Coordinate secretion of acid hydrolases in rat bile. Three lysosomal glycosidases, beta-glucuronidase (EC 3.2.1.31), beta-galactosidase (EC 3.2.1.23), and N-acetyl-beta-glucosaminidase (EC 3.2.1.30) have been investigated in bile that was freshly collected from rats through a complete bile fistula. Assay conditions have been established on the basis of appropriate kinetic studies. The biliary excretion patterns for these enzymes were found to vary considerably from rat to rat during the 24-h collection period. In a given animal, however, the three hydrolases were excreted in parallel and showed a gradual increase in activity with time, most marked after 10- 12 h of collection. 24-h biliary outputs of the three hydrolases averaged congruent with3% of their respective contents in total liver, and bile diversion had no effect on hepatic glycosidase activity or total protein content. Other enzymes known to be associated primarily with mitochondria, endoplasmic reticulum, and cell sap were also detected in bile, generally in smaller amounts. The biliary excretion of the plasma membrane markers, alkaline phosphodiesterase I and 5'-nucleotidase, however, was comparable to that of the lysosomal hydrolases. Biliary excretion of total protein was relatively constant and corresponded to 3.0% of the total hepatic protein content per day, whereas biliary bile acid secretion decreased during the first 12 h and then remained constant. Exocytic bulk discharge of hepatocyte lysosomes is proposed as the most likely mechanism for the biliary excretion of lysosomal enzymes. These results call attention to the possible pathophysiologic significance of biliary excretion of hepatic lysosomal contents as a means of residue disposal.


Inferior temporal lesions do not impair discrimination of rotated patterns in monkeys. Ablation of inferior temporal cortex in the rhesus monkey produces a visual discrimination learning deficit. The severity of this deficit has often been found to be a function of task difficulty. This report concerns a type of visual discrimination problem that, although difficult, is not sensitive to inferior temporal lesions. Monkeys with anterior, posterior, and complete inferior temporal lesions were repeatedly unimpaired or only slightly impaired in learning to discriminate a pattern from the same pattern rotated 90 degrees or 180 degrees; yet they were very severely impaired in learning equally or more difficult discriminations of two different patterns. This demonstration that discrimination of orientation of patterns is relatively spared after inferior temporal lesions helps specify the pattern-recognition processes that require inferior temporal cortex.


Contrasting effects of lateral striate and superior colliculus lesions on visual discrimination performance in rhesus monkeys. Rhesus monkeys with lesions of lateral striate cortex, monkeys with superior colliculus lesions, and unoperated monkeys were tested for retention of a preoperatively acquired pattern discrimination. The three groups of monkeys were then tested in two-choice, color, color discrimination tests, one involving varying degrees of stimulus-response (S-R) separation and the other, administered several months later, involving various directions of S-R separations. The monkeys were also tested in a series of two-choice pattern discriminations, following each of which they were tested for relearning when the patterns were masked with bars or circles. The monkeys with lateral striate lesions were moderately retarded in retention of the pattern discrimination, whereas those with superior colliculus lesions were not. The monkeys with colliculus lesions, but not those with lateral striate lesions, were impaired in both S-R separation tests, which demonstrates that their deficit was not transient or solely due to a difficulty in shifting the gaze in one direction. The lateral striate monkeys, unlike those with colliculus lesions, were dificient in relearning discriminations between masked patterns. These findings suggest that superior colliculus and striate cortex may be involved in two different aspects of attention: respectively, shifting attention (and orientation) from one spatial locus to another and maintaining attention on fixated stimuli. Alternative interpretations of the effects of the lesions, based on their retinotopic loci, are discussed.


Roles of gonadotropins and releasing hormones in hypothalamic control of lordotic behavior in ovariectomized, estrogen-primed rats. Intrahypothalamic effects of gonadotropins (luteinizing hormone and follicle-stimulating hormone), thyrotropin-releasing hormone (TRH), and luteinizing-hormone-releasing hormone (LRH) on lordotic behavior were evaluated in ovariectomized (OVX) rats maintained at different receptivity levels. Under conditions of low receptivity in which LRH has been shown to enchance mating behavior, medial preoptic area (MPOA) infusions of luteinizing hormone (LH) caused significant depressions in the lordotic response, whereas LH infusions into the arcuate ventromedial area (ARC-VM) had no sigificant effect. Follicle-stimulating hormone (FSH) infusions into either area did not alter the behavioral response. In the second experiment, in which OVX rats were primed with higher doses of estrone to maintain high ptrinfusion receptivity, MPOA or ARC-VM infusions of either LH or TRH were shown to depress lordotic behavior significantly, whereas neither LRH nor FSH was observed to inhibit the behavioral response. A third experiment evaluated the effects of LH, FSH, and TRH on LRH-facilitated mating behavior. Infusions of LRH into either the MPOA or the ARC-VM significantly enhanced mating behavior, whereas the addition of either TRH or LH to the LRH infusates abolished this response. The addition of FSH to LRH infusates neither enhanced nor depressed the behavioral response to MPOA or ARC-VM infusions of LRH. The antagonistic effects of LH and TRH on LRH-facilitated mating behavior were correlated with previous observations of antagonistic effects on hypothalamic unit activity and monoamine metabolism. The antagonistic interrelation between LRH and LH may represent a mechanism for the activation and coordination of sexual receptivity with ovulation.


Color categories in macaques.

Color categories in macaques. This experiment investigated whether macaque monkeys partition the photic spectrum into the same four basic hue categories that humans do, i.e., blue, green, yellow, and red. Monkeys were trained to respond in the presence of one chromatic stimulus and were tested, in extinction, for generalization to others. In extinction, the monkeys responded at similar and high levels to stimuli that fell in the same basic human hue category as the training stimulus and at similar and much lower levels to stimuli that fell in a different human hue category from the training stimulus. It was concluded that macaques and humans categorize the spectrum in a similar fashion.


Evaluation of laryngeal cartilages by computed tomography. Computed tomography of the larynx was performed in 21 patients and 3 anatomic specimens for evaluation of laryngeal cancer. Special attention was directed to the appearance of the laryngeal cartilages in both the normal and abnormal examinations in order to establish criteria for cancerous invasion. The problem presented by the normal nonuniformity of density of the cartilages is discussed in detail.


Patterns of brain perfusion: dynamic computed tomography using intravenous contrast enhancement. A representation of cerebral perfusion is obtained using cranial computed tomography following the rapid bolus injection of iodinated intravenous contrast medium. This technique permits the definition of four gross patterns of perfusion (normal, circulatory arrest, hypoperfusion, and hyperperfusion). This classification is affirmed in an experimental model of cerebral infarction in the baboon. The limitations of dynamic imaging using computed tomography are reviewed.


Dynamic computed tomography study of the brain. The dynamic computed tomography (CT) study of the brain consists of the rapid intravenous injection of 49 ml of radiographic contrast material in 7 sec followed by serial 5 sec CT scans with interscan times of only 1 sec. The data from these scans can be reprocessed to create 12 segmented images in 35 sec. When small samples of four to six pixels of cortex are examined by the cursor, sharp rises of 20 to 25 CT units (500 scale) are seen on the time--density curves. Samples of white matter are usually no more than 2 CT units. When larger cursor samples of 220 to 255 pixels including cortex and white matter are examined, the time--density curves represent a combination of these two patterns. Comparison between symmetrical areas in the two hemispheres generally shows parallel curves in controls. The studies provide high resolution cerebral perfusion images. In ischemia secondary to ipsilateral carotid stenosis, there is depression of the up-slope and a depressed and late peak. Infarctions show a flat perfusion curve. Several patterns in brain tumors are illustrated. The study, simple to perform, adds significant perfusion information to the previously static CT examination of the brain.


Sensitization to nonvenom contaminants in a venom preparation. An individual is described who appeared to be sensitive to nonvenom contaminants in a venom preparation. His IgE antibodies, measured by the immediate direct skin test and the radioallergosorbent test (RAST), reacted with a yellow jacket venom preparation obtained by "washing" of venom sacs. With yellow jacket venom obtained by electriral stimulation, there was a skin test reaction of equivocal significance and no serum antibodies were detected by the RAST. Moderate reactions were also found with yellow jacket body extracts. In contrast, sera obtained from patients with yellow jacket sting anaphylaxis showed strong reactions with the electrically stimulated venom preparation and only a few reacted with the body extract. In additional studies, the patient's serum reacted with yellow jacket extracts devoid of venom and a variety of hornet and wasp extracts. Analyses of the two yellow jacket venoms by gel diffusion using rabbit antisera showed the presence of body proteins in the venom obtained by venom sac "washing." Subsequent history revealed the presence of insect nests in the roof of the patient's bedroom, perhaps the source of inhalant exposure and sensitivity. This case history demonstrates the need for venom extracts that do not contain potentially sensitizing extraneous material.


Irritable bowel syndrome and dietary fiber. The effects of an increase in dietary fiber include: (a) More frequent stools, (b) more voluminous stools, (c) an alteration in the fecal flora, (d) an increase in fecal sterol excretion, and, it appears likely, (e) a reduction in intraluminal pressures in the sigmoid colon. Epidemiologic data comparing global differences in prevalence of certain diseases with the fiber content of diets suggest that there may be a relationship between the two. With a certain amount of post-ad hoc reasoning, it can be shown that some of the known effects of fiber could account for differences in disease prevalence between populations. The prevalence of irritable bowel syndrome is so high that one is forced to concede the very real possibility that the environment, including the diet, may be responsible for symptoms that might not otherwise exist. It remains to be seen whether a marked increase in dietary fiber will prevent the symptoms of irritable bowel syndrome. It seems fairly certain that, given the preoccupation most of these individuals have with their bowel movements, the large bulky stools resulting from a high-fiber diet satisfy a very basic emotional need to "have a good BM" (meaning large), but whether this leads to better health remains to be proved.


[External fetal electrocardiography, the maternal effort test and the oxytocin challenge in cases of fetomaternal rhesus isoimmunisation (author's transl)]. The authors present seven pregnant patients with Rh immunisation. Four of them were of mild degree and three were severely inmunised. All of them had an exercise tolerance test as well as an oxytocin test to evaluate the fetal response to maternal exercise and uterine contractility. The classification utilized for evaluating the fetal response to exercise and its clinical implications are presented. It is concluded that the exercise test is of diagnostic and prognostic value regarding chronic distress during pregnancy and the oxytocin test may help to decide wether the fetus will tolerate the uterine contractions of parturition, and if surgical intervention might be contemplated if during a monitorized labor severe fetal heart changes occur.


