Increased safety and specificity in the thin-layer chromatographic identification of marihuana.

We are reporting what we believe to be a safe, rapid TLC system and spray for the identification of marihuana. The use of Fast Blue 2B salt greatly enhances the specificity of the TLC analysis of suspected marihuana samples.


The rapid determination of cocaine and other local anesthetics using field tests and chromatography.

The application of field tests and chromatography to the detection of cocaine and some other local anesthetics that have been used to adulterate cocaine is described. Initial screening of samples by field tests, followed by concurrent TLC and GC, enables rapid identification of these compounds to be achieved. In particular, the use of flow-programmed GC shortens the time for analysis compared with conventional GC and requires negligible equilibration time between consecutive runs [12]. The method gives reliable quantitative data.


An efficient method to eliminate streaking in the electrophoretic analysis of haptoglobin in bloodstains.

A bloodstain extraction procedure that improves the analysis of haptoglobin in dried bloodstains has been developed. The streaking of electrophoresis gels caused by deteriorated hemoglobin can be eliminated by incorporating chloroform in the bloodstain extraction procedure. The method is easier to execute than previously published techniques for eliminating the adverse effects of deteriorated hemoglobin on the analysis of haptoglobin. Bloodstains up to two years old were correctly phenotyped in haptoglobin by this method.


radE, a new radiation-sensitive locus in Dictyostelium discoideum.

Dictyostelium discoideum strain M28, which has been used widely in genetic studies, was found to carry a radiation-sensitive mutation. This allele, termed rad-100, was recessive in heterozygous diploids and mapped in linkage group III. Complementation analysis and survival studies on strains carrying rad-100 suggested that this allele defines a new radiation-sensitive locus in D. discoideum, and this locus has been designated radE. radE strains were moderately sensitive to ultraviolet light (D10 90 J m-2) and slightly sensitive to 137Cs gamma rays D10 255 krad). radE strains also exhibited increased sensitivity to killing by N-methyl-N'-nitro-N-nitrosoguanidine but not by other alkylating agents such as ethyl methanesulphonate or methyl methanesulphonate. The frequency of spontaneous methanol-resistant (acrA) mutants was approximately the same in cultures of radE and radE+ strains. However, when amoebae of these strains were irradiated with ultraviolet light, the frequency of induced mutants was significantly lower in cultures of the radE strain. Furthermore, when amoebae of wild-type strain NC4 were plated in the presence of caffeine after ultraviolet-irradiation, the survival curves were very similar to the curves obtained for amoebae of radE strains in the presence or in the absence of caffeine. These results suggest that the radE100 mutation and caffeine interfere with an error-prone DNA repair pathway in D. discoideum.


Characterization and persistence of actinophage RP2 isolated from Streptomyces rimosus ATCC 10970.

While searching for true lysogens among oxytetracycline-producing Streptomyces rimosus strains, free phage particles were detected and isolated from a liquid culture of S. rimosus ATCC 10970 (R7). The actinophage, designated RP2, appears to be a typical temperate DNA phage producing turbid plaques on the sensitive strain S. rimosus R6. Electron microscopic examination of RP2 lysates showed that it belongs to group B of Bradley's morphological classification. The rate of RP2 adsoprption at 28 degrees C appeared to be low. The length of the latent period was about 6 h and the average burst size about 120 phage particles. The lysogenic nature of the host-virus system described was established on the basis of the following characteristics: spontaneous lysis frequency of 2 X 10(-6) per cell, resistance to curing with phage-specific antiserum, spontaneous curing frequency of less than 0.05% and immunity to superinfection with the homologous phage. Clear-plaque mutants of RP2, which failed to lysogenize sensitive cultures, arose at a frequency of 10(-5).


Regulation of 2-oxoglutarate dehydrogenase synthesis in Citrobacter freundii by traces of oxygen in commercial nitrogen gas and by glutamate.

Glutamate induced the synthesis of 2-oxoglutarate dehydrogenase 50-fold during anaerobic growth of Citrobacter freundii and, in the absence of glutamate, this enzyme was even more active in cultures sparged with N2/CO2(95:5, v/v). Enzyme synthesis was partially repressed when the inlet gas was passed through heated copper but totally repressed when the inlet gas was passed through alkaline pyrogallol and reduced benzyl viologen (a treatment which would remove CO2 as well as O2). Fumarate hydratase activity also decreased but alcohol dehydrogenase and the sum of the succinate dehydrogenase and fumarate reductase activities increased when residual O2 was removed from the sparging gas. Soluble cytochromes a1 and c552.5 were detected in rigorously anaerobic cultures. Thus traces of O2 which contaminate commercial compressed N2 are sufficient to induce 2-oxoglutarate dehydrogenase synthesis and to affect significantly the synthesis and incorporation of respiratory chain components into the cytoplasmic membrane.


Factors affecting aflatoxin production by Aspergillus parasiticus in a chemically defined medium.

The optimum levels of sucrose, (NH4)2SO4, MgSO4, KH2PO4 and ZnSO4 for aflatoxin production in a chemically defined medium have been established. The last two were found to be essential for fungal growth and aflatoxin production. The effect of various carbon sources on aflatoxin production was tested using the defined medium. Asparagine was found to be essential for aflatoxin production. Very little aflatoxin was produced in the absence of asparagine with any of the other inorganic nitrogen sources tested. Supplementation with yeast extract, Casamino acids, Casitone and peptone increased the aflatoxin yield, but omission of asparagine led to decreased aflatoxin yields even when complex nitrogen sources were present. Asparagine could be replaced by aspartic acid or alanine.


Biochemical differentiation in large colonies of Enterobacter cloacae.

Large colonies of Enterobacter cloacae which were about 700 micrometer thick were frozen in liquid nitrogen and sectioned horizontally. The sections were disrupted and several oxidative enzymes were assayed in the crude unfractionated homogenates. In the top 120 micrometer of the colonies the specific activities of the enzymes were high and characteristic of aerobically adapted cells. Cells nearer the base of colonies had very low enzyme activities.


Studies on the temperature-dependent DNA replication of the herpesvirus of the turkey in chicken embryo fibroblasts.

The development of cytopathogenic changes in chicken embryo fibroblasts infected with the herpesvirus of the turkey, strain PB-THVI, and the release of virus particles into the supernatant of infected cultures is accelerated at temperatures higher than 37 degrees C and is fastest at 41 degrees C, the normal body temperature of chickens. The growth rate of HVT in CEF cultures was followed by determination of the number of virus genome equivalents within infected cells at various time intervals p.i. A temperature-dependent increase in the amount of virus DNA per infected cell could be detected, which is highest at 41 degrees C. At all temperatures tested (34, 36, 41 and 43 degrees C) the number of virus genome equivalents per infected cell ultimately reaches the same level. In the course of infection, virus DNA in CEF cultures at 37 and 41 degrees C becomes associated with the cellular DNA, as determined by neutral CsCl gradients of the total cellular DNA and hybridization of each fraction with 32P-labelled virus-specific complementary RNA. Association of virus to cellular DNA occurs earlier at 41 than at 37 degrees C. However, the same proportion (45%) of the total virus DNA appears ultimately to be associated with cellular DNA at both temperatures. A temperature-shift of CEF cultures infected with PB-THVI from 41 to 37 degrees C 24 h p.i. resulted in the same replication kinetics of virus DNA as was found at 41 degrees C.


