Immunohistochemical localization of human carbonic anhydrase isoenzyme C. Methods for immunohistochemical localization of human carbonic anhydrase isoenzyme C (HCA C) with indirect fluorescent antibody and immunoperoxidase techniques are described. Both methods revealed large amounts of this "high activity" isoenzyme in the mucosae of human stomach and appendix. With the indirect immunofluorescent method the presence of the enzyme in human erythrocyte cytoplasm was also demonstrated. Correlations of present findings with those obtained with the traditional histochemical methods for demonstration of carbonic anhydrase activity are discussed.


Experimental degeneration of primary afferent terminals in the cuneate nucleus of the monkey (Macaca fascicularis). Six monkeys (Macaca fascicularis) were used for the present study. In animals which survived for 2-6 days after section of C5 to T1 dorsal roots, at least four types of degenerating afferent terminal were observed - electron-dense, electron-lucent, neurofilamentous and flocculent. The electron-dense degeneration was the most common and was seen as early as 2 days after rhizotomy. The neurofilamentous type was the second commonest and was found predominantly in the 3 days' survival material. The electron-lucent and flocculent types were less commonly encountered. Since the profiles exhibiting neurofilamentous hyperplasia showed varying degrees of electron density it is suggested that this type of degeneration progresses to the electron-dense type with time. The present study also showed that the primary afferent terminals in the cuneate nucleus of the monkey are mostly large and that they contain round vesicles. They are commonly found within synaptic complexes in which they are presynaptic to dendrites of various sizes, and are themselves postsynaptic to smaller axon terminals containing flattened vesicles. Degenerating terminals forming isolated synapses were less commonly seen. No dorsal root axon terminals formed axosomatic synapses.


Conformational changes of yeast tRNAPhe and E. coli tRNA2Glu as indicated by different nuclease digestion patterns. The susceptibility of yeast tRNAPhe and Escherichia coli tRNA2Glu to digestion by nucleases Tl and Sl are examined in a variety of environments, and the results are interpreted in view of the available three-dimensional structural information. Significant differences are found in the digestion pattern of the two tRNAs using the guanosine-specific Tl nuclease. In particular, differences are seen due to varying the type of salts in the environment. However, the Sl nuclease results on the two tRNAs do not differ greatly. E. coli tRNA2Glu is known to exist in two different conformations. Nuclease digestion results are presented revealing differences which make it possible to draw some inferences about the structural differences in these two conformations. In carrying out these analyses, the tRNA molecules are labeled either by putting 32P at the 5'-end of the molecular or by adding 32P-labeled pCp at the 3'-end. It is found that both yeast tRNAPhe and E. coli tRNA2Glu have modified Tl nuclease digestion patterns when pCp is added at the 3'-end of the molecule.


A high molecular weight DNA polymerase from Drosophila melanogaster embryos. Purification, structure, and partial characterization. The DNA polymerase of early embryos of Drosophila melanogaster has been purified to near-homogeneity. The purified enzyme gave a single, catalytically active protein band after polyacrylamide gel electrophoresis, under nondenaturing conditions. Four polypeptides with molecular weights 43,000, 46,000, 58,000, and 148,000 were resolved when this band was electrophoresed under denaturing conditions. At high ionic strengths, the DNA polymerase had a sedimentation coefficient of 8.7 S, a Stokes radius of 78 A and frictional ratio of 1.81, parameters that yield a molecular weight of 280,000. The purified DNA polymerase possessed no detectable endo- or exodeoxyribonuclease, ATPase, or RNA polymerase activity. Using an "activated" DNA template-primer, the enzyme had a pH optimum of 8.5. It was stimulated by (NH4)2SO4, KCl, and to a lesser extent, NaCl. A divalent metal cation was absolutely required; MgCl2 stimulating activity 7-fold more than MnCl2. It was inhibited by low concentrations of N-ethylmaleimide and Aphidicolon. Thus the DNA polymerase of D. melanogaster resembles most closely the alpha-DNA polymerases that have been purified from mammalian cells.


The rabbit pulmonary monooxygenase system. Immunochemical and biochemical characterization of enzyme components. The enzymatic components of the rabbit pulmonary monooxygenase system, cytochromes P-450I and P-450II and NADPH-cytochrome P-450 reductase, are immunochemically distinct proteins. In pulmonary microsomes, the N-demethylation of benzphetamine, amino-pyrine, and ethylmorphine, and the O-deethylation of 7-ethoxycoumarin are dependent only on cytochrome P-450I, and the hydroxylation of coumarin is apparently catalyzed by both cytochromes. Cytochrome P-450II is immunochemically distinct from the major forms of hepatic cytochrome P-450 induced by phenobarbital or 3-methylcholanthrene, whereas cytochrome P-450I is indistinguishable from the former on the basis of physical and catalytic as well as immunochemical characteristics. Pulmonary and hepatic NADPH-cytochrome P-450 reductases also have identical physical, catalytic, and immunochemical properties. The lack of response of the lung monooxygenase system to phenobarbital, therefore, is apparently not due to an inability of the lung to synthesize the enzymes induced by phenobarbital in the liver. The relatively high proportion of cytochrome P-450I in the lung appears to be responsible for the higher rates (per nmol of P-450) of N-demethylation that have been observed in rabbit pulmonary as compared to hepatic microsomal fractions.


Laminin--a glycoprotein from basement membranes. We have isolated a large noncollagenous glycoprotein, laminin, from a mouse tumor that produces basement membrane. The protein consists of at least two polypeptide chains (Mr = 220,000 and Mr = 440,000) joined to each other by disulfide bonds. Laminin and type IV collagen are major constituents of the tumor. Laminin is distinctly different from fibronectin, another component of basement membranes, in amino acid composition and immunological reactivity. Pepsin digestion of laminin releases a large, cystine-rich fragment which retains most of the antigenicity of the original protein. Immunological studies using purified antibody against laminin show that it is produced by a variety of cultured cells. In addition, these antibodies react with the basement membranes of normal tissues, suggesting that this protein or an immunologically related protein is a constituent of the basement membranes of these tissues.


Steroid induction of delta-aminolevulinic acid synthase and porphyrins in liver. Structure-activity studies and the permissive effects of hormones on the induction process. Quantitative aspects and structure-activity relationships of the inducing effects of natural steroids on delta-aminolevulinic acid (ALA) synthase and porphyrins have been investigated in monolayer cultures of chick embryo liver cells maintained in a serum-free medium as well as in the chick embryo liver in ovo. Many 5 alpha and 5 beta metabolites of neutral C-19 and C-21 hormones and hormone precursors stimulated porphyrin formation and ALA-synthase induction in the cultured liver cells as we have previously described. In these inducing actions a number of 5 beta epimers (A:B cis) were found to be more potent than their corresponding 5 alpha epimers (A:B trans). The structure-activity relationship between 5 beta and 5 alpha steroid epimers with respect to ALA-synthase induction in culture was also found to prevail with respect to induction of this enzyme in chick embryo liver in ovo. Hemin in concentrations of 2 x 10(-7) M inhibited steroid induction of porphyrin formation, and CaMgEDTA enhanced the responsiveness of the cultured liver cells to steroids by approximately 10 times. The addition of insulin, or insulin plus hydrocortisone or insulin plus hydrocortisone plus triiodothyronine, was important for the maintenance of protein synthesis and essential for maximal expression of the ability of steroids to induce porphyrins and ALA-synthase in the "permissive" effect which insulin, hydrocortisone, and triiodothyronine exert on allylisopropylacetamide induction of porphyrins and ALA-synthase also extends to the induction process which is elicited by natural steroids. These findings also strongly suggest that the regulation of hepatic porphyrin-heme biosynthesis by endogenous as well as exogenous chemicals is significantly influenced by the internal hormonal milieu.


Activation and regulation of ribulose bisphosphate carboxylase-oxygenase in the absence of small subunits. Ribulose 1,5-bisphosphate carboxylase from Rhodospirillum rubrum requires CO2 and Mg2+ for activation of both CO2, both the carboxylase and oxygenase activities are stimulated by 6-phoshpo-D-gluconate, fructose 1,6-bisphosphate, 2-phosphoglycolate, 3-phosphoglycerate, NADPH, and fructose 6-phosphate. The carboxylase activity is not activated by ribose 5-phosphate. The substrate, ribulose bisphosphate, neither activates nor inhibits the CO2 and Mg2+ activation of this enzyme. Activation by CO2 and Mg2+ is rapid and results in increased susceptibility to active-site-directed protein modification reagents. Because the R. rubrum carboxylase-oxygenase is a dimer of large subunits and contains no small subunits, these results suggest that the effector binding sites of the higher plant enzyme may also be found on the large subunit.


The nucleotide sequence of a cloned Drosophila arginine tRNA gene and its in vitro transcription in Xenopus germinal vesicle extracts. The DNA sequence of a cloned gene specifying Drosophila tRNA2Arg has been determined. Transcription of this gene in extracts from Xenopus germinal vesicles yielded a precursor RNA containing a seven-nucleotide leader sequence at the 5' side of the mature tRNA with pppGp as the 5'-terminal nucleotide. Transcription continues for at least eight nucleotides past the 3' terminus of the coding region of the gene and terminates over a stretch of seven T residues. The isolation of the primary transcript was facilitated by incorporation of 5'-(gamma-S)GTP into the 5' terminus of the precursor RNA and by its retention on mercury-agarose. The Drosophila tRNAArg gene does not contain an intervening sequence nor the C-C-A sequence corresponding to the 3' terminus of the mature tRNA. The nucleotide sequence of Drosophila tRNAArg formed in the in vitro system differs only in four positions from that of mouse tRNAArg.


Regulation and state of aggregation of Bacillus subtilis prephenate dehydratase in the presence of allosteric effectors. Prephenate dehydratase from Bacillus subtilis was found to exist in three states of aggregation. A high molecular weight (210,000) species was fully active and the catalytic activity was unaffected by the effectors methionine or phenylalanine. Low concentrations of phenylalanine caused dissociation to a Mr = 55,000 dimer. Heating to 32 degrees C also caused dissociation, but cooling and adding substrate or methionine favored association. When no effectors were present the enzyme eluted from Sephadex columns as a monomer. Both methionine and phenylalanine shifted the equilibrium from the inactive monomer to the active dimeric enzyme. In the presence of a saturating methionine concentration, the dimer possessed the same high activity as did the 210,000-dalton form. Phenylalanine inhibited the dimer, but not the higher molecular weight form. A model involving only three types of sites (catalytic, association-activation, and inhibition) is consistent with the data. It is proposed that phenylalanine is the preferred metabolite for binding both effector sites on the dimer; it binds the association-activation site with higher affinity than the inhibition site, but binding at the latter site has a greater effect on the catalytic rate. Methionine, like phenylalanine, has a hydrophobic side chain but is accommodated only at the association-activation site.


