketosteroids and creatinine can be explained with reference to environmental factors.


The effect of unilateral castration on plasma and testicular testosterone in rabbits from birth to 60 days. Male rabbits were hemigonadectomized every 10 days from 1 to 50 days of age and were sacrificed 10 days after the operation. Non-operated controls were sacrificed at the same stages. Plasma and testicular testosterone were quantified by radioimmunoassay. Compensatory testicular hypertrophy was not observed in hemicastrated rabbits. Hemicastration has variable effects on testicular and plasma testosterone levels as a function of age. When it is performed at birth, there is a significant increase in testicular and plasma concentrations of testosterone. Similar results are obtained if the operation is performed at 30 days. Unilateral castration performed at 10, 20 and 50 days reduces testicular and plasma testosterone levels. When hemicastration is performed at 40 days or at the adult age, testicular and plasma testosterone levels are not modified.


Effect of changing the frequency of conditioning tetanus on depressor responses evoked by stimulation of the aortic nerve. The effect of changes in frequency of the conditioning tetanus on the magnitude of the testing depressor response was studied in rabbits anaesthetized with urethane. Conditioning and testing stimulations were applied to the same aortic nerve. The duration of the conditioning tetani was set at 3 and 60 sec and the interval between stimulations amounted to 40 and 120 sec. At the testing interval of 40 sec the increase in frequency both of short and long conditioning tetani reduces the magnitude of the testing response which attains a minimum at frequency of about 30 cycles/sec. Conditioning stimulations of higher frequency are gradually less effective and cause the testing response to increase. Similar depression is observed at the testing interval of 120 sec but only following long-lasting conditioning tetanus. Short conditioning trains at the testing interval of 120 sec facilitate the testing response. The frequency of the conditioning stimulation which produces the greatest reduction of the depressor response indicates the range of control exerted by the conditioning tetanus over the testing blood pressure effect. The size of this control is determined by the lowest level of depression and the highest value of facilitation of the testing response.


Akaline phosphatase: activity and variation in human diploid fibroblasts. A method is introduced for the assay of alkaline phosphatase in homogenates of cultured human skin fibroblasts. In a first group of 11 strains, a four- to fifteen-fold increase of enzyme activity is consistently observed following a period of starvation. In the remaining 31 cell-strains similar specific activities of alkaline phosphatase are found irrespective of medium changes. In regularly fed cultures, an inverse exponential correlation between the specific activity of alkaline phosphatase and the age of the donor has been detected.


[The effect of a synthetic anticholinergic agent, N-methyl hyoscine methyl sulfate, on the exocrine pancreatic secretion of the alert dog]. The inhibitory action of N-methyl hyoscine methyl sulphate (N-methyl scopolamine, an anticholinergic drug) has been studied on the exocrine pancreatic secretion stimulated by secretin + caerulein on the conscious dog provided with Thomas cannulae. The dose-response curve shows an "all or nothing" effect on protein secretion since 0.38 microgram/kg. The inhibition of water and bicarbonate secretion is only observed from 12 micrograms/kg. The inhibitory effect of this drug was much greater than that obtained with similar molar quantities of atropine and no central effect has been observed.


Inhibitory activity of alpha 1-antitrypsin in infants with idiopathic respiratory distress syndrome. In two groups of neonates serum concentrations of alpha-1-antitrypsin (1-AT) were determined by means of radial immunodiffusion, and 1-AT inhibitory capacity determinations were made using benzoyl-1-arginine-p-nitroanilide as a substrate. There were 66 children in the first group of neonates with idiopathic respiratory distress syndrome (IRDS), and 82 healthy children in the second group with birth weight and gestational and post natal ages similar to those of neonates with IRDS. In the latter, 1-AT concentration and inhibitory activity levels were lower than those found in healthy children; these differences were statistically significant. These findings explain both the retardation in the lysis of the hyaline membrane and the fact that a decrease in serum 1-AT levels elicits lung tissue damage by the action of proteolytic enzymes contained in leucocytes.


Effect of chlorophyll-a, fluorouracil, and pituitrin on experimental acute pancreatitis. Intraductal administration of enterokinase in rats produced hyperamylasemia and acute hemorrhagic pancreatitis. The experimental pancreatitis and hyperamylasemia could be prevented by the concomitant intraductal injection of fluorouracil, pituitrin, or chlorophyll-a. The clinical implication of the study is that these agents, if given intraductally, may be useful in the prevention of iatrogenic hyperamylasemia and acute pancreatitis that may occur after endoscopic retrograde pancreatocholangiography.


Blockade of rapid axonal transport. Effect of intraocular pressure elevation in primate optic nerve. After acute intraocular pressure (IOP) elevation, an induced disturbance of rapid axonal transport at the optic nerve head began within three hours at the IOP levels tested. The accumulation of radioactive label at the scleral lamina cribrosa increased with time of IOP elevation. There was a 60% decrease in the amount of transported material in the optic nerve, tract, and lateral geniculate body (LGN). Detailed analysis suggests that this decrease is not due to a simple slowdown of transport, but results from a total block of rapid transport in some axons, with no impairment in other axons. This total blockade of rapid transport by elevated IOP in involved axons differs from the apparent slowdown of transport in experimental papilledema, and the difference may explain the response of ganglion cells to the two conditions.


Toxic effects of dental amalgam implants. Optical histological and histochemical observations. Dental amalgam and porcelain control discs were implanted in the subcutaneous tissue of rats. Amalgam implants exerted a toxic effect caused by release of metal ions which was characterized by prolonged inflammation, delayed granulation and disordered collagen formation. Dental porcelain was found to be a suitable biocompatible control which provided a bulk similar to that of dental amalgam but induced only an uncomplicated granulation response.


Fetal-maternal macrotransfusion--a study of 400 postpartum women. The results of postpartum fetal cell counting in 400 Rh-negative patients delivering Rh-positive infants are presented and correlated with the mode of delivery and the ABO blood group relationship of baby and mother. Manipulative procedures, especially Caesarean section, are shown to increase the incidence of macrotransfusion. To make the most economical use of the new 125 microgram doses of anti-D gamma globulin, quantitation of fetal-maternal transfusions is shown to be vital if the existing level of protection from rhesus sensitization is to be maintained or increased.


Immunofluorescence studies of anti-T cell antibodies and T cells in systemic lupus erythematosus. Selective loss of brightly staining T cells in active disease. Peripheral blood T cells from patients with systemic lupus erythematosus (SLE) and age- and sex-matched controls were studied by flow microfluorometry by using SLE anti-cell antibodies and fluorescein conjugated antibody to human IgM. Brightly staining cells were reduced in a number of patients with active SLE. Analysis of SLE T cells separated on a discontinuous stractin gradient indicated a preferential loss of cells, especially brightly staining cells, from one fraction. Quantitation of this phenomenon indicated that an average of greater than 90% of the brightly staining T cells from that fraction was lost in active SLE. This preferential loss of a subpopulation of T cells in patients with active SLE may be responsible for many of their immunologic abnormalities.


Detection of intermediate complexes by evaluation of the difference between gamma-globulin and IgG concentrations. A simple analysis based on techniques readily available in the routine clinical laboratory has been presented for identification of patients with moderate and high levels of intermediate complexes. If the gamma-globulin concentration determined by serum protein electrophoresis minus the IgG concentration determined by radial immunodiffusion exceeds 1.0 gm/dl, intermediate complexes of IgG should be present in concentrations greater than 0.5 gm/dl. In principle, any disease association with sufficient polymerization of IgG might cause an abnormal result with this analysis. Furthermore, sera from patients with liver disease and marked beta-gamma-bridging on electrophoresis may also give abnormal values.


Subtypes of i demonstrated by the use of atypical Ii cell types and inhibition studies. Haemagglutination, absorption and elution studies with several anti-i sera and atypical Ii red cell types and Oh cells were carried out and three subtypes of specificity were demonstrated. The fourth i subtype was demonstrated by an inhibition method using human serum and amniotic fluid as a source of i substance.


Binding of bleomycin to DNA: intercalation of the bithiazole rings. At pH 5.5, binding of bleomycin relaxed supercoiled ColE1 DNA without breaking it. Binding of tripeptide S, a fragment of the drug containing the bithiazole rings, also relaxed and then recoiled supercoiled DNA, at pH 5.5 and at pH 8.0, where bleomycin is normally active. The unwinding angle was 12 degrees. Both compounds lengthened linear DNA by 3.1 A per molecule bound, and linear dichroism (303--315 nm) of bleomycin bound to linear DNA oriented in an electric field indicated the presence of a chromophore making an angle of 59--61 degrees with the helix axis. These results strongly suggest that bleomycin binding to DNA involves intercalation of the bithiazole rings. In 0.1 M Na Cl at pH 8, supercoiled ColE1 DNA was broken at a rate 50% greater than relaxed closed circular ColE1 DNA. Since supercoiling increases the affinity of DNA for intercalators, this result suggests that intercalative binding is involved in bleomycin-induced breakage of DNA.