Regulation of imm gene expression in bacteriophage T4-infected cells.

Two polypeptides (imm-a and imm-b) which are not induced by an immunity mutant T4Dimm2 but by a wild-type strain T4D were identified by sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis. Their mol. wt. were 77 000 and 45 000, respectively. These polypeptides exhibited a similar kinetic pattern of synthesis. Within a few minutes p.i. the primary phage established the system that inhibited imm gene expression of superinfecting phage. This was shown by measuring both the phenotypic expression of immunity and the synthesis of imm gene polypeptides. The expression of two other immediate-early genes, namely genes s and 30, and early gene 33, was not affected by primary infection.


Replication and interaction of virus DNA and cellular DNA in mouse cells infected by a human adenovirus.

C57 black mouse cells infected with human adenovirus type 5 (Ad5) produced large amounts of early virus proteins, small amounts of late virus proteins and less than 0.2 infectious units (i.u.)/cell of infectious virus. Many cells died but the cultures recovered. Virus DNA and cellular DNA were synthesized. Some Ad5 DNA sedimented with cell DNA in alkaline sucrose, but virus DNA was rapidly lost from the culture after recovery and none of 28 unselected cloned survivors contained detectable amounts of virus DNA or antigens. Ad5 ts36 was temperature-sensitive for virus DNA replication in mouse cells, but ts125 was detective at 32.5 degrees C as well as at 39.9 degrees C. No difference was detected in the percentage of virus DNA that sedimented in alkali with cell DNA, in mouse cells infected by Ad5 ts+, ts36 or ts125 at 32.5 or 39.9 degrees C. All parts of the virus genome were equally represented in virus DNA that sedimented with cell DNA, in mouse cells infected by Ad5 ts+ or ts36 at either temperature.


Sequence analysis of lactosamine type glycans of individual membrane proteins of Semliki Forest virus.

3H-fucose and 14C-glucosamine labelled glycopeptides of the individual membrane proteins E1, E2 and E3 of Semliki Forest virus could be sequentially digested with alpha-neuraminidase, beta-galactosidase, N-acetyl-beta-glucosaminidase, alpha- and beta-mannosidase, N-acetyl-beta-hexosaminidase and finally with alpha-fucosidase. The degradations of the virus glycopeptides proceeded in the same way as stepwise digestions of reference glycopeptides of the lactosamine type obtained from IgG and alpha 1-acid glycoprotein. This suggests that all three membrane glycoproteins of Semliki Forest virus contained glycans with a monosaccharide sequence characteristic for lactosamine type oligosaccharides. The number of both distal and proximal N-acetyl-glucosamine residues was estimated to be usually two. According to exo- and endo-glycosidase digestions, fucose seemed to be attached to the innermost N-acetyl-glucosamine unit.


Morphological irregularities in Dane particle cores.

Electron microscopy of hepatitis B antigen has revealed Dane particles with abnormal morphology. These aberrant Dane particles contain incomplete cores. They were seen in large numbers in the serum of an HBsAg-carrying renal dialysis patient and were less numerous but invariably present in other Dane particle-containing sera --12 from asymptomatic carriers, 2 chronic hepatitis patients and 1 patient with acute hepatitis. All 16 sera were shown to contain HBeAg.


Isolation and characterization of a homogeneous DNA-protein complex from adenovirus type 2 virion.

Adenovirus DNA-protein complex purified by sedimentation on a sucrose gradient containing 4 M-guanidine hydrochloride was found to contain other virion proteins in addition to the terminal protein of mol. wt. 55000. In this report, we describe a simple and rapid method for the isolation of a homogeneous DNA-protein complex. The procedure involves gel electrophoresis of the complex on agarose in the presence of sodium dodecyl sulphate. DNA was found to migrate into the gel with a single protein of mol. wt. 55000 tightly attached to it. Restriction enzyme cleavage analysis of the DNA-protein complex shows that the protein is associated with the two terminal fragments.


Newly synthesized influenza virus proteins are transported from the nucleus.

Newly synthesized nuclear NP, M and NS1 proteins migrated from the nucleus over a period of several hours. Rates of transport were higher for NP than for M or NS1 and suggest that nuclear NP may play a role in multiplication.


Structural polypeptides of Orthopoxvirus: their distribution in various members and location within the virion.

The structural polypeptides of accepted species and recently isolated members of the genus Orthopoxvirus have been examined by SDS-polyacrylamide gel electrophoresis. The viruses shared many polypeptides but some differences were found. The viruses could be divided into a vaccinia group (including buffalopox, 'Lenny' and MK-10), an ectromelia group (including elephant virus and Moscow virus), cowpox, camelpox and monkeypox. Minor differences were found in the polypeptides of monkeypox virus strains from human and monkey outbreaks. Controlled degradation of virions showed that the polypeptides which enabled the viruses to be differentiated were located in the surface and sub-surface layers. The cores of the viruses all gave the same complex polypeptide pattern.


Cytoplasmic and nuclear input virus RNPs in influenza virus-infected cells.

Chicken fibroblasts and MDCK cells were infected with influenza virus labelled with either 3H-uridine or 14C-amino acids, and the location in infected cells and properties of input virus-labelled structures were studied. Input virus RNA and protein were found in the cytoplasm of nuclei 1 h p.i. A part of the intranuclear parental structures was associated with chromatin while the other part could be extracted from nucleoplasm by 0.16 M-NaCl and represented free ribonucleoprotein (RNP) particles. These RNPs sedimented in glycerol velocity gradients at 40 to 70S, very similar to cytoplasmic RNPs, but differed distinctly from them in buoyant density. The bulk of cytoplasmic RNPs after fixation with formaldehyde banded in CsCl at 1.34 g/ml while nucleoplasmic RNPs banded at 1.39 or 1.41 g/ml. RNPs isolated from virions and infected cells contained the NP polypeptide which was revealed by SDS-PAGE analysis as a double band. The ratio of the two bands varied in cytoplasmic and nucleoplasmic RNPs, the lower band being dominant in cytoplasmic but not in nucleoplasmic RNPs. In addition, cytoplasmic RNPs were phosphorylated. The possible significance of intracellular RNP modifications for virus replication is discussed.


Replication cycle of Newcastle disease virus in three host cells of different permissiveness.

Various degrees of permissiveness for NDV have been described in different cell lines. In the present work three systems were investigated: bovine kidney (MDBK) cells, chick embryo (CE) cells and mouse L cells producing 50 to 100 p.f.u., 2 to 10 p.f.u. and less than 0.1 p.f.u./cell, respectively. Analysis of the radioactive virus messenger RNAs (vmRNAs) accumulating in actinomycin D-treated cells throughout the infection cycle revealed that, except for the absence of one 18S vmRNA species in L cells, all vmRNA components were formed in the three cell types, although variations occurred in their total and relative amounts from one cell type to another. Kinetic studies of vmRNA synthesis confirmed the absence of one 18S vmRNA species in L cells and also showed that the labelling rate of this vmRNA component is higher in CE cells. Virus proteins synthesized in the infected cells were labelled with 14C-amino acids and analysed by polyacrylamide gel electrophoresis. All the major NDV polypeptides were formed as expected in both CE and MDBK cells but only traces were detected in L cells. In contrast in a cell-free translation system from wheat germ, all of the NDV major proteins were synthesized using RNA extracted from the three infected cell types. Moreover, the total radioactivity incorporated into NDV proteins was twice as great with CE cell RNA and five times as great with L cell RNA than with equivalent quantities of RNA from MDBK cells.