Quantitative analysis of intermediary metabolism in Tetrahymena. Cells grown in proteose-peptone and resuspended in a defined nutrient-rich medium. Tetrahymena pyriformis were grown to early-stationary phase and resuspended in a defined mixture containing glucose, fructose, ribose, glycerol, acetate, pyruvate, bicarbonate, glutamate, and hexanoate, with only one substrate labeled with 14C in any flask. Incorporation of label into CO2, glycogen, RNA, alanine, glutamate, glycine, lipid glycerol, and lipid fatty acids was measured 20, 40, and 60 min after the start of the incubation. To develop a model suitable for quantitative analysis of the data, it was necessary to join two preceding models, one for carbohydrate-metabolizing cells and one for acetate-metabolizing cells, eliminating the over-simplified sections of each. Equations were written and programmed for a digital computer to allow computation of the amount of label expected to be incorporated into any of the products measured for any given set of steady state flux values in the metabolic network. The model formed by simply joining the two preceding models did not yield satisfactory agreement with the complete data obtained in the present study, although each partial set of data could be fit well by the appropriate partial model. Analysis of the ways in which the model failed to yield good fits to the data indicated that another pool of P-enolpyruvate, of pyruvate, and of acetyl-CoA had to be added at the junction of the two models. The presence of such poolte into fatty acids as compared to the incorporation of label from glucose into fatty acids. A new model was therefore constructed which differed from the preceding model only in its structural organization at the level of P-enolpyruvate, pyruvate, and acetyl-CoA. The model is consistent with all known information on the compartmental structure of metabolism in Tetrahymena, on enzyme localization, and on the enzyme complement of this cell. Over 70 measurements of label incorporation into products were made at each time. These, plus a large number of "limit" measurements which constrain any possible solutions, were in sufficient excess of the 39 independent flux values to permit a stringent assessment of the model. A set of flux values was found which yielded a good fit to the data. These flux values therefore provide a quantitative description of metabolite flux in the intact cell during the slow adaptation to the nine-substrate mixture. The rates of utilization of glucose, fructose, glycerol, and ribose were in the ratio of about 10:1:0.33:0.16, i.e. fairly similar to the ratio observed with carbohydrate-metabolizing cells. Initial flux through phosphofructokinase is about 160 nmol/10(6) cells.h, increasing over 3-fold during tje jpir incubation. Initial flux through fructose-1,6-diphosphatase is about 110 nmol/10(6) cells.h and also increases almost 3-fold during the incubation. Thus net flux is glycolytic and increases 4-fold during the hour with a large amount of futile cycling at this step...


Biosynthesis of two lysosomal enzymes in macrophages. Evidence for a precursor of beta-galactosidase. We have purified beta-galactosidase and beta-glucuronidase from macrophages of thioglycollate-treated mice using concanavalin A chromatography and immunoprecipitation. The apparent molecular weight of the beta-galactosidase subunit, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, changed during a long term pulse-chase experiment. Following a 1-h pulse with [3H]leucine, radiolabel was present exclusively in an Mr = 82,000 form. However, after a 3-h chase in medium containing unlabeled leucine, most label migrated at Mr = 63,000, and at 24 h, all label was in the Mr = 63,000 form. Electrophoresis of peptides produced by cyanogen bromide cleavage of immunoprecipitates demonstrated structural similarities between precursor and mature forms. A mutation in the mouse, which is known to depress the rate of synthesis of beta-galactosidase in many cell types, proportionately decreased incorporation of [3H]leucine into both the Mr = 82,000 and 63,000 forms. Therefore, by kinetic, structural, and genetic evidence, the large molecular weight beta-galactosidase is a precursor of mature macrophage enzyme. No precursor of the Mr = 75,000 subunit of beta-glucuronidase was detected.


Failure of bromocriptine, dopamine, and thyrotropin-releasing hormone to affect prolactin secretion by human decidual tissue in vitro. To determine whether the secretion of PRL by human decidual tissue in vitro is influenced by factors which inhibit or stimulate pituitary PRL secretion, explants of decidual tissue were incubated in media containing bromocriptine, dopamine, or TRH at concentrations known to affect pituitary PRL secretion in vitro. The quantities of PRL secreted by the explants exposed to these factors were compared with amounts secreted by explants incubated in control medium. Bromocriptine in concentrations ranging from 1.5 x 10(-10) to 1.5 x 10(-7) M did not inhibit PRL secretion over a 3-day period and dopamine in concentrations ranging from 5 x 10(-5)-10(-9) M did not inhibit PRL secretion over a 4-h period. TRH in concentrations ranging from 10(-9)-10(-3) M did not stimulate PRL secretion. These results suggest that the mechanism of PRL secretion by decidual tissue in vitro is different, at least in part, from the mechanism of pituitary PRL secretion.


Comparison of a slide coagglutination technique with the Minitek system for confirmation of Neisseria gonorrhoeae. The Phadebact gonococcus test was performed on 314 bacterial isolates to compare the coagglutination technique with the Minitek system for the confirmation of Neisseria gonorrhoeae. Direct testing, using colonies of N. gonorrhoeae mixed with the Phadebact gonococcus test reagents, produced noninterpretable results in many cases. However, an alternative method, described by the manufacturer, of heating several colonies in distilled  water for 20 min before testing alleviated most interpretation difficulties. Complete agreement was achieved with both the Phadebact and Minitek tests for all 236 isolates of N. gonorrhoeae. Of the 78 nongonococcal isolates tested with Phadebact, two isolates gave false-positive results, whereas one isolate was reported as noninterpretable. The Phadebact gonococcus test appears to be a simple and rapid alternative to confirmatory biochemical procedures for N. gonorrhoeae.


Rapid serogroup identification of Neisseria meningitidis by using antiserum agar: Prevalence of serotypes in a disease-free military population. Nasopharyngeal cultures from 414 Marines were plated directly onto antiserum agar containing the antibiotics vancomycin, colistin, and nystatin for meningococcal isolation and serogroup identification. Meningococci were isolated from 267 Marines, giving a carrier prevalence of 64.5%. A total of 58% of the isolates could be placed into serogroups; of these 22.3% were group B, 4.7% were group C, 25.7% were group Y, 24.3% were group W135, and 23.0% were group 29E. No serogroup A organisms were recovered. Serotyping by agar gel double diffusion was performed on 148 strains. More than 70% of these strains were nontypable, and the disease-associated serotype 2 was present only in two group Y isolates. The same 148 isolates were also classified by major outer membrane protein patterns after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Polyacrylamide gel electrophoresis pattern IV was most common among all serogroups. The data demonstrate the effective use of antiserum agar for meningococcal surveillance and document the frequency of specific serotypes and polyacrylamide gel electrophoresis types among carrier isolates obtained from a nonrecruit military population.


Microimmunodiffusion test for nocardiosis. Sera from 71 patients with culturally proven nocardiosis were tested for precipitins against a pool of Nocardia asteroides and N. brasiliensis culture filtrates and against antigens from the supernatant of homogenized N. asteroides cells. A human nocardiosis case serum was used as a reference. Sera from 56 of the 71 cases were reactive with either the culture filtrate antigen, the homogenate antigen, or both antigens, resulting in an overall sensitivity of 79%. Sera from 35 of the patients (49%) were positive with the homogenate antigens, and 28 (39%) showed bands of identity with the reference serum. Sera from 50 nocardiosis cases (70%) were positive with the pooled culture filtrate antigens, and 29 (41%) produced bands of identity with the reference serum. Of 89 sera from patients with various systemic mycotic diseases, tuberculosis, or actinomycosis, 24 (27%) were positive with the nocardial homogenate antigens and 4 (4.5%) showed precipitin bands of identity. Thirty-five of the 89 sera (42%) were positive with the nocardial culture filtrate antigens, and 6 (6.7%) showed bands of identity. The majority of sera demonstrating false-positive reactions were from tuberculosis and actinomycosis cases. One of seven sera from well individuals produced a precipitin band with the culture filtrate antigen, but this was not a band of identity with reference serum. These antigens did not distinguish antibodies from patients with N. asteroides, N. brasiliensis, or N. cavia infections.


Natural infection of dogs on Cape Cod with Rickettsia rickettsii. Four isolates of rickettsiae from sick dogs on Cape Cod, Mass., were serologically identical to isolates of Rickettsia rickettsii from human patients with Rocky Mountain spotted fever. The antigenic analysis used the indirect fluorescent-antibody test and antisera prepared in mice to each of the isolates and to reference strains of R. rickettsii and Rickettsia montana. Serological responses of infected dogs were specific for R. rickettsii, although antibodies to R. montana were also detected in the sera of most of the canines.


Application of fluoroimmunoassay to cerebrospinal fluid immunoglobulin G and albumin. Special solid-phase fluoroimmunoasssay protocols were used to measure the amount of immunoglobulin G (IgG) and albumin in 1,511 samples of cerebrospinal fluid. The fluoroimmunoassay is an inhibition test conducted on the surface of a plastic probe, whose fluorescence is inversely related to the concentration of spinal fluid IgG or albumin. A microcomputer interfaced with the fluorometer calculates the sample IgG or albumin from a calibration curve based on standard cerebrospinal fluid values. The test and interpretation take less than 90 min. Correlation coefficients for over 100 cerebrospinal fluid samples tested by both fluoroimmunoassay and radial immunodiffusion were: IgG, 0.90 (slope, 1.04); albumin, 0.95 (slope, 0.99). The within-run precision (coefficient of variation) was: IgG, 4.4%; albumin, 6.3%. Run-to-run precision on a midrange sample was: IgG, 7.7%, albumin, 11.7%. These findings establish the simplicity, speed, and precision of the modified fluoroimmunoassay system for specific cerebrospinal fluid proteins.


Atrial septal thickness and area in normal heart specimens and in those with ostium secundum atrial septal defects. This anatomic study suggests that an understanding of the thickness of the atrial septum should allow better visualization of most portions of the blade-shaped atrial septum during cross-sectional echocardiography, both in normal subjects and in patients with ostium secundum atrial septal defects. The thickness measurements also demonstrate that difficulties in imaging the atrial septum are a function of resolution limits of existing echocardiographic equipment.