Laser-Raman investigation of phospholipid-polypeptide interactions in model membranes. The interaction of aqueous dimyristoyl-phosphatidylcholine liposomes with the polypeptides gramicidin A, poly-L-lysine, valinomycin, and gramicidin S was investigated by means of laser-Raman spectroscopy. Auxiliary data were obtained with differential scanning calorimetry. Studies were carried out over the temperature range of 0--50 degrees C, encompassing the gel phase, the transition region, and the liquid crystalline phase of the liposomes.  Conformational changes in the phospholipid molecules were investigated by measuring the intensity of the 1062-cm-1 Raman band which is assigned to C-C stretching vibrations of trans segments. Three different types of phospholipid-polypeptide interactions were indicated by the observed Raman data. They are interpreted as (a) orderly penetration of the phospholipid bilayer by a hydrophobic polypeptide; (b) polar interactions involving primarily the head groups of the phospholipid; and (c) disorderly hydrophobic binding between a polypeptide and the hydrocarbon domain of the phospholipid.


A dimer--dimer binding region in beta-galactosidase. alpha Complementation in beta-galactosidase is the restoration of enzyme activity by addition of the alpha donor CNBr2, from amino acid residues 3--92 of the polypeptide, to inactive M15 protein from the lacZ deletion mutant strain M15. M15 protein lacks residues 11--41 and is a dimer; the active complex, like native beta-galactosidase, is tetrameric [Langley, K. E., & Zabin, I. (1976) Biochemistry 15, 4866--4875]. A dimer--dimer binding region in beta-galactosidase has been identified by proteolytic and immunologic studies of alpha-complementation. Proteolytic experiments were carried out with trypsin. Treatment of native beta-galactosidase with trypsin, followed by reaction of the mixture with cyanogen bromide, yields intact CNBr2 as measured by its ability to complement M15 protein. Active CNBr2 is not obtained when urea-denatured beta-galactosidase is treated in the same way. Therefore the segment corresponding to CNBr2 is apparently buried within the folded protein. Immunologic experiments were carried out with antibodies against CNBr2, tryptic peptide T8 (residues 60--140), and CNBr3 (residues 93--187). Anti-CNBr2 and anti-T8 bind to M15 protein but not to beta-galactosidase, indicating that this area is exposed in the dimer. Anti CNBr2, but not anti-T8 or anti-CNBr3, inhibits the formation of alpha-complemented enzyme. These results indicate that an early part of the sequence, within the segment corresponding to CNBr2, is involved in dimer--dimer interaction.


Photoaffinity labeling of the catalytic site of prenyltransferase. Three photoreactive substrate analogues, o-azidophenethyl pyrophosphate, p-azidophenethyl pyrophosphate, and 3-azido-1-butyl pyrophosphate, have been synthesized as site-directed probes to label the catalytic site of prenyltransferase. Due to the relatively poor affinity of p-azidophenethyl pyrophosphate and 3-azido-1-butyl pyrophosphate for the enzyme, only o-azidophenethyl pyrophosphate (aryl azide) was utilized for photoaffinity labeling. This aryl azide has a UV absorption maximum at 250 nm. In the absence of activating light, binding studies demonstrate that the o-aryl azide competes for binding with both the natural substrates, isopentenyl pyrophosphate and geranyl pyrophosphate. More than 90% enzymatic activity is lost when enzyme is irradiated in the presence of the aryl azide as compared to irradiation in the absence of the azide, and the protein loses its capacity for substrate binding in direct proportion to photolabeling. A stoichiometry of 2 mol of affinity label covalently bound per mol of enzyme dimer was established with [1-3H]-o-azidophenethyl pyrophosphate. Since there are two catalytic sites per enzyme dimer, the o-aryl azide appears specifically to label the enzyme at its catalytic sites. Additional evidence that the reagent was specific for the catalytic site came from the observation that farnesyl pyrophosphate afforded complete protection against photoinactivation, while isopentenyl pyrophosphate provided partial protection. Gel isoelectric focusing verified this stoichiometry and indicated that the labeled enzyme has a more acidic isoelectric point than the native enzyme.


Association of bacteriorhodopsin with lipid-impregnated filters. Evidence for fusion of bacteriorhodopsin-containing vesicles with the lipid phase of the filter. Bacteriorhodopsin vesicles were associated with cellulose-nitrate filters impregnated with a solution of phospholipids in hexadecane. The generation of (photo)potentials upon illumination of the filter was studied in the absence and presence of ionophores, phospholipase A2, EDTA or polyene antibiotics. From these experiments the following conclusions are drawn. 1. Upon illumination of the filter, bacteriorhodopsin pumps protons into aqueous compartments located in the filter. 2. These aqueous compartments possibly do not originate from the compartments enclosed by the bacteriorhodopsin vesicles. Evidence is obtained that aqueous compartments are present in the surface layers of the lipid-impregnated filters. 3. The results are explained most easily by a mechanism, whereby fusion occurs between the vesicles and the lipids of the filter.


A study of gramicidin using deuterium oxide. The single channel conductivity of the gramicidin channel has been measured for all the alkali ions using both H2O and 2H2O as a medium. Significant changes in conductivity with medium have been observed in all cases except lithium.


The enzymatic hydrolysis of the phosphate ester bond in some thionucleotides. We prepared the 5'- and 3'-O-phosphorothioate esters of the antitumor agent O2 : 2'-anhydro-1-beta-D-arabinosylcytosine. We also included in this study esters of 2'-thio-2'-deoxycytidine, namely, 2'-S-dCyd-2' : 3'-P, 2'-S-dCyd-2'-P, and 2'-S-dCyd-3'-P, along with natural nucleotides. These compounds were subjected to the action of Escherichia coli alkaline phosphatase, potato acid phosphatase, and bovine pancreatic ribonuclease A. The data were analyzed by Lineweaver-Burk plots to obtain Km and KI values. Only 2'-S-dCyd-2'-P was a substrate for alkaline phosphatase; the anhydro-araCyt phosphorothioates were good competitive inhibitors, while 2'-S-dCyd-3'-P did not associate with the enzyme. Acid phosphatase hydrolyzed all four monoesters investigated, including the S-phosphorothioate. The cyclic phosphorothioate, 2'-S-dCyd-2' : 3'-P was neither hydrolyzed by, nor associated with, ribonuclease A. ORD spectroscopy was also used in an attempt to relate the structural features of analogs to the peculiarity of their hydrolysis.


[Interactions between human blood serum inhibitors and native and modified dextran proteinases--terrilytin and trypsin]. Interactions between native terrylytin and trypsin and their derivatives modified by water-soluble dextrans on one hand and human blood serum inhibitors on the other, were studied. It was shown that modification of the enzymes results in changes in the type of their inhibition by blood serum due to a decrease of affinity of polymeric enzyme forms for alpha 2-macroglobulin and alpha 1-antitrypsin. The inhibition constants for native and modified forms of terrylytin and trypsin were calculated. The effects of steric and electrostatic factors on the interaction between inhibitors of blood and polymeric forms of proteinases are discussed.


[Affinity alkylation of E. coli RNA-dependent DNA polymerase with TTP gamma-amidate derivatives]. TTP gamma-benzylamidates are shown to act as competitive inhibitors of poly(dT) synthesis catalyzed by E. coli RNA-dependent DNA polymerase. The KM value for TTP as well as KI values for the gamma-analogues have been determined. TTP gamma-4-(N-2-chloroethyl-N-methyl-amino)benzylamidate is shown to be an effective affinity reagent for this enzyme.


[Characteristics of myelin basic protein from dog spinal cord]. Some characteristics of the myelin basic protein from dog spinal cord were studied. The basic protein has a molecular weight of 18,000; the isoelectric point is 9.5. The amino acid composition of the protein was determined. Electrophoresis in 15% polyacrylamide gel revealed the heterogeneity of the basic protein. The EAE-activity of the basic protein was tested on guinea pigs.


[Inhibition of compound 48--80 induced histamine liberation from mast cells by triton X-100]. Triton X-100 at concentrations preceding those which liberated histamine, produced dose-dependent inhibition of compound 48/80-induced histamine release from rat mast cells. Triton X-100 (0.00002 1/1) depleted ATP content in the mast cells and blocked compound 48/80-induced histamine release. The inhibition of compound 48/80-induced histamine release and depletion of the ATP content in the mast cells was reversed by glucose (10 mmole). It is concluded that inhibition by Triton X-100 of histamine release induced by compound 48/80 is dependent on inhibition of energy production.


Experimental irradiation of human corpse skin with pulsed laser ray. Irradiation of skin with one pulse of a neodymium laser with an emission energy of 940 to 2190 J and a density of 52,220 to 273,750 J/cm2 leads to the formation of damage with characteristic macroscopic and microscopic features. The injuries that appeared were funnel-shaped cavities up to 2 mm deep, from 2 to 8 mm in dimaeter, and with a crater-like rim 1 to 2 mm wide. Tissue on cross section was pinkish-brown, as a rule. Histologically, there were sharply defined signs of coagulative necrosis, and a typical, distinct, micromorphological picture was present that is characteristic of electric marking: nuclei and cells at all layers of the epithelium that were elongated into "brush" or "swirl" shapes, coagulation of the epithelium, the formation in it of honeycombed cavities, swelling, and detachment of the endothelium and vessels from the skin proper. Skin stained with black ink, iodine, hematoxylin, or eosin is damaged by laser ray more intensely than unstained.