Sensitivity in horseradish peroxidase neurohistochemistry: a comparative and quantitative study of nine methods. Nine currently available methods for HRP neurohistochemistry have been compared with each other on matching tissue sections from four rats and four rhesus monkeys. The nine methods investigated in this report are the diaminobenzidine (DAB) procedures of LaVail JH and LaVail MM (J Comp Neurol 157:303, 1974), of Adams JC (Neuroscience 2:141, 1977) and of Streit P and Reubi JC (Brain Res 126:530, 1977); the benzidine dihydrochloride (BDHC) procedures of Mesulam M-M (J Histochem Cytochem 24:1273, 1976) and of De Olmos J and Heimer L (Neurosci Lett 6:107, 1977); the o-dianisidine (O-D) procedure of De Olmos J (Exp Brain Res 29:541, 1977); the p-phenylenediamine dihydrochloride and pyrocatechol (PPD-PC) procedure of Hanker JS et al., (Histochem J 9:789, 1977) and the tetramethyl benzidine (TMB) procedures of Mesulam M-M (J Histochem Cytochem 26:106, 1978) and of De Olmos J et al. (J Comp Neurol 181:213, 1978). Quantitative comparisons were based on counts of retrogradely labeled perikarya. The extent of anterograde transport and the size of the injection site were also compared at a more qualitative level. The results indicate that one TMB procedure (Mesulam M-M, J Histochem Cytochem 26:106, 1978) is distinctly superior to each of the other eight procedures in the number of labeled perikarya that it can demonstrate. Furthermore, these differences are statistically significant at better than the 0.05 level of confidence. Differences in sensitivity are most evident when the perikarya contain small quantities of transported HRP. The same TMB method also demonstrates more anterograde transport and a larger injection site than all the other procedures. If less sensitive procedures are employed, afferent or efferent connections that are clearly demonstrated by this TMB procedure are either underestimated or completely overlooked. It is suggested that sensitivity in HRP neurohistochemistry is determined by multiple factors which include the method of fixation, post-fixation storage, the choice of chromogen, the incubation parameters, the type of HRP enzyme that is administered, and the postreaction treatment.


Preservation of alveolar type II pneumocyte lamellar bodies for electron microscopic studies. Simultaneous fixation with glutaraldehyde and osmium tetroxide, followed by an uranyl acetate (UA) treatment before dehydration and embedding (Hirsch and Fedorko 1968) ensures a very good preservation of lamellar bodies (LB's) as well as of the cellular membranes in type II pneumocyte. The uranyl acetate treatment appeared to be the most efficient step of the procedure. The morphological aspect of lamellar bodies after such a preparation was similar to that observed after freeze-etching of lipid retaining methods. Moreover, the Hirsch-Fedorko procedure is very simple and can easily be used for routine ultrastructural and radioautographic studies. On the other hand, it appeared that the uranyl acetate phospholipid "complex" is very sensitive to the pH of chemical solutions used after sectioning. The "complex" is variously dissolved by alkaline solutions, photographic developers or stains. The best preservation of ultrastructure was obtained with neutral or acidic developers and acidic stains.


Alteration of immunoglobulin-bearing lymphoma cells by fixation. Four large cell lymphomas known to be monoclonal B-cell proliferations were studied with immunofluorescent and immunohistochemical methods for the detection of kappa- and lambda-light chains. Frozen sections of lymphoma tissues as well as formalin and B-5-fixed tissues embedded in paraffin were studied. Both immunofluorescent and immunohistochemical methods gave similar results on frozen sections; however, a number of discrepancies were noted between the results obtained on fixed tissues and those obtained on frozen tissues. In an effort to identify a fixative which did not alter immunoglobulin (Ig), mouse lymph nodes were fixed in different fixatives before Ig detection; but all of the fixatives tested destroyed the Ig present on normal cortical B lymphocytes. Immunoglobulin-bearing normal and neoplastic lymphocytes are better detected on frozen sections than on paraffin sections after routine fixation.


Dynamics of natural selection on a lethal fourth chromosome of Drosophila. Twelve-generation study of experimental populations of D. melanogaster. The dynamics of natural selection on a lethal fourth chromosome of Drosophila melanogaster was studied in replicated half-pint bottle and cage populations over 12 generations. Population numbers fluctuated widely in all populations, but there was no association between fluctuation in numbers and change in lethal frequency. In the bottle populations the lethal heterozygote frequency decreased to a low of 0.04 to 0.05 and then increased to 0.18 to 0.30, suggesting that the selection coefficients were not constant. In the cage populations heterozygote frequency decreased to about 0.35 to 0.40. In the cage populations the data suggest that the lethal chromosome is overdominant for both viability and fertility selection.


Simultaneous visualization of two antigens in the same tissue section by combining immunoperoxidase with immunofluorescence techniques. A simple method was developed whereby immunoperoxidase and immunofluorescence techniques were applied in consecutive steps to demonstrate the presence of two antigens in the same tissue section. This method was applied in three model, two antigens were shown: a) each (gastrin and pepsinogen II) inside one of two different cell types (gastrin (G) and antral peptic cells), b) each (kappa or gamma light chains) inside different cells of the same type (plasma cells); also, both (kamma and gamma light chains) inside the same cell (Reed-Sternberg cell), and c) both (pepsinogen I and II) inside the same cell (chief cell of oxyntic glands). The results could be viewed and photographed either simultaneously, when the antigens were in different cells, or sequentially, when the antigens were in the same cells.


[Qualitative and quantitative examinations of the corpus mamillare in the ontogenesis of male Tupaia belangeri and SPF cats]. The cyto-, myelo- and fibrilloarchitectonics of the Corpus mamillare were described in 33 male Tupaia belangeri at an age of 36 to 536 days of ontogenesis and in 37 male SPF-cats at an age of 39 to 584 days of ontogenesis. The fresh volumes of the Nucl. medialis and the Nucl. lateralis corporis mamillaris of Tupaia belangeri were determined and the growth of their fresh volumes described with the 6-parametric growth-function Likewise, the nuclei of SPF-cat were determined and these data were analysed with the logistic growth function The Nucl. medialis corporis mamillaris of Tupaia belangeri has a maximal fresh volume of 2.2 mm3 on the 82nd day of ontogenesis after which it procentually reduces by 16% to 101% of the ideal volume (1.9 mm3) on the 141st day of ontogenesis. The Nucl. lateralis corporis mamillaris reaches its maximum volume of 0.1 mm3 by the 90th day of ontogenesis, procentually reduces by 10% to 101% of the ideal volume (0.09 mm3) on the 169th day of ontogenesis. The half value times are 59 days of ontogenesis for the Nucl. medialis corporismamillaris and 48 days of ontogenesis for the Nucl. lateralis corporis mamillaris. In SPF-cat the Nucl. medialis corporis mamillaris reaches 99% of the ideal volume (11.7 mm3) on the 195th day of ontogenesis and shows a half value time of 86 days of ontogenesis. On the 229th day of ontogenesis the Nucl. lateralis corporis mamillaris reaches 99% of the ideal volume (0.57 mm3y and has a half value time of 83 days of ontogenesis.


"The diencephalon of Ptilocercus lowii (pen-tailed tree-shrew)". The pen-tailed tree-shrew (Ptilocercus lowii) has been regarded by Le Gros Clark (1926), and Martin (1968) to possess more primitive morphological and to a certain extent, neuro-anatomical characteristics than the other tree-shrew species belonging to the sub-family Tupaiinae. Investigations have been carried out on the diencephalon of Ptilocercus lowii which has not been completely done by Le Gros Clark and other workers since 1926, to find out whether the diencephalon has actually undegone any phylogenetic changes that should be more advanced than those of the Insectivora or remain more primitive than that of the Tupaiinae. The diencephalic structure that show the most significant phylogenetic features which differe Ptilocerecus from Tupaia are observed in the anterior, dorsolateral and ventrolateral thalamic groups, and the geniculate bodies. The nucleus anteroventralis is, by comparison, much smaller and poorly differentiated from the nucleus anteromedialis. The nucleus anterodorsalis is also comparably large and does not reach the dorsal surface of the thalamus as it does in the primates. The midline nuclei do not show any significant differences, except the notably larger size of nucleus parataenialis in Ptilocercus. The nucleus mediodoralis is small and indifferentiated although it does show some incipient signs of cellular differentiation. The nucleus centralis lateralis is well developed; its inferior (ventral) part is larger than its superior part which is the better developed of the two parts in the tupaiids and primates. The nucleus centrum medianum appears to be very small and forms only a lateral extension of the nucleus parafascicularis. The ventrolateral thalamic group basically consists of nuclei ventrales anterior, lateralis and posterior without intermediate or transitional zones as found in Tupaia and Primates. The nucleus pretectalis is the largest and the best developed of all the elements of the posterior thalamic nuclear group. The lateral geniculate nucleus is not differentiated into laminae like that in Tupaia and Primates; it is merely a homogeneous structure that appears to be smaller in size that the pregeniculate and medial geniculate nuclei. The medial geniculate nucleus is large in proportional size of the whole thalamus and appears to be well differentiated cellularly into several small parts. No remarkable changes are noted in the epithalamus, subthalamus and hypothalamus. However, the nucleus ventromedialis appears to be the most outstanding structure in the infundibular region of the hypothalamus. The mammillary region is of a simpler construction in Ptilocercus than in Tupaia; it does not protrude from the ventral surface of the hypothalamus. In the light of these findings, the phyletic status of Ptilocercus in the family Tupaiidae is discussed.


Heterogeneity of binding of human IgA subclasses to protein A. The ability of human IgA myeloma immunoglobulins to interact with protein A-containing Staphylococcus aureus was examined. Some IgA1 and IgA2 immunoglobulins bound to S. aureus although others of both subclasses failed to do so. These results were obtained by using both direct binding of radiolabeled immunoglobulins to S. aureus and with inhibition-type assays. Binding was dependent on the Fc fragment of IgA since there was no binding to S. aureus by an F(ab')2 fragment of IgA1. Nonprotein A-containing bacteria did not bind these immunoglobulins and isolated protein A interacted with radiolabeled immunoglobulins. This strongly suggested that protein A was responsible for the observed binding to S. aureus. These data indicate, in contrast to previous reports, that there is no simple relationship between IgA subclass and the capacity to bind to protein A.


A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. We have isolated a subclone of the mouse myeloma cell line P3-X63-Ag8 that does not express immunoglobulin heavy or light chains. This clone X63-Ag8.653 can be used for efficient fusion with antibody-forming cells to obtain hybrid cell lines producing pure monoclonal antibodies. Screening of hybrid cell lines for specificity and immunoglobulin classes was done with a modified enzyme-linked immunosorbent assay.