The quadratus lumborum muscle: a possible source of confusion in sonographic evaluation of the retroperitoneum. The quadratus lumborum muscle has been noted to appear as a hypoechoic structure on sonograms of the abdomen. On occasion its appearance may simulate a pathologic fluid collection such as an abscess or retroperitoneal hematoma. Illustrative examples of this confusing appearance, as well as ways of avoiding this potential pitfall, are presented.


The fluid-filled stomach: a new sonic window. Gastric air and peristalsis usually are obstacles to ultrasound imaging of the tail of the pancreas. Fifteen normal volunteers were scanned transversely in the supine position before and after intravenous administration of glucagon (1 mg) and oral administration of water. In 13 of 15 subjects glucagon produced a dilated gastric fundus that retained fluid for 30--60 min, thus allowing good imaging of the pancreatic tail in 10 subjects and fair imaging in 2 subjects. This method of creating a gastric sonic window should prove to be valuable in ultrasound imaging of the pancreas.


Detection of persistent left superior vena cava by two-dimensional contrast echocardiography. One hundred seventy adult patients with possible congenital or valvular heart disease underwent contrast two-dimensional echocardiographic examination as part of a precatheterization evaluation. Persistent left superior vena cava was detected in 5 patients, each of whom demonstrated an abnormally large coronary sinus. Injection of echocardiographic contrast material from a peripheral left arm vein resulted in early opacification of this structure before other right-side chambers, thus suggesting abnormal venous drainage. Persistent left superior vena cava was confirmed in all 5 patients at the time of catheterization and/or surgery.


Prenatal diagnosis of fetal lymphatic system abnormalities by ultrasound. Antenatal ultrasound diagnoses of gross lymphatic system abnormalities were made in three fetuses. Although the diagnosis was made in only 1 fetus at a stage early enough in pregnancy to allow selective termination, knowledge of the abnormality in the remaining 2 fetuses proved to be valuable for subsequent management of those pregnancies. It is stressed that the extent of the abnormality must be carefully assessed because of the possibility of corrective surgery should the lesion be small and that the parents must be given detailed counseling before any definitive measures are taken.


Ultrasound findings in liver hydatid cysts. Eighty-one liver hydatid cysts evaluated by gray-scale ultrasound were accumulated in our experience over a period of 2 years. The echographic evidence in accordance with our experience can be categorized as follows: (a) nonsuppurative hydatid cysts in which echo-free area within the fine homogeneous structure of the organ provide the basic clue to the diagnosis; then the echographic appearance of the wall offers further hints to the identity of solitary, multiloculated, and multiple cysts; (b) suppurative hydatid cysts in which, depending on the intensity of secondary microbial infection, the echo-free area turns to a low, medium, or high level of echoes while holding its constitutional pattern. This report is designed to present the various ultrasound patterns of hydatid disease of the liver and show how ultrasound may be most effective in achieving the correct diagnosis.


Echocardiographic aortic ejection area as a reflection of left ventricular stroke volume. Systolic cusp separation of normal aortic valves has been shown by echocardiography and by angiography to be related to left ventricular stroke volume. In this study, stroke volume was compared with echocardiographic area between the aortic cusps in systole (aortic ejection area). An excellent correlation was observed (r = 0.93) over a wide range of stroke volumes, even in the presence of mitral regurgitation. Stroke volume appears to correlate more closely with aortic ejection area than with angiographic or echocardiographic measurements of peak systolic cusp separation.


Upright exercise echocardiography.

Upright exercise echocardiography. A simple technique for obtaining high-quality echocardiograms of the left ventricle during upright bicycle exercise is described. The use of this method has resulted in left ventricular dimension data that are consistent with the results of studies in conscious, previously instrumented dogs during running. Careful subject selection is important, however, as only 1 of 5 normal subjects was found suitable for these studies. Therefore, we believe that upright exercise echocardiography is feasible in selected subjects and provides useful physiologic information about left ventricular performance during exercise.


Thalamic projections to area 17 in a prosimian primate, Microcebus murinus. Electrophysiological recording of single neurons was used to describe the representation of visual space in area 17, and the technique of retrograde transport of horseradish peroxidase (HRP) was applied to relate these results to projections from the thalamus in the prosimian primate Microcebus murinus. The visuotopic organization of area 17 was found to resemble that of other primates. On the dorsal surface, the border of area 17 corresponds to the representation of the vertical meridian. Proceeding medially across the surface the location of receptive fields descends along the vertical meridian, while moving caudally receptive fields progress temporally. Most of the dorsolateral surface is devoted to central vision and corresponds to a well developed area centralis. Following HRP injections in striate cortex, columns of labeled cells were found in the dorsal lateral geniculate (dLGN) extending orthogonally across all six layers. These columns run in a general ventrodorsal and caudorostral direction, parallel to a line connecting the cellular discontinuities corresponding to the optic disc. These discontinuities are present in magnocellular layer 1 and parvocellular layers 5 and 6, thus receiving from the nasal retina of the contralateral eye. The representation of the vertical meridian is situated in the ventromedial portion of the dLGN, and the monocular field is represented in the dorsal extremity. Anterior dLGN projects to the calcarine fissure (lower field) and posterior dLGN projects to the ventral surface of the cortex (upper field). Extrageniculate input to area 17 was found to originate from the pulvinar. HRP labeled cells were located in two distinct divisions of this nucleus, the cytoarchitecture of which is described. In addition, projections to occipital cortex were found to arise from the intralaminar nuclei.


Endocytic activity of subependymal microglial cells in the toad brain: a cytochemical study of peroxidase uptake. A population of microglial cells that rapidly incorporate extracellular material introduced into the ventricular system has been identified just beneath the ependyma of all four cerebral ventricles in the toad (Bufo marinus). In untreated tissue these cells appear to be scattered, possess few processes and have an elongate shape with their long axes lying parallel to the ventricular surface. Their most distinctive ultrastructural features are nuclei containing clumps of chromatin, cytoplasmic dense bodies and single strands of granular endoplasmic reticulum. When horseradish peroxidase (HRP) is perfused through the ventricular system and the tissue processed using the DAB cytochemical method, the cells change shape and incorporate HRP into cytoplasmic structures. Even after very short perfusion periods (2-5 minutes) cells become rounded, the surface is ruffled and pseudopodia develop that contain characteristic flocculent material. Reaction product for HRP is contained in plain and coated vesicles, tubules, vacuoles and long structures composed of two closely apposed membranes. At these early times, relatively few multivesicular bodies and dense bodies contain reaction product, but when the cells are viewed at longer time periods after the ventricular perfusion of HRP an increasing proportion of the multivesicular bodies and dense bodies contain reaction product. By 320 minutes reaction product is found almost exclusively in these two organelles. In addition, many pseudopodia containing dense bodies with peroxidase activity are found in the neurophile; some, but not all, can be traced from the subependymal microglial cells. The cell bodies have resumed their flattened shape. When compared to the subependymal microglial cells, other brain cells--oligodendrocytes, astrocytes, ependymal cells and neurons--contain relatively little reaction product at short time intervals; only by 320 minutes are moderate amounts of HRP present. Because of the position of the microglial cells and their ingestive capacity, it is suggested that they function to protect the brain from foreign substances entering from the CSF.


The raphe nuclei of the rabbit brain stem. The raphe nuclei of the rabbit brain stem were found in the midline and adjacent reticular formation of the medulla, pons, and mesencephalon. Nuclei raphe obscurus, pallidus, and magnus were located in the medulla. Nucleus raphe pontis and the caudal portion of nuclei raphe dorsalis and centralis superior were present in the pons. The rostral portion of nuclei raphe dorsalis and centralis superior, and nuclei linearis caudalis and intermedius were present in the msencephalon. Wings of neurons extended from the midline clusters of raphe neurons into the adjacent reticular formation. These wings of neurons contained serotonergic perikarya which were cytoarchitecturally indistinguishable from the midline neurons. A detailed localization of these nuclei is presented in atlas form. These raphe nuclei contained heterogeneous populations of neurons which varied in the size, shape and density of the cell bodies. In addition, the dendritic branching, specific orientation of dendrites, and appearance of spines were distinct for each of the raphe nuclei. Individual raphe nuclei often contained several subpopulations of neurons characterized by unique spatial configuration and orientation. The main morphological similarities of the raphe nuclei are location in or adjacent to the midline, the presence of serotonergic cell bodies in all raphe nuclei except the linear nuclei, and heterogeneous cell populations.


Excitatory synaptic ensemble properties in the visual cortex of the macaque monkey: a current source density analysis of electrically evoked potentials. The spatio-temporal distributions of excitatory synaptic ensemble activities in A17 and A18 of the visual cortex of the macaque monkey have been investigated. The synaptic activities were elicited by electrical stimulation of the primary efferents and were localized by applying the current source density analysis to the intracortically recorded field potentials. The principal results are as follows: 1. In A17, two groups of activity, evoked by fast and slow afferents, respectively, were distinguishable. 2. The fast afferents induced monosynaptic activity in layer IV C alpha and layer VI, disynaptic activity in layer IV C alpha and in the supragranular layers and trisynaptic activity in layer IV B. 3. The slow efferents induced monosynaptic activity in lower layers IV C beta and layer VI, disynaptic activity via strong connections in upper layer IV C beta, further disynaptic activity in layers III and IV B and trisynaptic activity in layers V A and II. 4. With the exception that the CSD data reveal more polysynaptic activity within layer IV, there is good agreement between the spatio-temporal distribution of synaptic activities and the cortical circuit diagrams proposed in anatomical studies. 5. In A18, activities from the slow and fast conducting afferent systems are revealed in layer IV, both most likely mediated by the monosynaptically activated target cells of A17. These activities are passed on to the supra-and infragranular layers. 6. In the lateral geniculate nucleus the safety factor of transmission is higher for activity conveyed by slow-than by fast-conducting retinal afferents. 7. The spatial distribution of monocularly evoked surface potentials failed to reveal the ocular dominance columns. 8. Comparison with the cat indicates that, with respect to the intracortical circuitry and LHN-transmission, there are more similarities between the fast-group activity in the monkey and the y-system in the cat and between the slow-group activity in the monkey and the x-system in the cat than vice versa.