The olivocerebellar projections to the flocculus and paraflocculus in the cat, compared to those in the rabbit. A study using horseradish peroxidase as a tracer. The projections from the inferior olive to the flocculus and paraflocculus in the cat have been mapped by means of the method of retrograde axonal transport of horseradish peroxidase. The findings show that the afferents to the flocculus are derived from the dorsal cap, ventrolateral outgrowth, the principal olive (the caudal parts of the ventral and dorsal lamella) and from the rostral part of the medial accessory olive. The fibres to the paraflocculus come from the caudal part of the principal and from the rostral part of the medial accessory olive. Details in the projections are seen from Figs. 1 and 2. Concerning some points the findings are at variance with those made in the rabbit by Hoddevik and Brodal, and suggest that there are species differences hitherto not known.


Evidence that 4S RNA is axonally transported in normal and regenerating rat sciatic nerves. Studies in regenerating goldfish optic nerves indicate that RNA may be axonally transported during optic nerve regeneration14,18,19. The present study was performed to determine if the axonal migration of RNA could be demonstrated during regeneration of the rat sciatic nerve. Rats, which had only the left sciatic nerve crushed 10 days earlier, were injected bilaterally with [3H]uridine into the spinal cord at segmental levels L5 and L6, thus labeling ventral horn cells giving rise to the sciatic nerve. Six, 14 and 20 days later rats were sacrificed by cardiac perfusion of saline followed by 10% formaldehyde. Formaldehyde-precipitable radioactivity, identified as [3H]RNA, was 4--5 times greater in the regenerating sciatic nerve compared to the normal nerve and moved without impediment beyond the point of the crush into the regenerating portion of the nerve. The axonal migration of free unincorporated labeled RNA precursors was also demonstrated, raising the possibility that the distribution of [3H]RNA along the sciatic nerve might be entirely extra-axonal; i.e., free [3H]uridine is taken up by Schwann cells from the axon where it is incorporated into [3H]RNA. This interpretation of the data would also result in the appearance of a proximodistal distribution of RNA associated radioactivity. To determine whether any sciatic nerve [3H]RNA was due to axonal transport, rats which had only the left sciatic nerve crushed 10 days earlier were injected bilaterally with [3H]uridine into the spinal cord. Fourteen days after injection, rats were sacrificed and radioactivity present in the nerve was confirmed as RNA by SDS polyacrylamide gel electrophoresis. Radioactivity in the various RNA species 14 days after intraspinal injection showed the following distribution: 28 + 18S RNA--normal 39.3% +/- 2.1; regenerating 45.4% +/- 1.6; 4S RNA--normal 43.0% +/- 1.3; regenerating 46.8% +/- 2.7. Similar characterization of sciatic nerve RNA 1 or 3 days following the intravenous administration of [3H]uridine gave the following distribution: 28 + 18S RNA--normal 72.4% +/- 3.0; regenerating 75.0% +/- 3.6; 4S RNA--normal 7.7% +/- 1.3; regenerating 10.7% +/- 0.8. The intraspinal injection of [3H]uridine would label Schwann cell RNA and, in addition, any species of intra-axonal RNA, while intravenous injections would label Schwann cell RNA and not axonal RNA. If 4S RNA is in the axon, one would predict relatively more labeled 4S RNA following intraspinal injections than following intravenous injections. The data demonstrate an enrichment of 4S RNA in both normal and regenerating rat sciatic nerve following the intraspinal but not following the intravenous injection of labeled precursor. Therefore, we suggest that 4S RNA migrates axonally in both normal and regenerating sciatic nerves of rats.


Immunological release of histamine and slow-reacting substance in domestic fowl. Immunological release of histamine from the whole blood of the sensitized chickens was demonstrated. Compound 48/80 (a potent histamine releaser) released histamine from isolated chicken lung. The sensitizing antigens (bovine albumin or horse plasma) did not release histamine from the lung. Bovine albumin and horse plasma released slow-reacting substance of anaphylaxis only from the lungs of adult domestic fowl and not from ileum or lungs of immature chickens.


Genetic background of Fuchs' heterochromic cyclitis. We studied the genetic background of 24 patients with Fuchs' heterochromic cyclitis (FHC). Each was given a careful eye examination which included family history and serological determination of HLA antigens. Two families each had 2 cases of FHC in the same family; in addition an ancestor in the second family possibly had FHC; in both families one healthy member had simple heterochromia. One patient with FHC had congenital uveal coloboma, one pigmentary retinal dystrophy, and four had cysts of the ciliary body. The frequencies of all HLA antigens in patients with FHC compared well with the frequencies in the controls. In a family in which HLA haplotypes could be derived, the patients with FHC showed different HLA haplotypes. We conclude that FHC has a hereditary basis but its immunological component is not genetically associated with the HLA system.


The neurological complications of brucellosis. Neurological complications of brucellosis have seldom been reported. Typical cases of meningoencephalitis, myelitis, and peripheral nerve involvement due to brucellosis, which have been collected over the past ten years, are described. Only four cases of extradural brucellar abcesses have been published previously. We now add four new observations. A CSF electrophoresis was done in the cases of meningoencephalitis and meningomyelitis and showed a marked increased of gamma globulin. This has never been reported previously.


Music therapy in palliative care.

Music therapy in palliative care. Initial observations regarding the use of music therapy at one hospital in the palliative care of patients with advanced malignant disease are presented. In the hands of a trained music therapist, music has proven to be a potent tool for improving the quality of life. The diversity of its potential is particularly suited to the deversity of the challenges - physical, psychosocial and spiritual - that these patients present.


Histocompatibility antigens as markers of abnormal iron metabolism in idiopathic hemochromatosis. To determine the frequency of HLA histocompatibility antigens in persons with idiopathic hemochromatosis and their usefulness as genetic markers of the disease, HLA typing for the A, B and C loci was carried out. HLA-A3 was found in 61% of 18 unrelated individuals with idiopathic hemochromatosis compared with 25% of 253 randomly chosen control subjects (P less than 0.001), and HLA-B7 was found in 50% and 22% respectively (P less than 0.025). Eighty-six members of seven families with idiopathic hemochromatosis were screened for abnormalities in iron metabolism with tests for serum iron concentration, transferrin saturation, serum ferritin concentration and iron content of the hepatocytes. Of the 14 persons selected for liver biopsy because of abnormalities detected by these tests, 8 had increased amounts of stainable iron in the hepatocytes. Body iron overload was subsequently demonstrated in six of the seven, who had undergone repeated phlebotomy. In sibships having one member with hemochromatosis, only 1 of 22 members had two haplotypes in common with the proband, whereas in sibships having more than 1 member with the disease 4 of 5 affected members had two haplotypes in common. HLA typing in families with hemochromatosis may provide a means of identifying persons at risk of acquiring the disease.


Superiority of adriamycin-containing combination chemotherapy in the treatment of diffuse lymphoma: a Southwest Oncology Group study. As a part of an ongoing prospective controlled trial, the Southwest Oncology Group compared the results of treatment of advanced non-Hodgkin's lymphoma with two CHOP regimens (cyclophosphamide, adriamycin, vincristine and prednisone with either low-dose bleomycin or BCG by scarification) to a COP regimen (cyclophosphamide, vincristine and prednisone) with low-dose bleomycin (COP-Bleo). The study design emphasized histopathology review and systematic restaging to define complete remission (CR). Confirmed rates of CR for 443 evaluable patients were 59% for 286 patients receiving the CHOP regimens and 59% for 157 patients receiving COP-Bleo. Rates of CR were higher for patients with nodular lymphoma (69%) compared to those with diffuse lymphoma (54%) (p = 0.005). For patients with nodular lymphoma there was no difference in CR rates according to treatment. For patients with diffuse lymphomas the CR rate was higher with the CHOP programs (58%) than with COP-Bleo (44%) (p = 0.10). Overall duration of CR and survival was significantly longer for patients with nodular lymphoma compared to diffuse lymphoma (p less than 0.01). At this time, remission duration and survival were similar regardless of induction regimen used in patients with nodular lymphoma. However, in patients with diffuse lymphoma, the duration of CR and overall survival were improved by treatment with the CHOP regimens compared to COP-Bleo (p = 0.02). Thus, in this controlled study we have demonstrated that initial combination chemotherapy employing the CHOP regimen was a superior remission induction therapy for patients with diffuse lymphoma.