Activation of complement by Schistosoma mansoni schistosomula: killing of parasites by the alternative pathway and requirement of IgG for classical pathway activation. Living Schistosoma mansoni schistosomula incubated with normal chicken, guinea pig, human, and monkey sera were killed after 4 hr contact at 37 degrees C. The following data indicate that this action is dependent on the activation of the alternative complement pathway (AP): a) the inactivity of RB, RD, and zymosan-treated serum against schistosomula; b) the partial activity of RD restored in FD; c) the full effect of the C4-deficient guinea pig, C2-deficient human, and the agammaglobulinemic human sera; d) the consumption of both the AP and FB after the incubation of NHS with schistosomula; e) the detection of C3d breakdown product during the contact of the C2-deficient human serum with these young parasites. Killing by serum was decreased as the immature schistosomes developed and was completely absent against 4-day-old lung schistosomula (LS). In other experiments, it was demonstrated that schistosomula, in the presence of IgG, were able to initiate complement activation also through the classical pathway (CP). However, the CP does not appear to play a role in the schistosomulicidal activity of complement. The in vivo relevance of these observations is considered.


Inhibition of the in vitro growth of Plasmodium falciparum. I. The effects of immune serum and purified immunoglobulin from owl monkeys. Sera from Aotus sp. monkeys (karyotypes II, III, and IV) which were immune to Plasmodium falciparum have been used to inhibit the in vitro growth of this human malaria parasite. Culture conditions used for the assays allowed 50- to 100-fold increases in the number of A+ erythrocytes infected in a 96-hr period in control cultures. Although normal monkey serum did not support growth as well as normal human serum, mixtures of normal monkey and human serum were found that did. Compared to such controls, as little as 3.5% immune monkey serum was found to cause approximately 56% inhibition in 4 days (2 replicative cycles). Purified globulin from immune monkeys inhibited 40% at 2 mg/ml and 75% at 7 mg/ml after a single replicative cycle. These data suggest that serum antibody is likely to play a major role in providing Aotus monkeys with protective immunity to P. falciparum.


A comparison of different enzyme-antibody conjugates for enzyme-linked immunosorbent assay. Calf intestinal alkaline phosphatase (CIAP)- and horseradish peroxidase (HRP)-linked goat antibody (Ab) conjugates have been prepared by two procedures. One procedure is by glutaraldehyde coupling of the proteins; the other is by intermolecular disulfide bond formation of the appropriately modified proteins. The conjugates, specific for rabbit IgG, were tested for their effectiveness as reagents in enzyme-linked immunosorbent assays for ragweed antigen E specific rabbit antibody. The CIAP-Ab conjugate prepared by glutaraldehyde coupling and the HRP-Ab conjugate made by disulfide bond formation were equally effective reagents for immunoassay, but the HRP-Ab conjugate obtained by glutaraldehyde coupling was definitely less useful than the other two conjugates. The assay procedure utilized an antigen E coupled paper disc as the immunosorbent and it was sensitive in the range of 0.5--100 ng per test. Inhibition of the enzyme-linked immunosorbent assay was used as a sensitive technique for measuring antigenic activity of antigen E or its derivatives.


Purification of antibodies to bacterial antigens by an immunoadsorbent and a method to quantify their reaction with insoluble bacterial targets. A combination of procedures was employed to develop a radioimmunoassay which quantified the binding of antibodies to antigens of either intact Propionibacterium acnes or to antigens of insoluble extracts derived from the bacteria. Reactive antibody populations were purified by use of bacterial immunoadsorbents which were prepared by coupling P. acnes to diethylaminoethyl cellulose. Binding of antibodies was detected with [125I]staphylococcal protein A ([125I]SpA) and optimal conditions for the assay defined by varying the amounts of antibodies, bacterial antigenic targets and [125I]SpA. In antibody excess, 100% of available [125I]SpA was bound by the target-antibody complexes. However, when antibody concentration was limiting, a linear relationship was demonstrated between per cent specific binding of [125I]SpA and antibodies bound to bacterial targets. These results were achieved only with immunoadsorbent-purified antibody populations and not with hyperimmune sera or IgG. The radioimmunoassay detected subtle antigenic differences and similarities between P. acnes, P. acnes extracts and a variety of unrelated microorganisms.


Measurement of antibodies to tubulin by radioimmunoassay. A solid-phase double antibody radioimmunoassay capable of measuring antibody to tubulin, the principal component of microtubules, is described. This assay is simple, combining sensitivity with specificity and also allowing determination of antibody subclasses.


Human secretory component. II. Easy detection of abnormal amounts of combined secretory component in human sera. A simple method, allowing easy detection of abnormally increased sIgA levels is described. It consists in quantitation of combined SC by gel double diffusion, using appropriate anti-SC immune sera. The technical conditions, locating the threshold of sensitivity of precipitation at about 25 microgram/ml, a value higher than that found in normal sera, were established. Comparison with other classical methods (SRID, ELISA and IHA) emphasizes the validity and simplicity of the technique which has shown convenient whenever a large number of sera have to be tested.


Elution of chicken Ig from fixed target cells. A method is described for preparing fixed chicken peripheral blood cells (PBC) for absorption of allo-antibodies and their subsequent elution. Glutaraldehyde fixation was satisfactory, and with 8 M urea elution haemagglutinating activity was preserved.


Heterogeneity and some properties of beta-galactosidase from newborn rat epidermis. Two forms of beta-galactosidase from newborn rat epidermis could be separated by DEAE-cellulose chromatography. Both enzymes showed similar enzymic properties. They had a pH optimum around 3.5--4.5 and the optimal temperature of these enzymes was approximately 60 degrees C. They were not affected by divalent cations, ethylenediaminetetraacetic acid(EDTA) and 2-mercaptoethanol(2-ME), while rho-chloromercuribenzoic acid (PCMB) was a strong inhibitor for each enzyme. These enzymes showed the same Km value (1.25 x 10(-4) M) towards 4-methylumbelliferyl-beta-D-galactoside. However they had different isoelectric points at pH 6.3 and 9.0, respectively. Six different forms of beta-galactosidase activity were found by using isoelectric focusing. When the crude extract was incubated with neuraminidase before electrofocusing, the acidic forms of the enzyme were largely lost and converted to more basic forms without loss of the total activity. This finding suggests the glycoprotein nature of newborn rat epidermal beta-galactosidase.


Synergy between carbenicillin and an aminoglycoside (gentamicin or tobramycin) against Pseudomonas aeruginosa isolated from patients with endocarditis and sensitivity of isolates to normal human serum. Isolates from the blood of 30 patients with endocarditis due to Pseudomonas aeruginosa were tested for synergy between carbenicillin and an aminoglycoside, either gentamicin or tobramycin, by in vitro checkerboard methods in modified (cation-supplemented) Mueller-Hinton broth. Twenty-five of the 30 isolates were affected synergistically. Whether given low (2.5--5 mg/kg) or high (8 mg/kg) doses of aminoglycoside along with 30 g of carbenicillin daily, all of the five patients infected with pseudomonads that were not synergistically affected were refractory to treatment with pseudomonads that were not synergistically affected were refractory to treatment with the carbenicillin-gentamicin combination, whereas the finding of synergy of carbenicillin with gentamicin (or tobramycin) did not assure a medical cure. Tests for synergy between carbenicillin and gentamicin yielded different results in Mueller-Hinton agar than in modified Mueller-Hinton broth. The majority (28) of 30 isolates of endocarditis-producing P. aeruginosa were resistant to the bactericidal effects of 50% pooled normal serum that had been freshly separated. One of the endocarditis-producing strains that was sensitive to 50% serum was resistant to 10% serum. However, sensitivity or resistance to freshly separated, pooled normal human serum did not predict the outcome of antibacterial therapy for pseudomonas endocarditis.


Species-specific resistance to antimocrobial synergism in Streptococcus faecium and Streptococcus faecalis. Combinations of penicillin with various aminoglycosidic aminocyclitols were tested against a collection of clinical isolates of Streptococcus faecium in vitro and were used to treat endocarditis caused by S. faecium in the rabbit model. S. faecium proved more resistant to penicillin than Streptococcus faecalis. Even more striking, however, was the resistance to in vitro synergism by combinations of penicillin and various aminoglycosides. At clinically achievable concentrations, penicillin-gentamicin was the only combination that was synergistic against all strains that were tested. Combinations of penicillin and streptomycin and penicillin and amikacin were synergistic only against those strains that were not highly resistant to streptomycin and kanamycin, respectively. Combinations of penicillin with kanamycin, tobramycin, sisomicin, or netilmicin failed to produce synergism against any of these strains. The possible clinical significance of these findings was verified by use of the rabbit model of endocarditis. Combinations of penicillin with gentamicin or streptomycin were synergistic in the therapy of endocarditis that was produced by a strain of S. faecium that did not have a high level of resistance to aminoglycosides. However, the combination of penicillin and netilmicin was no more effective than penicillin alone.


In vivo platelet retention in human bleeding-time wounds. I. Normal subjects and patients with platelet dysfunction. PRB was measured in standardized skin puncture wounds. Platelets in wound blood samples collected in EDTA-heparin-containing micropipettes were counted electronically following gravity separation of plasma. The relationship between the wound platelet count (expressed as percent of the venous blood platelet count) and time was found to be linear during the first 3 min of bleeding. The slope of the linear regression line for percent of venous blood platelet count vs. time was 20.8 +/- 8.9 (mean +/- S.D.) in 31 normal subjects. Nine of 10 patients with significant, nonpharmacologic platelet dysfunction were found to have markedly decreased PRB. The determination of PRB was found to be reproducible, and control studies excluded platelet clumping and sedimentation of platelets as factors which significantly influenced the results. In contrast to previous observations, the results of this study indicate that platelets are removed from circulating blood at an increasing rate during the first 3 min of bleeding in normal subjects. Furthermore, a significant degree of EDTA-reversible clumping of platelets normally occurs during bleeding. The method described permits an accurate assessment of the quantitative and dynamic aspects of platelet participation in the arrest of bleeding. The determination of PRB holds promise in the evaluation of the biological significance of in vitro evidence of platelet dysfunction, and it may be useful in the evaluation of patients with hemorrhagic and thromboembolic disorders.