The pathogenesis of infection in mouse brain by Mount Elgon bat virus.

Mount Elgon bat virus given intracerebrally readily killed mice up to 12 days of age (1 to 4 p.f.u/LD50). Virus in doses of 10 to 10(6) p.f.u. killed 40 to 80% of weanling mice of both sexes and up to 20% of adult females and 40 to 80% of adult males. Clearance of brain infectivity in the resistant mice coincided with the appearance of circulating virus neutralizing antibody which occurred earliest in the adult female mice. Immunosuppression of adult mice with cyclophosphamide resulted in the death of all the virus-inoculated mice of both sexes. Virus in these mice reached tenfold higher titres and their brains contained 30- to 60-fold more interferon than the brains of infected mice not given cyclophosphamide. Interferon was apparently without effect on the outcome of infection in the brains of resistant mice and its synthesis reflected the extent of virus growth.


The influence of arginine starvation on the synthesis of virus high molecular weight DNA in HeLa cells productively infected by adenovirus type 5.

In culture cells productively infected by adenovirus a high mol. wt. form of DNA is synthesized which is known to represent, at least in part, virus DNA integrated into cellular DNA. We found that the synthesis of this high mol. wt. DNA and the other DNA size classes can strongly and differentially be influenced by altering the metabolic state of the cells. The effects of different rates of cell growth were tested in this respect as well as arginine deprivation as opposed to application of complete growth medium. Synthesis of virus high mol. wt. DNA and unit genome length DNA is enhanced in actively growing as compared to resting Ad5-infected HeLa cells. Under arginine deficiency, in resting Ad5-infected HeLa cells, integration of virus DNA sequences into cellular DNA is almost totally suppressed whereas virus unit genome length DNA is still synthesized. This differential effect is interpreted by the hypothesis that the formation of virus high mol. wt. DNA is a synthetic process that is independent of the unit size virus DNA replication, but coupled to the synthesis of a special form of a special form of cellular DNA that is less effectively shut off by the infection than cellular DNA in general.


The structural and infected cell polypeptides of influenza B virus.

Structural and virus-induced infected cell polypeptides of several strains of influenza B virus were examined by high resolution polyacrylamide gel electrophoresis and shown to be directly analogous to those of the influenza A viruses. Eight structural polypeptides, P1, P2, P3, HA1, HA2, NA, NP and M were observed in purified virus and at least two additional polypeptides, HA and NS could be detected in infected MDCK cells. The three P proteins  plus NP were shown to be associated with RNA-dependent RNA polymerase activity and HA, HA1, HA2 and NA were shown to be glycosylated. Like the influenza A viruses, migrational differences of some of the infected cell polypeptides could be observed between different B strains. Investigation of a time course of virus replication failed to show any temporal control of protein synthesis in the infected cell.


The immune response of the mouse to lymphocytic choriomeningitis virus. I. Circulating antibodies.

A search was made in LCM virus-immune mice for virus-specific antibodies. With the help of an L-cell plaque assay, neutralizing antibody was readily detected. There were no essential differences between mouse strains, but marked differences existed between virus strains. Whereas the inoculation of either large or small doses of WE strain virus led to the early production of considerable concentrations of neutralizing antibody, in the case of E-350 strain virus, high doses were required and a much longer time interval had to elapse before the threshold of detection was attained. In addition to neutralizing antibody, LCM virus-infected mice produced sensitizing antibody (detected by the enhancing effect of an anti-mouse Ig antiserum on the ability of a serum to reduce virus infectivity) and complement-fixing antibody. Previous failures to detect neutralizing antibody in LCM virus-immune mice might have been caused by properties of the chosen virus, but in many instances lack of a suitable assay host is a more likely explanation.


[Hereditary demyelinating diseases - the leukodystrophies].

Schilder's disease and Greenfield's metachromatic leucodystrophy are hereditary demyelinating diseases, which are probably metabolic and lysosomal, although the responsible enzyme is not yet known for all.


[The use of Bayes theorum in genetic counseling in cases of Duchenne muscular dystrophy].

Bayes' law or theorem (1763) allows the expression of a posterior probability of heterozygosity for an X-linked gene, from two different kinds of information, namely: 1. the prior probability for the mother of an isolated case of Duchenne muscular dystrophy: --to be a carrier by mutation of the gene in one of her parents, or by segregation from earlier generations; --to be herself the origin of the mutation; 2. conditional probabilities, taking into consideration the existence of this woman's normal brothers, sons or maternal uncles and the serum creatine-kinase levels in the possible carrier(s) of the mutant gene. In some situations, these calculations give a recurrence risk which is lower than expected at first and allows sometimes to reassure anxious consultants on their genetic risk.


[Distal renal tubular acidosis: report of 3 cases].

Three observations of R.T.A. with nerve deafness are reported. Case 1 and 2 concern consanguinous brothers whose parents are not affected, which confirm the syndrom as an autosomal recessive entity. The third, sporadic, case relates to a 13-year-old non consanguinous girl. Metabolic abnormalities and renal evolution with nephrocalcinosis was such as in Albright disease. However a progressive nerve deafness makes distinction. The authors underline the importance of this sometimes difficult distinction for genetic counseling.


[Meckel's syndrome: an unusual pedigree].

The authors report the case, in the same family, of two children with the Meckel's syndrome, of two mis-arriages (one foetus with anencephalia, the other one with occipital meningocele), and of two still-born children (one with anencephalia, the other one with occipital meningocele). The genetic problems raised by this unusual genealogy are discussed.


[Genetic counseling in consanguinous marriages].

The authors report their own experience as regard to the genetic counseling for consanguineous marriages. They observe that outpatients still marry and procreate despite the medical advices.


Dane particles and associated DNA-polymerase activity in saliva of chronic hepatitis B carriers.

To investigate furhter the problem of salivary transmission of type B hepatitis, salivas free of blood contamination from three HBsAg-positive carriers with chronic active hepatitis were examined by CsCl equilibrium density gradients and electron microscopy (EM). In the CsCl gradient HBsAg of whole salivas distributed in a band centered at 1.19gm/cm3 with a clearly defined shoulder at 1.24 gm/cm3; the HBsAg was found mainly in the mucous component. On EM examination, fractions from the 1.19 gm/cm3 peak contained spherical HBsAg particles of 22 +/- 3 nm diameter, whereas in the 1.24 gm/cm3 shoulder Dane particles 43 nm in diameter with 28 nm cores were found. Specific hepatitis B virus associated DNA-polymerase activity also was found in the 1.24 gm/cm3 shoulder where the Dane particles occurred and was absent from the saliva of healthy controls. When salivas were incubated for three hours at 37 degrees C the total amount of HBsAg diminished and the 1.24 gm/cm3 shoulder disappeared, probably as a result of endogenous degradation of the Dane particles and the free HBsAg. These findings clearly indicate that the hepatitis B viral particle is present in the saliva of chronic HBsAg carriers with active disease and further confirm that saliva is an important vehicle of infection.