Changes in serum alpha 1 antitrypsin, alpha1 acid glycoprotein and beta 2 glycoprotein I in patients with malignant hepatocellular carcinoma. Serum alpha 1 antitrypsin, alpha 1 acid glycoprotein and beta 2 glycoprotein I concentrations were determined in 36 patients with malignant hepatocellularcarcinoma, 30 with cirrhosis and 35 with hepatitis by quantitative immunoelectrophoresis. Serum alpha 1 antitrypsin and alpha 1 acid glycoprotein levels were significantly higher in patients with hepatocellularcarcinoma than in those with cirrhosis (p less than 0.001) or hepatitis (p less than 0.001). Elevated levels of alpha 1 antitrypsin were found in 88.9% of patients with hepatoma compared to 23.3% of patients with cirrhosis and 28.6% of patients with hepatitis. Raised levels of alpha 1 acid glycoprotein were also found in 80.6% of patients with hepatoma compared to 20% of patients with cirrhosis and in only 5.7% of patients with hepatitis. beta 2 glycoprotein I levels were similar in the three conditions and therefore not useful for differential diagnosis. In monitoring the progress of tumor growth alpha 1 antitrypsin and alpha 1 acid glycoprotein levels were found to increase during the growth phase. Measurements of these two glycoproteins are suggested for differential diagnosis of these liver diseases, as tumor markers for the detection of hepatocarcinoma, and for the monitoring of the progress during treatment.


Immunogenicity of chemically induced murine colon cancers. The antigenicity and immunogenicity of three colorectal carcinomas induced in BALB/c mice by 1,2-dimethylhydrazine or N-methyl-N-nitrosourethan were studied. All tumors were readily transplantable. Two of these tumors metastasized when transplants reached sufficient size. All tumors were found to be immunogenic in the strain of origin, and all tumors were shown to contain unique tumor-specific transplantation antigens in cross-protection experiments. The use of these tumors as an animal model for studies of adjuvant immunotherapy and chemoimmunotherapy is suggested.


Studies with murine LPC-1 plasmacytoma using [6-14C]arginine. [16-14C]Arginine ([6-14C]Arg) was used as an in vivo pulse label to study BALB/c murine LPC-1 plasmacytoma synthesis and secretion of its tumour-associated M component (IgG2a, kappa). With this isotope, an eight- to ten-fold enhancement in the labelling of the gamma globulin region and ten-fold reduction in the albumin labelling were observed. Production and secretion of the M component was detected (within 30 min) after cell transfer. Only mice which received tumour cells showed significant labelling in the gamma globulin region 24 hr after isotope injection. The labelling behaviour of the tumour M component correlated with the administered cell dose. The peak heights of radioactivity in the gamma region increased with increments in cell number. When the percentage radioactivity diverted into M component was plotted as a function of cell dose, a linear relationship was noted. This study demonstrates the feasibility of using [6-14C]Arg as a tool to follow the newly synthesized tumour-associated protein, and provides a means of estimating tumour cell number.


Types of thalamo-cortical relay neurons in the anteroventral nucleus of the cat. A combined horseradish peroxidase--Golgi study. Neurons displaying a thalamo-cortical projection were marked by means of the retrograde transport of horseradish peroxidase (HRP), and the labeled elements were compared with neurons impregnated by the Golgi technique. Injections of HRP into the posterior area of the limbic cortex resulted in its uptake by various anterior thalamic nuclei, especially the anteroventral nucleus. HRP-positive cells are characterized by their position, dendritic orientation, and the shape and size of their somata. On the basis of the combined HRP-and Golgi-analysis three different types of thalamo-cortical relay neurons can be distinguished.


Fibroblast interferon in man is coded by two loci on separate chromosomes. We have examined viral and poly(rl):poly(rC) induction of interferon synthesis in several human, mouse and Chinese hamster cell lines, and in hybrids derived from the fusion of such cells. We observed species and cell-type differences in inducer effectiveness and in the kinetics of interferon production. In some cases, parental characteristics are preserved in somatic cell hybrids, and in other cases, the expression of the donor phenotype is modulated by the epigenetic state of the recipient cell. Mapping studies in human/mouse and human/Chinese hamster hybrids indicate that there are at least two structural genes for human fibroblast interferon. Chromosomes 2 and 5 each contain genetic information for the synthesis of fibroblast interferon. Gene dosage experiments indicate that one gene is on the long arm of chromosome 2 and another is on the short arm of chromosome 5. Leukocyte interferon genes could not be mapped to these chromosomes, but this negative result could be influenced by the epigenetic state of the hybrid cells.


A defined subgenomic fragment of in vitro synthesized Moloney sarcoma virus DNA can induce cell transformation upon transfection. The longest DNA molecules synthesized by endogenous reverse transcription in detergent-permeabilized Moloney murine sarcoma virus (Mo-MSV) virions (clone G8-124) are double-stranded DNA molecules of 5,8 kilobase pairs (kbp). This DNA species has been purified by sedimentation of total in vitro synthesized Mo-MSV DNA through neutral sucrose gradients. A physical map of the positions of the cleavage sites for a series of restriction endonucleases has been derived for this 5.8 kbp DNA. Mo-MSV DNA synthesized in vitro was found to induce morphological transformation of NIH-3T3 mouse fibroblasts upon transfection. The foci had a morphology indistinguishable from that of Mo-MSV-induced foci, and the induced transformed phenotype was stable. The 5.8 kbp double-stranded DNA (dsDNA) purified by agarose gel electrophoresis also induced focal transformation. Furthermore, gel-purified, restriction endonuclease-generated fragments of 5.8 kbp dsDNA containing the region from 2.8--4.9 kbp on the physical map of Mo-MSV DNA were able to induce foci. In contrast, endonuclease-generated DNA fragments lacking this region on the map were unable to transform cells upon transfection. When transformants derived by transfection with 5.8 kbp dsDNA were infected with Moloney murine leukemia virus (Mo-MLV) helper virus, Mo-MSV was rescued from a small portion of these cells, suggesting the establishment of the complete viral genome in these cells. One Mo-MSV DNA fragment, spanning 2.8--4.9 kbp on the physical map, was generated by cleavage of 5.8 kbp DNA with endonucleases Hind III + Sal I and currently represents our maximum estimate for the size of the transforming region of the Mo-MSV genome. This fragment includes the Mo-MSV sequences which are found in the DNA of uninfected mouse cells.


[Study of the Gc protein in several French population samples : genetic polymorphism by isoelectrofocusing and quantitative results]. The authors describe the results obtained in an examination by isoelectrofocusing of more than 1,000 serum samples from various regions in France. The autochthonous populations of the north side of the Pyrénées are characterized by a high frequency of the gene Gc2 and a very low frequency of the Gc1F gene. This distinguishes them from the other groups studied (southwestern, northwestern and eastern France). The quantitative analysis of the data emphasizes the difference. These results can only be explained by the interdependance of the genetic and metabolic factors.


[Antigenic determinants of beta-LPH, beta-MSH, alpha-endorphin, ACTH and alpha-MSH revealed by anti-beta-endorphin in neurons of the human infundibular nucleus]. Comparison of adjacent serial sections of the tubero-infundibular region of Human adult hypothalamus demonstrates that the same perikarya, axons and terminals are stained both with anti-beta-endorphin and anti 17-39 ACTH antisera. The most immunoreactive of these neurons are also revealed with anti alpha-endorphin, anti alpha and beta-MSH, anti-1-24 ACTH and anti beta-LPH. These results suggest that neurons of the infundibular nucleus can store and probably secrete peptide similar to propiocortin or fragment(s) of this molecule.


The influence of nasal respiration on the function of the auditory tube. The importance of treating disturbances of nasal ventilation before any surgical intervention on the ear is stressed in many papers. It is said that ventilation of the auditory tube is not normal when respiration through the nose is handicapped. Based on these considerations, we have investigated by tympanometry the influence of abnormalities of nasal respiration measured by rhinomanometry on the auditory tube function. There was no significant relation between osseous occlusion of nasal ventilation or a swelling of the nasal mucous membranes or conchas and a dysfunction of the tube. The middle ear pressure varied within the normal range. This indicates that treatment of impaired nasal respiration before an operation on the ear is only necessary when an ascending nasal infection is feared or if there is nasopharyngeal disease.


Regional mapping of the HLA on the short arm of chromosome 6. A detailed gene marker study was performed on a partial 6p trisomic child resulting from a balanced maternal translocation t (2;6) (p 2505; p2105). HLA typing and mixed lymphocyte reaction showed that the breakpoint on chromosome 6 was located within the HLA gene cluster, allowing an accurate location of the D determinants. Localization of the P blood group locus within the region 6 p 2105 to 6 p ter was excluded.


Prenatal diagnosis of polycystic kidneys and encephalocele (Meckel syndrome). Two unrelated families are presented with repeated occurrences of a congenital syndrome of which the main stigmata were polycystic kidneys and occipital encephalocele (Meckel syndrome). Prenatal diagnosis, followed by interruption of pregnancy, was performed in one case. The diagnosis was based on an increase of amniotic alpha-fetoprotein (AFP), and on the mode of growth and cell types of cultured amniotic cells. In another similarly examined case the diagnosis was suspected, but the parents did not wish the pregnancy to be interrupted. The child was stillborn and malformed. AFP values are presented and discussed in relation to the observed malformations. Neural tube defects are associated with an increase of AFP in amniotic fluid, but, as in normal pregnancies, the values decrease with increasing gestational age. On the other hand, kidney malformations seem to be associated with AFP values which remain high or even increase with increasing gestational age.


