Quantification of computer analyzed serial EEGs from stroke patients. While considerable studies have been conducted by others in automation of the EEG, this method does appear to hold promise for quantitative evaluation and automation of the EEG in the CVA patients or those with similar neurologic deficits. Patients were followed up to 1 year post-ictus. The computerized evaluation of serial records in the CVA patients showed trends which were similar to the percent disability of the patients who were graded neurologically. The patients included in the study were those with middle cerebral artery, carotid and vertebro-basilar defects. A substantial increase in the low frequency activity with a concomitant decrease in high frequency activity was observed in the CVA patients. The results observed in these patient examples demonstrate the utility and efficacy of one method of automated analysis of serial EEGs. The parameters defined by this study have proven to be consistent and effective measures. Since these parameters are objective, they are not affected by patient histories and hence, can be utilized to track the ongoing EEG activity and the accompanying clinical state regardless of the disease entity or the course of therapy pursued. The addition of automated objective quantitative measures to EEG analysis provides a dimension not currently available to the clinician.


Amplitude changes in the visual evoked cortical potential with backward masking. A series of three experiments examined backward visual masking effects and visual evoked cortical potential correlates of such masking. In Experiment I, which was concerned with the effects of sequential sets of like stimuli (grids), it was found that when backward masking occurred, it was accompanied by decreased visual evoked potential (VEP) amplitudes. Experiment II replicated the first experiment using different stimuli (letter B's) and several new subjects. Again, backward masking was accompanied by decreased VEP amplitudes. In Experiment III, which examined the effects of sequential sets of unlike stimuli (B's and grids), it was found that when earlier stimuli differed in configuration from later stimuli, there was an absence of backward masking and VEP amplitude changes also failed to appear. Thus, when backward masking did not occur, no changes in the VEP were observed. The results are discussed in terms of interactions between excitatory and inhibitory activities produced at the visual cortex by the earlier and subsequently presented stimuli. Various concepts regarding the visual system mechanisms involved in backward masking, including integration and interruption hypotheses, may help to explain the complexity of findings obtained thus far in studies using VEP correlates of backward visual masking.


Distribution of skin potential responses according to states of sleep during the first months of life in human babies. Sixty polygraphic records were made: 16 in newborns and 44 in infants aged from 1 to 25 weeks. 1. The stage of development of the central nervous system is better defined by criteria such as spindle maturation or skin potential response (SPR) distribution according to stages of sleep, then by chronological age; SPR prevalence during "quiet" sleep ("adult" distribution) is not apparent before 12 weeks of age. Classification according to the presence or absence of spindles during quiet sleep shows that: (a)in the group without spindles (1-6 weeks old), SPRs are more numerous during "active" sleep than in newborns; (b)in the group with sporadic spindles (3-11 weeks old) the SPR mean frequency is about the same in "active" and in "quiet" sleep; (c)SPR prevalence during "quiet" NREM sleep ("adult" distrbution) appears clearly in the group with spindles present throughout "quiet" sleep (10-25 weeks old). On the contrary, no relation was found between SPR distribution and the mode of onset of sleep. 2."Active" REM sleep seems to mature earlier than "quiet" NREM sleep. Our studies show that, in active sleep, SPRs increase significantly during the 1st month of life; after that, their frequency does not change any more. On the contrary, SPRs in quiet sleep increase throughout the whole period studied. This increase is also parallel to sleep spindle maturation.3. During transitional periods SPR frequency is between that of active and that of quiet sleep. There is no significant difference between SPR frequency during the first and the second sleep cycles.


Human sleep and 5-HTP. Effects of repeated high doses and of association with benserazide (RO.04.4602). On account of the conflicting data concerning the action of 5-HTP on human sleep, repeated high doses were administered to three healthy subjects. The paradoxical sleep time and percentage tend to show a decrease during the 2nd week of treatment, followed by a rebound effect after the end of the treatment. In two subjects who initially had a small amount of slow wave sleep (SWS), the treatment induced an increase in SWS that persisted after the end of the treatment. Similar results were obtained after administration of 5-HTP together with RO.04.4602.


A warning device which indicates the stoppage of paper in an ink-writing recorder. A simple and inexpensive warning device is described which indicates when the paper has stopped moving beneath the pens of an inkwriting polygraph. The device operates on the principle that the electrical resistance of the paper under the recorder pen is markedly reduced when the paper stops moving and is made wetter by the ink. A buzzer sounds within a few seconds of the paper stopping and so reduces the possibility of failing to record otherwise irretrievable information, for example during all-night recordings of the EEG in sleep research.


Evoked potential analysis: on-line signal optimization using a mini-computer. An automated computer-based system is described for the analysis of evoked potentials. All procedures are carried out in real-time A small computer performs the following functions; timing of random stimulus presentation, rejection of artifact contaminated responses, collection of digital data, computation of averaged evoked responses, computation of the Wiener filter, storage of the filtered and unfiltered averages and display of the resultant averages. The Wiener filter as described by Walter (1969) and Doyle (1975) is used to improve the estimate of the evoked potential by discriminating against frequencies likely to be contaminated with noise. The defining equation for the Wiener filter states that information at any frequency is to be weighted by the ratio of the power known to be in the signal (response) at that frequency over the corresponding power known to be in both the signal (response) and the noise (background EEG) at the same frequency. The technique requires the computation of the Fourier transform for each response in order to produce the power spectra necessary for the Wiener filter. Earlier reports dealing with this technique have usee large computers to analyze the evoked potential data off-line. The system described here allows for greater routine utilization of this powerful technique and the concomitant automated rejection of artifact contaminated responses. Highly improved estimates of the evoked potential are resultant using a minimal number of stimuli.


Seizure moitoring: a new tool in electroencephalography. A seizure monitoring system, based on telemetry and computer techniques, has been developed to provide a reliable means of recording the patient's spontaneous seizures. It allows a patient's EEG to be monitored for hours or days with 16 channels of EEG from surface, sphenoidal or stereotaxically implanted depth electrodes, from any ward or room within the hospital. The EGG is delayed in time by up to 4 min by a mini-computer so that either the patient can push a button when he experiences his aura or others can push the button when they observe his seizure. Since the delayed EEG is written out, the preictal, ictal and postictal are recorded on paper, without many pages of uneventful interictal information. During the past 16 months, the seizure monitoring system has been used as a clinical service to examine patients with surface electrodes (42), sphenoidal wires (25) and depth electrodes (7) during 146 recording sessions for over 3200 h of monitoring time while over 200 seizures have been recorded.


Diazepam modification of evoked and spontaneous lateral geniculate activity. The effects of diazepam on evoked and spontaneous activity of dorsal lateral geniculate nucleus (dLGN) principal (P) and inhibtory (I) cells were examined in rats. In the majority of P cells tested both spontaneous and evoked activity were suppressed following diazepam treatment with these effects being altered little by a pentylenetetrazol (Metrazol) challenge. In two P cells, which were offresponding to photic pulse stimulation, evoked and spontaneous activity was enhanced by diazepam but again these effects were altered little by the Metrazol challenge. I cell spontaneous activity was suppressed by diazepam and augmented by the Metrazol challenge; however, post-stimulus activity was relatively unaffected by either treatment. These results were discussed in terms of support for a functional re-evaluation of the rat dLGN.


Androgen of the human prostate.

Androgen of the human prostate. The relationship between the concentrations of total and apparent free testosterone in the plasma and the levels of testosterone (T), dihydrotestosterone (DHT) and 5 alpha-androstan-3-alpha, 17 beta-diol (DIOL) in 13 benign hypertrophic and 6 carcinomatous prostates was studied. The androgen concentration within both types of glands was nearly 4-fold that in the blood but bore no direct relationship to the blood level. About 75% of the androgen in the tissue was DHT. The most striking finding was that, in spite of a 25.5% less concentrated pool of apparent free testosterone in the blood, the level of T and its metabolites in the cancer tissue was 29% above that in the samples of benign hypertrophic prostate.


Immunochemical study of the peptidoglycan of gram-negative bacteria. The specificity of antibodies directed against the peptidoglycan of gram-negative bacteria was studied. The peptidoglycans of Proteus vulgaris, Escherichia coli, Moraxella glucidolytica, Neisseria perflava, give identical precipitin reactions. By means of inhibition studies with various peptidoglycan subunits and synthetic peptides, it was shown that the antibodies are essentially directed against the peptide moiety of the peptidoglycan: L-Ala-D-Glu (L)-mesoA2pm-(L)-D-Ala, that the peptide reacts better with antibodies when it is not cross-linked, and that the C-terminal portion-meso-A2pm-D-Ala of the peptide is immunodominant. These results explain the immunological identity of the peptidoglycans of gram-negative bacteria, which possess the same peptide subunit. Only weak cross-reactivity was observed with the peptidoglycans of gram-positive bacteria (Streptococcus faecium, Micrococcus lysodeikticus, Corynebacterium poinsettiae) where meso-diaminopimelic acid is replaced by L-lysine or L-homoserine. However, the peptidoglycan of Bacillus megaterium which possesses the same peptide subunit as gram-negative bacteria, gives only a reaction of partial identity with these bacteria. This result suggests the presence on the peptidoglycan of gram-negative bacteria, of other undefined antigenic determinants.


Dinitrophenol, dicoumarol and pentachlorophenol as inhibitors and parasite substrates in the ATP phosphoribosyltransferase reaction. Adenosine-triphosphate phosphoribosyltransferase from Escherichia coli is inhibited by dicoumarol and pentachlorophenol in competition with ATP. Ki was approximately 60 muM for dicoumarol and 50 muM for pentachlorophenol. Carbonylcyanide m-chlorphenylhydrazine did not seem to have any kinetic effect. Dicoumarol is bound to the extent of 6 sites per enzyme hexamer with a dissociation constant Kd of 50 muM. Dicoumarol and pentachlorophenol partly prevent the binding of ATP and AMP to the transferase. The reverse reaction is inhibited by dicoumarol and pentachlorophenol without changes in [s]0.5 for phosphoribostladenosine trophosphate. Dicumarol, dinitrophenol and pentachlorophenol diminish the yield of phosphoribosyladenosine triphosphate in the transferase reaction apparently by acting as parasite substrates; carbonylcyanide m-chlorophenylhydrazone had no effect.