Large-scale efficacy trials of hepatitis B vaccines in the USA: baseline data and protocols.

The protocols of three placebo-controlled, randomized, double-blind clinical trials to assess the efficacy of an inactivated hepatitis B vaccine in dialysis patients and staff, and in male homosexuals are described. Based on the projected attack rates of HBV in the controls, the vaccine efficacy and rates of loss to follow-up, over three thousand participants followed for at least twelve months are required to prove the efficacy of the vaccine. The results of longterm baseline studies in these populations are also reviewed.


Rat-brain pyruvate kinase: purification and effects of lithium.

Purified pyruvate kinase was prepared from pooled brains obtained from untreated rats. Its properties suggest that it is similar to type 'M' pyruvate kinase. Lithium inhibition was demonstrated at pharmacologically significant lithium concentrations (7%-12% at 2 mmol 1-1 Li) and this was similar in character to that previously seen in rabbit muscle pyruvate kinase, namely noncompetitive with respect to phosphoenol pyruvate, K+, and Mg2+ but competitive with ADP.


Molecular specializations of the axon membrane at nodes of Ranvier are not dependent upon myelination.

Nodes of Ranvier from normal and 'dystrophic' mice have been examined with quantitative freeze-fracture electron microscopy. Regions of nodal, paranodal and interparanodal axolemma of normal fibres are clearly distinguishable on the basis of particle size distributions in electron micrographs of freeze-fractured replicas. Protoplasmic fracture faces of normal nodes of Ranvier, contain approximately 40% 100 A particles and about 25% elongated particles 150 by 250 A. Paranodal and interparanodal membranes contain a more uniform distribution of smaller diameter particles. 'Dystrophic', mice of the 129/ReJ-Dy strain have a genetic defect of Schwann cell development and myelinogenesis. Axons of the sciatic and deep peroneal nerves in dystrophic mice, which appear to be normally myelinated, possess approximately the same distributions of particles as axons in normal mice. However, in affected regions of the ventral and dorsal roots, Schwann cell wrappings may be missing, creating heminodes of Ranvier where the myelination terminates or begins again. At such heminodes, there is a circular band of axonal membrane which bears particles of sizes and packing densities similar to that found at normal nodes. High voltage electron microscopic examination of 0.25--1 micron thick sections from these hemi-nodal regions reveals the presence of a filamentous layer beneath the particle-rich membrane. In addition, completely amyelinated regions of root axons contain particle patches having size-density distributions similar to that of both normal and hemi-nodal membranes. Thus, the nodal membrane displays a characteristic particle-size distribution profile. The occurrence of this particle profile does not appear to be dependent upon the presence or absence of Schwann cells. These observations suggest that the functions subserved by the numerous particles at the node of Ranvier are not dependent upon myelination for their local differentiation within the axonal membrane.


The alkaloids of Delphinium cashmirianum.

Dephinium cashmirianum Royle (Ranunculaceae) has yielded the new base cashmiradelphine (12), together with the known alkaloids anthranoyllycoctonine  (9), lycaconitine (15), avadharidine (17), lappaconitine (4), and N-deacetyllappaconitine (7). Pyridinium chlorochromate oxidation of lycoctonine furnished the new aldehyde lycoctonal (11). The arrhythmogenic and heart rate effects of several of these diterpenoidal alkaloids have been measured on the isolated guinea atria. Lappaconitine was arrhythmogenic at 10(-4)M concentrations. But in contrast to the reference drug aconitine, lappaconitine did not increase the heart rate. In anesthetized rabbits injected with lappaconitine, N-deacetyllappaconitine, and lappaconine up to 1 mg/kg, cardiac arrhythmia was quickly observed. Even up to 5 mg/kg, the other substances were non-arrhythmogenic.


Improved antitumor activity by modification of nogalamycin.

Nogalamycin, an antitumor antibiotic, has been converted to a series of analogs by removal of the carbomethoxy group at C-10 and replacement of the neutral sugar at C-7 by other groups. Removal of the carbomethoxy group to give disnogamycin (6) followed by acidic alcoholysis gave pairs of isomeric 7-alkoxy compounds differing in configuration at C-7. Treatment of 6 with trifluoroacetic acid followed by nucleophiles gave a series of analogs having substituents at C-7 with a configuration at C-7 opposite to that of nogalamycin. Among the analogs prepared, 7-con-O-methylnogarol (7) is a highly active antitumor agent.


The relationship between some measures of synaptic ultrastructure as a function of distance from the soma on lamprey reticulospinal neurons.

Synaptic junctions located on the dendrites of lamprey (Petromyzon marinus) reticulospinal neurons labelled with intracellularly-injected horseradish peroxidase were studied. The normal ultrastructure of the synaptic junctions was defined and several quantitative measures made from each junction in order to test the hypothesis that distally-located synapses are ultrastructurally different from those located at proximal dendritic sites. A total of 820 contacts from one neuron and 279 from a second neuron ranging from 20 to 340 microns from the soma were quantified. The vast majority of the presynaptic endings contained round, clear-cored vesicles and formed an asymmetrical membrane differentiation with the postsynaptic dendrite. A small fraction of the population contained flattened or pleomorphic vesicles and these synapses were equally distributed with respect to distance from the soma. Many of the terminals contained a few large dark- and clear-cored vesicles. Four quantitative measures of each synaptic contact were made. These included vesicle number, length of differentiated membrane, vesicle area and terminal area. Four ratios relating the different quantitative measures were also calculated. Each ratio or measurement from the synaptic junctions was plotted as a function of distance from the soma to determine if differences existed at any distance. It was found that synaptic junctions are uniformly similar and that distal junctions did not differ significantly (P greater than 0.05) from those at proximal dendritic sites. It is concluded that if distal synapses do compensate for their remote location they do this is some other way, possibly by increasing the number of synaptic contacts made by each presynaptic axon.


Comprehensive survey of indigenous Iraqi plants for potential economic value. 1. Screening results of 327 species for alkaloids and antimicrobial agents.

Three hundred and twenty-seven indigenous Iraqi plant species were screened for their antimicrobial activities and for the presence of alkaloids. The plants represent 221 genera and 49 families. Ethanolic extracts were tested for the presence of alkaloids with both Mayer's and Dragendorff's reagents. Of these, 146 species, belonging to 109 genera distributed among 32 families, showed positive alkaloid tests to both reagents. Of these, 26 genera have not been previously reported in the literature as containing alkaloid-bearing plants. The extracts were also tested for their in vitro antimicrobial activity by an agar dilution-streak method against six economically significant microbes. Five species were highly active, while another 90 species showed weak activity against one or more microorganisms.


Morphological changes in afferent vestibular hair cell synapses during the postnatal development of the cat.