Visual thalamocortical connections in sheep studied by means of the retrograde transport of horseradish-peroxidase. In order to study the visual thalamocortical connections in the sheep, horseradish peroxidase (0.3--0.5 microliter of a 30% solution) has been injected in the gyri marginalis, ectomarginalis medius pars medialis, ectomarginalis medius pars lateralis and ectosylvius caudalis. The results show that: (1) the dorsal lateral geniculate nucleus (LGNd) projects to the former three gyri. Dorsal parts of the LGNd project to caudal areas, whereas its ventral parts project to rostral areas of these gyri; medial parts of the LGNd project to the gyrus ectomarginalis medius pars lateralis, while lateral parts project to the gyrus marginalis; (2) the medial interlaminar nucleus (MIN) or pars geniculata pulvinaris of Rose ('42b) projects to the caudal part of the gyrus marginalis and to the gyrus ectomarginalis medius pars lateralis; (3) the pulvinar proper of Rose (PUL) projects to the caudal part of the gyrus ectosylvius caudalis whereas the rostral part of this gyrus receives input from the medial geniculate body. In relation to Rose's cytoarchitectonic study of the cortex of sheep ('42a) the present study has shown that the LGNd projects to both the area striata (gyrus marginalis + gyrus ectomarginalis medius pars medialis) and area occipitalis (gyrus ectomarginalis medius pars lateralis) of Rose, that the gyrus marginalis and the area occipitals receive a second projection (from the MIN), and that the PUL projects beyond the area occipitalis to the area parietalis of Rose.


Subcortical projections of six visual cortical areas in the owl monkey, Aotus trivirgatus. Subcortical projections of six visual cortical areas (Areas 17 and 18, the Middle Temporal, Dorsomedial and Medial Areas, and the Posterior Parietal Region) in the owl monkey, Aotus trivigatus, were investigated with autoradiographic methods following injections of tritiated proline. No contralateral projections were demonstrated. While some brainstem structures received input from all six subdivisions of cortex, each cortical area appeared to exhibit its own unique pattern of subcortical projections. All six cortical areas were found to project to the superior and inferior divisions of the pulvinar, reticular nucleus of the thalamus, pretectum and superior colliculus. Other subcortical targets of one or more visual cortical areas were the basal ganglia, claustrum, zona incerta, one or more of the intralaminar nuclei, lateral posterior nucleus, pregeniculate nucleus, dorsal lateral geniculate nucleus, and pontine nuclei. Furthermore, details of corticofugal projections to the dorsal lateral geniculate nucleus, pretectum and superior colliculus varied with the cortical area studied. The projections to the reticular nucleus, pregeniculate nucleus, dorsal lateral geniculate nucleus, the inferior and a portion of the superior division of the pulvinar and the superior colliculus were found to be topographically organized. The targets of the subcortical projections were compared with those of the retina, as revealed by autoradiographic methods following tritiated proline injections of the eye and were found to overlap to varying extents in the superior colliculus, pretectum and dorsal lateral geniculate nucleus and to be segregated in the pregeniculate nucleus. The results substantiate the validity of previous studies in the owl monkey that suggest that the visual cortex is subdivided into several functionally distinct areas; and illustrate the complexity of corticofugal influence on visual processing.


The inferior pulvinar complex in owl monkeys: architectonic subdivisions and patterns of input from the superior colliculus and subdivisions of visual cortex. Patterns of connections with other visual structures and architectonic characteristics were used to subdivide the inferior pulvinar complex of owl monkeys into three distinct nuclei termed the central inferior pulvinar, IPc, the medial inferior pulvinar, IPm, and the posterior inferior pulvinar, IPp. IPc occupies about 70%; IPm about 20%, and IPp about 10% of the inferior pulvinar complex. Encapsulating fiber bands distinguish the boundaries of the three nuclei. IPm is also identified by a much greater packing density of neurons than IPc and IPp. Both IPp and IPc receive input from the superior colliculus, but the terminations in IPp are denser. Visual cortical Areas 17, 18, MT, DM, M and PP (Allman and Kaas, '76) project to IPc and IPm in patterns that indicate that central vision is represented dorsorostrally and peripheral vision ventrocaudally in both nuclei. Terminations in IPm from Area MT are particularly dense. None of these visual areas projects to IPp. Rather, input to IPp appears to originate in cortex rostral to Area MT in the temporal lobe.


The striate projection zone in the superior temporal sulcus of Macaca mulatta: location and topographic organization. In the rhesus monkey, the caudal portion of the superior temporal sulcus (STS) receives a direct projection from lateral striate cortex, the striate are representing central vision. The present study was undertaken to determine whether STS also receives a direct projection from areas of striate cortex representing peripheral vision, with the intent of defining the entire striate projection zone in STS as well as providing information regarding a possible topographic organization within this secondary visual area. A series of five rhesus monkeys was prepared with unilateral lesions of lateral, posterior, or medial striate cortex, such that, collectively, the lesions in the series included all of striate cortex with little or no invasion of prestriate cortex. The monkeys were sacrificed seven days after surgery and their brains were processed by the Fink-Heimer procedure. An analysis of the distribution of terminal degeneration within STS indicated: (1) All areas of striate cortex project to a restricted region along the caudal portion of STS. The ventral limit of this region can be demarcated by an imaginary line connecting the ventral tips of the lunate and intraparietal sulci; from this limit the region extends dorsocaudally for approximately 12 mm to the point at which STS frequently bifurcates, sending one spur forward into the inferior parietal lobule. (2) Within this portion of STS there is an orderly mapping of the visual field; progression from central vision to the far periphery is represented by a progression down the posterior bank of STS and continuing along the entire floor, or insula-like portion, of the sulcus. (3) Projections from striate cortex to STS terminate predominantly in layer IV and the deep part of layer III. (4) There is a distinctive pattern of myelination contained within the striate projection zone of STS. These anatomical findings concerning the striate projection zone of STS in the rhesus monkey are remarkably similar to those that have been described for the middle temporal visual area (MT) in New World monkeys, and thus support earlier proposals that the two areas are homologous.


Cells of origin of long descending propriospinal fibers connecting the spinal enlargements in cat and monkey determined by horseradish peroxidase and electrophysiological techniques. The cells of origin of the long descending propriospinal tract (LDPT) in the cervical enlargement were studied in cat and monkey by using the retrograde transport of horseradish peroxidase (HRP). Their distribution was confirmed electrophysiologically in cat by recording their antidromic action potentials. In cats and monkeys unilateral injections of HRP were made into the gray matter of the lumbosacral enlargement, but there was some spread to the contralateral side. In cats labeled somas were found in greatest numbers in lamina VIII and medial lamina VII, bilaterally. Labeled cells also were found bilaterally in laminae I, IV--VI, and X, but few were in IV and VI. Those in lamina V were usually in the lateral part of the lamina near the reticulated region. The cross-sectional areas of 20 neurons from each of laminae I and V--VIII were measured. Cells in lamina I were smallest and the largest were in VII and VIII. In cats with the spinal cord hemisected between the injection site and the cervical enlargement containing the somas, the bilaterality of the LDPT neurons in laminae VII and VIII was confirmed anatomically and physiologically. Contralaterally projecting neurons in laminae VIII and medial VII constituted a majority of LDPT cells in those laminae. The LDPT neurons in the dorsal horn appeared to project mainly ipsilaterally, but the number of labeled dorsal horn cells in these preparations was small. The distribution of antidromically localized cells of the LDPT was found to be in good agreement with the anatomical results. Their conduction velocity was 59 +/- 22 m/s (mean +/- s.d., n = 245). Histograms of the conduction velocity by laminae are given. In monkey the distribution of labeled somas was similar to that in the cat, except that the concentration of labeled somas in the ventral horn was more medially and dorsally located. Labeled somas were found bilaterally in laminae I, IV--VIII, and X, but more appeared to be ipsilateral to the side of the injection, especially in the dorsal horn. The bilaterality of the LDPT in the monkey was not tested with hemisections of the spinal cord. Neurons of the LDPT are ideally situated for conveying sensory information from the forelimb for eliciting reflexes in the hindlimb, as has been observed after stimulating afferents in the forelimb, and for coordinating, in general, motor functions between the two pairs of limbs.


The effect of calcitonin in vitro on tooth germs in protein-energy malnourished rats. The effect of calcitonin on the development of incisor and molar tooth germs was studied in normal and nutritionally deprived newborn rats. Calcitonin affected the calcium uptake of nutritionally deprived molar tooth germs in the early postnatal period; this interaction was not seen in the later postnatal period, however.


Calcium-specific precipitation of dentin phosphoprotein: a new method of purification and the significance for the mechanism of calcification. Calcium, but not magnesium or strontium ions, specifically induce the precipitation of dentin phosphoprotein, which is reversibly solubilized by EDTA. This finding is very useful in pruifying dentin phosphoprotein of either a free or matrix-bound type. Bovine dentin phosphoproteins, thus isolated, contain a minimal amount of contaminant protein, unlike previous preparations.


Bromocriptine induced pregnancy in two cases of euprolactinemic hypothalamic amenorrhea. Two euprolactinemic women with hypothalamic amenorrhea, previously unsuccessfully submitted to clomiphene citrate therapy, were treated with bromocriptine. PRL secretion was studied in basal conditions and under dynamic tests: TRH and chlorpromazine. Serum FSH, LH and 17-beta-estradiol were determined before and during the treatment. Both patients conceived, and one delivered a healthy baby at term. Bromocriptine appears to be an effective drug for treating women with hypothalamic amenorrhea, particularly those unresponsive to clomiphene.


Plasma immunoreactive thyrotropin releasing hormone (TRH) values in normal newborns. The study reported here extends investigation on the pituitary thyroid axis in newborn infants, including the assay of plasma immunoreactive TSH levels at different intervals after delivery. Blood samples were collected at birth and after 30, 60, 120 minutes, 6, 24 and 48 hours. Plasma TRH levels were also estimated in normal adult subjects and pregnant women. No significant difference was observed with regard to sex, pregnancy or age, except for a marked increase in newborn infants after delivery. Plasma TRH values, already moderately high at birth (mean 46 pg/ml, range 34-57) reached rapidly a peak of 78 pg/ml (range 60-93) 30 minutes after delivery, decreased rapidly between 30 minutes and 2 hours post-partum, then fell gradually to normal range at 24 hours. A comparison of plasma TRH and TSH levels measured simultaneously suggests that the acute TSH surge at delivery is mediated by TRH secretion.


The effect of TRH on the release of TSH, PRL and GH in man under basal conditions and following methysergide. Pretreatment with methysergide, a blocker of serotoninergic receptors, significantly reduced the TSH response to TRH in six male volunteers. The TRH-induced PRL release was also slightly decreased, while plasma GH levels were not significantly modified by TRH either during the control period or after methysergide. These results indicate that alterations of central nervous system monoaminergic pathways may modify the hypophysiotropic effects of TRH.