Paromomycin and dihydrostreptomycin binding to Escherichia coli ribosomes. Paromomycin binds specifically to a single type of binding site on the 70-S streptomycin-sensitive Escherichia coli ribosome. This site is different from that of dihydrostreptomycin since paromomycin binds to streptomycin-resistant ribosomes and sine dihydrostreptomycin does not compete for paromomycin binding. Paromomycin binding, unlike dihydrostreptomycin binding, is independent of changes in ribosome concentration but influenced by magnesium ion concentration. Moreover, paromomycin does not bind to the 30-S subunit of the streptomycin-sensitive ribosome, except in the presence of dihydrostreptomycin, which probably induces the conformational changes necessary for a paromomycin binding site. This induction does not occur with streptomycin-resistant ribosomes. Neither antibiotic binds to the 50-S subunit. In general, binding of the one antibiotic increases the number of sites available for binding of the other. Both antibiotics exhibit marked non-specific binding at high antibiotic/ribosome ratios. Competition studies have enabled the classification of other aminoglycosides according to their ability to compete for the paromomycin and dihydrostreptomycin binding sites. Derivatives structurally related to paromomycin compete for its binding, the degree of competition being related to antibacterial activity, but do not compete for dihydrostreptomycin binding; they, on the contrary, increase the number of dihydrostreptomycin binding sites. Neither gentamicin nor kanamycin derivatives, which induce a high level of misreading, nor kasugamycin and spectinomycin, which do not induce misreading, compete for paromomycin or dihydrostreptomycin binding sites. Other sites may be involved in the binding of these aminoglycosides and in inducing misreading.


Biosynthesis of gramicidin S with the aid of dipeptides by gramicidin S synthetase. Dipeptides L-phenylalanyl-proline, D-phenylalanyl-proline, prolyl-valine, valyl-lysine, lysyl-leucine and leucyl-phenylalanine, derived from the sequence of gramicidin S, are substrates of the gramicidin S synthetase. When any of these dipeptides are used to replace the two corresponding amino acids in the reaction assay, cyclodecapeptide antibiotic synthesis occurs, and requires the whole multienzyme system. Active esters, like the thiophenyl and p-nitrophenyl esters of D-phenylalanyl-proline are unable to promote gramicidin S biosynthesis with the gramicidin S synthetase system or with the heavy enzyme alone.


Editorial: Chromosomes and human neoplasms. Achievements using new staining techniques. Numerical and structural chromosome aberrations are frequently found in neoplastic cells. As demonstrated by the new chromosome banding techniques these aberrations are not random, but tend to show a specific occurrence. A model example is the leukemias where many cytogenetical investigations have been done to date. In leukemia chromosome analysis serves the following purposes: to identify a neoplastic process, to confirm and strengthen the hematological diagnosis, for the early diagnosis of transformation from a chronic leukemia into its blastic phase and for following up the clonal evolution of a leukemic cell line. In the discussion of chromosomes and neoplasms it must be mentioned that individuals demonstrating chromosomal instability and some trisomic patients show a greater tendency toward the development of a malignancy. Malignancy is primarily a cellular phenomenon caused by a disturbance in cellular regulation, whose fine events are not known. Therefore the exact role of the chromosomes in neoplastic processes cannot be stated. From experimental investigations it appears that the affected chromosomes carry cell growth regulating factors and also that a specific aberration is the result of the action of a specific agent.


Inhibition by 5-hydroxytryptamine of anaphylactic histamine release from bovine granulocytes. A suspension of granulocytes from calves sensitised to horse plasma released histamine when exposed to the antigen. Histamine release was strongly inhibited by 5-hydroxytryptamine in a dose-related fashion. 5-HT is known to be released from bovine lungs in vitro, and may regulate the release of histamine in vivo during anaphylaxis.


Degradation of encephalitogenic protein in aerobic ascorbic acid solutions. Aerobic ascorbic acid solutions are capable of extensively cleaving the peptide chain of the myelin basic protein. Cleavage occurred most readily with C-terminal to aspartic acid, serine, threonine, glutamic acid and leucine residues.


Localization of bovine pancreatic polypeptide (BPP)-like immunoreactivity in rat pancreatic monolayer culture. Antiserum to bovine pancreatic polypeptide (BPP) has been used for immunofluorescent staining in the light microscope. With this technique it is possible to detect the presence of specific cells in monolayer culture from neonatal rat pancreas which contain BPP or a closely related peptide.


Preparation of specific antiserum against Rana esculenta pre-alpha lens crystallin. Specific antiserum against Rana esculenta lens pre-alpha crystallin was prepared in a rabbit by injecting antigen-antibody precipitate of this crystallin obtained from immunoelectrophoresis of esculenta total soluble lens proteins against homologous antiserum.


Hemagglutination of neuraminidase-treated human erythrocytes by Leishmania enriettii infected guinea-pig sera. Sera from guinea-pigs infected with the protozoan parasite Leishmania enriettii showed higher hemagglutination (HA) titres for neuraminidase treated human erythrocytes than those of normal guinea-pig sera. This HA activity was associated mostly with the 19S fraction of the immune serum and could be absorbed out with an antigenic fraction of the parasite membrane. Antigenic determinants involved in this HA reaction consisted of, at least, beta-D-galactosyl or lactosyl residues.


2-Phenylethanol, a presumed sexual stimulant produced by the male cabbage looper moth, Trichoplusia ni. A sex pheromone produced by male cabbage looper moths, Trichoplusia ni (Hübner), has been isolated from the genital scent brushes and identified as 2-phenylethanol. It is shown conclusively to elicit specific behavioural responses in the female (such as wing vibration and abdominal elevation), as determined by a novel behavioural laboratory bioassay. This is taken as further evidence that the male pheromone of T. ni acts as a sexual stimulant (aphrodisiac) prior to mating. 2-Phenylethanol represents the first identification of a genital scent brush pheromone in the family Noctuidae, and of a male pheromone in the subfamily Plusiinae.


[Autoantibody to luteinizing hormone in patient with systemic lupus erythematosus (author's transl)]. One hundred microgram of luteinizing hormone releasing hormone was intravenously injected into 19 female patients with systemic lupus erythematosus and plasma luteinizing hormone levels before and after the injection were determined with a method of solid phase radioimmunoassay. In 4 patients among them, the basal levels of luteinizing hormone were abnormaly high. A question about the existence of autoantibodies to luteinizing hormone in the patient's plasma raised to explain the reason for the high level of luteinizing hormone. The results from 3 experiments for solution of this question were as follows. 1) The patient's plasma was proved to have a binding activity with labeled luteinizing hormone. 2) This binding activity is specifically inhibited by purified luteinizing hormone. 3) This binding activity was located in gamma-globulin fraction. From the results, the existence of autoantibodies to luteinizing hormone in the 2 patients' plasma was concluded.


Avian infectious bronchitis vaccine: primordial or derived virus? Vaccine formulations of virulent, modified/attenuated, or inactivated mono- or multivalent types of chicken embryo-propagated avian infectious bronchitis virus (IBV) at different passage levels and administered by non-parenteral routes have been used for immunization of chickens against the disease. Two doses of vaccines are generally used: 1) high level virus of lessened virulence, antigenicity and immunogenicity as "derived" from its parental virus via serial passage in and selective adaptation  to the chicken embryo, and 2) low passage level virus with the above properties more closely related to the greater potency of the "primordial" parental virus in nature. The efficacy of vaccines with contemporary or candidate virus has been interpreted from a variety of criteria among which are: 1) clinical response, residence and dissemination of virus, 2) in vivo and in vitro local tissue response of tracheal epithelium, 3) protection against virulent homologous or heterologous types of IBV, and 4) induced secretory and humoral antibody. Vaccines have been economically useful and of epizootiological value but the greatest difficulty is the uncertainty of protection because of the changing antigenic, immunogenic and other properties of existing virus and of newly emerging types. With a complex virus such as IBV, the opportunities for natural selection and evolution of new or modified antigens via association with antibody of immune or of partially immune hosts are as enormous as antigenic lability resulting from fragmentation of the primordial genome under natural  and/or artificial environmental stress. The present day IBV might be progeny from a unique pool of primordial genome via mutation and host-induced variation, or even persistence of primordial virus as an infinitesimal portion of the population.


Cellular localization of intestinal calcium-binding protein in pig duodenum. The gamma-globulin fraction of a rabbit antiserum against porcine intestinal calcium-binding protein (CaBP) was used in an immunoperoxidase method to study CaBP localization in porcine duodenal tissue. Specific immunostaining, indicative of the presence of CaBP, was noted within the cytoplasm of the duodenal epithelial cells. No CaBP was detected in goblet cells or in the subepithelial layers. When the specific antibody was replaced by either nonimmune rabbit gamma-globulin or when the specific antibody was preadsorbed with excess CaBP, no positive immunostaining was seen. Our studies lend support to the hypothesis that CaBP may function in intracellular calcium transport.


Blood glycoprotein levels in diabetes mellitus. Five glycoproteins have been measured in the blood of 145 diabetic patients with and without clinical evidence of complications. Patients with diabetic complications have higher glycoproteins levels particularly when expressed as a ratio to serum albumin levels. In 32 pairs of patients matched for age, sex, body weight, duration and treatment of diabetes, significantly higher haptoglobin, fibrinogen and caeruloplasmin levels were associated with the presence of diabetic complications, but blood glucose levels were not significantly different. Beta-lipoprotein levels were positively correlated with age and alpha2-macroglobulin levels with the duration of clinical disease, but the type of antidiabetic therapy administered did not significantly alter glycoprotein levels. It is suggested that rising levels of certain glycoproteins in the blood of diabetic patients may indicate the development of diabetic vascular complications, but a prospective study is required before it can be decided whether this change predates the clinical appearance of the complications.


[Cardiac myxoma: anatomic and clinical description of four cases (author's transl)]. Four cases of cardiac myxoma diagnosed among 68 solid intracavitary cardiac formations are described. The myxomas (all unexpected autopsy findings of the left atrium) are illustrated by their clinica, macroscopic, histologic and histochemic characteristics. The differences between heart myxomas and thrombi are briefly outline and discussed.