In the course of postnatal development in the cat, there is a decrease of about 93% in the total number of synaptic bodies (synaptic balls and synaptic bars) in type I hair cells. In type II hair cells, there is no change in the number of synaptic balls. Simultaneously, the length of specialized neuroepithelial contact increases by approximately 300% during type I hair cell maturation. Only the synaptic bars displaying a polylamellar ultrastructure persist in the type I hair cells of the adult animal. It is suggested that the afferent vestibular synapses of the type I hair cell are transformed during ontogeny.


Thyroid-hormone concentrations after radioiodine therapy for hyperthyroidism.

Fourteen hyperthyroid patients (11 men, three women), ages 28-66, were followed with serial measurements of serum thyroid hormone levels for 1 mo after therapy with I-131. Twelve patients had diffuse toxic goiters (25--70 g in size); two patients had multinodular glands (40--100 g). The patients were taking no antithyroid medications; ten patients were treated with propranolol. All patients received the equivalent of 5000 rad, except the two with multinodular glands, who received larger doses. There was no consistent pattern of serum T4 and T3 levels after the I-131 therapy. For the entire group, there was no significant increase of the mean serum hormone concentration. One group (three patients) had a mean T4 increase of 28% and a T3 increase of 91% above baseline at Days 10--11. Seven patients had minimal increases of hormone levels at Days 2--3, and a third group (four patients) had no increase of thyroid hormones after I-131 therapy. The patients with no rise in hormone concentrations had smaller goiters than the other groups. There was no correlation of the dose of radioactive iodine, or of the initial hormone concentration, with the rises or declines of T4 and T3 levels after I-131 therapy. Radioiodine therapy caused no significant increase of serum T4 and T3 concentrations in the majority of patients.


Myocardial accumulation of labeled phosphate in malignant pericardial effusion.

Recognition of the presence of a malignant pericardial effusion is important because of the insidious onset and life-threatening potential. This report presents data on three patients with metastasizing carcinoma of the breast, all admitted with a presumptive clinical diagnosis of malignant pericardial effusion. Two of the three revealed a diffuse concentration of Tc-99m PPi within the myocardium. In these cases the diagnosis was confirmed by identification of malignant cells in the pericardial fluid. We suggest that the use of labeled phosphate agents may allow early recognition of myocardial involvement in patients with disseminated neoplasm that results in malignant pericardial effusion.


Xenon-133 retention in hepatic steatosis-correlation with liver biopsy in 45 patients: concise communication.

This study presents the results of comparison of hepatic fat content with hepatic xenon retention in 45 patients. The degree of hepatic Xe-133 retention was measured during pulmonary ventilation studies. The amount of hepatic steatosis was graded 0 to 4+ on histologic liver sections obtained by needle or surgical biopsy. There was agreement between the amount of hepatic xenon retention determined scintigraphically and the degree of steatosis determined histologically. These results suggest that Xe-133 retention in the liver provides a simple means of evaluating fatty infiltration of the liver. The potential of this technique as a noninvasive means of investigating hepatic fatty infiltration is discussed.


Gated and cinematic perfusion lung imaging in dogs with experimental pulmonary embolism.

To determine how pulmonary respiratory motion affects detection of pulmonary emboli, 11 dogs had routine lung scans and gated or cinematic perfusion images after undergoing autologous experimental pulmonary embolism. Six dogs had routine six-view perfusion studies, plus end-inspiratory and end-expiratory gated perfusion studies performed with a physiologic synchronizer set to 80% threshold. Five other dogs had three-view ungated and cinematic (post., LPO, RPO) perfusion images. Cinematic studies were acquired by synchronizing a camera-computer system to the Harvard respirator that ventilated the dog. Before death, all animals received i.v. India ink to outline pulmonary perfusion defects, and postmortem lung dissection verified sites of emboli. An ROC curve analysis of randomized perfusion studies showed that end-inspiratory gated images yielded true-positive rates 5--10% higher than ungated images at any given false-positive rate. Lesion detection by cinematic studies was comparable to detection by ungated images, but detection by end-expiratory images was worse. End-inspiratory gated imaging may be useful as an occasional adjunct to routine perfusion lung imaging.


Potential anticancer agents. XIV. Isolation of spruceanol and montanin from Cunuria spruceana (Euphorbiaceae).

Montanin (1) and spruceanol (2), two quite different diterpenes, were found to be responsible for the cytotoxic and antitumor activity of the root and root bark of Cunuria spruceana (Euphorbiaceae). The structure of spruceanol (2) was deduced from spectral interpretation and chemical correlation with 12-methoxycleistanth-8,11,13-trien-3-one (4).


The in vitro stability of [131I]o-iodohippurate.

The kinetics of [131I]o-iodohippurate (OIH) decomposition were evaluated to develop an accelerated technique for the prediction of radiopharmaceutical stability. Decomposition due to temperature and self-absorbed radiation was characterized simultaneously. Using the proposed technique, the extent of OIH decomposition was calculated a priori to be 7.6% over a 28-day period when extrapolated to a specific concentration of 2 mCi/ml and a storage of 25 degrees C. Extended storage under these same conditions yielded a mean decomposition value of 8.0% over an equivalent time interval. The techniques developed offer a rapid means of accurately assessing the stability of new radiopharmaceuticals as they are developed.


Regional ventilatory clearance by xenon scintigraphy: a critical evaluation of two estimation procedures.

Estimates of ventilatory clearance are usually made by inspecting xenon washout images. Quantitative computer procedures have been described that produce regional clearance rates, yet their accuracy is not well established. We define a mathematical model for scintigraphic ventilation data based on 96 clinical studies, and with this model we test the accuracy of two procedures used to estimate ventilatory clearance. The least-squares curve-fitting technique for both washin and washout data has the same accuracy as a modified Stewart-Hamilton method (A/H) that uses washout data alone. Both procedures demonstrate relative errors of less than 5% and coefficients of variation of 10-20% when regions with equilibrium count rates of 3 cps and clearance times between 10 and 90 sec are examined. Because the A/H procedure is preferred for its simplicity and speed, we analyze two of its main sources of error: early washin/washout termination and background activity. To measure regional ventilation by the A/H procedure, we recommend: (a) washin and washout periods at least three times the largest clearance time of clinical interest; b) a regional equilibrium count rate of at least 3cps; and c) a 25- to 50-sec average of the equilibrium count rate.


Can the extent of coronary artery disease be predicted from thallium-201 myocardial images?

The accuracy with which the extent of coronary artery disease can be predicted from stress thallium-201 myocardial images has been assessed in 81 patients with chest pain. Whereas the appearance of the myocardial images was both a sensitive means of detecting coronary artery disease (images abnormal in 43 of 47 patients with abnormal coronary arteriograms) and specific in excluding it (images normal in 31 of 34 patients with normal arteriograms), there was poor correlation between the extent of disease predicted from the Tl-201 images and the findings at arteriography. It is concluded that although stress Tl-201 myocardial imaging is a useful method for the noninvasive diagnosis of coronary artery disease, it cannot be relied upon to predict the number of abnormal vessels.


Reliability of gated heart scintigrams for detection of left-ventricular aneurysm: concise communication.