Unchanged thyrotropin and prolactin responses to thyrotropin releasing hormone after indomethacin treatment. Experimental effects of prostaglandin synthetase inhibitors have been considered in the literature as a clue to the possible interactions of prostaglandins with the hypothalamic releasing hormones at the pituitary level. Some results of the administration to man of these drugs are apparently in contrast with the in vivo and in vitro animal data. The present investigation deals with the comparison between the thyrotropin releasing hormone (TRH) effect on prolactin and thyrotropin when the hormone was administered intravenously at doses of 50, 100 and 200 microgram respectively to three groups of six men (aged 22 to 30 years), before and on the sixth day of indomethacin administration (50 mg orally at 6-hour intervals). No significant change in the releasing hormone effect was observed either in the case of prolactin, where TRH caused a consistently similar release of the hormone at every dose employed, or in the case of thyrotropin, where a dose-dependent releasing effect was obtained before and after indomethacin treatment.


The effect of prednisone on serum thyrotropin, thyroxine and triiodothyronine concentrations in hypothyroid patients. The effect of exogenous prednisone on serum thyrotropin (TSH), thyroxine (T4), and triiodothyronine (T3) concentrations was investigated in four patients with non-functioning thyroid glands receiving T4 replacement therapy. Orally administered prednisone, in a dose of 20 mg each day for nine days, resulted in a significant decrease in mean serum TSH levels (p less than 0.01) without significant changes in levels of serum T4, T3, and thyroxine binding globulin (TBG). These findings suggest an inhibitory action of relatively low pharmacologic doses of prednisone on TSH release without changes in circulating thyroid hormone concentrations or inhibition of the peripheral conversion of T4 to T3.


Health care cost containment in West Germany. The West German health care system has experienced a serious escalation of expenditures since about 1970. For a variety of reasons, many features of the national health insurance program that might have provided restraints on costs were gradually eliminated. Most notably, some restrictions on the supply of physicians and hospital facilities were ended, as was an earlier system of fixed budgeting for ambulatory care services. In addition, legislative and judicial decisions have continually expanded both the benefits and the standard of care that must be provided to publicly insured patients. In 1977, the government passed a Health Care Containment Act which ordered several measures to curb costs: prospectively negotiated ceilings on expenditures for physicians' services, dentists' services and prescription drugs; strengthening of utilization review; composition of a unified fee schedule; small increases in cost sharing and limitations on insurance benefits; and some changes in financing. The reform efforts seem to have had a significant effect, though it is still too early to tell exactly what caused the declining growth rate of health expenditures, and whether the new trend will persist.


Measles in monkeys: an epidemiological study. This study describes aspects of measles in non-human primates. Monkeys infected before importation are shown to produce non-immune offspring in captivity in England. The high antibody titres found in most recently imported monkeys decline slowly during captivity in England. While measles is often fatal to monkeys, we have described an outbreak in which a number of symptomless infections occurred. Histological examination of fatal cases produced evidence of infection in the wall of the urinary bladder in one monkey. The close similarity between measles in humans and monkeys has been confirmed. It is considered that the study of infection among the latter may have significance for the former.


Growth agglutination and growth inhibition tests in the diagnosis of brucellosis. Growth agglutination and growth inhibition tests were established for the diagnosis of Brucella abortus infection. The former involves the agglutination of living organisms while the latter is a bactericidal test. Using mouse, guinea-pig, rabbit and bovine serum it was shown that the growth agglutination test is approximately ten times, and the growth inhibition test one hundred times, more sensitive than the conventional tube agglutination test. It is suggested that these techniques may be of assistance in diagnosing bovine brucellosis in situations in which the tube agglutination test results are suspected of being falsely negative.


Studies on the Fc gamma receptor of the murine macrophage-like cell line P388D1. II. Binding of human IgG subclass proteins and their proteolytic fragments. Binding studies of human IgG proteins to murine P388D1 cells indicated that they bind to an apparently homogeneous Fc receptor population. The association constant was 0.89 x 10(6)M-1 at 22 degrees C and was comparable to the binding affinities of homologous murine IgG2a and IgG2b. The number of receptor sites was found to be approximately 6 x 10(5)/cell. Fc gamma 1 and Fc gamma 3 fragments bound with an affinity comparable to that of the parent proteins. The P388D1 receptors could discriminate between the human IgG subclasses; the relative cytophilic activity was IgG3 greater than IgG1 greater than IgG4 and IgG2 was devoid of binding activity. Fragments corresponding to the C gamma 2 and C gamma 3 domains of human IgG1 were both unable to bind to the P388D1 receptors either alone or in equimolar combination. This suggests that the cytophilic site may be formed cooperatively by interaction between the two domains. The integrity of the hinge region appeared to be essential for full expression of cytophilic activity since reduction of the hinge-region disulfides in both human IgG1 and its Fc fragment markedly decreased their binding affinity. In addition, a mutant IgG1 molecule lacking the hinge region was significantly less cytophilic than its normal counterpart.


Two new recombinant H-2 haplotypes, one of which juxtaposes Kb and Ik alleles. Two new recombinant H-2 haplotypes have been detected and established as congenic resistant lines on the C57BL/10 background. On the basis of serologic testing and immunoprecipitation analyses, the sublocus composition of the first recombinant haplotype, H-2bq1 (B10.MBR) has been shown to be KbIkDq, and that of the second recombinant, H-2sq3 (B10.SQR) to be KsIsSsDq. The occurrence of the Kb I-Ak juxtaposition after a recombination between H-2b and H-2m contrasts with the almost uniform failure to observe H-2b/H-2k recombinants in previous studies. This finding and the occurrence of a second recombination event involving the same chromosome soon after the first in our studies may imply that recombination within H-2 is not generally a random event. The B10.MBR line has proved useful in the production of specific anti-H-2Kb and anti-I-Ab antisera previously quite difficult to obtain contamination by anti-Ia or anti-H-2K antibodies, respectively.


T lymphocyte-mediated suppression of myeloma function in vitro. I. Suppression by allogeneically activated T lymphocytes. Alloreactive cells generated by in vitro stimulation of C57BL/6 (H-2b) spleen lymphocytes with irradiated MOPC 315 or MOPC 104E(H-2d) cells were shown to lyse 51Cr-labeled myeloma targets at high effector:target ratios under conditions of inefficient cell contact, the alloreactive cells cause variable and frequently minimal lysis of myeloma targets but markedly suppress antibody secretion even by viable myeloma cells. The suppressor cells are radioresistant T cells lacking I-J subregion-encoded surface determinants; their precursors are insensitive to cyclophosphamide; suppression is H-2 specific and not mediated by secreted factors; and the suppression is blocked by Cytochalasin B, a known inhibitor of T cell-mediated cytolysis. These properties are typical of cytolytic T lymphocytes (CTL) and not of defined suppressor T cells, suggesting that inhibition of myeloma function probably represents a pre-lytic effect of the alloreactive CTL, although a CTL-like suppressor cell effect cannot be definitively excluded. These results are discussed with reference to the possible relationships between suppressor and cytolytic T lymphocytes.


In vitro adherence of lymphocytes to unfixed and fixed high endothelial cells of lymph nodes. Rat thoracic duct lymphocytes (TDL) bind selectively to venules lined by high endothelial cells (HEV) when overlaid onto glutaraldehyde-fixed frozen sections of lymph nodes. This report describes the characteristics of TDL binding to HEV in unfixed frozen sections and compares this reactivity with that observed after fixing sections with different reagents. We found that TDL bound to unfixed HEV and that the pattern of adherence to such sections was identical to that observed when using glutaraldehyde-fixed tissue. Fixation of the sections with glutaraldehyde, however, enhanced the binding reaction. This effect was also observed when sections were treated with the diimidoester, dimethylsuberimidate (DMS) but not when methanol or formaldehyde was used. Since glutaraldehyde and DMS are each bifunctional cross-linking reagents, the results suggest that in vitro HEV adherence was facilitated under conditions in which the endothelial binding sites were present in an aggregated form.


Purification and characterization of a single chain precursor C3-protein (pro-C3) from normal human plasma. Fresh human plasma was treated with proteinase inhibitors and passed through an immunoadsorbent column of Sepharose anti-C3 globulin. The insolubilized C3 was eluted with 5 M guanidine and, after extensive dialysis, was reduced with 0.2 M 2-mercaptoethanol and electrophoresed on SDS-polyacrylamide gels. The bulk of the eluted C3 dissociated into two major protein bands, m.w. 125,000 and 75,000 corresponding to the alpha- and beta-chains of C3. In addition, a nonreducible single polypeptide chain (SPC) with a m.w. value of 197,000 +/- 2,000 similar to the apparent m.w. of unreduced C3 was consistently present. SPC has been purified by elution from SDS (SPC) and found to remain a single polypeptide chain upon re-electrophoresis on SDS gels in the presence of 0.2 M 2-mercaptoethanol. The purified SPC reacted with antisera to denatured C3, C3alpha, and C3beta chains. Additionally, antisera to SPC, also reacted with denatured C3, C3alpha-, and C3beta-chains, revealed a reaction identity between SPC and C3, and detected partial identity between SPC and C3alpha- as well as C3beta-chains. This suggested that SPC and C3 are antigenically related. The amino acid compositions and tryptic peptide maps of SPC and C3 were similar. Based on these findings, it is suggested that SPC must represent a single-chain precursor C3 (pro-C3) in plasma that escaped post-synthetic proteolytic cleavage into a two-subunit chain C3 molecule.


Pretreatment of plastic Petri dishes with fetal calf serum. A simple method for macrophage isolation. We have developed a simple method which can recover the highly purified macrophages or monocytes in suspension from mouse peritoneal exudate cells and human perpheral blood mononuclear cells. Plastic Petri dishes coated overnight with heat-inactivated fetal calf serum (FCS) selectively bind macrophages and monocytes. The adherent macrophages and monocytes are easily removed by incubation in phosphate-buffered saline containing 0.2% ethylenediamine tetraacetate (EDTA) and 5% FCS, and recovered as a cell suspension with greater than 95% purity. A small number of isolated cells can restore the mitogenic response to phytohemagglutinin (PHA-P) of macrophages-depleted lymphocytes and can lyse 51Cr-labeled target cells in an antibody-dependent cell-mediated cytotoxicity system. Thus, the method should be valuable for studies of various functions of macrophages and monocytes from different immune tissues of man and animals.