In vivo platelet retention in human bleeding-time wounds. II. Effect of aspirin ingestion. PRB was studied in normal human subjects before and after aspirin ingestion. Aspirin ingestion resulted in a prolongation of individual bleeding times greater than 2.4 min (greater than 2 S.D. beyond the group mean before aspirin) in 62.5% of 48 paired studies. The relationship of platelets retained vs. time was linear during the first 3 min of bleeding before and after aspirin. The mean PRB decreased from 22.1 +/- 9.2 to 9.6 +/- 8.6 (p less than 0.001) after aspirin ingestion. Subjects whose bleeding time was prolonged greater than 2.4 min had a significantly higher mean PRB before aspirin and a significantly greater mean decrease in PRB after aspirin than those whose bleeding time was prolonged less than or equal to 2.4 min. Aspirin ingestion reduced the number of EDTA-irreversible clumped platelets present in wound blood approximately 50% during the second and third minute of bleeding, but large numbers of EDTA-reversible platelet clumps were observed in wound blood before and after aspirin. Although platelet retention was significantly decreased during the first 3 min of bleeding after aspirin, the percent of venous blood platelets present in wound blood just prior to the arrest of hemorrhage was equal before and after aspirin. These observations indicate that aspirin prolongs the bleeding time by decreasing platelet clumping and slowing the rate of platelet thrombus formation in severed blood vessels. The presence of platelet clumps in wound blood after aspirin ingestion indicates that alternative mechanisms of platelet aggregation, independent of the arachidonate pathway of prostaglandin synthesis, proceed in vivo unaltered by aspirin.


Determination of C1q in human sera.

Determination of C1q in human sera. All detectable C1q in serum is precipitated with the euglobulin fraction, and all other detectable hydroxyproline-containing protein in serum is excluded from this fraction. Since C1q contains 4.3% hydroxyproline, an estimate of its content in serum can be calculated by multiplying the hydroxyproline content of the euglobulin fraction by 23.3. The mean value of C1q in five normal male subjects determined by this method was found to be 63 microgram/ml as compared with 144 microgram/ml when determined by RID, indicating that the latter procedure seriously overestimates C1q in serum. C1q content of serum from patients with RA was found to be elevated by both procedures.


Mouse alloantibodies capable of blocking cytotoxic T-cell function. I. Relationship between the antigen reactive with blocking antibodies and the Lyt-2 locus. In an attempt to produce allonatibodies to cytotoxic T-cell receptors, hyperimmune anti-lymphocyte antisera have been raised in mice of various strain combinations, and have been tested for their ability to block allogeneic cell-mediated lymphocytotoxicity (CML) in the absence of complement at the T killer cell level. Most of the sera failed to show any significant and reproducible inhibitory effects. However, among C3H anti-B10.BR antisera, some sera were found to be capable of significantly inhibiting CML. This effect was attributable to antibodies reacting with the killer population rather than the target cells, because the sera inhibited B10 anti-C3H CML but not C3H anti-b10 CML. Among mouse strains tested, A/J, BALB/c, B10, and B6 strains were sensitive to the inhibitory effect of the sera whereas AKR, CBA, C3H, and DBA/2 strains were insensitive. The sensitivity of killer cells to the inhibitory effect correlated well with the strain distribution of the Lyt-2.2 antigen. In the presence of complement, these same sera were toxic to 100% of spleen cells of AKR, BALB/c, B10, and DBA/2 strains, with comparable cytotoxic titers. Thus, the inhibitory activity of the sera could not be explained by nonspecific effects of high-titered antibodies. To study the relationship between the antigen(s) responsible for the blocking effect and Lyt-2-linked genes, killer cells from Lyt-2 congenic strains were tested and conventional anti-Lyt-2.2 antisera were raised in an appropriate congenic strain combination. Killer cells from B6, but not from B6.Ly2.1 animals, were significantly sensitive to the blocking effects of the inhibitory C3H anti-B10.BR sera. The conventional anti-Lyt.2.2 sera did produce CML blocking, although there was no apparent correlation between such blocking and the anti-Lyt-2.2 cytotoxic titer. These results thus indicate that the target molecules responsible for blocking of killer cells are encoded or regulated by genes that are closely linked to or identical with Lyt-2.


Genetic control of the immune response to collagen. II. Antibody responses produced in fetal liver restored radiation chimeras and thymus reconstituted F1 hybrid nude mice. The level of antibody produced in response to calf skin collagen in mice is influenced by genes which are closely linked to the I region of the H-2 major histocompatibility complex. This influence is shown to be expressed during lymphoid maturation by testing the antibody responsiveness to collagen in two types of chimeric mice. First, high responder and low responder parental strain mice were lethally irradiated and restored with fetal liver cells from (high X low responder) F1 mice. These F1 leads to parent chimeras exhibited an immune response phenotype characteristic of the irradiated parental strain animals, establishing that H-2 determinants of the host affect antigen responsiveness. Second, (high X low responder) F1 congenitally athymic (nude) mice were restored with fetal thymus transplants from either high or low responder parental strain mice. After a period of maturation these mice were shown to be competent for a T-dependent IgG response to SRBC. The responsiveness to collagen in these mice was characteristic of the parental strain thymus donors, indicating that the expression of H-2 determinants in thymic tissue during lymphoid maturation influences the antibody response phenotype expressed by mice.


Intracellular localization of Rickettsia tsutsugamushi in polymorphonuclear leukocytes. Rickettsia tsutsugamushi (Gilliam strain) was serially propagated in BHK-21 cell cultures and incubated with guinea pig peritoneal polymorphonuclear leukocytes to study the ultrastructural features of rickettsial uptake and entry into the leukocytes. Significant numbers of rickettsiae were phagocytized selectively by these leukocytes within 30 min. About one-half of these rickettsiae remained sequestered in phagosomes but the other one-half were free from the phagosome and localized directly in the polymorphonuclear leukocyte cytoplasm. Various stages of rickettsial release from the phagosomes were observed. Once free within the polymorphonuclear leukocyte cytoplasm, the rickettsiae were preferentially localized in the glycogen-packed areas which are devoid of lysosomes and other cytoplasmic organelles. This study indicates that rickettsiae phagocytized by polymorphonuclear leukocytes can escape from the phagosome into the cytoplasm.


Intrastrand self-complementary sequences in Bacillus subtilis DNA. Intrastrand self-complementary sequences have been isolated from the DNA of Bacillus subtilis by hydroxyapatite (HA) chromatography following thermal renaturation of strands separated by chromatography on methylated albumin kieselguhr (MAK). The instrastrand structures derived from the MAK H strand (HA HII) were biologically active showing transforming activity for a wide variety of markers, as well as hybridization to both pulse-labelled and ribosomal RNA. Removal of regions of single-strand DNA with S1 nuclease did not significantly alter the biological activity of the self-annealed molecules. The overall efficiency of transformation and hybridization of the intrastrand self-annealing DNA was low suggesting that many sequences in the population are neither active in transformation to prototrophy nor transcribed into RNA.


Genetic and biochemical studies of mutants of Penicillium chrysogenum impaired in penicillin production. Seventy-eight mutants of Penicillium chrysogenum strain NRRL 1951, that were impaired in penicillin production, were isolated following treatment with various mutagens. Twelve that yielded about 10% of their parental penicillin titre were studied in detail. Analyses of heterozygous diploids formed between them revealed the existence of at least five complementation groups with respect to penicillin production--V, W, X, Y and Z. Most mutants belonged to group Y. A biochemical investigation of the intracellular peptides in strains representing the five groups demonstrated the absence of the tripeptide alpha-aminoadipoylcysteinyl-valine from mutants of groups X, Y and Z. Extracts of mutants of groups W, Y and Z were able to catalyse a penicillin acyl-exchange reaction, a mutant of group V showed only a trace of activity and mutant from group X completely lacked this ability.


Effect of piperacillin on D-alanine carboxypeptidase activities from Pseudomonas aeruginosa. Membrane-bound D-alanine carboxypeptidase activity from Pseudomonas aeruginosa is very sensitive to inhibition by piperacillin.


Sodium dodecyl sulphate--polyacrylamide gel electrophoresis of cell-surface proteins as an aid to the identification of the Bacteroides fragilis group. The outer-membrane complexes from 40 strains of Bacteroides, representing eight of the species included in the Bacteroides fragilis group, were released by EDTA treatment. The component polypeptides were examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis on slab gels. Within a species (biotype) the patterns produced indicated marked similarities in the structures of the surface proteins among the strains examined. The patterns produced by strains belonging to different species, however, showed fewer similarities. An unknown organism could therefore be identified to species level using this technique and a few selected biochemical tests.


The effect of transition metal ions on the resistance of bacterial spores to hydrogen peroxide and to heat. The presence of 10 microM-Cu2+ increased the lethal effect of hydrogen peroxide on spores of Clostridium bifermentans but not on those of Clostridium sporogenes PA 3679, Clostridium perfringens, Bacillus cereus or Bacillus subtilis var. niger. Cu2+ at 100 muM also increased the lethal effect of heat on spores of C. bifermentans but not on those of B. sutilis var. niger. The rate and extent of Cu2+ uptake by spores of C. bifermentans and B. subtilis var. niger were similar, but examination of unstained sections of spores by electron microscopy suggested that Cu2+ is bound by the protoplasts of spores of C. bifermentans but not of B. subtilis var. niger.


Genetic instability of sporulation-associated characters in a Bacillus subtilis mutant: analysis of the segregation pattern and genetic studies. A Bacillus subtilis mutant which formed dark-brown 'medusa' (M) colonies was obtained. It sporulated at a high frequency, overproduced extracellular protease during sporulation and possessed a high genetic instability with a complex segregation pattern. Segregation was maintained after repeated re-isolation of single M colonies. The major wild-type-like class of segregants (B) was stable, sporulated normally and produced normal amounts of protease. Occasionally segregants were obtained which produced extremely high amounts of protease, sporulated poorly, formed transparent colonies and were either highly unstable (TD) or stable (TDst). Rarely B(D) (stable, normal sporulation and protease overproduction) and W and T (both stable and asporogenous) segregants were produced. The M phenotype was transmitted as a single factor by transformation but not by transduction. The results of transduction experiments suggest the presence of two mutations, ScoC and ScoD. It is proposed that this new segregating system in B. subtilis may result from tandem duplication of part of the bacterial chromosome.


Genetic instability of sporulation-associated characters in a Bacillus subtilis mutant: relationship between sporulation, segregation and the synthesis of extracellular enzymes (kinetic studies). In the genetically unstable, protease-overproducing 'medusa (M) strains of Bacillus subtilis, segregation of stable, wild-type-like B cells occurred mainly during sporulation. After the end of the exponential growth phase, a small fraction of M cells sporulated quickly and formed M spores, while the majority of the cells, after a 'critical period', gave rise to B segregants which sporulated after a delay. Segregation occurred without cell division. Delayed sporulation, segregation and protease overproduction are related. Similar but more complex results were obtained with the highly unstable TD strains. Sporulation and the kinetics of protease overproduction were also followed in several stable segregants. Depending on the strain, either the rate of protease production or both the rate and time course were affected. The results are interpreted in terms of sequential activation and de-activation of sporulation genes. The production of the alkaline and the neutral proteases was, in general, under common genetic control. In some strains alpha-amylase was also overproduced.