Gated heart scintigrams and angiocardiograms were performed on 138 patients with coronary artery disease. Scintigraphy detected 61 of 64 left-ventricular aneurysms demonstrated by angiography. The scintigram correctly identified all 54 apical and anteroapical aneurysms and one inferior aneurysm, but missed one of six anterior and two of three posterobasal aneurysms. In 74 patients with angiograms negative for aneurysm, the scintigram was also negative in 72, with two false positives. Overall accuracy of gated heart scintigraphy for the detection of aneurysm was 96% (133/138). Cardiac surgery was performed on 76 patients, and the angiographic findings were confirmed in all cases. Scintigraphic findings were confirmed in 74 of 76 patients. Among ten patients scintigraphed before and after aneurysmectomy, problems were first recognized by the scintigram in three cases. Gated heart scintigraphy is recommended as a screening procedure for suspected left-ventricular aneurysm because of its high reliability in the apical and anterior portions of the heart, where most of the aneurysms occur.


Comparison of wall motion and regional ejection fraction at rest and during isometric exercise: concise communication.

The detection of regional abnormalities of left-ventricular wall motion provides strong evidence for the presence of coronary heart disease. In 129 patients undergoing coronary arteriography, the relative value of radionuclide angiographic assessment of wall motion was compared with computer-generated regional ejection fraction, at rest and during handgrip exercise. Wall motion was determined by superposition of computer-derived end-diastolic and end-systolic perimeters. Relative regional ejection fraction was determined using a computer-generated 16-color isocount image that permitted a quantitative assessment of zonal contribution to ejection fraction. Of the 129 patients, coronary arteries were normal in 31 and diseased in 98. Of 24 patients with single-vessel disease, wall-motion abnormalities were present, at rest or during exercise in 15, whereas regional ejection fraction detected 20 patients. Seventy-four patients had multivessel disease. Of these, wall-motion abnormalities occurred in 52 but regional ejection fraction was abnormal in 69 (p less than .01). Overall, sensitivity was 67% by wall motion and 91% by relative regional ejection fraction (p less than .001). Specificity was 94% by wall motion and 87% by regional ejection fraction (not significant). Thus, radionuclide angiographic assessment of regional ejection fraction during handgrip exercise is both highly sensitive and specific for coronary heart disease and significantly enhances detection of coronary heart disease compared with wall-motion assessment, with little loss in specificity.


Comparison of the liver's respiratory motion in the supine and upright positions: concise communication.

The effect of the upright position, compared with the supine position, in reducing respiratory motion during liver scintigraphy has been measured in a group of 51 randomly selected patients. Little improvement in spatial resolution can be expected from simply standing the patient upright, and analog motion correction is recommended as much more effective in minimizing respiratory motion artifacts in liver scintigraphy.


Radioimmunoassay of hair for determining opiate-abuse histories.

Heroin and morphine metabolites can be detected in hair with the use of commercially available radioimmunoassay reagents and with minor sample preparation. Hair samples obtained from morphine-treated mice and heroin users contained nanogram levels of the drug per milligram of hair (single human hair). The results of the hair analyses for all subjects admitting the use of heroin were positive, whereas the results of only 30% of thin-layer chromatographic urinanalyses of these same subjects were positive. In addition, differences in drug concentration for sections of hair near the scalp and near the distal end correlated with the length of time the drug had been used. These results exemplify the potential advantages of the use of hair analysis over urine and serum analyses in terms of accessibility, sample stability, and long-term retention of information.


The biodistribution of Br-77-labeled estrone: visualization of tissues containing increased estrogen receptors.

The biodistribution of a lipid-soluble brominated female sex hormone, Br-77-labeled estrone, has been determined in dogs and rabbits. Following i.v. or intragastric administration, Br-77-labeled estrone localized in the gallbladder bile within 20-25 min. Fifty-six percent of the injected activity was found in the bile at 90 min. Breast tissue was imaged by scintillation camera in three female dogs with postpartum breast enlargement at 24 hr following i.v. injection. Analysis of digitized images revealed 50% more counts per pixel in stimulated female breast tissue than was in surrounding tissue. Increased Br-77 activity in the breast was associated with the presence of increased concentrations of estrogen receptors.


"Circumferential profiles:" a new method for computer analysis of thallium-201 myocardial perfusion images.

A method for computer analysis of thallium-201 scintigrams is described, in which the left-ventricular activity is measured along radii constructed from the center of the left ventricle (LV) to each point on the LV circumference. Data are then displayed graphically as a "circumferential profile" of normalized activity against radial location. Thallium defects are identified and scored by comparison of the profile curve with empirically determined normal limits. In patients with coronary artery disease, defect scores were found to be quantitative and reproducible, and to agree generally with subjective visual analysis.


Radioiodinated DNA as potential tumor-imaging agent.

Radiolabeled heat-denatured DNA and some related nucleic acids localize in tumor tissue following i.v. administration. Retention of radioactivity by tumors is longer than for other organs and localization of tumor tissues is possible by scintigraphic techniques 8-24 hr after injection.


Improvement of pulse-mode photographic images in MDS computer systems.

Users of scintigraphic computer systems manufactured by Medical Data Systems (MDS) frequently experience textured distortion of oscilloscope images when scans are produced by Z-axis modulation. The source of this malfunction has been identified and a way to eliminate the problem is described. The modification is simple and inexpensive. The availability of Z-axis modulation significantly enhances the capabilities of MDS systems.


Labeling efficiency and stomach concentration in methylene diphosphonate bone imaging.

When Tc-99m-labeled methylene diphosphonate was used 4.75 hr after reconstitution, stomach concentration was seen in the majority of patients. The number of patients with stomach concentration increases with time and is associated with the increasing amount of product breakdown. The reconstituted labeled MDP should not be used after 4-5 hr if stomach concentration is to be avoided.


Gallium-67 citrate scanning--a new adjunct in the detection and follow-up of extrapulmonary tuberculosis: concise communication.

Gallium-67 citrate scans were obtained in 11 patients considered at risk for extrapulmonary tuberculosis. Radiographic and bacteriologic studies were performed routinely and tissue biopsy selectively. Of five patients with proven extrapulmonary tuberculosis, there were three with renal tuberculosis, one with Pott's disease, and one with peritoneal tuberculosis. The Ga-67 scan correctly predicted presence or absence of active extrapulmonary foci in all 11 patients. Follow-up scans correlated well with clinical response to therapy. The diagnosis of extrapulmonary tuberculosis is often overlooked because of nonspecific symptoms and frequent lack of concurrent lung involvement. Scanning with Ga-67 citrate offers a reliable and simple means of screening patients at risk and of monitoring response to treatment.


A transferrin-mediated uptake of gallium-67 by EMT-6 sarcoma. I. Studies in tissue culture.