Spontaneous pneumothorax as a result of intensive cytotoxic chemotherapy. Spontaneous pneumothorax occurring as a complication of the antitumor effect of cytotoxic chemotherapy has been reported in occasional cases of osteogenic sarcoma. Its occurrence in other tumors has not been described. This report describes two cases of this complication in patients with germinal tumors and discusses possible pathophysiologic mechanisms.


Long survival in patients with bronchogenic carcinoma complicated by superior vena caval obstruction. When superior vena caval obstruction complicates cancer of the lung; the prognosis is grave; 1 percent of such patients survive for one year. Improved palliation is reported from many radiation therapy centers using higher initial dose fractions and tumoricidal doses. We now report the findings in three patients with histologically confirmed bronchogenic carcinoma who had superior vena caval obstruction and who survived for nine years (two patients) and seven years (one patient). Aggressive therapy with irradiation seems to provide better palliation and may occasionally be associated with remarkably long survivals in this normally highly lethal disease.


False negative biopsy in Pneumocystis carinii pneumonia. Unlike most pneumonias, the diagnosis of Pneumocystis carinii pneumonia is based solely on identifying organisms by stain, usually with methenamine-silver. Because of technical problems involved with adequate staining, control samples usually are done concurrent with tissue specimens to be examined. Lung containing fungi often is used as a control. We recently observed false-negative biopsy specimens in a case of P carinii pneumonia where the Pneumocystis organism failed to stain with methenamine-silver on several occasions, although fungal controls were positive. This report emphasizes the importance of using P carinii as a control whenever attempting to diagnose P carinii pneumonia.


Chromosomal isolabelling caused by three rounds of synthesis in late replicating regions. Isolabelling only occurs in CHO cells that have been allowed to replicate for more than 2 but less than 3 cell divisions in the presence of BrdU. The isolabelling is confined to late replicating regions of the chromosomes. The staining patterns obtained indicate that BrdU was incorporated three times in these regions and that the isolabelling did not come from the segregation of label in polynemic chromosomes.


EEG feedback training of epileptic patients: clinical and electroencephalographic analysis. To evaluate the clinical efficacy and mechanisms underlying EEG feedback training of epileptic patients, 5 adult patients with poorly controlled seizures were studied for 4--10 months during which quantitative analysis of seizures, the EEG, and serum anticonvulsant levels was conducted. Sustained seizure reduction did not occur during the first 4--5 weeks in which feedback signals were presented randomly in relation to the EEG. When feedback was then made contingent upon central 9--14 c/sec activity, seizures declined by 60% in 3 patients. Power spectral analysis showed upward shifts in EEG frequency, decreases in abnormal slow activity, and enhancement of alpha rhythm activity as a function of contingent training, but no specific EEG change was associated with seizure reduction in all patients. No evidence was obtained for the hypothesized involvement of a 'sensorimotor rhythm' or motor inhibition in the training effects. The lack of effect in two patients could not be attributed to insufficient training, lack of motivation, or to differences in seizure classification. A second phase of research showed that continued laboratory training was both sufficient and necessary for maintaining clinical and EEG effects. Results indicate that: (1) significant seizure reductions can occur with EEG feedback training which are not related to placebo effects, non-specific factors or to changes in medication; (2) EEG changes associated with such training can best be described as 'normalization'; (3) continued clinical investigation of EEG feedback training as a non-pharmacological adjunct to conventional therapy appears justified.


Evaluation of frequency-following potentials in man: masking and clinical studies. The frequency-following potential (FFP) can have applicability in the assessment of hearing-impaired subjects only if it can be shown that its generation is initiated by neurons which have low best frequencies (2.0 kHz or lower). This study presents results from five subjects with high frequency hearing losses and three subjects with normal hearing. Using 500 Hz tone bursts in the presence of continuous noise of various configurations, it has attempted to determine how in normal hearing subjects, the amplitude and latency of the FFP may be affected. Recordings of the FFP in the presence of noise and wave forms from hearing imparied subjects provide evidence that the FFP is initiated in the cochlea largely by neurons whose best frequencies are 2.0 kHz or lower. Hearing impaired subjects may exhibit 'deviant' responses to tone bursts. These FFP responses may be related to peculiarities of the hearing loss and provide, therefore, a potential means for assessing the temporal viability of the low frequency channels of the auditory neuraxis.


Quantitative assessment of photic driving in renal failure. Photically driven EEGs were recorded from patients with renal disease using photic stimulation at integer rates between 3 and 12 flashes/sec. Changes in the structure of the power spectrum of the potentials produced by this stimulus paradigm are described as a function of a patient's clinical state. The basic overall findings are that (1) harmonic activity is attenuated while activity below the fundamental driving frequency is increased as a patient's clinical condition deteriorates, and (2) that these effects are substantially reversed and controlled by means of dialysis and renal transplantation.


Visual evoked potential in man: early oscillatory potentials. Short latency visual evoked oscillatory potentials to bright light stimulation were recorded from the scalp of 15 normal human adult subjects. The onset latencies of these potentials recorded over anterior frontal and posterior scalp regions were 9--17 msec and 13--24 msec, respectively. The frequency of the oscillations was about 100 c/sec. These potentials were widespread in their distribution over the scalp but were most prominent at midline and parasagittal recording locations. Like similar potentials recorded in animals, it seems that these potentials arise in both subcortical and cortical visual structures. The mechanism underlying the generation of these potentials and their possible functional significance are discussed.


[Temporal organization of paroxysmal discharges in the child. II. Polygraphic recordings during sleep]. A study of the paroxysmal activities in the course of all night sleep was carried out on 6 children aged 6 to 12 years, who were previously recored by means of biotelemtry during the day (cf. Findji et al. 1978). The discharges were analysed and quantified in the same way as in daytime but here in relation to the organization of sleep. The latter shows serious disturbances by comparison with normal children or children who only have sleep problems. The global quantifications (average densities of discharges per stage) do not, in our population, evidence any effect of facilitation or diminution which would be specific to a given stage. Comparison of the chronological evolutions of the densities of paroxysmal activities with the different stages of sleep shows that the connections between these phenomena can be either nil, or very close, according to the subject, and in one case, according to the time of night. This raises the problem of the heterogeneity of the 'states' gathered under the same codification of stages and shows the multiplicity of factors which can interfere with the modulation of the discharges, even during sleep. The analysis of long duration day or night records underlines the extreme complexity of the temporal organization of paroxysmal discharges, in which both hypotheses formulated at the beginning of this study seem to find partial support: internal modulation connected to more or less stable biological rhythms, and the role of environmental factors and the subject's reactions, which more or less mask the intrinsic rhythms.


Changes in P3 waves with event repetition: long-term effects on scalp distribution and amplitude. Event-related brain potentials to tachistoscopically presented events were recorded from adults 24--36 years of age. Subjects counted the number of target events (P = 0.12) randomly interposed in sequences of background events (either P = 0.88 or P = 0.76). In some sequences, slides bearing the letter A were targets and those bearing the letter B were backgrounds; the reverse was true in the other sequences. Also interposed (P = 0.12) in these sequences of targets and backgrounds were one of two types of non-target events: in some sequences non-targets were slides (termed dims) bearing any one of the letters C to Z and having one-tenth the luminance of the target and background slides, while in the others, they were slides bearing 'novel' patterns, each consisting of a different, quasi-random, unrecognizable color pattern. The scalp amplitude distribution of P3 waves to novels were initially frontal, but, with repeated presentations of novel events, they became parietal. The scalp distributions of P3 waves to targets and dims were both initially parietal and did not change with repeated presentations of target and dim events. P3 amplitudes to targets were unaffected by repeated presentations of target events. However, P3 amplitudes to dims and novels were affected. P3 amplitudes to dims decreased linearly at Fz, Cz and Pz with repetition of dims. P3 amplitudes to novels decreased at Fz and increased at Pz with repetition of novels. An abbreviated analysis of possible effects of sequential event probabilities on P3 amplitudes and latencies to novels, dims and targets was performed and no effects were found. It is suggested that P3 waves may be affected by at least 4 factors: the facility with which events may be categorized by subjects, the importance attached to events (via task instructions, subjective assessment of the contents of events, etc.), a priori category probability, and sequential event structure.


[EEG spectral analysis in normal children between the ages of 6 to 16. Choice and validation of the most significant parameters]. Numerous parameters have been proposed in order to describe the EEG power spectrum. Within the scope of an EEG study on the normal child between 6 and 16 years of age, the following problem had to be solved: given a group of 19 variables measured on 232 subjects in 5 situations, which of these variables best describe the population under examination. The algorithm proposed to answer this question is based on principal component analysis on the correlation matrix. Only 4 parameters are necessary to describe the EEG of a normal child of 6 to 16 years--these are Hjorth's parameters (activity, mobility and first order complexity) and alpha band power. They allow the observation of individual differences and the quantification for a given subject of the reaction to eyes opening, sensitivity to hyperventilation or even reactions that are difficult to perceive visually such as those resulting from doing arithmetic or vocabulary tests.