Effect of bacteriophage C5 on ultraviolet light survival in Pseudomonas aeruginosa. Bacteriophage C5 of Pseudomonas aeruginosa is able to reactivate ultraviolet (u.v.)-irradiated phage E79 in coinfection experiments and decrease the u.v.-sensitivity of a host-cell reactivation deficient mutant. These properties suggest that phage C5 has a gene(s) which is involved in the repair of u.v.-damaged DNA. The isolation of two u.v.-sensitive mutants of C5 supports this hypothesis.


Isolation of atypical lipopolysaccharides from purified cell walls of Pseudomonas cepacia. Wall fragments were prepared from two strains of Pseudomonas cepacia and from P. aeruginosa, and their contents of readily extractable lipid, pronase-digestible protein and lipopolysaccharide were measured. Lipopolysaccharide extracted from P. cepacia, although biologically active, contained no detectable 2-keto-3-deoxyoctonic acid, but contained phosphate, rhamnose, glucose, heptose and hexosamine in concentrations comparable to those found in P. aeruginosa.


Regulation of the interferon system: evidence that Vero cells have a genetic defect in interferon production. A clone of Vero cells was isolated and shown to be totally unable to synthesize interferon and insensitive to the toxic effect of poly(rI).poly(rC) treatment. Cells of this clone and mouse L cells were fused by treatment with polyethylene glycol or Sendai virus. Hybrid cell clones were isolated following selection  in medium containing hypoxanthine, thymidine and ouabain. The hybrids were sensitive to the antiviral effect of poly(rI).poly(rC) and synthesized mouse, but not primate, interferon. It is proposed that in Vero cells, the gene for interferon synthesis is defective or absent.


Susceptibility of various cells treated with interferon to the toxic effect of poly(rI).poly(rC) treatment. Various cells were treated with interferon and then exposed to polyriboinosinic-polyribocytidylic acid complex [poly(rI).poly(rC)]. With mouse L cells, there was a marked cytotoxic effect from low doses of interferon and poly(rI.poly(rC), whereas chick embryo cells showed an effect only after high doses. When primate cells (LLC.Mk2,BSC.B, Vero and human embryo cells) were treated with human or monkey interferon, poly(rI).poly(rC) was not cytotoxic.


Established cell line sensitive to influenza C virus. Various strains of influenza C virus grew productively in an established line of monkey kidney cells (LLCMK2) without prior adaptation. When trypsin was added to the medium, higher virus yields were obtained than in other cell cultures. All influenza C virus strains tested formed well defined plaques under the agar overlay medium containing trypsin. Infectivity determined by plaque assay in LLCMK2 cells was higher than that determined by amniotic inoculation of fertile hens' eggs.


Mechanism of persistence of rubella virus in LLC-MK2 cells. LLC-MK2 cells chronically infected with two strains of rubella virus, HPV-77 and Thomas, have been examined over several months to find out the mechanism of persistence. Evidence is given for the presence of defective particles in these cultures by finding virion RNA which sedimented at 12S instead of the 40S typical of the fully infectious virus. A 'provirus' DNA copy of the rubella virus genome was not detected by methods which included filter hybridization and in situ hybridization, or by treatment of the chronically infected cells with mitomycin C, antinomycin D or 5-bromodeoxyuridine. In addition, the chronically infected cells contained RNA-dependent RNA polymerase activity, but no RNA-dependent DNA polymerase activity.


Demonstration of hepatitis B e antigen (HBeAg) in association with intact Dane particles. Mild detergent treatment (0.1% Sarkosyl-0.1% beta-mercaptoethanol) of Dane particle-rich fraction from human serum resulted in the release of core particles together with HBe antigen activity when examined by the reversed passive haemagglutination method. Furthermore, when the core particles isolated by the above procedure were exposed to stronger detergent (1% Sarkosyl-0.1% beta-mercaptoethanol), additional HBe antigen activity was released only from intact core particles with DNA polymerase activity and not from empty core particles.


Lipid-containing bacteriophage PR4: structure and life cycle. The structure of the purified lipid-containing phage PR4 was studied electron microscopically using thin sectioning, negative staining and freeze-fracturing techniques. The lipid layer was located inside a rigid capsid and had a bilayer structure. The innermost dense core was probably composed solely of DNA since no major structural proteins were missing in empty phage particles lacking DNA. During infection, phages were attached to the cell wall of the host. They apparently inject their DNA through the cell envelope. The lipid layer of the phage might play an active role in the injection process. The maturation of the phage capsid takes place within the nuclear region of the cell, whereas intact phages with DNA were always seen in the cell periphery.


Morphology and morphogenesis of a new paramyxovirus (PMV 107). The morphology of the virions and nucleocapsids of paramyxovirus 107 (PVM 107) and the replication of the virus were investigated by electron microscopy. The virions and nucleocapsids exhibited the same structural properties as other paramyxoviruses. Nuclecapsids were found in the nucleus and cytoplasm of infected bovine embryonic lung (BEL) cell cultures. A similar situation has been described for the morbilliviruses measles, SSPE, distemper and rinderpest. Alignment of nucleocapsids beneath the plasma membrane and budding of PMV 107 in the productive BEL cell infections were also similar to the morbillivirus-infected cells. In a line of monkey cells (CV1) persistently infected with PMV 107 only cytoplasmic nucleocapsids could be demonstrated. On the basis of its morphology and morphogenesis it is suggested that PMV 107 should be classified as a paramyxovirus. Since nucleocapsids could also be found in the nucleus of infected BEL cells the morphogenesis of PMV 107 closely resembles that of viruses of the morbillivirus group.


[Mucopolysaccharidesis Type VII resulting from beta-glucuronidase deficiency. Report of one family]. The patient is a north african female, fourth born child in a family with consanguinity. Facial dysmorphia, clubfeet, swollen extremities and heel borne ponctuate calcifications are observed soon after birth. beta glucuronidase activity is very low in serum, leukocytes and skin fibroblasts. At 18 months, gorwth and psychomotor development are normal. Flat facies and dorsolumbar cyphosis are striking. There is no clinical sign of storage disease, neither ocular or cytologic (blood, bone-marrow) abnormalities. Squeletal abnormalities are predominant on cervical and lumbar column and pelvis. Urinary excretion of chondroitin 4 sulfate and chondroitin 6 sulfate is increased. A 4 year old sister is affected. Facial dysmorphia, mild squeletal abnormalities are observed. Again, growth is normal and there is no symptom of storage disease. Enzymic expression of parent heterozygotism is marked in serum studies, but less marked in leukocytes and fibroblasts. The last two children are heterozygotes. At the time of a 5th pregnancy, enzymic activity studies of amniotic fluid and amniotic cells have shown that the foetus was an inaffected female. This child was normal at birth.


Astrovirus infection in volunteers.

Astrovirus infection in volunteers. An extract and a filtrate prepared from feces of a child with mild gastroenteritis were shown by electron microscopy to contain numerous astrovirus particles and were given to eight volunteers by mouth. One subject developed diarrheal illness and concurrently shed large amounts of astrovirus in feces, and one other had mild constitutional symptoms with a lower level of virus shedding. Nine other volunteers were given fecal filtrate from the volunteer with diarrhea, and astrovirus shedding subsequently occurred in two of them. The syndrome accompanying virus shedding appeared distinct from that associated with the "W" agent in previous experiments. Thirteen of 16 astrovirus-inoculated subjects subsequently developed a rise in titer of the homologous antibody in serum. It was concluded that astrovirus causes a transmissible infection that is of low pathogenicity for adults. Immunofluorescence of human embryo kidney cells inoculated with astrovirus and shown by electron microscopy to contain 28 nm virus-like particles was used both to detect virus in feces and to assay astrovirus antibody.


OC43 strain-related coronavirus antibodies in different age groups. Serum antibodies against human coronavirus OC43 in different age groups were measured by complement fixation (CF), haemagglutination inhibition (HI), radial diffusion haemolysis-in-gel (HIG), and solid-phase radioimmunoassay (RIA) methods. Antigen grown in suckling mouse brain was used in all tests. Results obtained by the CF and HIG tests, and the RIA, were in good agreement with regard to the presence or absence of antibodies. Similar results were also obtained with the HI test if nonspecific haemagglutination inhibitors were first removed by treatment with phospholipase C and only titers of 1:20 or greater were considered positive. Children 6--23 months of age (n = 45) were without measurable coronavirus antibodies in all four assays. A rapid increase in the prevalence of antibodies then occurred in subsequent age groups, and practically all persons 6 years of age or older were found to have OC43 antibodies as measured by the HIG test or the RIA. The mean antibody levels determined by these two methods continued to increase, however, up to the age group of 10--14 years. This increase in antibody levels after the initial antibody incidence plateau may be due to boosting effects caused by related coronavirus strains, since OC43 antigens are known to cross-react with antibodies induced by other human coronaviruses. Taken together, these data suggest that OC43 virus, or an antigenically related coronavirus strain, is very common in Finland.


Autonomic innervation of the mammalian penis: a histochemical and physiological study. The distribution of adrenergic and cholinergic nerves was studied in penile tissue from rat, rabbit, cat, monkey and man. Glyoxylic acid histofluorescence and acetylcholinesterase-positive fibers were found in all tissues examined, but histofluorescent, presumably adrenergic fibers predominated. In general, except in the rabbit, more nerve fibers of both types were found in the corpus cavernosum (CC) than in the corpus spongiosum/penile urethra (CS). The rabbit penis showed slightly more adrenergic fibers in the CC than in the CS and more cholinergic nerves in the CS than in the CC. The CC of the monkey demonstrated an alpha receptor mediated contractile response to norepinephrine (NE) stimulation and no response to acetylcholine (ACh) in an in vitro muscle bath.


Effects of plumbous ion on guanine metabolism. The enzyme guanine aminohydrolase (guanase) is inhibited by low levels of Pb2+. The inhibition is noncompetitive and the Ki is 3.0 X 10(-6) M. The only other heavy metals that are inhibitory at low concentrations are Ag+, which is 36% more, and Hg2+, which is about 50% less inhibitory than Pb2+. The inhibition of guanase by Pb2+ and Hg2+ is synergistic and the inhibition of the enzyme was readily reversed by EDTA. The relationship of these studies with guanase and to the etiology and treatment of saturnine gout, which appears in humans suffering from lead poisoning, is discussed.


Pattern sensitive epilepsy: a case report. A child with pattern sensitive epilepsy is reported. She had both absences and generalised seizures. The latter occurred spontaneously but the former followed concentration on patterned materials. These self-induced seizures appeared to have an affective component. The EEG showed generalised atypical spike and wave discharges both in the resting state and during intermittent photic and pattern stimulation. The patient responded to sodium valproate.