We have studied the in vitro uptake of gallium-67 by exponentially growing EMT-6 sarcoma cells in long-term tissue culture. In this system, the addition of transferrin to the medium was required before an appreciable cellular uptake of Ga-67 occurred. The transferrin effect was complex, with an initial stimulation to a peak cell-to-medium ratio of 8--10:1 at low concentrations of transferrin (0.2 mg/ml), followed by a gradual decline in uptake as transferrin in the medium was increased further. EMT-6 tumor-cell uptake of Ga-67 was probably mediated by a specific cellular receptor for transferrin. Scatchard analysis of the EMT-6 cellular binding of human transferrin labeled with iodine-125 indicated a cellular receptor with affinity for transferrin of 5 X 10(6) l/mole and abundance of 500,000 receptors per cell. Over the experimental range of transferrin concentration in the medium, the observed uptake of Ga-67 was closely correlated with the degree of formation of Ga-67-labeled transferrin and the fraction of transferrin bound to the cellular receptor (N = 69, r = 0.86, p less than 0.0001).


A transferrin-mediated uptake of gallium-67 by EMT-6 sarcoma. II. Studies in vivo (BALB/c mice): concise communication.

The EMT-6 sarcoma-like tumor of BALB/c mice can be grown as a solid subcutaneous transplantable tumor in vivo or as a monolayer culture in vitro. We have studied the uptake of gallium-67 by this tumor growing subcutaneously on the backs of 6-week-old BALB/c mice. After i.v. administration of Ga-67 citrate, tumor uptakes were as high as any others reported for mouse tumors. Also, for unknown reasons, there was appreciable reduction in tumor uptake with increasing amounts of Ga-67 citrate, even in the microcurie range. Furthermore, when mouse serum is prelabeled with Ga-67 and then injected, the EMT-6 uptake is greater than with Ga-67 administered as citrate (p less than 0.02). We believe that the finding of avid Ga-67 uptake in vivo helps to establish this unique in vivo/in vitro tumor system as a valid experimental model for studies regarding the mechanism of Ga-67 accumulation by neoplastic tissue.


Detection of a false left ventricular aneurysm by first-pass radionuclide ventriculography.

A false left ventricular aneurysm complicating an inferior myocardial infarction was successfully identified by computerized first-pass radionuclide ventriculography using a multicrystal gamma camera. The aneurysmal chamber exhibited paradoxical systolic expansion, a pattern of contraction confirmed by contrast cineangiography. Because of the propensity of false aneurysms to rupture, early noninvasive firm diagnosis is desirable and may be accomplished by first-pass radionuclide angiography.


The gallium "bone scan" in acute leukemia.

A 15-year-old boy with acute leukemia had a gallium-67 scan that was virtually identical to his technetium-99m pyrophosphate bone scan, except for lack of renal visualization. The quality of the radiopharmaceutical was assured by the normal appearance of gallium scan performed in another patient on the same day and with the same radionuclide batch. This extensive osseous uptake was probably due to bone-marrow replacement by leukemia cells and is a pattern that should be recognized as indicating a diffuse marrow-infiltrating disease.


DL-[Carboxyl-11C]tryptophan, a potential agent for pancreatic imaging; production and preclinical investigations.

In animal studies, DL-[carboxyl-14C]tryptophan [DL-Try(C-14)] showed a high specificity for the pancreas, which suggested the potential of DL-[carboxyl-11C]tryptophan [DL-Try(C-11)] for clinical pancreatic inaging. The blood clearance and tissue uptake of the amino acid were very rapid, and no carrier effect was observed through a dose of 5 mg/kg. None of three transplanted hamster pancreatic adenocarcinomas that we studied showed a selective uptake of DL-Try(C-14) by the tumor, and none of the three enzymatic regimens investigated gave significant enhancement of the pancreatic specificity. Commercial L-Try(C-14) gave slightly better pancreatic specificity than the analogous racemic compound but without enough improvement to warrant attempts at optical resolution. DL-Try(C-11) was synthesized in amounts up to 325 mCi using a rapid, high-temperature, high-pressure modification of the Bücherer-Strecker amino acid synthesis. Yields ranged from 30--60%, and a total of 40 min was required for synthesis and chromatographic purification. DL-Try(C-11) thus appears to have significant potential as a clinical pancreas-imaging agent, particularly when used in conjunction with positron computerized transaxial tomography.


The distribution of the muscarinic acetylcholine receptor antagonists, quinuclidinyl benzilate and quinuclidinyl benzilate methiodide (both  tritiated), in rat, guinea pig, and rabbit.

The distribution of [3H]quinuclidinyl benzilate and its methiodide salt was determined in rat, guinea pig, and rabbit. Accumulation in the myocardium of up to 2% of the injected dose per gram of tissue was obtained with both compounds, providing heart-to-blood ratios of approximately 30 and heart-to-lung ratios of approximately 4. The accumulation in the heart was blocked (89%) by preinjection of atropine. The distribution of tritium in rabbit heart corresponds to the muscarinic receptor densities determined in vitro. Calculation of the theoretical maximum for the bound-to-free ratio, based on in-vitro equilibrium binding isotherms, resulted in ratios in reasonable agreement with the experimental results. Because of the high accumulation in the heart with low serum concentration, we conclude that the methiodide salt of quinuclidinyl benzilate represents an ideal parent structure for the design of a receptor-binding gamma-emitting radiopharmaceutical for imaging of the myocardium.


Labeling of red blood cells with Tc-99m after oral administration of SnCl2: concise communication.

In vivo labeling of red blood cells with Tc-99m was possible after prior oral administration of SnCl2, both in rats and human volunteers. Absorption of oral SnCl2 was low but sufficient for more than 95% labeling efficiency. Prior i.v. administration of stannous chloride is known to induce in vivo labeling of red blood cells with pertechnetate. We have observed that such labeling is possible even after oral administration of stannous chloride. Nearly 95% of the circulating radioactivity and 93.7% of the administered radioactivity was in RBCs 30 min after i.v. injection of 99mTcO4- in rats that were fed 5 mg of stannous chloride (3.13 mg Sn2+ ion) 2 hr before injection. Red blood cells from four human volunteers could bind pertechnetate, both in vitro and in vivo, after oral administration of 100 mg of SnCl2. We have obtained a blood-pool image of the human heart by labeling the RBCs in vivo by this method. We have also studied various parameters affecting the in vivo binding of RBCs with Tc-99m--such as the amount of orally administered SnCl2, the time of injection of radionuclide after oral SnCl2, and the optimum time for the imaging.


Release of gamma-aminobutyric acid from isolated brain synaptosomes during semicarbazide-induced convulsions.

The in vivo effect of semicarbazide (SC) and aminoxyacetic acid (AOAA) upon gamma-aminobutyric acid (GABA) levels in the synaptosomal fraction, and GABA release from the same fraction were studied in the mouse. The convulsive dose of SC reduced the GABA content in synaptosomes, and when the convulsions were protected by pretreatment with AOAA, the reduced GABA content in synaptosomes rose to or above the normal level. Moreover, the SC treatment decreased the GABA releases from synaptosomes both in a Ca2+-free Ringer's solution and in a high K+ Ringer's solution. When the convulsions were protected by pretreatment with AOAA, the decreased GABA releases in both the conditions rose to or above the normal levels. Therefore, it is suggested that the decrease in GABA release from the nerve endings, because of the decrease of GABA content in the same compartment, is possibly an important factor in the onset of some kinds of convulsions.


Developmental and convalescent changes of the anemia caused by excess methionine in the rat.