EMG activity of cricothyroid and chin muscles during wakefulness and sleeping in the sleep apnea syndrome. The authors studied the direct and integrated EMG activities of the cricothyroid (CT) and chin muscles in 7 patients with the sleep apnea syndrome. They noted: (1) A tonic activity with phasic inspiratory reinforcements in the CT during wakefulness. (2) A decrease in the tonic activity without any modification of the phasic inspiratory reinforcement during sleep. (3) A decrease or even disappearance of the phasic activity during sleep-induced hypopneas. (4) A complete cessation of both the tonic and the phasic activities of the CT during obstruction apneas: resumption of respiration is marked by the reappearance first of the phasic inspiratory activity and secondly of the tonic one. (5) In some patients similar activities are recorded in the chin muscles during wakefulness, sleep and sleep apnea. These results favor possible obstruction of the upper airway at the laryngeal level: the conjunction of inhibition of the inspiratory activity of the laryngeal muscles with the subsequent closing of the glottis and inspiratory depressure could produce an obstruction.


Electroretinographic responses in Lafora disease. In 3 patients with Lafora disease specific ERGs were observed resembling closely those seen in the Schubert-Bronschein type of night blindness. In contrast to essential hemeralopia, where the b-wave is completely lacking a gradual recovery of the b-wave was recorded in Lafora disease. The recovery occurred within about 15 min of dark adaptation. ERG examinations may have diagnostic significance in patients with myoclonic epilepsy.


A fast-recovery electrode amplifier for electrophysiology. To overcome the problem of amplifier 'blocking time' due to stimulus artifact, a fast-recovery electrode amplifier is developed. The design of this amplifier and its application to a clinical EMG unit is discussed.


An immunological approach to the study of protein conformational heterogeneity: its application to growth hormone. The study of multiple forms of a hormone due to changes in its secondary or tertiary structure (conformational heterogeneity) has been restricted to methods of physical chemistry which require relatively large quantities of purified preparations and has precluded such studies on circulating hormones. The present report describes an immunological approach to the study of GH conformational heterogeneity based on the simultaneous use of two homologous RIAs: one which recognizes porcine (p) GH in its native conformation (pGH RIA) and a second (pB RIA) for a 55-residue S-aminoethylated cyanogen bromide fragment (pB) of pGH (corresponding to residues 126-180 of human GH). In the pGH RIA, pB was nonreactive, whereas pGH displacement of [125I]iodo-pB from anti-pB serum required about 10(3) M excess and was nonparallel to pB. Treatment of pGH with urea plus mercaptoethanol (ME), which would be expected to affect the conformation of the GH molecule, increased its immunoreactivity 68-fold in the pB RIA and its dilution curve became parallel to that of pB. Similar treatment of porcine pituitary extract increased its immunoreactivity 97-fold in pB RIA with a corresponding slope change. In contrast, urea plus ME decreased pGH and pituitary extract immunoreactivity by 84% and 92%, respectively, in the pGH RIA. The pB RIA/pGH RIA ratio of immunoreactivity for pGH and for pituitary extract increased 795-fold and 569-fold, respectively, after combined treatment, whereas lesser increases were observed after urea or ME treatment alone. In conditions where alterations in GH conformation were suspected (highly purified, aged human GH preparations which were no longer satisfactory as radioiodinated tracers for radioimmunoassay), calculation of the pB RIA/hGH RIA ratio of immunoreactivity indicated increased values. Based on these results, a model is proposed in which unfolding a native hormone, with concomitant exposure of the antigenic determinant of pB, hidden within pGH, leads in increased immunoreactivity in the pB RIA and decreased immunoreactivity in the pGH RIA. In this model, the pB RIA/pGH RIA ratio of immunoreactivity would thus serve as an index of the state of hormonal unfolding.


Somatostatin induces histamine secretion from rat peritoneal mast cells. Cyclic somatostatin strongly stimulated secretion of histamine from rat peritoneal mast cells. The energy, temperature and calcium dependence of this effect indicate that the action of somatostatin on the mast cells is similar to that of the classic mast cell secretagogue compound 48/80 and is to induce exocytosis.


Studies on the mechanism of goitrogenic action of diphenylthiohydantoin. Diphenylthiohydantoin (DPTH) is a potent goitrogenic compound and produces goiters in rats. Like methimazole, DPTH depresses plasma T4 and T3 concentrations and elevates plasma T4 and T3 concentrations and elevates plasma TSH concentration. Unlike methimazole, however, DPTH does not suppress thyroidal radioiodine uptake and thyroid hormone synthesis, although the monoiodotyrosine to diiodotyrosine ration is elevated by DPTH. DPTH does not inhibit thyroidal radioiodine release or augment the degradation of thyroid hormone. DPTH depresses an increase of plasma T4 and T3 in thyroidectomized rats maintained on T4 or T3 by augmenting fecal excretion of hormones. In addition, DPTH decreases conversion of T4 to T3 in vitro. It is suggested that DPTH is a unique goitrogen which acts at two different extrahyroidal sites, viz. fecal loss of thyroid hormone and conversion of T4 to T3.


Pituitary basophils from immature male and female rats: distribution of gonadotrophs and thyrotrophs as studied by unit gravity sedimentation. Isolated pituitary cells from 14-day-old female, 14-day-old male, and adult male rats were separated into several fractions by unit gravity sedimentation. With preparations from 14-day-old animals, the percentage of basophils increased from top to bottom of the gradient. The bottom fractions contained large basophils with a purity attaining almost 90% in the female and 75% in the male preparation. The same fraction prepared from adult male rats was composed of over 80% of large acidophils. As identified by immunostaining, the majority of the female large basophils were gonadotrophs, whereas in 14-day-old males about 1/3 of these basophils were thyrotrophs. The relative proportion of medium and small sized gonadotrophs and of thyrotrophs was also higher in the 14-day-old rats than in the adults. An estimate of the distribution of FSH and LH among the gonadotrophs was made by comparing the number of cells reacting with anti-FSH, with anti-LH, and with a mixture of both antisera. The data suggested that irrespective of cell size, the majority of the gonadotrophs in 14-day-old rats contained both FSH and LH. In most fractions, the occurrence of cells with only one hormone was more apparent in the males than in the females. In adult rats, cells with only FSH were most abundant in the fraction with large gonadotrophs. The distribution of hormone content, as measured by RIA, was highly characteristic for each animal group. In 14-day-old females, more than 70% of the recovered hormones was in the large gonadotrophs. In 14-day-old and adult males, substantial amounts of hormone were also recovered from medium sized gonadotrophs. The present findings suggest a relationship between developmental changes in pituitary hormone secretion and changes in pituitary cell type distribution.


Effect of intraventricular administration of anti-somatostatin gamma-globulin on the lethal dose-50 of strychnine and pentobarbital in rats. Effects of intraventricular injection of sheep anti-somatostatin gamma-globulin (anti-SSG) on strychnine-induced seizures, strychnine LD50, and pentobarbital LD50 were examined in male rats under light ether anesthesia. Ten microliters of anti-SSG given 2 h earlier significantly decreased the duration of strychnine-induced seizures as compared with that in the control rats pretreated with normal sheep gamma-globulin (NSG). This effect of anti-SSG seemed to be specific, as there was no difference in seizure duration between sheep anti-LHRH gamma-globulin (anti-LHRHG)- and NSG-pretreated rats. Survival rates in anti-SSG-pretreated rats after injection of strychnine and pentobarbital were significantly larger (P less than 0.01 and P less than 0.05, respectively) than those in the control rats receiving NSG. The administration of anti-SSG resulted in 26.7% and 22.9% increases in the LD50 of strychnine and pentobarbital, respectively. These results indicate that endogenous somatostatin in the cerebrospinal fluids and/or the periventricular tissue nodulates the response of the central nervous system to strychnine and pentobarbital in rats.