Influence of steroid-receptor complexes on transcription by human hypertrophied prostatic RNA polymerases. The effects of 5alpha-dihydrotestosterone-receptor complexes on transcription in human hypertrophied prostate tissue were studied in a cell-free system reconstituted from the various subcellular fractions prepared from specimens of the diseased gland. Two major RNA polymerase species were isolated from human hypertrophied prostate. These were designated A and B and were distinguishable by their preference for divalent cations and their sensitivity to salt and alpha-amanitin. Moreover, RNA polymerase B, but not RNA polymerase A, could effectively transcribe a prostate chromatin template. Any enzyme activity endogenous to some chromatin preparations was shown to be characteristic of RNA polymerase B. 5alpha-Dihydrotestoterone-receptor complexes were transferred into prostatic chromatin both steroid- and tissue-specifically. The association of steroid-receptor complexes with chromatin produced changes in template activity and increased the transcription of the chromatin by exogenous and endogenous RNA polymerase B. With a number of specimens, however, there was considerable variation in accessible cytoplasmic receptor sites, uptake of steroid-receptor complexes by chromatin preparations, the template activity of the chromatin and its response to steroid-receptor stimulation. Nevertheless, the transcription characteristics of human hypertrophied prostatic chromatin appear to be influenced by steroid-receptor complexes, and the extent of the response to added complexes would undoubtedly be governed by pre-existing complexes having had an earlier effect.


Immune response in fulminant viral hepatitis, type B. Serial serum samples from the time of exposure until fatal outcome in 3 patients with fulminant viral hepatitis, type B, were examined for the presence of the antigens associated with hepatitis, type B, and their corresponding antibodies. The titers of hepatitis B surface antigen (HBsAg) were found to decrease by greater than 50% before death. Antibody to surface antigen (anti-HBs) was not detectable in any sample. Patterns of antibody to core antigen (anti-HBc)), HBsAg subtype "e" antigen, and anti- "e" were unremarkable, and could not be distinguished from those that might occur in many self-limited cases of hepatitis, type B. A rise in alpha-fetoprotein before demise suggests that late but inadequate liver regeneration occurred in these patients.


[Identification of simian chromosomes by the differential staining method with Romanovskii-Giemza stain. II. Papio hamadryas]. By means of differential staining with the Romanovsky - Gimsa dye the distribution of G-bands was studied in chromosomes of bone marrow cells and of cultured peripheral blood lymphocytes in Papio hamadryas. All the homologous chromosome pairs were identified. It was shown, as compared Papio hamadryas karyotypes with that of Macaca mulatta, that patterns of most chromosomes differentiation were similar. Certain differences between these species were observed in the character of differentiation of chromosomes 1 and 5 and also in parameters of chromosomes 2 and 21 (X).


[Effect of substrate and small doses of cortisone on the induction of Tryptophan 2,3-dioxygenase (author's transl)]. A number of enzymes are induced by steroid hormones. In this paper the reaction of tryptophan 2,3-dioxygenase is further analyzed. In particular we show in which way the substrate and low doses of cortisone cause an induction. 1) For the induction of tryptophan 2,3-dioxygenase in adrenalectomized rats by 2.5 mg cortisone/kg, the presence of the substrate is necessary as well. Under these conditions an induction of the enzyme can already be registered in the presence of 12.5 mg L-tryptophan/kg. 2) In animals treated before with cortisone, the enzyme maximum appears 30 min after L-tryptophan injection, The enhancement of enzyme activity in animals which are treated with 2.5 mg cortisone/kg before is blocked by actidione only until 30 min after L-tryptophan injection. 3) Experiments with antibodies in animals treated with a low dosis of cortisone show that L-tryptophan acts mainly via enzyme degradation or the saturation with the coenzyme hematin, respectively.


Banding pattern observed in human chromosomes by the modified BSG technique. A successful modification of the BSG technique to reveal C and R bands simultaneously in human chromosomes is described. Conventional air dried preparations were treated first with 0.1 N HCl for 30 min at room temperature, then denatured in freshly prepared 3% aqueous solution of Ba(OH)2SH2O for 10 min at 50 degrees C. After rinsing, the slides were incubated for 1 h at 60 degrees C in 2 X SSC, and stained with Giemsa. The striking intense staining pattern could be observed in chromosome No. 19. The factors involved in the present technique were analyzed changing the concentrations of the reagents and the treatment time. It was evident that R, T and C bands correspond to a progressive destruction of the chromosome structure mainly by the Ba(OH)2SH2O solution.


CT banding of human chromosomes: description of the banding technique and some of its modifications. A technique is described for staining centromeric areas and reverse, mainly telomeric bands in human chromosomes. With this "CT" technique karyotyping of C-banded metaphases is possible without previous or subsequent use of other banding methods. The method consists of an alkaline pretreatment at 60 degrees C with Ba(OH)2, followed by salt incubation in 2 X SSC at 60 degrees C and staining with the cationic dye "Stains-all". In a series of experiments the influence of the variables in the procedure was studied, with the following main results: 1) Ba(OH)2 treatment alone and subsequent staining produces a distinct reverse banding pattern in which the secondary constriction of chromosome 9 is positive. 2) The 2 X SSC incubation in the CT procedure causes the Ba(OH)2 induced reverse bands to become weaker; the centromeric regions, however, become very prominent. 3) If the temperature of the 2 X SSC treatment is raised to 85 degrees C, the CT technique results in a specific staining of the short arm regions of some probably variant acrocentric chromosomes. The interphase nuclei of individuals possessing such acrocentrics usually show very distinct chromocentres after this treatment; in the polymorphs these chromocentres are often situated along the nuclear membrane. The mechanisms which may form the basis of the staining results obtained, and the possible significance in human cytogenetics of the techniques described, are discussed briefly.


Q-banding of human chromosomes after BUdR and BCdR treatment. Two different Q patterns were found in BUdR and BCdR treated chromosomes of human lymphocyte cultures: X-type pattern, in which Giemsa and quinacrine banding both are reversed; Y-type pattern, in which Q-banding remains conventional in spite of reverse G-banding. Possible mechanisms of these findings are discussed.


Chromosome analysis on 930 consecutive newborn children using quinacrine fluorescent banding technique. Chromosome analysing using quinacrine fluorescence was performed on 930 consecutive newborn infants. The total incidence of major chromosome aberrations including numerical changes of the sex chromosomes, and structural changes of autosomes, was 0.54%. Incidence of XYY (0.4%) and XXY (0.2%) were relatively higher as compared to other studies. About 0.75% of the newborn infants were found to have a variable bright fluorescent band located on the proximal area of the short arm (p11) rather than on the proximal long arm (q11) of chromosome No. 3. Attempts were also made to record the variable fluorescent regions on 7 autosomes and the Y chromosome.


Alpha-fetoprotein concentration in cord blood from twins and from a set of quadruplets--a case of superfetatio? Alpha-Fetoprotein has been determined in cord blood fron mono- and dizygotic twins and in a case of quadruplets. The possibility of superfetatio has been evaluated by the clinically calculated gestational ages and by the alpha-fetoprotein levels in cord blood, especially in a case of quadruplets.


The influence of particulate carriers and of mineral oil in adjuvants on the antibody response in rabbits to human gamma globulin. A single inoculation of rabbits with human gamma globulin, administered with polystyrene latex particles and mineral oil as adjuvant, gave rise to at least the same level and duration of antibody formation as could be achieved with classical Freund's adjuvant made "complete" with killed mycobacteria.


Immunological release of histamine from bovine leucocytes. Unusual adrenergic modulation. Sensitized bovine granulocytes release histamine when exposed to specific antigen. In comparison with in vitro systems in several other species, the modulation of this release by adrenergic agents is unique. Beta-Adrenoceptor stimulation potentiates (rather than inhibits), whereas alpha-adrenoceptor stimulation inhibits (rather than potentiates) histamine release. Adrenaline, which is generally considered to be a physiological antagonist of the anaphylactic reaction, potentiated histamine release in this study. Dopamine, which is present in high concentration in bovine mast cells, was without effect. The results are discussed in terms of the possible role of granulocytes in bovine hypersensitivity.


[Blood regeneration after blood substitution with dextran 60 or hydroxyethyl starch of an identical volume]. Male rats of the Wistar strain survived an isovolaemic blood volume replacement down to a haematocrit value of 20% using commercial dextran 60 or hydroxyethyl starch. No significant changes of the blood volume have been observed. 14 days after treatment, the blood values were found to be practically within the normal range. As early as 3 days after the blood exchange, a significant increase in reticulocytes has been noted which reached its maximum on the 7th day. In both series the blood regeneration was identical.


Isolation from human gastric juice of an antigen closely related to the carcinoembryonic antigen. A carcinoembryonic antigen-like antigen(s) (CELIA) was isolated from normal human gastric jucie by immunoadsorbents and gel chromatography. The isolated CELIA reacted identically to CEA in immunodiffusion testa against anti-CEA and anti-CELIA. Other experiments suggested subtle differences between CELIA and CEA. Two out of six CELIA preparations tested in immunodiffusion did not react in a completely identical manner with CEA.In gel filtration, CELIA eluted slightly after CEA. Absorption of anti-CEA with CELIA resulted in antisera with greatly reduced reactivity to CELIA, but retained anti-CEA activity. Such antisera may be helpful in attempts to increase the cancer specificity of the CEA test.


Detection of human tumor-associated antigens by the leukocyte migration in agarose assay. Twenty-three of 36 (64%) lung cancer patients, 19 of 36 (54%) melanoma patients and 18 of 27 (66%) sarcoma patients tested in the leukocyte migration in agarose assay against soluble extracts of histologically similar tumors showed significant inhibition of leukocyte migration. Reactivity to extracts of dissimilar tumors was low. Sera of only 1/13 (7%) lung cancer patients, 2/19 (10%) melanoma patients and 7/21 (33%) sarcoma patients were inhibited by extracts of histologically dissimilar tumors. Only 7-9% of cancer patients reacted to paired extracts of normal tissue from the tumor donors. An average of 13% of sera from normal controls reacted to tumor extracts. Stage of disease and mode of therapy appeared to have little effect on overall reactivity in this assay, although the number of patients within the various categories was small for purposes of statistical analysis. The leukocyte migration in agarose assay shows a sensitivity and specificity to tumor-associated antigens comparable to that of the older capillary tube method in general use and may facilitate performance of migration inhibition. This assay may not be useful as a prognostic test due to the lack ofcorrelation with stage of disease and treatment modality. However, its high specificity and economical use of tumor antigen suggest applications in tumor antigen purification. The use of soluble tumor antigen preparations may make it possible to purify these antigens further to increase specificity and reactivity.