Complement fixation for study of placental-type alkaline phosphatase. Preparations of human placental alkaline phosphatase differing in specific enzyme activities were compared by microcomplement fixation assays using monospecific antisera. While both specific enzyme activity and complement fixation units increased 15,000-fold upon purification, the ratio between these units remained constant. Separation of an alkaline phosphatase preparation into 'A' and 'B' forms by ampholine isoelectric focusing indicated that these forms also possessed the same ratio of immunoreactive enzyme protein to enzyme activity. The correspondence of complement fixation units with specific enzyme activity indicates that complement fixation with monospecific antisera can be used to analyze structural differences among alkaline phosphatase isoenzymes.


The quantitation of alternative pathway complement function by timed lysis assay. A simple timed lysis assay is described for quantifying haemolytic complement activity in human serum. Classical pathway complement function was determined by measuring the time taken to lyse 50% of a standard suspension of antibody-coated sheep erythrocytes; the time required for 50% lysis of a standard rabbit erythrocyte suspension was similarly used to evaluate alternative pathway function. Because target erythrocytes prepared on different days gave slightly different 50% lysis times, it was necessary first to construct a series of calibration curves for converting 50% lysis times into CH50 U/ml. For this purpose, a range of dilutions of the standard human serum, of known haemolytic activity, was tested against erythrocytes prepared on 10 separate occasions. The standard serum was subsequently included with each batch of unknown sera and used to select the appropriate calibration curve for direct conversion of the 50% lysis time into CH50 U/ml. Eleven samples of normal human serum were tested by both the timed lysis assay and by the dilution methods of Mayer (1971) (classical) and Platts-Mills and Ishizaka (1974) (alternative pathway). Comparable results were obtained in all cases.


A simple electrophoretic technique for the estimation of complement C3 conversion: specific application to the investigation of anaphylactoid response to I.V. agents. A simple electrophoretic technique employing commercially available agarose films is described for the routine estimation of plasma complement C3 conversion. This technique has particular value in the investigation of anaphylactoid responses in patients following the administration of intravenous hypnotic drugs, plasma substitutes or radio-contrast media.


Differential radiolabelling of lymphocyte membrane alloantigens and immunoglobulins: variation of H2O2 concentration during lactoperoxidase catalyzed cell surface radio-iodination. The effect of H2O2 concentration on lactoperoxidase catalyzed cell surface radio-iodination and subsequent isolation of Murine splenic lymphocyte Ia, H-2K and Lyb-3 surface antigens and membrane immunoglobulins was studied. For most membrane polypeptides analyzed 0.3 mM H2O2 proved to be optimal for the recovery of radiolabelled antigens from detergent lysates of labelled cells by immunoprecipitation. Marked variations among surface antigens and membrane immunoglobulin polypeptide chains were observed for the iodination and recovery of these proteins above and below the optimal peroxide concentration. The results suggest that cell surface radio-iodination conditions should be standardized to the requirements of the particular membrane protein being studied. The differential iodination and recovery of discrete membrane components above and below optimal conditions may prove useful in the analysis of surface membrane protein structure and membrane association.


Fibro-osseous lesions of the cranio-facial bones. A brief review of cranio-facial fibro-osseous lesions is given and 15 cases of fibrous dysplasia and ossifying fibroma are presented with clinical, radiological and histomorphological findings. After histological verification of the fibro-osseous nature of the process, 5 lesions were diagnosed on an radiological basis as fibrous dysplasia exhibiting diffuse (blending) margins, while the diagnosis of ossifying fibroma was given to 10 radiologically sharply-demarcated lesions. Both groups of lesions presented the same histomorphological features, although to a varying degree. Spheroidal calcifications, however, were identified in ossifying fibroma only, but not in all of these. Thus, the differential diagnosis of fibrous dysplasia versus ossifying fibroma rests on a radiological criterion after the histopathologist has verified the fibro-osseous nature of a lesion. The observation times varied from 1 1/2 to 34 years. Two cases of fibrous dysplasia and 3 cases of ossifying fibroma recurred. In one case of fibrous dysplasia an osteosarcoma developed 33 years after irradiation. Pain was a common symptom in the present material.


Assay and properties of 18-hydroxylation of endogenous and exogenous corticosterone in rat adrenals. Evidence for heterogeneity of 18-hydroxylase activity. A mass fragmentographic technique for assay of 18-hydroxylation of labeled (exogenous) and unlabeled (endogenous) corticosterone in adrenal mitochondria and in reconstituted cytochrome P-450 systems has been developed. An extract of an incubation of [14-14C]corticosterone is subjected both to thin-layer radiochromatography and to mass fragmentography (as O-methyloxime-trimethylsilyl ether derivative). In the latter procedure the ions at m/e 605 and 607 (specific for the derivatives of unlabeled and labeled 18-hydroxycorticosterone, respectively), at m/e 591 and 593 (specific for the derivatives of unlabeled labeled aldosterone, respectively) and at m/e 548 and 550 (specific for the derivatives of unlabeled and labeled corticosterone, respectively) were followed through the gas-liquid chromatography. From the ratio between the peaks obtained in the mass fragmentography and from the percentage conversion of [4-14C]corticosterone obtained in the thin-layer radiochromatography, the amount of endogenous and exogenous 18-hydroxycorticosterone and aldosterone could be calculated. The effects of time, enzyme, and substrate concentration of 18-hydroxylation were studied and optimal conditions for assay were determined. Under most conditions, the ratio between labeled and unlabeled 18-hydroxylated products was about constant, indicating that labeled and unlabeled corticosterone were not in equilibrium. It was ascertained that the 18-hydroxycorticosterone and aldosterone formed in the incubations were derived from corticosterone. [4-14C]18-Hydroxydeoxycorticosterone was not converted into aldosterone or 18-hydroxycorticosterone. In vitro studies with different 18-hydroxylase inhibitors (spironolactone, canrenone, and canrenoate-K) and studies with rats pretreated with KCl in drinking fluid suggest that 18-hydroxylation of corticosterone is catalyzed by an enzyme system different from that catalyzing 18-hydroxylation of deoxycorticosterone.


A gene necessary for late sperm function in Drosophila melanogaster. A male-sterile mutant, ms(1)7, of Drosophila melanogaster is defective in post-ejaculatory sperm function. The mutant gene is located at one of at least five male fertility loci in 18F through section 20 of the polytene X-chromosome. It is proposed that the ms(1)7+ gene product modifies a component of the sperm head. A nearby gene may be functionally related to ms(1)7+.


Laboratory studies of self-reinforcement (SR) phenomena. Fifty-three empirical laboratory studies of self-reinforcement (SR) published between 1962 and 1977 are discussed and critiqued. Studies are organized under two major headings--infrahuman and human--and are summarized in relation to the acquisition, motivational properties and comparative effectiveness of SR procedures. Each experiment is rated in terms of the degree to which the procedures reported represent actual SR operations, and in terms of overall experimental validity. The reviewed literature shows that SR behavioral patterns can be readily acquired through modeling or direct external reinforcement; and affected by a variety of training, individual difference, procedural, and task factors; and, once acquired, have sometimes been shown to possess functional reinforcing properties, and to be generally as effective as external reinforcement procedures in maintaining and strengthening conditional or target responses. A variety of methodological and theoretical issues pertaining to SR research are discussed, and the relationships between SR and contemporary radical operant and social learning psychology are discussed.


The impact of psychiatric intervention on patients with uncontrolled seizures. There is much evidence that emotional stess can trigger both neurogenic and hysterical seizures in susceptible patients. We reviewed out experience with 37 patients whose seizures appeared to be precipitated at times by emotional stress and had not been controlled by anticonvulsant medication alone. Approximately 70 per cent of patients demonstrated substantial improvement in seizure control after psychiatric treatment and maintained this improvement during follow-up. The findings of this study suggest that patient characteristics associated with better prognosis include normal intelligence, partial (as opposed to generalized) neurogenic seizures, a diagnosis of hysterical seizures, a less severely abnormal EEG, and being hypnotizable. After psychiatric treatment, 32 per cent of patients had their anticonvulsant medication reduced and another 16 per cent had it discontinued.


Single case study. Complementary effects of phenelzine and psychotherapy in long term treatment of depression. A case report is described wherein the monoamine oxidase inhibitor phenelzine was administered for 10 months at different doses. Drug treatment in the initial part of the study was double blind. Weekly psychotherapy was instituted at the point of symptomatic recovery. At a reduced dose,  in month 3, the patient experienced a relapse in depression. While platelet monoamine oxidase inhibition was greater than 80 per cent the patient was well, but at the point of relapse, inhibition was 14 per cent. Clinical ratings at relapse (Beck and SCL-90 scales) revealed greater readiness by the patient to report psychological discomfort compared with the original interview. The combined effects of psychotherapy and pharmacotherapy were felt to be responsible for this change. However, psychotherapy in this form and duration did not prevent relapse, which depended upon maintaining an adequate dose of phenelzine.


Responses of baboon cerebral and extracerebral arteries to prostacyclin and prostaglandin endoperoxide in vitro and in vivo. The responses of baboon cerebral and extracerebral arteries to prostaglandin endoperoxide (PGH2) and prostacyclin (PGI2) were investigated on isolated arteries and in vivo by serial angiography. Both PGH2 and PGI2 could produce dose-dependent contraction or relaxation of isolated arteries. PGH2 induced relaxation was indicative of prostacyclin synthetase activity, the enzyme which converts PGH2 to PGI2. In isolated arteries tested one to four hours post mortem only the vertebral artery showed prostacyclin synthetase activity. Thus PGH2 induced contraction of cerebral arteries may be indicative of a physiological function. Vasomotor tone may in part be the result of a balance between PGH2 constriction and PGI2 dilatation. In vivo PGI2 infusion caused pronounced and prolonged dilatation of cerebral arteries, which lasted longer than the cardiovascular changes. As PGI2 is the most potent cerebral vasodilator drug tested, it may be of clinical use in the treatment of cerebral vasospasm.


Effect of degree and duration of protein energy malnutrition on peripheral nerves in children. Nerve conduction studies were conducted in 67 children to assess the effect of malnutrition on the peripheral nervous system. Significant reduction in nerve conduction velocities was demonstrated in severe protein energy malnutrition and ongoing long-term malnutrition.