Role of the serum estrogen-binding protein in the control of tissue estradiol levels during postnatal development of the female rat. The role of the serum estrogen-binding protein (EBP) in the control of tissue estradiol levels during postnatal development of the female rat was examined. The estradiol-binding capacity of serum from the 1-day-old rats far exceeded the physiological level of estradiol in serum. The binding capacity decreased exponentially during the first 5 weeks of life to reach the low adult level at about the time of vaginal opening on day 37. From these observations one would predict that EBP would bind estradiol in the serum of the neonate, thereby preventing tissue uptake of the hormone. As the levels of EBP decline with advancing age, there should be a corresponding shift in the distribution of estradiol from serum to tissues. We have taken in vivo and in vitro approaches to evaluate these proposals. Female rats of various ages (1 day to 1 yr old) were sacrificed 1 h after [3H]estradiol injection and the radioactivity in serum and tissues was determined. During the first 11 days of life, the concentration of [3H]estradiol in serum was greater than the concentration of this hormone in estrogen-sensitive (uterus) and insensitive (lung, cerebral cortex, and diaphragm) tissues. Tissue to serum ratios of [3H]estradiol increased progressively between 13-34 days and then plateaued at about the time of puberty (37 days of age) at levels which were 50- to 150-fold greater than those observed in the neonate. The increase in tissue to serum ratios of [3H]estradiol during postnatal development probably resulted from the decline in serum EBP, since injection of neonatal serum into 28-day-old rats reduced tissue to serum ratios of [3H]estradiol to levels which were similar to those observed in 16-day-old animals. To determine the effects of EBP on uterine uptake of estradiol in vitro, uteri from 21-day-old rats were incubated with [3H]estradiol and serum obtained from rats of various ages. As the concentration of serum EBP declined with advancing serum donor age, there was a corresponding increase in the uterine uptake of [3H]estradiol. These results suggest that the decline in EBP is responsible for the progressive increase in tissue to serum ratios of estradiol during the first 5 weeks of life. It is suggested that the increase in tissue to serum ratios of estradiol between days 13-37 postpartum is an important factor in the initiation of estrogenic events during postnatal sexual maturation in the female rat.


Inhibition by various steroids on dihydrotestosterone binding to androphilic protein in human benign prostatic hypertrophy. The effect of various steroidal compounds on the binding of the androphilic protein to dihydrotestosterone in the cytosol of tissues of human benign prostatic hypertrophy was examined. Androgens, as well as estrogens and gestagens showed an inhibitory effect on the binding. Non-steroidal anti-androgens were revealed to be weak inhibitors on the binding. Two androphilic proteins were observed in Sephadex G-200 chromatography in fractions eluted at the void volume and in fractions appearing at the site of IgG, and the rate of inhibition on the binding of both fractions by various steroids was compared. The rate of inhibition by various compounds was generally higher in the IgG fraction than in the void volume fraction. When the ligand and inhibitors were incubated with the cytosol prior to fractionation by Sephadex chromatography, rate of inhibition was lower than that obtained when the ligand and inhibitors were reacted with fractions after chromatography. Implications of the difference observed in these two experiments are not clear at this moment. The results obtained by the protamine precipitation experiment were almost the same as those by the experiment using the extract of the unfractionated acetone-dried cytosol, therefore, the protamine procedure does not seem to precipitate the tissue specific androphilic protein specifically.


SGC (small granule chromaffin) cells in the mouse adrenal medulla: light and electron microscopic identification using semi-thin and ultra-thin sections. Adrenal glands of the mouse, fixed either in glutaraldehyde followed by osmium tetroxide or in a mixture of potassium dichromate and glutaraldehyde, and embedded in Epon 812, were investigated by light and electron microscopy. An argentaffin reaction was applied to semi-thin sections for light microscopy and to ultra-thin sections for electron microscopy. Since the mature secretory granules in the Small Granule Chromaffin (SGC) cell were argentaffin and were mainly located along the cell membrane, this cell was clearly distinguishable under the light microscope both from the A (adrenaline) cell whose secretory granules were non-argentaffin and from the NA (noradrenaline) cell whose cytoplasm was rich and was filled with large, strongly argentaffin granules. Chromaffinity of the SGC cell was demonstrated under the light microscope. The SGC cell was intensively stained with toluidine blue without revealing metachromasia. It was demonstrated at the EM level that not only the secretory granules but also the synaptic-like vesicles in the SGC cell contained argentaffin substances. Possible functional relationship between the secretory granules and the synaptic-like vesicles was discussed.


Aberrant peptide hormones in patients with urogenital carcinomas. A series of 229 patients with urogenital carcinomas were investigated for aberrant peptide hormone activities. Serum TSH and prolactin were frequently measured in elevated levels and showed some relation to the stage of disease. Ectopic production of beta-HCG was not observed in any of the cases, thyroid and steroid hormones did not exceed the normal ranges.


Combined surgery and chemotherapy for retroperitoneal metastases of testicular carcinomas other than pure seminoma. The results of treatment in 210 patients with testicular carcinomas other than pure seminoma are analysed. Patients with negative retroperitoneal nodes had a very good prognosis following lymph node dissection alone. Patients with positive nodes did much better after adjunctive chemotherapy than after post-operative irradiation. Patients with advanced disease showed significant improvement with intensive chemotherapy combined with surgery. Presently retroperitoneal lymphadenectomy combined with intensive chemotherapy seems to be the treatment of choice for metastatic testicular carcinomas.


Significance of the acetylation phenotype and the therapeutic effect of procainamide. In order to estimate the relative anti-arrhythmic effect of procainamide and N-acetyl-procainamide, 18 randomly selected, patients with arrhythmia were divided into two groups; the first was treated with Pronestyl in the first half of the investigation period, followed by Duretter in the second half, and the second group began with Duretter and terminated with Pronestyl. The concentrations of procainamide and N-acetylprocainamide were measured twice a day during the steady state part of each treatment period. The acetylation phenotype of the patients was determined with sulfadimidine, and compared with the relative serum concentrations of procainamide and N-acetylprocainamide. N-acetylprocainamide was found to antagonize the action of procainamide.


Anatomical and functional aspects of the associative projections from somatic area SI to SII. 1. Electrophysiological and morphological (retrograde axonal transport of horseradish peroxidase, HRP) experiments have been carried out in the cat in order to study the associative projections from area SI to ipsilateral SII. 2. Microelectrode recordings were performed in the forepaw focus of SII both in normal (64 units) and in SI-undercut (51 units) cats. 29.6% of the neurons recorded in the unoperated and 29.4% of those collected in the operated cats were excited by electric stimulation of the ipsilateral SI (forepaw focus). In both preparations almost all such units were endowed with large (either contra- or bilateral) receptive fields (RF). Cell population recorded in the SI-undercut cats showed no significant impairment to peripheral stimuli and/or changes in the size of the RFs. 3. From the forepaw focus of SI, 150 units have been recorded and tested by stimulation of the homologous focus of the ipsilateral SII. Eight of them were fired antidromically and thus identified as association cells. Their RFs were very small and located only in the digits of the contralateral forepaw. 4. Both single or multiple HRP injections were performed in SII. Retrogradely labelled cells were found in the ipsilateral SI. The great majority of association cells are pyramids and dwell mainly in layer III. In spite of the large diffusion of the exogenous reaction product in the injected SII and of the presence of retrogradely labelled cells anywhere in the ipsilateral thalamic VB complex, the distribution of association cells is unequal throughout SI since they strongly predominate in the digit zone of the forepaw representation.


Accumulation of Phosvel in adipose tissue of hens. Phosvel, an organophosphorus pesticide, was stored in the adipose tissue of hens after they were given daily a single oral dose. The concentration of Phosvel in fat was related to the size of the daily dose.


Sensitivity of human lymphocytes to bleomycin increases with age. The sensitivity of human peripheral blood lymphocytes to bleomycin and mitomycin-C was assessed by measuring the inhibition of phytohemagglutinin stimulated proliferation. The sensitivity to bleomycin, and not to mitomycin-C, increased with the age.


Axoplasmic transport in regenerating limbs of Ambystoma maculatum larvae. Axoplasmic transport of 3H-leucine labelled molecules from spinal cord segments into regenerating larval salamander limbs was observed. However, labelled molecules were not observed in cells of the regeneration blastema.


Interphase studies with a simplified method of silver staining of nucleoli. A simple silver staining method is presented providing a rapid and reliable technique for the selective staining of nuclear structures synthesizing ribosomal RNA (18S and 28S RNA).


[Dysfunctional uterine bleeding. Hormonal management with chlormadinone acetate]. In 138 patients with uterine dysfunctional bleeding a hormonal treatment with chlormadinone acetate, was given. The dosage varied according to four therapeutic stages: hemostatic, maintenance, consolidation and observation. Patients were placed in three groups: Group I, adolescents, Group II, reproductive life, Group III, premenopausal. In 51.9% of the patients there was endometrial hyperplasia, proliferative in 26.6% and secretory in 14.1%. Good results were obtained in all four therapeutic stages.


Vitamin B12 binding protein as a tumour marker for hepatocellular carcinoma. Grossly raised levels of tumour related vitamin B12 binding protein, reflected by rises in serum vitamin B12 and unsaturated vitamin B12 binding capacity (UBBC), were found in three of 44 patients with hepatocellular carcinoma. All three were HBsAg negative and had normal serum alpha fetoprotein levels. The patients did not have underlying cirrhosis and the tumours contained characteristic intracellular inclusions. In the first patient the UBBC level fell during a partial remission induced by adriamycin therapy and in the second patient UBBC levels rose with progression of her disease. In the third patient serum B12 binding protein levels fell after tumour resection. Assay and subsequent monitoring of serum vitamin B12 and UBCC may prove valuable in the assessment and follow-up of some patients with hepatocellular carcinoma whose alpha fetoprotein levels are normal.