Interactions of murine leukemia virus (MuLV) with isolated lymphocytes. III. Alterations of splenic B and T cells in Friend virus-infected mice. Lymphoid tissues of mice infected with murine leukemia virus (Friend) (MuLV-F) were examined for the presence of cellular markers of MuLV-F infection. The Friend virus-associated cell membrane antigen (FVMA) and the virus group-specific antigen (GSA) were detectable on cells from the spleen and, to a lesser degree, on cells from the bone-marrow. In contrast, neither FVMA nor GSA was found in cells from the thymus. Alterations in the B-cell and T-cell spleen populations of MuLV-F-infected mice were then studied. The proportion of Ig-positive cells declined from the initial 45% (in non-infected controls) to about 10% after 2 weeks of infection. A similar decline of theta-positive cells was noted. However, complement-bearing cells (EAC rosettes) declined even more rapidly and became undetectable in the second week after infection. The treatment of spleen cells from MuLV-F-infected mice with anti-FVMA serum plus complement in vitro reduced the number of detectable Ig-positive cells, specifically, whereas the number of theta-positive cells remained unchanged. Furthermore, B and T cells from spleens of infected mice were separated on an affinity column with anti-Ig antibody-coated beads. The initial cell suspension contained about 45% FVMA-positive cells, about 40% Ig-positive cells and about 40% theta-positive cells. Ig+ cells were retained on the column. The theta-positive cell fraction was collected in the eluate and contained very few FVMA-positive cells with some "null" cells. Most of the FVMA-positive cells were retained on the column, which strongly suggested that they were B cells. These results confirm the previous experiments which showed the selective infections of purified splenic B cells by MuLV-F in cultures.


Murine neuroblastoma clones with varying degrees of C-type virus expression. Three tissue culture clones of neuroblastoma cells derived from the C-1300 tumor in strain A/J mice were found to contain type-C virus-specific gas antigen. None of the clones spontaneously released mouse-tropic type-C viruses although one clone, N-4, spontaneously released a xenotropic virus. Two clones, NB-2A and N-4, could be induced by treatment with 5-iododeoxyuridine and dexamethasone phosphate to produce B-tropic type-C virus, but clone N-18 failed to release either ecotropic or xenotropic virus under several different induction conditions. Karyotype analysis did not reveal specific chromosome deletions in clone N-18.


Serologically detectable specific and cross-reactive antigens on the membrane of a polyoma virus-induced murine tumor. With the aid of an assay measuring complement-dependent cytotoxocity mediated by syngeneic antibodies, we performed a serological analysis of surface antigens of a polyoma-virus-induced murine tumor (SEYF-a). In vivo propagated SEYF-a ascites tumor cells expressed a specific membrane antigen in addition to various other cross-reacting antigens. Among these we could identify at least four separate specificities. Two of these were present on MuLV-induced lymphoma cells, the first on Moloney-virus-induced YAC cells and the second on Gross-virus-induced GHA cells. The third cross-reacting antigen was detected on EL-4 cells. At least one additional specificity was present on two methylcholantthrene-induced murine sarcomas. Normal syngeneic lymphoid cells were insensitive to cytotoxicity mediated by the anti-tumor antisera. Quantitative and perhaps also qualitative differences between that antigenic expression of in vivo propagated on cultured SEYF-a cells were indicated. These studies show that hyperimmune sera produced in syngeneic mice against transplanted tumors may contain a considerable number of antibody specificities, only some of which are specific for the tumor. Furthermore, the results also suggest that polyoma-virus-induced tumors may possess individually distinct antigenic specificities, over and above the known cross-reacting TSTA or TSSA type antigen.


Specific anti-fibroblast cytotoxicity of antibodies to fibroblast surface antigen. An antiserum to SF antigen was shown to be selectively cytotoxic to two human fibroblast lines, while two tumor cell lines (one melanoma and one Hela cell line) were resistant. The selective cytotoxicity was also demonstrable in mixtures of fibroblasts and tumor cells. The cytotoxic effect was abrogated by admixture of human serum containing SFA.


Dissociation and denaturation behaviour of sesame alpha-globulin in sodium dodecyl sulphate solution. The effect of anionic detergent, sodium dodecyl sulphate, on the major protein, alpha-globulin of sesame seed (Sesamum indicum L.) has been investigated by gel filtration, sedimentation velocity, viscosity, optical rotation, difference spectra and fluorescence measurements. The detergent causes dissociation of the protein first and then denaturation. In the detergent concentration range of .175-4.0 X 10(-"3) M four components are observed in the ultracentrifuge. The specific rotation of the protein increases with the detergent concentration above 2.5 x 10 (-3) M detergent suggesting conformational change; above 8 X 10(-"3) M detergent the value of -[alpha] does not change. The reduced viscosity etared however, increases above .25 X 10(-3) M detergent and does not attain a plateau value. The difference spectrum of the protein indicates that both tryptophan and tyrosine groups have been affected by the detergent. The fluorescence intensity decreases and the maxima shifts towards red in the detergent solution resulting in an "isoemissive point" at 355 nm. The double difference spectra in sucrose-detergent protein system show that below 5-0 X 10(-3) M detergent, the difference absorption and fluorescence spectrum result from the binding of the detergent near the chromophoric groups and are not due to conformational change. Binding studies by equilibrium dialysis indicate the presence of 50 binding sites in the protein and binding constant of 3-0 X 10(3).


Peter Shaffer's Equus--a psychoanalytic exploration. In this paper on Equus, an attempt is made to explore from an analytic point of view the psychodynamics of the protagonist, Alan, and his therapist, Dysart. Both the preoedipal and oedipal aspects are discussed. The main thesis of my remarks are aimed at revealing how the author seized upon a real event, namely the blinding of horses by an English lad, and has woven this into his own fantasies. It is emphasized that the playwright in addition employed various sophisticated ego functions that enabled him to express these fantasies in Greek mythological terms (the oedipal myth and the voyage of Agamemnon). The final work then comprises a true creative effort, since, as I mention in my introductory remarks on the creative process, it employs both primary and secondary process thinking and symbolism. The final synthesis represents an eloquent mythopoetic expression and combination of id and ego (autonomous ego functions).


A cross-cultural test of the Freudian theory of circumcision. Freud's thesis that circumcision is a symbolic substitute for castration as a result of the Oedipus conflict was tested by examining 111 societies (see the note to Table 1). The results show that circumcision is likely to be found in societies in which (1) the son sleeps in the mother's bed during the nursing period in bodily contact with her, and/or (2) the father sleeps in a different hut, reducing his influence on the son as a superior and competing male. Since these two factors seems to intensify the son's oedipal attachment to the mother, the data are compatible with Freud's theory. Bettelheim's theory of circumcision is less fully formulated, and only when we assume that males significantly reduce their vagina envy after puberty rites is his theory compatible with the data.


Study of the lysosomes by vital stains in normal keratocytes and in keratocytes from macular dystrophy of the cornea. The acridine orange is an excellent vital stain for the "in vivo" study of the lysosomal dynamics in cultured keratocytes. In normal keratocytes the lysosomes are mostly grouped around one or both nucleus poles. In keratocytes from macular dystrophy of the cornea there is a hypertrophy of the lysosomal system, which in vain tends to compensate for the consequences of the enzymatic deficiency.


The dynamics and location of axonal transport blockade by acute intraocular pressure elevation in primate optic nerve. Axonal transport in primate optic nerve axons was studied by autoradiography, scintillation counting, and electron microscopy under conditions of short-term intraocular pressure elevation. With elevation of intraocular pressure to 30 mm. Hg below mean arterial blood pressure, blockage of transport was detected within 2 hours by autoradiography and within 1 hour by electron microscopy. The earliest buildup of radioactively labeled protein and ultrastructurally visible cellular organelles was within the scleral lamina cribrosa. The degree of blockage, judged by amount of label buildup, increased with time. Some transported material traversed the lamina cribrosa despite pressure elevation at the level tested. Reversal of transport blockade occurred rapidly after normalization of intraocular pressure.


Chromosomal studies of human prostatic tumors in vitro. Chromosomal studies were performed on primary cultures from human prostatic adenomas and carcinomas. All cells from both tumor types showed diploid chromosomal counts with a normal human karyotype. Chromosomal analysis was also performed on cells of the permanent epithelial line EB 33 derived from a human prostatic carcinoma in this laboratory. In contrast to the primary cultures, hypotriploid chromosome numbers were found during continuous culture for more than 1 year. The range of chromosome numbers increased with time in culture and was reduced to the original model number of 64 by only one animal passage ("nude" mice). Feulgen fluorometry on different passages of the permanent cell line EB 33 showed tetraploidy. This discrepancy to the hypotriploid chromosome number can be explained by different Feulgen hydrolytic properties of the different cell types.


Effect of candicidin and colestipol on the testes and prostate glands of BIO 87.20 hamsters. The oral treatment of groups of aged BIO 87.20 male hamsters, an inbred line of Syrian hamsters that develops spontaneous cystic prostatic hypertrophy, with either the heptaene macrolide, candicidin, or the bile acid-sequestering anion exchange resin, colestipol, resulted in a significant reduction of prostate size and weight. The effect was most prominent in the high dosage (75 mg per kg per day) candicidin group and moderate in the high dosage (400 mg per kg per day) colestipol group. It was observed microscopically that the extreme dilation of the prostatic acini of the untreated control prostate glands was partially prevented by the treatment, resulting in much less distended prostatic acini. Concomitantly, there was a notable decrease of the total cholesterol content of the entire prostate gland and the plasma of the high dosage candicidin group. The relationship of certain aspects of sterol metabolism and the possible prevention of cystic prostatic hypertrophy in this new experimental animal disease model is discussed. The effect of these drugs on the nutritional state of the animal and on the synthesis and metabolism of androgens from the testis are questions that remain to be answered.


Defective cell-mediated immunity in carcinoma of the prostate. Cell-mediated immunity was assayed in 18 patients with a histologic diagnosis of carcinoma of the prostate and in 10 control patients with benign prostatic hypertrophy. In vivo cell-mediated response was assayed by intradermal skin testing to a battery of five delayed hypersensitivity antigens. In vitro cell-mediated immunity was studied by lymphocyte transformation using both heterologous and autologous "serums" and nonimmune rosette formation. Lymphocyte transformation studies were carried out using both specific antigens (Dermatophyton and streptokinase-streptodornase) and a nonspecific mitogen (phytohemagglutinin). Nonimmune rosette formation was performed under numerous temperature ranges. A defect in cell-mediated immunity was found in patients with carcinoma of the prostate. Skin tests demonstrated a significantly increased incidence of energy in cancer patients. Mitogen-induced lymphocyte transformation was significantly depressed when autologous serum from patients with carcinoma of the prostate was utilized in the procedure. It is concluded that defect in cell-mediated immunity does exist in patients with carcinoma of the prostate. This defect is expressed in vivo and in vitro and seems to be mediated by a serum factor.