Composition of geniculostriate input ot superior colliculus of the rhesus monkey. 1. In order to examine the composition of the geniculostriate input to the superior colliculus, microelectrode recordings were undertaken in this structure of the rhesus monkey while parvocellular or magnocellular laminae of the LGN were reversibly inactivated by injecting minute quantities of lidocaine or MgCl2. 2. The inactivation of magnocellular laminae disrupted the visually driven activity of most cells in the topographically corresponding areas of the colliculus, but not in the superficial retinotectal recipient zone. The inactivation of parvocellular lamina had no effect on the visually driven activity of collicular cells. 3. Several controls were carried out to rule out the possibility of intervention with fibers of passage. We ascertained that the LGN injections did not affect the direct retinotectal pathway by comparing the effect of such inactivation with the effect produced by reversibly cooling visual cortex. These two manipulations yielded similar results: cells in the most superficial regions of the superior colliculus were unaffected by both cortical cooling and by magnocellular injections, while below this region the response of collicular cells was reduced or eliminated in both cases. 4. These results suggest that the indirect visual pathway to the superior colliculus via cortex is activated selectively by the broad-band system, which is relayed through magnocellular LGN. The color-opponent system does not appear to have a corticotectal input sufficient to drive collicular cells independently.


Prophylactic phenytoin in severe head injuries. We are reporting a retrospective study of 62 patients whose head injury was sufficiently severe to cause a high probability of posttraumatic epilepsy. Of 50 patients treated with phenytoin, 10% developed epilepsy of late onset. Twelve patients not treated with phenytoin but who had head injuries of equal magnitude had a 50% incidence of epilepsy. These data from a highly selected group of patients with severe head injuries confirm the bias that treatment with phenytoin decreases the incidence of posttraumatic epilepsy.


Utilization of L-alanyl-L-tyrosine by nephrectomized rats when infused as part of a total parenteral nutrition regimen. L-Alanyl-L-tyrosine is well utilized as a tyrosine source in parenterally fed rats. Such utilization may depend upon filtration of peptide into the glomerular filtrate, reabsorption into renal epithelial cells, hydrolysis to component amino acids in or at the surface of epithelial cells, and release of component amino acids to the blood. Bilaterially nephrectomized rats were infused with a parenteral solution providing L-alanyl-L-[U-14C]-tyrosine at 0.5 mmoles/kg over a 2 hour period to test this hypothesis. Despite the absence of kidneys, peptide did not accumulate in plasma or tissues. Plasma and liver tyrosine and alanine levels increased significantly over values noted in animals infused without peptide. One-quarter to one-third of the infused radioactivity was released as 14CO2, with the remainder found in the tissues. Between 15 and 51% of radioactivity in individual tissues was free tyrosine, the remainder was incorporated into protein. Isolation of this protein, acid hydrolysis and simultaneous radioactivity-amino acid analysis demonstrated that 94 to 99% of the radioactivity in protein was tyrosine. The data indicate good utilization of alanyl-tyrosine by nephrectomized rats when administered as part of a total parenteral nutrition regimen.


Phagocytosis and trypsin-resistant glass adhesion by osteoclasts in culture. Osteoclasts were cultured in vitro and tested for the functions normally associated with cells of the mononuclear phagocyte system. They were found to ingest glutaraldehyde-fixed red cells and latex, but not opsonised or complement-coated sheep red cells. Osteoclasts are glass-adherent and resist removal by trypsin. The implications of these findings for the identity of the osteoclast precursor cell and its mechanism of fusion are discussed.


The comparative pathology of the primate colon. This survey has revealed further close similarities between the pathology of certain colonic diseases in humans and primates. The changes taking place in acute inflammations of varying severity confined to the lamina propria are virtually identical as are those seen in the healing process. In primates chronic inflammatory processes caused by protozoa resemble those in humans very closely. Microherniation of mucosal glands through the muscularis mucosae were found in a wide range of species. Evidence pointed to such herniations playing an important role in the spread of inflammatory diseases from the lamina propria into the submucosa by affording the aetiological agent access through the muscularis mucosae. No case of adenocarcinoma was found in this survey and the cause for the striking difference in incidence of this disease in human and primate colons is discussed.


Use of the serum bactericidal titer to assess the adequacy of oral antibiotic therapy in the treatment of acute hematogenous osteomyelitis. Serum bactericidal titers against Staphylococcus aureus were measured in 63 children who were receiving mafcillin or methicillin intravenously, or dicloxacillin, penicillin, or cephalexin orally. The SBTs obtained following unit does of 25 mg/kg of dicloxacillin, 35 mg/kg of penicillin, or 25 mg/kg of cephalexin with probenecid were comparable to those seen following intravenous doses of 40 mg/kg nafcillin or methicillin. Twenty-two children with acute hematogenous osteomyelitis proven or presumed to be due to S. aureus were treated intravenously until point tenderness and fever had resolved, and then with oral therapy. The mean duration of intravenous therapy was 14 days. Oral doses were adjusted so that a peak SBT of greater than or equal to 1:16 and a trough SBT of greater than or equal to 1:2 were obtained in most children. No recurrences occurred. The SBT proved to be a practical means of assessing the adequacy of oral therapy in children with infections due to S. aureus.


Xerophthalmia, protein-calorie malnutrition, and infections in children. All children with evidence of xerophthalmia who were admitted to an inpatient facility for treatment of severe protein-calorie malnutrition and its complications were compared to other inpatients with severe PCM but free from signs or symptoms of vitamin A deficiency. Xerophthalmic inpatients were older and had more severe hypoproteinemia than inpatients without eye signs, and were more undernourished by anthropometric criteria than a comparison group matched for age, sex, and type of PCM. Children with more severe eye lesions were more retarded in growth than those with minimal ocular signs. All inpatients had high rates of bacterial infections, regardless of their vitamin status. However, xerophthalmic children had a highly significant increase in the rate of positive urine cultures. Mortality rates were similar in all study groups.


Scanning electron microscopy of ciliary zones of the ciliate protozoa in the large intestine of the horse. The surface structure of the ciliary zone in 13 species of ciliates found in the large intestine of the horse was observed by scanning electron microscopy. In Holophryoides ovalis many fine depressions considered to be a result of phagocytosis or pinocytosis in the naked cytostome were noticed. In Blepharocorys spp. a distinct section was present between the portion with cilia and that without cilia. It was not present, however, in some species of the family Buetschliidae, such as Bundleia postciliata and Didesmis spp. The species of Entodiniomorphida had a lip around the ciliary zone with cilia forming synciliary tufts. In Spirodinium equi and Tetratoxum unifasciculatum the ciliary zone revolved counter-clockwise in an en face view. Some differences in the surface structure of the ciliary zone between the entodiniomorphid and spirotrich ciliates are discussed.


The role and safety of early postoperative feeding in the pediatric surgical patient. The authors report their experience with early postoperative feedings in a variety of pediatric surgical patients utilizing a needle catheter jejunostomy. A small caliber catheter, similar to that used for antecubital central venous cannulation, is inserted in the antimesenteric border of the jejunum providing a subserosal tunnel. A purse-string suture is placed around the catheter and then it is secured to the abdominal wall, and the catheter is brought out through the abdominal wall via a needle puncture wound. Our experience with 27 insertions in 25 patients ranging in age from 1 day to 17 yr, has demonstrated the ease of placement and feasibility of immediate postoperative feeding. Patients were generally started on a dilute elemental diet through the jejunostomy within 12 hr of the operative procedure. Patients were administered between one and 3.5 g of protein kg/day and between 45 and 100 cal/kg/day depending on age and weight. Duration of treatment with enteral nutrition ranged from 10 to 150 days. There was no catheter-related complications utilizing the technique described. Technical details of catheter placement and protocol for administration of early postoperative feedings are discussed. The ability to provide nutritional support via the gut has obviated the need for total parenteral nutrition in the majority of these patients.


Prostaglandin prodrugs III: Synthesis and biological properties of C9- and C15-monoesters of dinoprost (prostaglandin F2 alpha). Methods are described for the synthesis of dinoprost C9- and C15-monoesters using protective groups. Esters at C9 were synthesized by acylation of dinoprost 11,15-bis(tetrahydropyran-2-yl)ether followed by acid-catalyzed protective group removal. Esters at C15 were synthesized by initial formation of the protected intermediate, dinoprost 9,11-n-butylboronate, followed by acylation and hydrolytic protective group removal. Many esters were active in vivo in the hamster antifertility screen. Plasma hydrolysis studies showed that the C15-esters were more readily cleaved than the C9-esters. In vivo studies in the rat showed that both the C9- and C15-esters resulted in urinary excretion of 5 alpha, 7 alpha-dihydroxy-11-ketotetranorprosta-1,16-dioic acid in amounts comparable to those obtained after dosing with dinoprost, indicating that ester hydrolysis occurred in vivo.


Short-term effects on Artemia salina of aponin and Gomphosphaeria aponina in unialgal cultures and in mixed cultures with gymnodinium breve. Previous investigations showed that the natural marine substance aponin, produced by the blue-gree alga Gomphosphaeria sponina, was cytolytic towards Florida's red tide organism, Gymnodinium breve. As part of a study of the aponin-alga biological interactions, the effects of unialgal (G. aponina) and mixed cultures (G. aponina + G. breve) on the viability of artemia salina were investigated. Two contrasting effects were observed in mixed cultures: survival of A. salina was promoted in low G. aponina populations, while enhanced toxicity of G. breve to A. salina occurred at higher culture populations. Unialgal G. aponina cultures exerted no adverse effects on A. salina. The apparent protective effect is thought to result from the observed change in G. breve morphology (motile to sessile); toxicity was the result of enhanced toxin release by cytolyzed G. breve cells. In dose-response studies, aponin exhibited no adverse effect on A. salina at concentrations (1 unit) that were deleterious to G. breve. However, at higher applied levels (4--6 units), mortality was substantial (greater than 70%) after 48 hr of incubation. Probit analysis yielded an apparent LD50 of 2.3 units, where 1 unit was that amount required to cytolyze 50% of the G. breve cells (2000 cells/ml) after 20 hr of incubation.