Warm fibers innervating palmar and digital skin of the monkey: responses to thermal stimuli. 1. Three hundred fourteen warm fibers innervating the glabrous skin of the monkey's hand were isolated by dissection in the median and ulnar nerves in two species, Macaca mulatta and M. nemestrina. Fiber samples in the two species were functionally similar and uniform in their properties. Their mean conduction velocity of 1.2 m/s (SD 0.5; n = 50) implies that these warm fibers were all unmyelinated. 2. A parametric study of the responses of warm fibers to near-rectangular warming and cooling pulses applied to glabrous skin was completed using 104 fibers. At a steady base-line skin temperature (T-base) of 34 degrees C all these warm fibers responded to warming pulses in the intensity range 0--8 degrees C with a simple, uniform discharge, which reached a peak rate of 1.5--4.0 s after the onset of stimulation; subsequent decay in this discharge rate had a time constant of 5--12 s and was virtually independent of the intensity of the warm pulse. The intensity function was linear for most fibers when the interstimulus interval was 60 s or longer. At a T-base of 29 degrees C, warm fibers were less responsive, but the temporal profile of the response was similar to that at a T-base of 34 degrees C in the intensity range 4--8 degrees C, and the intensity function was again linear. 3. At a T-base of 39 degrees C the intensity function of each warm fiber was complex. Most fibers responded briskly to warming pulses of 2--4 degrees C: the response to more intense warming pulses, particularly when the skin temperature rose above 45 degrees C, was structured and reproducible, but varied greatly among different fibers. With some the discharge evoked was of very high frequency for a few seconds, and then ceased. More than 80% of the sample of warm fibers did not discharge at all in response to warming pulses, which raised the skin temperature to 50 degrees C or above. 4. The responsiveness of warm fibers to warming pulses was dependent on previous stimulation when the interstimulus interval was less than 60 s. This temporal suppression was precisely structured and was examined quantitatively for trains of warming pulses, each lasting 4.0 s and presented every 10 s. The pattern of suppressive interaction was similar in form to that previously reported for cold fibers innervating palmar skin. 5. A quantitative study of the receptive fields of individual warm fibers demonstrated a spatiotemporal response pattern, which is best described in terms of a focal receptor zone less than 1 mm in diameter surrounded by thermally conducting skin. The skin's thermal conductivity is paramount in determining the warm-fiber's receptive-field characteristics. 6. The responses of warm fibers to cooling pulses and to warming ramps are described.


Coding of incremental changes in skin temperature by single warm fibers in the monkey. 1. Experiments were designed to answer the question: how well does a single warm fiber innervating the glabrous skin of the monkey's hand resolve incremental changes in the intensity of near-rectangular warming pulses applied to the fiber's receptive field? 2. In these experiments the measure of the warm fiber's capacity to resolve incremental changes in the intensity of successive warming pulses was termed the discriminable stimulus increment (DSI). The DSI is defined as that incremental difference in the intensity of a pair of warming pulses that could be resolved correctly, with a probability of 0.75, by comparing the fiber's responses to these two stimuli. In the specified conditions of the experiment, DSI = 0.67 sigma delta tau/(dR/dI) where sigma delta tau is the standard deviation of the difference in responses of the fiber to pairs of stimuli, and dr/dI is the fiber's sensitivity to incremental stimulus change. (dr/dI) was experimentally determined as the mean rate of change of the fiber's responses to incremental changes in the intensity of the warming pulse. 3. The DSI, as defined above, assumes that the basis for differentiating the stimuli in each pair was that the larger response in the fiber was in each instance generated by the more intense stimulus. A more general form of the DSI was also developed and used to examine the effects on intensity resolution of different discrimination rules that the brain might use. 4. In the experimental analysis the response measure of each warm fiber was the cumulative impulse count over successively longer segments of the stimulus period. With short integration intervals the DSI was high (i.e., intensity resolution was poor), but typically the DSI fell to a plateau level within 2.0--2.5 s of the onset of the warming stimulus. 5. The DSI was measured on 23 warm fibers in Macaca nemestrina for warming pulses with intensities of 0, 2, 4, 6, and 8 degrees C, at T-base levels of 29, 34 (near normal temperature of palmar skin), and 39 degrees C. For most observations the intensity resolution possible from the responses of single warm fibers, measured over this wide variety of stimulus conditions, was less than is achieved by the human observer trained to differentiate comparable warming pulses applied to the skin of the thenar eminence.


Excitation of primate spinothalamic neurons by cutaneous C-fiber volleys. 1. The responses of spinothalamic tract cells in the lumbosacral spinal cords of anesthetized monkeys were examined following electrical stimulation of the sural nerve or the application of noxious thermal and mechanical stimuli to the skin on the lateral aspect of the foot. 2. The spinothalamic tract neurons were classified as wide dynamic range (WDR), high-threshold (HT), or low-threshold (LT) cells on the basis of their responses to mechanical stimuli. 3. All of the WDR and HT spinothalamic tract cells tested responded to volleys in A- and C-fibers. However, strong C-fiber responses were more common in HT than in WDR cells. 4. The responses atributed to C-fibers were graded with the size of the C-fiber volley. The latencies of the responses attributed to C-fibers indicated that the fastest afferents involved had a mean conduction velocity of 0.9 m/s. The responses remained after anodal blockade of conduction in A-fibers. 5. Temporal summation of the responses of spinothalamic tract cells was demonstrated both to brief trains of stimuli at 33 Hz and to single stimuli repeated at 1- to 2-s intervals. The latter phenomenon is often called "windup." 6. The responses of several spinothalamic tract cells to noxious heat pulses could still be elicited during anodal blockade of conduction in A-fibers. Similarly, it was possible to demonstrate an excitatory action of noxious mechanical stimuli despite interference with conduction in A-fibers by anodal current. 7. The cells investigated were located either in the marginal zone or in the layers of the dorsal horn equivalent to Rexed's laminae IV-VI in the cat. The cells were generally activated antidromically from the caudal part of the ventral posterior lateral nucleus of the thalamus.


Responses of primate spinothalamic neurons to graded and to repeated noxious heat stimuli. 1. The responses of primate spinothalamic tract cells innervating the glabrous skin of the foot to noxious thermal stimuli have been examined. 2. Of the 41 cells studied, 98% responded to noxious thermal stimuli. Heating the cutaneous receptive field with a series of stimuli from 35 to 43, 47, and 50 degrees C produced a graded increase in discharge rate. The responses were characterized by an onset, which occurred after the temperature change had either slowed or stopped, an acceleration in the discharge up to a peak, and then an adaptation to a new base-line level. The time constants of adaptation were faster than those reported for C polymodal nociceptors. 3. No systematic differences were found in the responses to noxious thermal stimuli of cells with wide dynamic range receptive fields and of cells with narrow dynamic range, high-threshold receptive fields. There were also no differences in the responses of cells located in the marginal zone and of cells located in the neck of the dorsal horn. 4. The relationship between peak frequency and final skin temperature with a 30 s stimulus duration can best be described by a power function with an exponent of 2.1. An increase in the stimulus duration to 120 s resulted in an increase in the exponent of the power function to 3.2. 5. Repetition of the series of 30-s heat stimuli resulted in an increase in peak frequency, total impulse count, and background activity. Repetition of stimuli having a duration of 120 s produced an increase in the peak frequency at 43 and 45 degrees C, a smaller increase at 47 degrees C, and a decrease at 50 degrees C. Background activity was increased by the lower temperature stimuli, but was decreased following higher temperature stimuli. 6. In six additional cells, the skin was heated with three consecutive presentations at each temperature level (43, 45, 47, and 50 degrees C) for 30 s. No change was observed in the peak frequencies of the responses to successive stimuli of the same intensity. However, the exponent of the power function relating the average peak frequency for the six cells to changes in skin temperature was 3.9. This exponent was larger than that seen when two series of graded heat stimuli of 120 s duration were used, indicating more sensitization despite the fact the total time of exposure to noxious heat was less. 7. A role for both high-threshold and wide dynamic range spinothalamic cells in transmitting nociceptive information to the diencephalon is postulated.


Reinnervation of glabrous skin in baboons: properties of cutaneous mechanoreceptors subsequent to nerve crush. 1. A total of 791 fibers were isolated from the ulnar nerves of five baboons. Over half of these were obtained from the right ulnar nerves subsequent to reinnervation following a nerve crush; the other fibers were obtained from the undamaged left ulnar nerves. 2. The conduction velocities in the proximal portion of the injured axons dropped below normal, and this reduction persisted until reinnervation appeared nearly complete. 3. The response properties of 65 cutaneous afferent fibers serving reinnervated glabrous skin were compared to 80 fibers from normal skin. 4. Of the afferent fibers reinnervating skin, the proportion judged to have abnormal response properties was not significantly greater than the proportion in normal skin. 5. After reinnervation, cutaneous rapidly adpating fibers displayed tuning curves characteristic of their submodality, while some cutaneous slowly adapting fibers could still be differentiated into type I and type II fibers. However, both types of slowly adapting fibers displayed an increased rate of adaptation and a lowered sensitivity to sustained displacements even at 5 mo following reinnervation. 6. In conclusion, the cutaneous mechanoreceptors in reinnervated glabrous skin regained response properties that allowed them to be assigned to the same submodalities found in normal skin. Submodality was recognizable at an early stage when the receptive field was still immature and when the threshold was elevated. With time, the receptive-field sizes and shapes returned to normal, the thresholds approached normal, and the remaining differences from normal cutaneous afferent fibers became minor.


Birthmarks with serious medical significance: nevocullular nevi, sebaceous nevi, and multiple café au lait spots. Birthmarks of potential medical significance were identified among 4,641 newborn infants: 1.1% had a nevocellular nevus and 0.3% had a sebaceous nevus; 1.8% of black infants, but no white infants, had three or more café au lait spots. The large nevocellular nevus is considered premalignant, but the significance of the small nevocellular nevus has not been established. The sebaceous nevus fades after birth but undergoes hyperplasia at puberty. Multiple café au lait spots occur less commonly in white than in black infants, and are therefore of greater concern as possibly an early sign of neurofibromatosis.


The immature monkey as a model for studies of bronchopulmonary dysplasia. The monkey is a potential model for BPD since there is considerable background information on the normal-developing lung, the prematurely delivered infant is viable, HMD can be produced, the infant is large enough to permit physiologic measurements, and it should be possible to test the effects of positive pressure, oxygen, and pharmacologic agents. Clearly further information is needed on the cellular and subcellular changes occurring during the acute and recovery stages of HMD. The monkey has already proven to be of value in this inquiry. Studies on mechanisms of altered lung repair by various injurious agents are needed, and will require an animal model as well as in vitro systems. Basic understanding of the pathogenesis of bronchopulmonary dysplasia with establishment of the relative importance of the contributing factors should help in our efforts to prevent or minimize chronic lung disease in the newborn infant.