In vitro cultivation of prostatic epithelium. A tissue culture procedure for in vitro cultivation of human prostatic tissue that utilizes the enzyme collagenase to disrupt the prostate specimen is described. Collagenase effectively disperses the tissue stroma into single cells which are easily separated from the largely intact glandular structures. The isolated glandular fragments then are used to initiate monolayers of prostatic epithelium in vitro.


Fluorogenic substrate turnover in single living cells. Intracellular diffusion properties and enzyme activities in single living cells can be analysed by means of fluorogenic substrates that diffuse into the cells where they are converted into a fluorescent product by an enzymic reaction. The reaction-kinetic analysis of this process as a system of consecutive reactions provides information on the diffusion of the substrate into the cells, on intracellular enzyme activities and on the efflux of the fluorescent product. Separation of diffusion and enzyme-mediated processes is obtained by inducing specific changes of the cellular membrane using gramicidin D. A model for the functional interpretation of the experimental findings is proposed. Application of this method as a viability test for freshly prepared and frozen platelets is discussed.


Cytochemical localization of transport adenosine triphosphatase in the ferret placenta. The distribution of transport adenosine triphosphatase in the ferret placenta was examined cytochemically by light and electron microscopy. The enzyme was detected in the syncytiotrophoblast but was absent from maternal tissues. It appeared to be associated with cytoplasmic processses on syncytiotrophoblast surfaces directly related to foetal or maternal capillaries. The functional significance of transport adenosine triphosphatase is discussed with reference to the transport of solutes between the maternal and foetal circulation across the trophoblast layer.


What do you think you are quantifying? An appraisal of histochemical methods in the measurements of the activities of lysosomal enzymes. The effect of a standard method of fixation (formalin-calcium 24 hr at 4 degrees C, followed by washing in gum-sucrose) on the activity of three lysosomal enzymes (N-acetyl-beta-D-glucosaminidase, acid phosphatase, beta-D-galactosidase) in the liver of three species (human, rabbit, lamb) was studied by biochemical methods, and the results were compared with staining intensities in histochemical preparations of the same tissues. The following conclusions were reached: (1) Fixation by formaldehyde changes the characteristics of the enzymes and makes comparisons between biochemistry and histochemistry difficult to interpret; (2) The intensity of staining in fixed or unfixed tissue sections bears no relation to absolute levels of enzyme activity; (3) Changes in staining intensity of a particular enzyme activity in a particular organ of a particular species prepared in a particular way are significant. Quantifying these changes is useful, as long as absolute values are not considered, and it is realized that it is only the difference that is being quantified.


The quantification of formazans in tissue sections by microdensitometry. I. The use of neotetrazolium chloride. This article describes the use of a microdensitometer for the measurement of formazan deposits in tissue sections. Some examples are given to illustrate the various applications of this technique in the assessment of glucose-6-phosphate dehydrogenase activity. These are (I) the separate measurement of the red half-formazan intermediate and purple diformazan of neotetrazolium, and the effect of incubation time on their production, (2) the measurement of activities in different regions of the liver lobule, and the selective effect of phenobarbitone, and (3) the measurement of enzyme activity in individual cartilage cells in normal and osteoarthrosis-prone animals. All activities can be expressed in absolute units as nmol hydrogen/mm3/hr, and thus compared with standard biochemical data. The activities obtained all fall within the range of published values for biochemical systems.


A model of the breakdown and removal of the chromatin components during Feulgen acid hydrolysis. A stochastic model of Feulgen hydrolysis is proposed. The model, constructed so as to embody the main features of chromatin structure, is simple enough to allow the calculation of extraction rates. Extraction rate curves generated by the model are compared with experimental curves obtained under various conditions (different fixatives and hydrolysis solutions). A good correspondence is found between the experiments and the predictions of the model.


The composition of stains produced by the oxidation of Methylene Blue. The composition of some stains produced by the oxidation of Methylene Blue has been studied by thin-layer chromatography. Various named methods for the production of Polychrome Methylene Blue, Azure A, Azure B, Azure C and Methylene Violet Bernthsen have been found to give complex mixtures of varying proportions of up to eleven dyes. Ten of these, namely Methylene Blue, Azure B, Azure A, sym-Dimethylthionine, Azure C, Thionine, Methylene Violet Bernthsen, Methyl Thionoline, Thionoline and Thionol, have been identified by their visible absorption spectra. The remaining dye could not be identified. When used on a laboratory scale, these methods give stains of constant composition independent of the batch of Methylene Blue. Stain composition as revealed in the present study has been compared with that previously indicated by other, less effective, analytical techniques. Reasons are presented why the latter give equivocal results.


Factors affecting the estimation of the relative amount of chromophore and chromophore area by the two-wavelength method of Patau and Ornstein. Factors influencing the calculation of the relative amount of chromophore and the chromophore area by the two-wavelength method are examined. The study was carried out with the help of models and further tested on Feulgen stained preparations. Except for certain restrictions the difference between the chromophore area as calculated from the two transmissions measurements and the chromophore area obtained by planimetry can be used as a guide for determining the proper measuring conditions, including the choise of the two wavelengths.


Distribution and properties of an adenosine triphosphatase in the tanycyte ependyma of the IIIrd ventricle of the rat. The distribution, histochemical properties and ultrastructural localization of adenosine triphosphate (ATP) hydrolyzing enzymes in the tanycyte ependyma of the third ventricle have been studied in female Wistar rats. Using a calcium-cobalt procedure and a lead capture technique, splitting of ATP could be demonstrated in perikarya and processes of tanycytes in the region of the ventromedial nucleus. The reaction showed no dependence on magnesium or sodium ions, did not occur with other monodi-, and tri-phosphates as substrates, and was inhibited by p-chlormercuribenzoate (PCMB) and sodium fluoride, but not by ouabain. With the calcium-cobalt method the highest intensity of reaction was found at pH 9.4, whereas the lead method gave optimal results at pH 6--8. At the ultrastructural level, the reaction product was found at the outer surface of the plasma membranes of tanycytes and reached its highest concentrations in the region of the region of the apical microvilli; From the findings it is concluded that splitting of ATP in tanycytes is due to a true ATPase. The enzyme might be involved in an active transport of substances by tanycytes.


Concanavalin A-perioxidase-diaminobenzidine (Con-A-PO-DAB)-alcian blue (AB): a reliable method for dual staining of complex carbohydrates. A method has been established for the dual staining of complex carbohydrates in light microscopy. It is a combined concanavalin A-peroxidase-diaminobenzidine (Con-A-PO-DAB)-alcian blue (AB) (pH 2.5) method, and with this method it is possible to color alpha-D-glucosyl and alpha-D-mannosyl residues and acidic groupings of complex carbohydrates in tissues brown and blue respectively. Histochemical experiments using histological sections with reactive complex carbohydrates and casein films containing carbohydrates of known chemical structure have substantiated the validity of the above significance of the dual staining. Thus, the present dual staining method is a reliable one and a new addition to a series of dual staining techniques hitherto employed in the light microscopic histochemistry of complex carbohydrates.


Ultrahistochemical study on the ruthenium red surface staining. I. Processes which give rise to electron-dense marker. The cell coat picture effect which is usually obtained with the conventional RR method, that is with the RR/OsO4 coupled reaction, is investigated. In this first paper, each of conceivable events which might take place between RR, OSO4 and cell surface membrane is discussed or studied. Various tests are carried out on ascites Ehrlich carcinoma cells and Zajdela ascites hepatoma cells treated with numerous chemical reagents, as also on a few pure proteins. The set of data supports the concept that the staining pattern is due to the combination in surface membranes of RR with a "colloidal-like" form of OsO2. The latter might occur during the formation of stable cyclic osmic acid diesters between OsO4 and membrane unsaturated lipids. A possibility by which the resulting marker is though also to be in a "colloidal-like" state is put forward. A next report will deal with this problem.


Ultrahistochemical study on the ruthenium red surface staining. II. Nature and affinity of the electron dense marker. The electron-dense marker which is thought to produce the ruthenium red surface staining is studied. This stain is prepared under conditions which should give its rise in cell surface membrane, and its nature and charge are tested electrophoretically and by measuring the turbidity, respectively. It is a positive colloid resulting from the recharging of colloidal osmium dioxide by RR polycations. Controls on the affinity are carried out by applying positive sol to gelled agarose sections containing hyaluronic acid, polyvinyl sulfate or polylysine. Controls are also carried out on ascites Ehrlich carcinoma and Zajdela ascites hepatoma cells subjected to prior enzymatic and chemical treatments. It is found that the osmium-RR system visualizes all acidic groups in the outer hydrophilic leaflet, that is the greater part of compounds in this external cell layer. A model is presented for the mechanism underlying its rise in cell surface membrane.


An evaluation of some complexing methods for the histochemistry of calcium. A selection of reagents capable of complexing with calcium in various ways was compared by applying them to tissue sections known to contain calcium in a variety of physiochemical states. Their histochemical potential was evaluated according to their sensitivity, specificity, accuracy of localisation and intensity of staining. Murexide, for light microscopy, and 8-hydroxyquinoline, for fluorescence microscopy proved to be the best overall reagents. They failed to demonstrate calcium oxalate, which was well shown by naphthalhydroxamic acid.


Conclusions about aminopeptidase in tissue sections from studies of amino acid naphthylamide hydrolysis. Catalytic properties (KM, Vmax) of aminopeptidase in pig kidney sections, in isolated membranes and in a solubilized purified form were investigated using amino acid 2-naphthylamides and 4-methoxy-2-naphthylamides. In the first case these properties were estimated on the basis of the stain intensity resulting from the coupling of product with Fast Blue B, in the latter two cases they were measured fluorometrically. The following observations were made: (1) In all three cases the substrate turnover was shown to be a direct function of time and enzyme concentration. (2) The values obtained for the solubilized and the membrane bound form were practically identical but differed from those found in tissue sections. (3) Each amino acid derivative had defined constants, but these were difficult to obtain in sections, especially if it was necessary, on account of poor solubilities, to use low substrate concentrations. (4) Hydrophilic amino acid derivatives were adsorbed to tissue membranes much less than hydrophobic ones. (5) Fast Blue B caused a non-competitive inhibition of enzymic activity. (6) Binding of antibody against pure aminopeptidase caused inhibition of the enzymic hydrolysis of all the naphthylamides. Thus, histochemical stain intensities per time and area derived from one substrate at a defined concentration are suitable for the determination of enzyme concentrations. However, no conclusions regarding the homogeneity of the enzyme in sections can be drawn by comparing the stain intensities obtained with different substrates in contrast to data from the inhibition of substrate hydrolysis by antibody.