[Clinical and biochemical follow up of Refsum's disease (author's transl)]. Within a family refered here, two sisters are presenting heredopathia atactica polyneuritiformis (Morbus Refsum) with the complete clinical, electromyographical and serological findings (increased phytanic acid level, accumulation of mono- and diphytanyl, triglycerides). In addition, some symptoms of Refsum's disease are clinically apparent in the eight year old daughter of one of the patients, but significant principal characteristics are absent (dissociation of the spinal fluid, retinopathia pigmentosa, increased phytanic acid level). The phytanic acid level in the serum of ten clinically normal blood relations (three of which are also obligatory heterozygotes) is normal. In both patients with manifest illness we found not only the usual spinal fluid changes with highly increased protein levels (gammaglobulin, IgA- and IgG-fraction) but also a remarkably large number of macrophages with vacuoles. Within almost two years of keeping to a strict diet with low phytanic acid and phytol content, both patients showed a distinct clinical improvement in parallel with a decrease of the phytanic acid level. Even a temporary increase of the phytanic acid level in one of the patients did not cause clinical relapse. Regarding the biochemical control of the course of the disease, the phytanic acid containing triglycerides proved to be a highly sensitive parameter.


[Formamidase--microheterogeneity, catalytic properties and inhibitors (author's transl)]. Formamidase from rat liver proved to be microheterogenous. After preparative isoelectric focusing in density gradient columns, two peaks of formamidase with identical substrate specificity were identified. By analytical focusing in thin layers of polyacrylamide or Sephadex G-75 SF, even five bands could be separated. Their isoelectric points were 4.75, 4.78, 4.82, 4.92 (main band) and 5.11, but their Michaelis constants did not differ significantly (54 to 62 mumol/l). An identical molecular weight of 34700 +/- 3200 for all bands was determined by disc electrophoresis. This value was confirmed by sedimentation analyses (so20,w = 3.00 S) and electrophoresis in the presence of sodium dodecyl-sulfate (Mr 34900 +/- 2300), which only gave a single band. The homogeneity was also confirmed by electrophoresis in the presence of 6M urea. Repeated disc electrophoresis of focusing under native conditions with single, isolated formamidases again resulted in different bands which were identified, not only by Coomassie Blue, but also by their hydrolytic cleavage of naphthyl acetate. Formamidase showed neither proteolytic nor asparagine-amidohydrolase activity and oligosaccharide conjugates were not detectable. Ampholytes, buffer ions, pH and peroxodisulfate did not affect the heterogeneity. "Initial burst" measurements with diethyl(4-nitrophenyl) phosphate yielded an equivalent weight of 36,300. Formylkynurenine reduced this inhibition very effectively. Thus, an extraordinary reactive serine residue appeared to be located in the catalytic site of formamidase. A participation of sulfhydrylgroups in the inactivating reaction of arsenite was excluded although two such groups were detected by 5,5'-dithiobis(2-nitrobenzoic acid). N-Bromosuccinimide reacted primarily with one of the nine tryptophan residues without loss of enzymatic activity, but a 18.6-fold excess of this reagent resulted in a complete loss of activity. The reaction rates of the most effective inhibitors and of the protective action of formylkynurenine were determined. Thus, formamidase must clearly be distinguished from typical serine esterases and proteases.


Inhibition of condensation of human Y chromosome by the fluorochrome Hoechst 33258 in a mouse-human cell hybrid. The fluorochrome Hoechst 33258 which binds preferentially to A-T base pairs, drastically inhibits the condensation of A-T-rich centromeric heterochromatin regions in mouse cell lines. The condensation of all other regions of these chromosomes is also inhibited to some extent. The human Y chromosome contains a large heterochromatic region, which is also rich in A-T base pairs. This chromosome is not affected by Hoechst 33258 in human leukocyte cell cultures. On the other hand, condensation of the multiple copies of human Y chromosome in the mouse-human cell hybrid RH-28Y-23 is inhibited and the chromosomes appear distorted in Hoechst 33258-treated cells.


Liposomes in immunology: further evidence for the adjuvant activity of liposomes. The immune response to HSA-phosphatidylcholine complexes administered to rabbits was not markedly enhanced when compared with the response to unmodified HSA. It was found in earlier work that HSA entrapped in liposomes (mainly composed of phosphatidylcholine) evoked a strong immune response under conditions where no response was detected to free HSA. The present results exclude the possibility that HSA-phosphatidylcholine complexes which may arise from liposome-encapsulated HSA may be responsible for the adjuvant activity of the liposome. The adjuvant activity of liposomes could also be established after administration of a liposome-associated strong antigen (BGG).


Localization of blood-group A and I antigenic sites on inside-out and rightside-out human erythrocyte membrane vesicles. Investigations of the fixation of 125I-labelled anti-A and anti-I antibodies onto rightside-out and inside-out membrane vesicles prepared from human A1 and OI erythrocytes, respectively, showed that both antibodies were bound to the rightside-out vesicles, giving clear evidence that blood group A and I antigenic sites are exclusively localized on the external surface of the membrane.


beta2-Microglobulin production by highly purified human T and B lymphocytes in cell culture stimulated with various mitogens. This study attempts to evaluate beta2-microglobulin production by highly purified (greater than 98%) peripheral and tonsil T and B lymphocytes cultured with various mitogens. beta2-Microglobulin was measured by the radioimmunoassay method. It was found that PHA and Con A markedly stimulated beta2-microglobulin production in cultures of T but not B lymphocytes. B lymphocytes were greatly activated, on the other hand, by Staphylococcus aureau Cowan I organisms cSpA), though the level of beta2-microglobulin production was less than that observed in PHA- and Con A-stimulated T lymphocytes. PWM only slightly increased beta2-microglobulin production of T lymphocytes, although the incorporation of [3H]-thymidine was highly enhanced. The highest level of beta2-microglobulin obtained with PHA or Con A was observed when the T/B lymphocyte ratio was between 90/10 and 80/20. These results lead to the conclusion that: (1) SpA is a specific mitogen for B lymphocytes, and its mitogenicity is independent of the presence of T lymphocytes, while PHA, Con A, and PWM are ineffective as stimulants of B lymphocytes; (2) the beta2-microglobulin producing ability of B lymphocytes is less than that of T lymphocytes, even when the lymphocytes are markedly activated; (3) the beta2-microglobulin production and DNA synthesis by T lymphocytes is markedly enhanced by the helper effect of B lymphocytes; (4) the level of beta2-microglobulin production reflects lymphocyte activation, especially in T lymphocytes stimulated with PHA or Con A.


Peripheral blood leucocyte response to coccidial infection: a comparison of the response in rats and chickens and its correlation with resistance to reinfection. The peripheral blood leucocyte (PBL) response to infection with two species of the intestinal protozoan Eimeria shows a remarkably similar pattern in both rats and chickens. Like resistance to these parasites, the PBL response to reinfection is species-specific and it coincides with the inhibition of development of the parasite and with cellular infiltration of the lamina propria. In primary infections, the numbers of PBL increased biphasically and significant changes were found in the count of polymorphonuclear cells (PMN), lymphocytes, and large mononuclear cells (LMN). A consistent pattern of two increases in PMN, some lymphopenia, and two increases in LMN was found in rats and chickens. The response to secondary infections was more rapid and much briefer, particularly in chickens in which significant changes were detected within hours of infection and consisted of an almost equal increase in PMN and in lymphocytes. This secondary response occurred only in animals challenged with the homologous species. It was not evoked by challenge with a related species which develops in approximately the same area of the intestine.


On the feedback regulation of humoral immune response. I. Evidence for 'B suppressor cells'. Evidence has been presented that complete and antigen-specific immune inhibition can be obtained by 'B suppressor cells'. Transfer of spleen cells from twice-immunized (SRBC) donors to untreated syngeneic recipients resulted in antigen-specific inhibition of the hosts' immune response. The cell responsible for this phenomenon could be shown to be the 7S-producing B cell; participation of T cells and macrophages could be excluded. After a second immunization of the donors, these B cells remained inhibitory for more than 20 weeks in the donors as well as in the recipients after transfer. Passively administered specific IgG antibody caused a similar inhibition of the hosts' immune response, which, however, lasted for less than 9 weeks only. The extent of inhibition caused by transfer of hyperimmune cells was parallel to the number of transferred 7S producing cells. Since it could be demonstrated that memory cells were present at times when the transferred cell material had lost its inhibitory potency, we concluded that inhibition is not caused by the mere presence of these cells. Since the transferred cells regained their inhibitory capacity after non-specific activation with LPS, we concluded that a product of such activated cells--most likely the specific 7S antibody--was responsible for the observed inhibition. Thus, it is demonstrated that B cells may serve as 'suppressor cells' in appropriate transfer experiments. It is, however, concluded that this effect is basically mediated by produced IgG and may in its mechanism be identical to the phenomenon of antibody-mediated regulation of humoral immune response.