Effects of systemic and intraventricular administration of cannabinoids on schedule-controlled responding in the squirrel monkey. The effects of a number of cannabinoids in squirrel monkeys trained to respond on a chain fixed-interval fixed-ratio schedule of food presentation were determined after intraperitoneal (i.p.) and intraventricular (i.v.t.) administration. The order of potency was (+/-)-9-nor-9 beta-OH hexahydrocannabinol, 11-OH-delta 9-tetrahydrocannabinol, delta 9-tetrahydrocannabinol (delta 9-THC), cannabinol and cannabidiol. (+/-)-9-Nor-9 alpha-OH-hexahydrocannabinol was inactive at doses up to 3 mg/kg i.p. and 0.1 mg/kg i.v.t. Although the order of potency was the same by both routes of administration, the i.v.t./i.p. potency ratio differed markedly. This demonstrates the importance of route of administration in assessing structure-activity relationships of cannabinoids and suggests that differences in penetration to the central nervous system may be an important determinant of behavioral activity. Although 11-OH-delta 9-THC was more potent than the parent compound delta 9-THC by both routes, the potency difference was less after i.v.t. administration. It was also demonstrated that metabolic conversion of [3H]delta 9-THC does not take place in squirrel monkey brain when administered i.v.t. which could account for the direct i.v.t. effects of delta 9-THC. These observations suggest that metabolic conversion of delta 9-THC in the liver is not necessary for its behavioral effects.


Responses of striate cortex cells to grating and checkerboard patterns. 1. Cells in visual cortex have been alternately considered as bar and edge detectors, or as spatial-frequency filters responding to the two-dimensional Fourier component of patterns. 2. The responses to gratings and to checkerboards allow one to test these alternate models: the Fourier components of a checkerboard pattern do not occur at the same orientation as the edges, nor do the checkerboard spatial frequencies correspond to the check widths. 3. Knowing the orientation tuning of a cell for gratings, one can precisely predict its orientation tuning to checkerboards from the orientation of the fundamental Fourier components of the patterns, not from the orientation of their edges. This was found for both square and rectangular checkerboards, and held for both simple and complex cortical cells. 4. Knowing the spatial tuning of a cell for sine-wave gratings, one can precisely predict its spatial tuning to square and rectangular checkerboards from the spatial frequencies of the fundamental Fourier components of the patterns, not from the widths of their checks. 5. When presented with checkerboards in which not the fundamental but the upper harmonics were within its spatial bandpass, a cell's orientation tuning was found to be predictable from the (quite different) orientation of the higher Fourier harmonic components, but not from the orientation of the edges. 6. Knowing a cell's contrast sensitivity for gratings, one can predict the cell's contrast sensitivity for checkerboards much more accurately from the amplitudes of the two-dimensional Fourier components of the patterns than from the contrasts of the patterns. 7. The orientation tuning, spatial-frequency tuning and responsiveness of cells to a plaid pattern were also found to be predictable from the pattern's two-dimensional Fourier spectrum. 8. Both simple and complex striate cortex cells can thus be characterized as two-dimensional spatial-frequency filters. Since different cells responsive to the same region in the visual field are tuned to different spatial frequencies and orientations, the ensemble of such cells would fairly precisely encode the two-dimensional Fourier spectrum of a patch of visual space.


An epidemiological study of neonatal and postneonatal mortality in Macaca radiata at the California Primate Research Center (1966-1973). This study evaluates neonatal and postneonatal survival of Macaca radiata to 180 days of age at the California Primate Research Center from 1966 through 1973. Of a total of 287 live births, infant mortality rates (IMR) for three different types of housing were significantly different: 12% for inside cages, 41% for an outside one-half-acre enclosure and 31% for outside corncrib cages. Experience of the female in raising an infant to weaning age was a significant determinant only for the outside housing IMR. Introduction of laboratory-reared females into the breeding colony made the maternal breeding experience an important factor.


Behavioral interaction in social-deprivation-reared Macaca mulatta: effects of cerebellar lesions on aggressive and afiliative behaviors. Maintaining isolation-reared adult female Macaca mulatta in a group enclosure resulted in rapid resocialization. The influence of cerebellar lesions on social behaviors was most marked on aggressive interactions and cage stereotype pacing and circling.


Studies with timed-pregnant squirrel monkeys (Saimiri sciureus). Timed-pregnant monkeys were produced in a large nonhabituated colony of Saimiri sciureus of Bolivian origin. In a colony of 373 females and 40 males, 277 females (74%) were considered to be inseminated, based on microscopic observation of sperm and/or detection of a coagulum (plug) in the vagina. Forty-six full-term progeny were delivered. The mean gestational period was 152.5 days (SD = 3.9 days). For continuous cohabitation, the median time to insemination was five days, with 75% of inseminations occurring within eight days. Pregnancy evaluation (mouse bioassay) indicated a high level of resorptions within the first 50 days of gestation. This may help explain the low birth rates reported for other nonhaibuated colonies.


Characteristics of the menstrual cycle in nonhuman primates. I. Similarities and dissimilarities between Macaca fascicularis and Macaca arctoides. Comparative studies of reproductive characteristics were carried out on a colony of Macaca fascicularis and M. arctoides. Seasonal differences were not significant between species, and conceptions occurred throughout the year. The occurrence of short cycles ('luteal phase defect') was found in 2.9% of all M. fascicularis cycles and 1.5% of all M. arctoides cycles. Long cycles (40-50 days) were found in 4.0% of all M. fascicularis cycles and 3.5% of all M. arctoides cycles. Gestation lengths averaged 172.4 and 165.3 days, respectively, for M. arctoides and M. fascicularis. Ovulation in M. fascicularis occurred at a day of ovulation/cycle length ratio of 0.48 +/- 0.08 compared with 0.48 +/- 0.09 for M. mulatta data previously published.


Placental radioangiography as a method for the study of uteroplacental blood flow in the rhesus monkey. Placental radioangiography was performed in lightly anesthetized, pregnant rhesus monkeys under close monitoring of vital signs. Two angiographies in the same animal performed at an interval of approximately 20 min consistently produced identical patterns but not at intervals of one or two weeks. Evidence is adduced that the parameters, calculated from the angiograms, represent indices of uteroplacental blood flow under the standardized conditions of the angiographic procedure described here. Thus, placental radioangiography constitutes a relatively simple procedure for comparative, semiquantitative studies.


A radioangiographic study of the effects of catecholamines on uteroplacental blood flow in the rhesus monkey. Two placental radioangiographies were performed with a 20-min interval in ten lightly anesthetized rhesus monkeys. Amniotic and aortic pressures, as well as the maternal ECG, were continuously monitored. Norepinephrine or metaproterenol was administered in a low dose which did not cause general cardiovascular effects, prior to the second angiography. Norepinephrine appeared to cause constriction and metaproterenol, dilatation of the uteroplacental vessels. These effects could be diminished or abolished by an alpha- or beta-adrenergic blocking agent, respectively, and thus would appear to be caused by stimulation of adrenergic receptors in the uteroplacental vasculature.


A twinning event in Macaca sylvanus of Gibraltar. Linear measurements and body proportions for a set of free-ranging Macaca sylvanus twins are presented. Their measurements are compared to a full-term perinatal dead female. The twins are dizygous and probably 28 days premature.


Isoimmunizations among marmosets to identify leucocyte antigens. Saguinus fuscicollis ssp., S.o. oedipus, and S. nigricollis marmosets were immunized with white blood cells (WBC) from S.f. illigeri marmosets. Of the three genetic donor-host combinations attempted, only an interspecies immunization of S.o oedipus marmosets resulted in antibody formation which was detectable by agglutination or cytotoxicity tests. Analysis of one of the antisera with a panel of WBC from S.f. illigeri marmosets indicated varying specificities, reactions ranging from strong, intermediate, to completely negative. That the serum was detecting genotypic as well as phenotypic markers was suggested by the similarity of reactions of WBC from co-twin pairs and in preliminary testing of individual families.


A longitudinal study of facial growth in Papio cynocephalus after resection of the cartilagenous nasal septum. The present study evaluates in Papio cynocephalus the effectiveness of varying the amount of nasal septum resected and the age at surgery in an attempt to develop a technique which may have clinical value for the human condition. The analysis of interdental dimensions by graphical techniques demonstrates that a critical age exists at which total septum resection leads to maximum growth arrest of the premaxilla. Comparisons of linear regression curves for operated and control animals confirm these findings and also demonstrate the existence of sexual dimorphism in premaxillary growth rates.


Rearing of conventional and gnotobiotic nonhuman primates (Pan troglodytes, Papio cynocephalus, Saguinus nigricollis). Rearing techniques for conventional and gnotobiotic nonhuman primates are described. Up to four months of age there was no significant difference in weight gain between conventionally and gnotobiotically reared chimpanzees or baboons. After four months, gnotobiotic chimpanzees exceeded their conventional counterparts in weight gain, whereas conventional baboons showed higher weight gain than gnotobiotic baboons. Gnotobiotic chimpanzees and baboons had significantly lower absolute numbers of neutrophils than their conventional counterparts, but the absolute numbers of lymphocytes were not different. The gnotobiotic rearing of marmosets is also reported.


Characteristics of the menstrual cycle in nonhuman primates. II. Ovulation and optimal mating times in macaques. An analysis of the results of 1,259 limited-duration matings was conducted on colonies of Macaca arctoides and M. fascicularis. Maximum conception occurred at a day of breeding/cycle length (DB/CL) ratio of 0.40--0.41 with a range of DB/CL ratios for successful matings from 0.39 to 0.44. These values are compared with published values for various endocrine parameters equated to cycle length.


Influence of glucocorticoid administration and inhibition on fetal baboon pulmonary maturity and the amniotic fluid L/S ratio. The pregnant baboon feto-placental unit responded to intra-amniotic dexamethasone by a rise in the lecithin-sphingomyelin ratio (L/S ratio) and accelerated pulmonary maturity with evidence of increased amounts of pulmonary surfactant. Metyrapone 11 beta-hydroxylase inhibition of corticoid synthesis prevented the expected rise in the L/S ratio without influencing the normal development of pulmonary maturity. The L/S ratio did not accurately reflect pulmonary maturity when metyrapone was administered to the pregnant baboon. Prudently, model systems such as the baboon should be used for more detailed studies of endocrine effects on pulmonary development because of the reported untoward effects of exogenous steroid administration on the human neonatal nervous system.


Chronic methods to study the release of catecholamines from the neostriatum in nonanaesthetized monkeys (Macaca mulatta). The present report describes two methods allowing the chronic collection of catecholamine (CA) released from the neostriatum in the nonanaesthetized monkey (Macaca mulatta). In both methods the monkeys were placed in a restraining chair. Indwelling electrodes allowed the correlation of behavioural observations with polygraphic recordings. The first method uses a special superfusing cup permanently implanted on the lateral surface of the caudate nucleus, which permits the demonstration of spontaneous and D-amphetamine-induced release of (3H)-dopamine (DA) newly synthesized from L-(3,5-3H)-tyrosine over a period of six days. In the second method the collection of unlabelled CA is carried out by a localized ventricular perfusion. A radio-enzymatic estimation enabled the measurement of unlabelled DA and noradrenaline. With this second method the animals were kept for 25 to 30 days.