[Azoindoxyl methods for the histochemical investigation of hydrolases. I. Lactase (lactase-beta-glucosidase complex) (author's transl)]. An azoindozyl method for the histochemical demonstration of lactase (lactase-beta-glucosidase complex) is described. The incubation medium consists of 5 mg 5-Br-4-Cl-3-indolyl-beta-D-fucoside (dissolved in 0.5 ml N,N-dimethylformamide) and 0.02 ml hexazotized prosaniline in 10 ml 0.1 M citric acid phosphate buffer, pH 6-6.5. By means of this method lactase can be exactly localized in the brush border of the enterozytes in the jejunum of suckling rats. Compared to the corresponding indigogenic method the azoindoxyl reaction proceeds faster and the reaction product is often precipitated more precisely.


Aldehyde-fuchsin staining of antral endocrine cells. Correlative studies have been made to identify the endocrine cells stained by Aldehyde-Fuchsin (AF) in the cat and human antral mucosa. D-cells weakly stained by AF after oxidation: this staining might be related to the disulphide group present in the molecule of Somatostatin (the hypothalamic hormone recently localized in D-cells). The gastrin-producing G-cells were on the contrary strongly stained (without oxidation) in tissues fixed in Helly-fluid. Histochemical investigations on the mechanism of such staining lead to the conclusion that it might be related to affinity of the stain for hidden acidic groups.


An alcohol-soluble Schiff's reagent: a histochemical application of the complex between Schiff's reagent and phosphotungstic acid. A schedule for staining partially hydrated PAS-positive structures using non-aqueous solutions has been devised. Tissues are dewaxed, taken down to 70% alcohol, oxidised for 10 min in a 1% w/v alcoholic solution of periodic acid, treated with an alcoholic solution of phosphotungstic acid-Schiff reagent complex (PTA-Schiff reagent) for 25 min, washed in alcohol, cleared in xylene and mounted in a synthetic medium. The PTA-Schiff reagent complex prepared from de Tomasi Schiff reagent by precipitation with PTA may be stored in the deep freeze for many months and dissolved freshly in alcohol for use. The PTA-Schiff reagent used as above allows staining of highly water soluble materials such as dextran. From blocking and digestion studies the mode of action seems similar to de Tomasi Schiff reagent. The partial hydration of the tissues prior to reaction was found to be essential for effective staining.


Polarization microscopic evidence for an oriented cytoplasmic structure in the "dark" variants of adrenalin-storing cells. A diffuse cytoplasmic birefringence confined to "dark" adrenalinstoring cells has been described. The main optical characteristics of the birefringence factor include: regular orientation of birefringence relative to the base-apex axis of cells; additive anisotropic staining with methods based on the principle of topo-optical staining reactions; dependence of birefringence on labile morphologic properties. On the basis of the capacity of the macromolecular matrix of chromaffin granules to form lamellar liposomal structures in vitro it has been proposed that a reorientation of molecular organization in the matrix of chromaffin cells is responsible for the observed optical phenomenon. The direction of birefringence was explained by a preferential direction of contractile forces acting during "dark" cell formation.


Cytochemical evaluation of the Guard procedure a regressive staining method for demonstrating chromosomal basic proteins. I. Effects of fixation, blocking reactions, selective extractions, and polyacid "differentiation". Appropriately fixed preparations stained by a modification of the Guard (1959) reaction for "sex chromatin" display selective staining of interphase chromatin and mitotic or meiotic chromosomes. This is a regressive staining method which seems to depend on the selective displacement of an acidic dye from less basic structures, and retention of the dye at more basic sites. The results obtained with the reaction can be controlled by the length of time that the preparations are "differentiated" in solutions containing phosphomolybdic and phosphotungstic acids (polyacids). After three- or four-hour exposures to polyacid solutions, all chromatin is stained. However, with longer differentiation, "condensed" chromatin can be stained preferentially. Of a number of fixatives investigated, only 10% formalin, ethanol-acetic acid (3:1), and Bouin's solution proved useful. Others resulted in diminished specificity or a total loss of selectivity. The most intense staining was obtained after formalin fixation. Less intense dyebinding was observed after fixation in 3:1 - probably due to extraction of some histone fractions-and the least amount of dye was bound in Bouin's-fixed chromatin - probably due to blockage of arginine residues by picric acid. The reaction was not affected by enzymatic removal of nucleic acids or the extraction of lipids. It was diminished by treatment with trypsin or weak acetylation, and it was completely prevented by strong acetylation, deamination, or extraction of basic proteins with HCl. The results presented suggest that the modified Guard (1959) procedure selectively demonstrates basic nucleoproteins. Further, by the use of regressive differentiation in polyacid solutions, the retention of dye in more condensed chromatin can be favored.


Cytochemical characterization of the modified Guard procedure a regressive staining method for demonstrating chromosomal basic proteins. II. Substitution of dyes for biebrich scarlet. A number of acidic dyes, including various fluorochromes, were substituted for biebrich scarlet in the modified Guard (1959) procedure, a regressive staining method which appears to demonstrate basic chromosomal proteins. These substitutions were made to test the possibility that dyes other than biebrich scarlet might provide advantages in sensitivity and/or contrast, or that more control could be exerted over the "differentiation" step in which solutions of phosphotungstic and phosphomolybdic acids (polyacids) are used. Of the dyes tested in this investigation, six were found to be especially suitable: procion brilliant blue, procion yellow, geranine G, brilliant sulfoflavine, eosin Y, and eosin B. While procion brilliant blue could be used as an absorption dye only, the other dyes were used more profitably as fluorochromes. The various dyes displayed considerable variability in the ease with which they could be displaced from substrates with polyacid solutions during the differentiation step. Procion brilliant blue, procion yellow, and geranine G were displaced gradually and thus resembled biebrich scarlet. In contrast, eosin B, eosin Y, and brilliant sulfoflavine were displaced more easily from all but the most highly condensed chromatin in substrates. Brilliant sulfoflavine yeilded exceptionally bright and nearly selective fluorescence of consensed chromosomes in division, "X" chromosomes of grasshopper spermatocytes, and sperm heads. Weak, but selective fluorescence was observed when monazo sulfonated dyes, including ponta chrome violet SW, eriochrome black, diamond red, and ponta chrome blue black, were substituted in the modified Guard procedure. Similar results were obtained with solochrome cyanin R. As expected, these dyes seemed to function as weakly acidic dyes.


[Results of bleomycin therapy in recurrent head and neck tumors (author's transl)]. 40 patients with previously treated recurrent head and neck carcinomas have been treated with Bleomycin. The total dose of drug administered ranged from 360-600 mg. In three patients, treatment had to be interrupted because of severe side effects of the drug. Temporary complete disappearance of tumor occurred in three patients, whereas incomplete tumor regression briefly occurred in one third of those patients treated. Survival time for all patients averaged three months. A possible immunosuppressive effect of Bleomycin is also discussed, as based on clinical observations noted in one of the patients treated.


[On the endogenous peroxidase in the spleen of swine (author's transl)]. We studied the distribution of endogenous peroxidase in the spleen of swine by modifications of the Graham and Karnovsky diaminobenzidine procedure. There is a peroxidatic activity in the majority of the ellipsoid cells (cells of the sheathed capillaries of Schweigger-Seidel), which is localized in the endoplasmic reticulum and the perinuclear cisterna. This staining is inhibited completely by aminotriazole and is rapidly destroyed even by low concentratoins of glutaraldehyde. Furthermore, the reaction is abolished after boiling of tissue sections or in the absence of H2O2. The macrophages of the red pulp and a minority of the ellipsoid cells are peroxidase negative. Our results are discussed in respect to some recent studies on the system of mononuclear phagocytes. It is suggested, that the enzyme active ellipsoid cells represent a special form of macrophages, enzyme histochemically related to Kupffer cells and resident peritoneal macrophages. The enzyme negative cells of the ellipsoids are probably fibroblasts.


Noninfectious canine arthritis: the inflammatory, nonerosive arthritides. Noninfectious, nonerosive arthritis was seen as an important manifestation of a number of chronic systemic diseases of the dog. Sixty-three dogs with this type of arthritis were seen at the Veterinary Medical Teaching Hospital during an 18-month period between 1973 and 1975. Of these dogs, 29 had systemic lupus erythematosus, 15 had arthritis in association with some chronic infectious disease process, and 19 had a similar type of arthritis, but without serologic evidence of systemic lupus erythematosus or any chronic infectious disease process.


Antibiotics derived from a mutant of Bacillus circulans. A mutant of Bacillus circulans, which produces butirosins only when 2-deoxystreptamine is added to the fermentation medium, was employed in the biosynthesis of antibiotics containing modified aminocyclitols. The blocked mutant converted 2,5-dideoxystreptamine into 5-deoxybutirosamine. Streptamine was incorporated into a complex differing from butirosin by an additional hydroryl at C-2.


Presence of rfe genes in Escherichia coli: their participation in biosynthesis of O antigen and enterobacterial common antigen. In Salmonella, ilv-linked rfe genes participate in the biosynthesis of the enterobacterial common antigen (CA) as well as of certain types of O antigen (serogroups C1 and L). rff genes, probably in the same cluster with rfe, are required for CA synthesis (P.H. Mäkelä et al., in preparation). Several Escherichia coli strains were studied to determine whether they also have rfe-rff genes that are involved in the synthesis of O antigen and CA, or of CA only. In a first approach, E, coli K-12 F-prime factors carrying the genes ilv and argH or argE and presumably rfe-rff genes were introduced into CA-negative Salmonella mutants that are blocked in CA synthesis because of mutated rfe or rff genes. All resulting ilv+ hybrids were CA positive. In recipients with group C1-derived rfb genes, the synthesis of O6,7-specific antigen was also restored. This result shows that E. coli K-12 has rfe and rff genes providing the functions required in the synthesis of CA and Salmonella 6,7-specific polysaccharide. By introduction of defective rfe regions from suitable Salmonella donors into E. coli O8, 09, and O100 strains, the synthesis of CA as well as of the O-specific polysaccharides was blocked. This indicates that in the E. coli strains tested the rfe genes are involved in the synthesis of both O antigen and CA. This suggestion was confirmed by the finding of E. coli rough mutants that had simultaneously become CA negative. In transduction experiments it could be shown that the appearance of the rough and CA- phenotype was due to a defect in the ilv-linked rfe region.