The attachment between tooth and gingival tissues after periodic root planing and soft tissue curettage. Utilizing a nonhuman primate model, a study was carried out to determine the nature of the attachment between the tooth and the gingival tissues following periodic root planing and soft tissue curettage. Under the conditions of this investigation, periodic root planing and soft tissue curettage combined with thrice weekly plaque control resulted in the formation of a long junctional epithelium with no new connective tissue attachment. In eight of the 22 experimental pockets, however, this procedure produced discontinuities or "windows" in the junctional epithelium. The coronal attachment of gingival tissues to the root surface (increased resistance to probing) commonly reported following root planing and soft tissue curettage appears to result from the formation of a long junctional epithelium rather than new connective tissue attachment.


Distribution and excretion in the rat and monkey of [82Br] bromocriptine. Radiolabeled bromocriptine was administered to rats and monkeys and its tissue distribution, rate of excretion and metabolism were determined. In both species, [82Br]bromocriptine is predominantly concentrated in the liver, metabolized and then excreted through bile. Extractable free drug is undetectable in blood after 30 min. After 2 hr, the estimated concentration of free drug in rat brain is 2 x 10(-8) M. Terminal phase half-lives of radiolabel excretion were 2.9 days for the rat and 27.3 days for the monkey. The retained organically bound 82Br is present in a form more polar than bromocriptine.


[Meiosis in Microsporidia: effects on biological cycles]. Synaptinemal complexes have been demonstrated in 7 microsporidian species belonging to 6 different genera (Gurleya, Thelohania, Pleistophora, Tuzetia, Baculea, Glugea). Thus, it can be presumed that a meiosis and consequently a karyogamy occur during their life cycle. Meisis occurs at the beginning of sporogony; therefore, karyogamy, must occur between spore and merogany, i.e. during the poorly known part of the life cycle. In the microsporidian species studied, with uninucleate spores and diplokaryotic merogony (Thelohania for instance), the 2 joined nuclei, each of them containing meiotic chromosomes, not only fail to fuse, but actually separate at the beginning of sporogony; afterwards, each of them undergoes meiosis. Their separation is accompanied by the appearance of an organelle whose structure and function are poorly understood. However, its structure resembles that of the kinetic center. The Nosema species studied do not have synaptinemal complexes; thus, their life cycle is difficult to understand: either karyogamy and meiosis occur during the unobserved part of the life-cycle, or sexual phanomena are absent altogether. In the latter case, the Nosema-type life cycle might be limited to vegetative multiplication which could be explained by the dimorphism theory of Microsporidia. It is shown also in the present study that the life cycle of Microsporidia does not involve haploid organisms which it might be thought to contain by comparing it with the cycles of sporozoa.


Henneguya adiposa Minchew (Myxosporida) in the channel catfish: ultrastructure of the plasmodium wall and sporogenesis. Wall ultrastructure and sporogenesis were studied in plasmodia of Henneguya adiposa Minchew which infects the channel catfish, Ictalurus punctatus (Rafinesque). Plasmodia were located among connective tissue bands of the adipose fin and were always separated from host fibrocytes by collagen fibers. The plasmodium wall consisted of a single unit membrane which was continuous with numerous pinocytic canals extending into the parasite's ectoplasm. The membrane was highly convoluted, producing an irregular parasite surface, and was covered by a fine granular coat of almost uniform thickness. Early sporogenic stages were located in a zone of cytoplasm rich in mitochondria, just interior to the zone of pinocytic canals. Later sporogenic stages, including mature spores, were concentrated in the center of the plasmodia. Sporogenesis began with the envelopment of one generative cell, the sporont, by a 2nd, nondividing, cell--the enveloping cell. The sporont and its progeny proceeded through a series of divisions until 10 cells were present within the enveloping cell. Once divisions were completed, the 10 cells became arranged into 2 indentical spore-producing units, each consisting of one binucleate sporoplasm and 2 capsulogenic cells, all surrounded by 2 valvogenic cells. Later stages of spore development indicated that capsulogenesis, valvogenesis and sporoplasm maturation occurred concimitantly.


Formation and fate of extranuclear chromatin bodies in Tetrahymena. The fate of extranuclear chromatin bodies (ECBs) formed by exclusion of macronuclear material at the time of karyokinesis was followed quantitatively in Tetrahymena pyriformis strain GL-I. In a logarithmic growth phase culture, 51% of the dividing cells produced one (43%) or more (8%) ECBs. Most of these gradually disappear before the next cell division, but approximately 13% are retained and carried into subsequent cell cycles. The random distribution of ECBs into anterior or posterior daughter cells, their staining and morphological characteristics, and their rapid loss in cells in starvation medium, all indicate that ECBs play no more of a role in cellular activity than that of an internally produced food vacuole.


Selective inhibition of flagellar activity in Chlamydomonas by nickel. Flagellar activity in the biflagellate chlorophyte Chlamydomonas reinhardtii is selectively inhibited by Ni2+ or by treatment with Ca2+-chelating agents. Inhibitions of swimming speed, geotaxis, phototaxis, and pattern swimming result from qualitative and quantitative losses in the activity of individual flagella and in the coordination of activity between the 2 flagella of each cell. Addition of Ca2+ (a) prevents inhibition and (b) restores normal flagellar activity in inhibited cells. Mg2+ is partially effective in reversal of inhibition. Other ions do not cause similar inhibition or reversal of nickel inhibition. The characteristics of inhibition and reversal suggest that the primary target for nickel is a component of the flagellar apparatus, and that this component uses Ca2+ to perform its normal function in the regulation of flagellar activity. A 2nd target for nickel is a Ca-requiring process specific to phototaxis (and not involved in the photophobic response).


[Results of treatment of breast cancer with a betatron (author's transl)]. The authors analyze the results of post-operative therapy with accelerated electrons from a betatron, more especially with pendular bi-energetic electrontherapy, applied to 459 patients with various stages of breast cancer. They stress that as well as the favourable survival rate, sequelae were rare, and there were very few local recurrences.


Distribution of gold in blood following administration of auranofin (SK&F D-39162). After oral administration of auranofin (SK&F D-39162), a new antiarthritic gold compound, to rats, dogs and humans, a major portion (approximately 50%) of the blood gold content was found to be associated with cellular components. This property is in marked contrast to that reported for gold sodium thiomalate and represents another of the many physical, chemical and pharmacological differences between these therapeutic gold compounds. Although the clinical significance of this property is not presently known, it is recommended that pharmacokinetic studies involving auranofin include the assessment of both blood and serum gold levels.


Inhibition of pokeweed mitogen-induced immunoglobulin production in humans by gold compounds. The effect of gold compounds on an in vitro model of antibody production was investigated. Peripheral blood mononuclear cells from healthy adult individuals were stimulated with pokeweed mitogen (PWM) and generation of immunoglobulin secreting cells (ISC) was assayed. Addition of gold sodium thiomalate 10 microgram/ml markedly inhibited mitogen-induced ISC production. The suppression of Ig production could not be explained by nonspecific lymphocyte toxicity or interference with the assay system. Inhibition of PWM responsiveness was mediated directly by the gold moiety and was not a function of the thiomalate. Furthermore, it was shown that maximum suppression of ISC generation occurred only when gold was added to cultures within the first 24 hours of incubation. This suggested that gold compounds inhibit an early step in B cell activation and differentiation rather than merely blocking Ig secretion by mature plasma cells.


Quantitation of gold lymphocyte binding during chrysotherapy. Carbon rod atomic absorption analysis (CRA) was applied to the quantiation of gold lymphocyte content (GLC) during chrysotherapy. Sensitivity and accuracy of CRA compared favorably with 195Au isotopic scintillation counting, circumventing the limitations and hazards of the latter in clinical applications. Picogram gold quantification of limited sample volume, less than 10 ml blood, e.g., 10(4)-10(5) lymphocytes (5 microliters) containing 5-10 pg was achieved. GLC after IM administration increased significantly in 60% of patients; for the remainder, GLC was observed to be independent of plasma gold content or cumulative dosage. GLC during auranofin administration (6 mg/day p.o.) approached values for IM gold despite significantly lower plasma levels. Unique sensitivity of CRA enables analysis of GLC under therapeutic conditions that could elucidate whether gold alters functional determinants.


Sequential observations of in vitro responses of lymphocytes to phytohemagglutinin in patients receiving gold therapy for rheumatoid arthritis. Forty-seven patients receiving gold therapy for rheumatoid arthritis were observed sequentially at 6-monthly intervals. When the disease was in remission, and remained so, the in vitro responses of lymphocytes to phylohemagglutinin stimulation tended to be normal. Improvement in the disease were associated with improvement in lymphocyte response and deterioration associated with depression of response. Our observations suggest that these changes are a reflection of disease activity and their relationship to gold therapy is indirect.


The amyloidosis of juvenile rheumatoid arthritis - comparative studies in Polish and American children II. Serum immunoglobulins, kappa and lambda light chain and C3 levels. The concentration of serum IgG, IgM, IgA, IgD, IgE and complement C3 as well as Kappa and Lambda free and bound light chains were determined in sera from 21 amyloidotic Polish children with juvenile rheumatoid arthritis (JRA), Polish and American children with JRA, and control children from a Polish population. Comparable concentrations of IgG, IgM, IgA, IgE and C3 were found in amyloidotic children when compared to Polish JRA children with active polyarticular disease. However, the amyloidotic Polish children with JRA had significantly higher levels of IgG, IgM, IgA, IgE and complement C3 fraction when compared to American JRA children. The levels of IgA and IgE were significantly elevated in amyloidotic and Polish JRA populations when compared to American JRA patients. Free immunoglobulin light chains were not found in the sera of any of the children tested in this study. However, in the patients with amyloidosis, significantly diminished levels of bound Kappa and Lambda light chains were found.


Gold in the treatment of rheumatoid arthritis -- why, when and how? The use of gold salts is reviewed and discussed in terms of rationale (Why), indications and side-effects compared to other disease suppressives (when), and different types of gold and their modes of administration (How).


Action of gold salts on the inflammatory response and inflammatory cell function. Information relating to the effect and mechanism of action of gold salts on acute and chronic inflammatory processes has been reviewed. Gold salts are effective in suppressing various aspects of the acute inflammatory process although large gaps exist in the knowledge of the mechanisms involved. Available evidence indicates that gold salts affect many biochemical pathways and that interaction with enzymes plays a major role in their pharmacology. Because they are more difficult to investigate, the action of gold salts on various important aspects of chronic inflammation have not yet received much attention.