Isolation and separation of toad bladder epithelial cells. The epithelium of the urinary bladder of Bufo marinus is composed of 5 cell types, i.e., granular (Gr), mitochondria-rich (MR) and goblet (G) cells which face the urinary lumen, microfilament-rich (MFR) and undifferentiated cells (Un) located basally. The epithelium was dissociated by collagenase and EGTA treatment. Fractionation of dispersed cells by isopycnic centrifugation on dense serum albumin solutions yielded 4 fractions: (i) a very light fraction (p approximately equal to 1.025) enriched in MR and MFR cells; (ii) a light fraction (p approximately equal to 1.045) enriched in vacuolated Gr cells; (iii) a heavy fraction (p approximately equal to 1.065) composed essentially of aggregated Gr cells, and (iv) a pellet (p approximately equal to 1.085) enriched in G and undifferentiated cells. Recoveries were based on cell counts and DNA measurements. DNA content per cell was 13.2 pg +/- 0.9 (n = 37). From 1 g fresh tissue, 62 +/- 5 x 10(6) (n = 10) cells were recovered before isopycnic centrifugation of which about 70% excluded Trypan blue. After centrifugation, 90 to 95% of the cells excluded the vital dye and approximately 3(9) x 10(6) cells were recovered from the gradient. Cell metabolism in each fraction was estimated by oxygen consumption measurements in absence or presence of ouabain, acetazolamide, and dinitrophenol. The consumption measurements in absence or presence of ouabain, acetazolamide, and dinitrophenol. The consumption was threefold higher in the very light and light fractions when compared to the heavy and pellet fractions. Ouabain sensitive oxygen consumption (QO2) represented 12 to 35% of the total O2 consumption depending on the cell fraction, and acetazolamide sensitive QO2 varied from -0.8% in the heavy fractions to 20% in the lighter fractions. DNP increased QO2 in all fractions by 20 to 50%. Finally, the cells were able to reaggregate and form junctional complexes upon addition of calcium to the medium.


Microfilament-rich cells in the toad bladder epithelium. Basal cells of the bladder epithelium of Bufo marinus have been found heterogenous and consist of microfilament-rich cells (MFR-cell) and undifferentiated cells (Un-cell). The MFR-cell, which represents approximately 20% of the epithelial cell population, lies between the epithelial layer lining the urinary space and the basement membrane; it extends under several epithelial cells by processes of varying widths and lengths which contact, via desmosomes, other MFR-cells, as well as cells in the superficial layer, i.e., granular and mitochondria-rich cells. The cytoplasm of MFR-cell is filled with intermediate filaments arranged in bundles which run parallel to the plane of the epithelium and no dense granules, typical of granular cells, have been detected. Strong immunofluorescence for actin is associated with cells which occupy the same basal position as MFR-cells. Undifferentiated cells have no contact via desmosomes with adjacent cells and their cytoplasm is filled with free ribosomes; they lack bundles of intermediate filaments and possess no specialized organelles. After a 4-hr pulse of 3H-thymidine, 1.5% of epithelial cells incorporate thymidine into nuclear DNA, out of which 3/4 are basally and 1/4 are apically located. Identification of cell types by electron microscopy reveals that approximately 10% of undifferentiated basal cells are labeled, whereas less than 0.1% of granular cells and no MFR-cells incorporate 3H-thymidine into DNA. When dissociated from the epithelium and separated by isopycnic centrifugation, MFR-cells possess a mean buoyant density of approximately 1.025, cosediment with mitochondria-rich cells and exhibit a strong immunofluorescence for actin. The function of MFR-cells remains unknown; however, they may play a role in cell coupling and responses to hormonal and physical factors.


Action of steroids on H+ and NH+4 excretion in the toad urinary bladder. This study was done to determine if steroid compounds will stimulate the urinary bladder of the toad to increase its capacity to acidify the urine and excrete NH+4. Aldosterone, 17 beta-estradiol, dexamethasone, pregnenolone, and cholesterol were tested on the bladder. All compounds tested were found to stimulate the rate of acidification by the bladder, above that of a paired control hemibladder. In contrast, only the steroids aldosterone and 17 beta-estradiol were found to stimulate NH+4 excretion in the bladder. Cycloheximide was found to block the action of aldosterone on the NH+4 excretion, but did not have a significant effect on the stimulation of acidification by aldosterone. We conclude that steroids stimulate H+ and NH+4 excretion in the toad urinary bladder. In addition, the NH+4 excretory system seems to be more specific to this effect than is the H+ excretory system.


Mechanism of ovarian carcinogenesis: effect of 7,12-dimethylbenz[a]anthracene administration on intrasplenic ovarian grafts in unilaterally ovariectomized C3HeB/Fe mice. A single oral administration of 7,12-dimethylbenz[a]anthracene (DMBA) 2 weeks after intrasplenic grafting of ovarian tissue in unilaterally ovariectomized C3HeB/Fe mice resulted in a high tumor incidence (47%) in the grafted tissue, with only 1 tumor (3%) in the orthotopic ovary. No tumors were seen in the control group (unilaterally ovariectomized mice given intrasplenic grafts of ovarian tissue without subsequent DMBA administration), nor did tumors develop in response to DMBA treatment in mice with both ovaries in situ and no grafted tissue in the spleen. The results indicated that some local change caused by the grafting procedure rendered the tissues more sensitive to the action of DMBA and/or more responsive to gonadotropic stimulation.


Purification of measles virus and characterization of subviral components. Purified measles virus was obtained from [35S]methionine-labeled cells infected at 33 degrees C and maintained in the absence of fetal calf serum. The pellet that was produced by a single high-speed ultracentrifuge spin of culture medium contained virus of purity sufficient for structural analysis. Purified virions contain seven polypeptides with estimated molecular weights of: L, 200,000; G, 80,000; P2, 70,000; NP, 60,000; A, 43,000; F1, 41,000; and M, 37,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Treatment of virions with 0.25% trypsin resulted in a less dense particle which lacked polypeptides G and F1. Solubilization of the viral membrane with the detergent Triton X-100 in low-salt buffer resulted in the loss of the G polypeptide, whereas in the presence of 1 M KCl, Triton X-100 also removed most of the M polypeptide. The nucleocapsids (p = 1.3) obtained from virions treated with Triton X-100 and 1 M KCl contained the L, P2, NP, and M polypeptides. Nucleocapsids isolated from the cytoplasm of infected cells were predominantly composed of the NP polypeptide with smaller amounts of either polypeptide P2 or novel polypeptides, related to NP, with estimated molecular weights of 56,000 to 58,000 and 45,000 to 46,000. A significant amount of polypeptide L was always found in association with nucleocapsids isolated either from virions or from the cytoplasm of infected cells. A membrane component containing the viral membrane polypeptides G, F1, and M was also isolated from infected cells. The data presented here thus suggest that L is an integral part of the nucleocapsid complex. In addition, 37,000-molecular-weight polypeptide (M) appears to have the function described for the matrix proteins of other paramyxoviruses.


Uptake of minute virus of mice into cultured rodent cells. The uptake of minute virus of mice into cells in tissue culture was examined biochemically and by electron microscopy. Cell-virus complexes were formed at 4 degrees C, and uptake of virus was followed after the cells were shifted to 37 degrees C. The infectious particles appeared to enter cells at 37 degrees C by a two-step process. The first and rapid phase was measured by the resistance of cell-bound virus to elution by EDTA. The bulk of the bound virus particles became refractory to elution with EDTA within 30 min of incubation at 37 degrees C. The infectious particles became resistant to EDTA elution at the same rate. The second, slower phase of the uptake process was measured by the resistance of infectious particles to neutralization by antiserum. This process was complete within 2 h of incubation at 37 degrees C. During this 2-h period, labeled viral DNA became progressively associated with the nuclear fraction of disrupted cells. The uptake of infectious virus could occur during the G1 phase of the cell cycle and was not an S phase-specific event. The uptake process was not the cause of the S phase dependence of minute virus of mice replication. In electron micrographs, virus absorbed to any area of the cell surface appeared to be taken into the cell by pinocytosis.


Urachal diverticulum in a 3-year-old boy. A case of a urachal diverticulum that presented as a right lower quadrant mass in a 3-year-old boy is reported. There was no urinary tract infection or obstructive uropathy in this patient. Classification of urachal anomalies is reviewed.


Salmonella in wildlife from Trinidad and Grenada, W.I. Forty-four of 219 animals from Trinidad and Grenada, W.I., yielded 20 serotypes of Salmonella, 16 of which are known to have been associated with human infection in the United States in recent years. Toads (Bufo marinus) provided the greatest number of isolates. Other carriers were mammals, vultures, lizards, a tree-frog and a cave cockroach.


Experimental infection of dogs with Sarcocystis from wapiti. Ten domestic dogs became infected with Sarcocystis when fed simple portions of heart, esophagus and diaphragm from a two-year-old female wapiti (Cervus canadensis). The prepatent period was 14 days in all exposed dogs; the patent period ranged from 8 to 20 days. Neither the 10 control dogs, nor two dogs fed sporocysts collected from the infected dogs passed sporocysts within the study period. Sporocysts averaged 16.5 by 11.1 micron in size.


Pathomorphologic findings in short-tailed voles (Microtus agrestis) experimentally-infected with Frenkelia microti. Following oral infection of Microtus agrestis with sporocysts of Frenkelia microti, transient focal necrosis and cellular infiltrations in the liver, hyperplasia of lymphoid organs, and inflammatory infiltrations in the heart, pulmonary veins, skeletal muscles and brain occurred during the first asexual multiplication period of the parasite in the liver. Frenkelia cysts were first observed in the brain 23 days after infection.


Parasite-related lesions in a bearded seal, Erignathus barbatus. A free-ranging adult male bearded seal (Erignathus barbatus) killed by polar bears was salvaged and examined at necropsy. Significant findings included diffuse intrahepatic bile duct fibrosis and chronic cholangitis; multiple nodules of chronic fibrosing pancreatitis; and gastric ulcers. Trematode eggs (Family Campulidae) were found in the pancreatic nodules. These eggs and the trematodes that produced them probably caused the pancreatic and liver lesions. Phocanema decipiens and Contracaecum osculatum were found in the stomach lumen; several P. decipiens were attached to gastric ulcers. Sarcocystis sp. is reported for the first time in a bearded seal.


Multilevel care. A Veterans Administration initiative in health care cost control. The Veterans Administration's (VA) Department of Medicine and Surgery, the largest of the civilian federal health care systems, is under continuous White House, congressional, and public pressure to provide greater accountability and increased cost-effectiveness for its $5 billion-plus annual budget. Responding to these pressures, the VA is evaluating in ten of its 172 medical centers a new system of resource allocation and financial management known as a multilevel care system. This is a major VA intiative with highly important implications for both the VA and the private health care sector.