Purification and properties of two deoxyribonucleases of Pseudomonas aeruginosa. A survey of the major deoxyribonucleases in Pseudomonas aeruginosa strain PAO was undertaken. Two activities predominated in Brij-58 lysates of this organism. These have been purified from contaminating nuclease activities, and some of their properties have been elucidated. The first was a nuclease that degraded heat-denatured deoxyribonucleic acid (DNA) to mono- and dinucleotides. The activity of this enzyme was confined to single-stranded DNA, and 100% of the substrate was hydrolyzed to acid-soluble material. The Mg2+ optimum is low (1 to 3mM), and the molecular weight is 6 X 10(4). The second predominant activity was an adenosine 5'-triphosphate (ATP)-dependent deoxyribonuclease. This enzyme had an absolute dependence on the presence of ATP Mg2+ concentrations of approximately 10 mM. Five moles of ATP was consumed for each mole of phosphodiester bonds cleaved. The acid-soluble products of the reaction consisted of short oligonucleotides from one to six bases in length. Only 50% of the double-stranded DNA was rendered acid soluble in a limit digest. The molecular weight of this enzyme is 3 X 10(5). The observation of these enzymes in P. aeruginosa is consistent with the possibility that recombinational pathways similar to those of Escherichia coli are operating in this organism.


Strain-specific variation in the protein and lipopolysaccharide composition of the group B meningococcal outer membrane. Variation in the protein and lipopolysaccharide composition of the meningococcal outer membrane may be due to either serotype differences or to changes in cultural conditions. There are 12 antigenically distinct serotypes of group B meningococci, and these are associated with distinct major outer membrane protein patterns on sodium dodecyl sulfate-polyacrylamide gels. In most strains the predominant outer membrane protein carries the serotype-specific determinant. Certain strains, when grown under similar conditions in different media showed an altered membrane composition. The type 2 strain, M986, grown in modified Frantz medium-A, had a reduced amount of the major 41,000-dalton protein while a 28,000-dalton protein predominated. The altered protein composition may be related to changes in cell metabolism as reflected by the pH of the medium after growth. Growth of the organism in Frantz medium-B caused a negligible drop in pH and the 41,000-dalton protein remained predominant. There was also variation associated with changes in the growth rate. Increasing the aeration caused a concomitant increase in growth rate and cell yield. We observed two quantitative changes in outer membrane proteins in four of seven strains examined: (i) where only a single major protein changed (three strains), and (ii) where an increase in one protein component was associated with a decrease in another protein (one strain). When the strains were grown in tryptic soy broth (Difco Laboratories, Detroit, Mich.) with either high or low aeration, the total protein in the outer membrane remained constant. In contrast, with high aeration there was a significant increase in lipopolysaccharide. These studies suggest that the cell surface proteins may be altered by the organism to meet a variety of environmental conditions.


Effects of phenethyl alcohol on Bacillus and Streptococcus. The activity of phenethyl alcohol (PEA) on Bacillus cereus, B. megaterium, and Streptococcus faecalis was studied by electron microscopy of thin sections and by the assay of intracellular K+ leakage. S. faecalis was unaffected by PEA at concentrations up to 0.5%, B. cereus was severely damaged by 0.5% PEA, and B. megaterium behaved intermediately. Important membrane ultrastructural alterations were observed in B. cereus cells treated with 0.5% PEA, namely the change in the geometry of the membrane profile from asymmetric to symmetric, the occurrence of prominent, complex mesosome-like structures, and membrane fracturing and solubilization. Protoplasts from B. megaterium were found to be quickly lysed by 0.5% PEA due to the disruption of the cytoplasmic membrane. The electron microscopic observations, together with the results of the study of the K+ efflux from B. cereus and B. megaterium, indicate that PEA primarily and directly damages the cytoplasmic membrane of sensitive bacteria. The breakdown of the permeability barrier probably is responsible for the observed bactericidal action of 0.5% PEA on B. cereus.


Reconstruction of intact gamma-globulin from S-sulfonated gamma-globulin in vivo. The conversion of the S-surfonate group in a peptide to a disulfide bond has been observed in vitro. This paper reports that the conversion of this group in human S-sulfonated gamma-globulin (S-GG) appeared to occur in vivo judging from the change in molecular weight of S-GG observed in the presence of sodium dodecyl sulfate (SDS) and the restoration of hemolytic activity.


A statewide biomedical communications network for South Carolina. In 1972, the Medical University of South Carolina was awarded a contract to establish 4 Area Health Education Centers in South Carolina. These centers, based in community hospitals, provide residency programs, clinical instruction for students, and continuing education programs for health professionals. In late 1974, contractual agreements between the Medical University of South Carolina's Library/Learning Resource Center and the Area Health Education Centers were negotiated to provide book and nonbook learning materials to all health practitioners in South Carolina. The history and the functions of the resulting network and evaluation of audiovisual and self-instructional learning materials procured and distributed by the network are described.


[Acid mucopolysaccharide detection on ultra-thin sections by alcian blue in dental tissues embedded in Epon]. Ultra-thin sections of dental tissues, fixed in glutaraldehyde and osmium tetroxide, embedded without previous demineralization in Epon, were contrasted the grids with an alcian blue solution after previous oxidation periodic acid or hydrogen peroxide. Correlations with histochemical results obtained with the optical microscope, other results obtained with ruthenium red and phosphotungstic acid and controls made after enzymatic digestion with chondroitinase AC suggest that this technique is specific for the detection of acid mucopolysaccharides in those calcified tissues studied.


Lipid vesicle-cell interactions. III. Introduction of a new antigenic determinant into erythrocyte membranes. The introduction of a new antigenic determinant, 2,4-dinitrophenyl-aminocaproyl-phosphatidylethanolamine (DNP-Cap-PE), into the surface membranes of intact human erythrocytes is described. Fresh cells were incubated in the presence of liposomes composed of 10% DNP-Cap-PE, 5% stearylamine, 20% lysolecithin, and 65% lecithin. Such liposome-treated erythrocytes are shown to be susceptible to immune lysis by anti-DNP serum in the presence of complement. Uptake of DNP-Cap-PE by erythrocyte membranes is also demonstrated by immunofluorescence using indirect staining with rabbit anti-DNP serum followed by fluroescein-conjugated goat anti-rabbit IgG and by electron microscopy using ferritin-conjugated antibody. Antigen uptake did not occur at low temperatures or from vesicles lacking lysolecithin and stearylamine. Fluorescence microscopy shows that the antigen-antibody complexes are free to diffuse over the cell surface, eventually coalescing into a single area on the cell membrane. Electron microscopy suggests that a substantial proportion of the lipid antigen is incorporated by fusion of vesicles with the cell membrane. There are indications that vesicle treatment causes a small proportion of cells to invaginate.


Mobility restriction in vivo of the heavy ribosomal subunit in a secretory cell. Analysis in insect (Chironomus tentans) salivary gland cells of labeled RNA as a function of time after precursor injection and its distance to the nuclear membrane, cytoplasmic zone analysis, could previously be used to demonstrate the presence of short-lasting gradients in newly labeled ribosomal RNA. Since the gradients were sensitive to puromycin, they are likely to be a result of diffusion restriction due to an engagement of the subunits into polysomes. In the present paper the possibility was explored of recording gradients that were caused by labeled subunits in puromycin-resistant associations, which, in all probability, involve the endoplasmic reticulum. It was found that labeled 28 S and 5 S RNA but not 18 S RNA were present in radioactivity gradients lasting for at least 2 days but less than 6 days. The gradients also remained during inhibition of RNA synthesis by actinomycin, and they were completely resistant to puromycin whether given in vivo or in vitro. The semipermanent gradients formed fhere offer a unique parameter for the in vivo study of conditions for formation and maintenance of heavy subunits in puromycin-resistant bonds. An explanation for these and previous results is that the light subunit, although restricted in movement by engagement to polysomes, is nevertheless free to exchange and spread between rounds of translation, whereas at least part of the heavy subunit population is bound to the endoplasmic reticulum and less free to spread. These results offer a good in vivo correlate to previous results on in vitro exchangeability of subunits.


[Sarcomas of the gallbladder].

[Sarcomas of the gallbladder]. The authors report two recent personal cases including one explored by retrograde cholangiography and discuss the main clinical, pathological characteristics of this rare disease. They emphasize the severity of the course which does not usually last more than a few weeks after the onset of the first signs.


Familial partial target organ resistance to thyroid hormones. A 30-year old woman with a history of recurrent goiter, who had undergone two partial thyroidectomies, is described. She presented with tachycardia, nervousness and a fine tremor of the fingers. Initially, she had normal serum thyroid hormone levels: thyroxine (T4 (D)) 11.6 MUG/100 ML, TRIIODOTHYRONINE (T3) 138 ng/100ml, normal levels of binding proteins and a very high serum thyrotropin (TSH), 98 muU/ml. During follow-up T4 (D) increased to 17.2 mug/100 ml, T3 increased to 277 ng/100 ml, while TSH decreased to 11 muU/ml. There was an exaggerated response of TSH to a peak value of 550 muU/ml after intravenous administration of 200 mug thyrotropin-releasing hormone (TRH). Administration of 60 mg prednisolone daily resulted in a blunting of the response to TRH. Administration of 50 mug T3 daily for 1 month resulted in a fall in serum TSH from 98 to 50 muU/ml. Later, when the serum TSH level had fallen spontaneously to 20 muU/ml, administration of 100 mug T3 daily for two weeks resulted in a fall in serum TSH to 5.3 muU/ml. Treatment with 20 mg carbimazole daily for 3 weeks resulted in a decrease in serum T4 levels with a concomitant increase of serum TSH. There was no evidence of pituitary enlargement and other pituitary hormone levels were normal. All the relatives studied (father, sister, three children) had elevated T4 levels with normal basal TSH values. It is concluded from this study that our patient presents evidence of partial resistance to thyroid hormones.


Comparison of androgen metabolites in benign prostatic hypertrophy (BPH) and normal prostate. 5 alpha-Dihydrotestosterone (DHT) and androstanediols (diols) have been measured in human prostate tissue. DHT levels in surgical specimens of prostate from 8 patients with BPH averaged 5.6 +/- 0.93 S.E. ng/g and were significantly greater than (P less than 0.01) values of 2.1 +/- 0.32 S.E. ng/g in 6 normal prostates obtained post-mortem from males less than 50 yrs old. Androstanediols averaged 2.3 +/- 0.35 S.E. ng/g in the BPH specimens compared to values of 10.2 +/- 2.4 S.E. ng/g in the normal prostates (P less than 0.01). This significantly higher (P less than 0.001) ratio of diols/DHT in the normal (5.1 +/- 0.93 S.E.) compared to the BPH prostate (0.45 +/- 0.08 S.E.) suggests that a decrease in 3-hydroxysteroid oxido-reductase, which converts DHT to diol, may be an important clue to the pathogenesis of BPH.


Isolation and characterization of a bacteriophage specific for Neisseria perflava. Six isolates from normal throat samples have been shown to contain phage active against Neisseria perflava. The phage isolates were similar in terms of host range, latent period, burst size, antigenic properties, morphology, and nucleic acid content. Neutralization studies with antisera demonstrated that the isolates exhibited a very high degree of serological relatedness. These results taken together suggested that the isolates represented a single strain of bacteriophage. This phage, which we have designated NP-1, exhibited a high degree of host specificity, attacking only one of the several strains of N. perflava tested and none of the other species tested. One-step growth experiments yielded minimum latent periods of approximately 35 min; average burst sizes varied from 34 to 63 plaque-forming units per cell. Electron micrographs revealed particles with heads averaging 75 nm in diameter and tails averaging 300 nm in length and 18 nm in diameter. The phage contained double-stranded DNA with a guanine plus cytosine content of 38%.


Section preparation of human marrow for light microscopy. Preparation of human marrow sections has been studied systematically in order to facilitate accurate identification of marrow cells. Both of the methods developed involve embedding marrow cores in methyl methacrylate. In one, acrolein fixation is followed by staining of deplasticized sections with eosine-y followed by azure II; in the other, neutrophilic forms are identified by their esterase-specific reactivity in marrow fixed with neutral-buffered formalin. These preparations are suitable for quantitative studies of marrow cellularity.


Effect of orally administered duodenal fluid susceptibility of newborn calves to an Escherichia coli challenge. This study was to determine if duodenal contents inoculated orally could alter symptomatic diarrhea produced by Escherichia coli [09:K(A)] in neonatal calves. Inoculum was obtained 2 h postprandially from duodenal-cannulated milk-fed calves. Thirty Holstein bull calves obtained within 3 h of birth were assigned alternately to a 3 X 2 factorial experiment. Calves received either 1) no E. coli challenge, 2) challenge at 12 h, or 3) challenge at 24 h each with or without 200 ml duodenal fluid orally with the first feeding of colostrum. Rectal temperature, urine output, milk intake, and fecal dry matter were recorded daily, and body weight and hematocrit every 3rd day for 1 wk. Gamma globulin of serum was determined at 2 h of age. Inocula and duodenal tissue and fluid of the last 12 calves were assayed for total anaerobic bacterica, anaerobic and aerobic lactobacilli, coliforms, and total aerobic bacteria. Inoculated calves exhibited lower incidence of diarrhea, greater urine output, lower hematocrit, and superior average daily gain as compared to uninoculated calves. Concentration of gamma globulin in serum of inoculated calves was subnormal. Bacterial populations of tissue and fluid at 7 days were not influenced by treatment.


An audio-tutorial aid for dietary instruction in renal dialysis. Four 10-min, color, video-tape cassettes, which include graphics, cartoon characters with dialogue, and live demonstrations, were developed to improve dietary adherence by patients receiving maintenance hemodialysis. The format of the tapes was directed primarily to the needs of educationally deprived patients. The tapes were assessed for efficacy following presentation to sixteen randomly selected patients in he University of Tennessee Artificial Kidney Center, Memphis. Highly significant, positive changes were revealed when post-test informational scores were compared with pre-test scores. In the behavioral scores, some positive changes were observed, although further reinforcement would be needed to achieve sufficient application of the information.


Management of chronic non-rheumatic pain in the elderly. Chronic non-rheumatic pain in the aged should be viewed in the context of physiologic and clinical alterations due to aging which may create diagnostic problems. In the elderly, the abnormality of the clinical presentation and the patient's failure to remember the time and the nature of the precipitating incident may be extremely misleading. Many clinical features may also be altered by the aging process itself. Thus the unwary physician is apt to be mistaken in establishing the etiologic basis for pain syndromes. Once a diagnosis has been achieved, it should be evaluated in the light of other age-related disturbances. Treatment should be as simple as possible. The prescription of combination preparations may involve use of a drug that is beneficial for one disorder but harmful for another. Aged persons display a marked tendency toward abnormal reactions to the usual drug regimens. A wise combination of treatment by medical means (including drugs), physiotherapy, and sometimes surgical procedures, usually is effective for the relief of this type of chronic pain.


Ultrastructural studies on the localization of neurohypophysial hormones and their carrier proteins. Neurophysin, vasopressin and oxytocin were localized in different portions of the supraopticohypophysial tract (SHT) using the unlabeled antibody enzyme technique at the ultrastructural level. In vasopressin-positive supraoptic perikarya, vasopressin and neurophysin were present in all neurosecretory granules. Within the zona interna of the median eminence, vasopressin and neurophysin were present in two populations of axons, one with granules of 1300-1500 A and one with granules of 900-1300 A. Following exposure of thin sections of median eminence to antiserum to neurophysin, reaction products were present in granules and in the extragranular cytoplasm in the axons with larger granules; in all other cases reaction product was confined to the granules. Vasopressin-positive fibers were also presented in large numbers of the zona externa of the median eminence and many terminated on the pituitary primary portal plexus. A few oxytocin fibers were present on the portal capillaries in the infundibular stalk. In the posterior pituitary all axon profiles were neurophysin positive. Neurophysin was present as both a granular and cytoplasmic pool. Vasopressin-containing axons account for 90% of the neuronal elements in the posterior pituitary and oxytocin for the remaining 10%. Findings on the subcellular distribution of these peptides are related to current theories on transport and release of neurohormones.


Immunocytochemistry of the pituitary glycoprotein hormones. The storage sites of the pituitary glycoprotein hormones were identified with the use of electron microscopic immunocytochemical techniques and antisera to the beta (beta) chains of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and thyroid-stimulating hormone (TSH). The TSH cells in normal rats is ovoid or angular and contains small granules 60-160 nm in diameter. In TSH cells hypertrophied 45 days after thyroidectomy, staining is in globular patches in granules or diffusely distributed in the expanded profiles of dilated rough endoplasmic reticulum. The gonadotrophs (FSH and LH cells) exhibited three different morphologies. Type I cells are ovoid with a population of large granules and a population of small granules. Staining for FSHbeta or LHbeta was intense and specific only in the large granules (diameter of 400 nm or greater). Type II cells are angular or stellate and contain numerous secretory granules averaging 200-220 nm in diameter. They predominate during stages in the estrous cycle when FSH or LH secretion is high. Type III cells look like adrenocorticotropin (ACTH) cells in that they are stellate with peripherally arranged granules. They generally stain only with anti-FSHbeta and their staining can not be abolished by the addition of 100 ng ACTH. In preliminary quantitative studies of cycling females, we found that on serial sections FSH cells and LH cells show similar shifts to a more angular population of cells during stages of active secretion. However, the shifts are not in phase with one another. Furthermore, there are at least 1.5 times more FSH cells than LH cells at all stages of the cycle. Our collection of serial cells shows that some cells (usually type I or II) stain for both gonadotropic hormones, whereas others (usually type II or III) contain only one.


Single-strand nuclease action on heat-denatured spermiogenic chromatin. The aim of this study was to compare the sensitivity of chromatin from representative cellular stages of spermiogenesis to a single-strandeded nuclease after heat denaturation. Thermal denaturation of chromatin was assayed in situ in fixed round, elongating and elongated spermatids and in testicular sperm from mice. Production of single-stranded deoxyribonucleic acid (DNA) at elevated temperatures was monitored by digesting chromatin with endonuclease specific for single-stranded DNA (S1 nuclease), staining the residual DNA with gallocyanin-chrome alum (GAC) and measuring the stain content by absorption cytophotometry. Changes in GCA staining were minimal over the temperature range of 22-90 degrees C in each cell type not exposed to nuclease. Staining of undigested cells decreased progressively with advancing cell maturity. Nuclease had no effect on the GCA content of round spermatids below 60 degrees C, but above this temperature there was a progressive decrease in GCA-stainable chromatin. Both round and elongating spermatid stages showed a significantly greater sensitivity to nuclease digestion than did more mature stages; sperm showed no effects of nuclease action below 80 degrees C. Progressive chromatin condensation and a concomitant decrease in the number of available DNA phosphate groups during spermiogenic cell maturation may be responsible for the observed decline in sensitivity to nuclease and decreased GCA staining. Thermal denaturation of round spermatids labeled with 3H-thymidine produced no change in autoradiographic mean nuclear grain counts, indicating no loss of thymidine-labeled DNA from the slides during denaturation. When round spermatids and sperm were hydrolyzed with hot tricholoroacetic acid before staining, both nuclear GCA content and autoradiograph grain count were partially reduced, indicating incomplete DNA removal. Almost complete loss of Feulgen-stainable material occurred in these cells and may be due to depurination and elimination of Feulgren-reactant aldehyde groups.


Concanavalin A-horseradish peroxidase bridge staining of alpha-1 glycoproteins separated by isoelectric focusing on polyacrylamide gel. The Coomassie Blue protein stain and the periodic acid-Schiff stain for glycoproteins are compared to a new method of staining glycoproteins resolved electrophoretically. The method utilizes a Concanavalin A-horseradish peroxidase sequence to visualize selectively glycoproteins with terminal or internal mannose or terminal N-acetylglucosamine. The method applied to characterization of M and Z allele products of alpha-l-antitrypsins separated by isoelectric focusing of polyacrylamide gels slabs have revealed differences in carbohydrate content of various components that were previously undetected.


Effect of stirring during fixation upon immunofluorescence. Results with distribution of albumin-producing cells in liver. When the immunofluroscent study on the distribution and the incidence of albumin-producing hepatocytes in the rat liver was performed by the method of Sainte-Marie, the number of positive cells showed various values (10-60%). It was surmised that when the permeability of the fixative was delayed, albumin had flowed out from the cytoplasm of the unfixed hepatocytes. By the simple means of constant stirring of the fixative using a magnetic stirrer, we accomplished rapid fixation and achieved results in which positive cells attained 100%. On the other hand, the incidence of positive cells decreased markedly when rats were fed a protein-free diet.