An experiment was performed to investigate the progressive changes of the anemia caused by excess methionine in rats, during the developmental and convalescent periods. Hemoglobin concentration, hematocrit value and red blood cell count were elevated in the initial stage of excess methionine feeding, they then gradually decreased. During the convalescent period, after ceasing administration of the excess methionine diet, symptoms of anemia gradually disappeared. The nitrogen efficiency ratio was markedly decreased by excess methionine feeding, however, it rapidly recovered after ceasing administration of the excess methionine diet. Iron deposited in the spleen also disappeared quickly during the convalescent period. Of the symptoms of anemia caused by excess methionine, the activity of delta-aminolevulinic acid synthetase (ALAS) in the bone marrow showed the most acute changes. As mentioned previously (1), although lowered globin biosynthesis due to amino acid imbalance could be considered as one of the important causes of the anemia and also would be one of the causes of the compensative elevation of ALAS activity, the possibility still remain that there are other unknown effects of excess methionine on ALAS activity in the bone marrow in this case.


An autoradiographic study of new fat cell formation in adipose tissue in adult mice during malnutrition and refeeding.

The renewal of adipose cells in adult mice has been autoradiographically studied. The number of adipose cells was diminished by eighty percent during malnutrition and the same number of adipose cells proliferated during the refeeding stage. The results of our study showed that adipose tissue, which had previously been believed to be stable in cell number, has the capacity for cell proliferation according to changes in nutritional status.


Effect of antivitamin B6 on regional GABA metabolism in mouse brain and its relation to convulsions.

The effects of administration of DL-penicillamine (PeA), thiosemicarbazide (TSC), semicarbazide-HCl (SC) as convulsants and pyridoxine (PN) as anticonvulsant on gamma-aminobutyric acid (GABA) content, glutamic acid decarboxylase (GAD) and gamma-aminobutyric acid transaminase (GABA-T) activities in cerebral cortex, striatum, diencephalon, mesencephalon, cerebellum and pons/medulla were investigated. The onset of convulsions induced by these convulsants coincides with the fall in GABA content and GAD activity in the mesencephalon area, and in contrast, the cessation of the convulsions by PN supplement coincides with the recovery in both the parameters. Aminooxyacetic acid (AOAA), a potent GABA-elevating agent showed an anticonvulsant property against convulsion by TSC for several hours after the injection of AOAA, but lost this property 16 hr after the treatment. The TSC administration 16 hr after the AOAA pretreatment significantly decreased the GABA content in all the regions, particularly in the mesencephalon and diencephalon areas, which had been elevated by the AOAA pretreatment, together with its ability to induce convulsion. FRom the above results it may be postulated that the critical drop of GABA level from a plateau to another lower level following the decrease of GAD activity in the mesencephalon area is an important factor in the induction of convulsion.


Assay of 25-hydroxyvitamin D3 in human plasma by high-performance liquid chromatography.

A method using preparative and analytical high-performance liquid chromatography (HPLC) is proposed for the assay of 25-hydroxyvitamin D3 (25-OH-D3) in human plasma. A constant volume (0.2--2.0 ml) of a plasma sample was saponified. The unsaponifiable matter was first applied to preparative HPLC using a column of the straight-phase type (Zorbax SIL) in order to separate a 25-OH-D3 fraction from lipophilic concomitants giving ultraviolet-absorbing noise. Then, the separated 25-OH-D3 fraction was applied to analytical HPLC using a column of the reversed-phase type (Zorbax ODS) in order to measure the content of 25-OH-D3 from the peak height. This is a revised method from Jones (1978): Clin. Chem., 24, 287--298). The results showed that the clean-up procedure by the first preparative HPLC was successfully performed because the peak corresponding to 25-OH-D3 on the chromatogram of the second analytical HPLC was not disturbed by any other interfering peaks. Moreover, recovery through the whole procedure was satisfacotry (about 100%) and the procedures of saponification and isolation of the unsaponifiable matter diminished the overload to the columns. These are the revised points of Jones' method. When two determinations were performed on 12 samples of plasma taken from normal adults in October, the values were 22.6 +/- 4.8 and 21.0 +/- 3.6 (mean +/- SD) ng/ml, respectively.


Effect of alpha-tocopherol on the binding of benzo(a)pyrene to nuclear macromolecules.

The binding of metabolically activated 14C-benzo(a)pyrene (BP) to rat liver nuclei was studied. The nuclear framework structure, termed the nuclear matrix, was found to bind BP preferably. alpha-Tocopherol inhibited the binding of BP to nuclear macromolecules in the presence of phenobarbital (PB)-induced microsomes. alpha-Tocopherol decreased the level of BP metabolites in nuclei, though it did not inhibit the activation of BP by PB-induced microsomes.


Tocopherol level in human blood cells.

The study was undertaken on alpha-tocopherol levels, separated from other analogs, in the red blood cells (RBC) of cord blood, premature infants, healthy children and adults as well as in pregnant women, and compared with the plasma levels. 1. The majority of tocopherol found in the RBC was localized in the membranes. 2. Only alpha-tocopherol was found in the RBC, while alpha- and gamma-tocopherol were found in the plasma. 3. Alpha-tocopherol level in the RBC changes during development in parallel with that in the plasma. However, changes in the level in RBC were smaller than those of the plasma. 4. The ratio of alpha-tocopherol level in the RBC to the plasma was higher in the cord blood and premature infants than in the children and adults. 5. With regard to the cases with high plasma tocopherol levels, a different finding was obtained on the RBC level between pregnant women and adults with a large amount of tocopherol. In the latter, high RBC levels were observed as the plasma levels were elevated, while in spite of a high plasma tocopherol level, the lowest RBC levels were found in the pregnant women.


Concentrations of thiamine and its phosphate esters in rat tissues determined by high-performance liquid chromatography.

Thiamine and its phosphate esters in rat tissues were determined by high-performances liquid chromatography after conversion to thiochrome and its phosphate esters as described previously (Ishii, K. et al. (1979) Anal. Biochem., 97, 191--195). Total thiamine concentrations determined in the brain, liver, heart and kidney were 6.39, 25.5, 15.7, and 14.9 nmol/g wet weight, respectively. In all the tissues tested, thiamine, its monophosphate, pyrophosphate, and triphosphate were found to be present at the rates of 1.2--7.7, 5.9--11.0, 85.7--90.0, and 0.7--1.6%, respectively. Although these percentages of thiamine phosphates were similar to those reported previously using different procedures, the percentage of thiamine triphosphate content in rat tissues was found to be fairly lower than that reported. The time required for the whole process of analysis averaged 90 min after isolation of the organs from the animal.


Effect of feeding amino acid mixtures on lipid transport from rat liver as measured by liver perfusion.

The present study examined lipid transport from the liver by application of a perfusion technique in situ in rats fed diets consisting of a low level Wheat- or Rice- or Miyazaki-pattern amino acid mixture. The net release of hepatic lipids was directly estimated from analysis of the perfusate continuously circulated through the liver. The net release of triacylglycerol from the liver into the perfusate was much more severely inhibited in the Wheat-pattern diet group, and more depressed in the Rice-pattern diet group than in the control group, and slightly reduced in the Miyazaki-pattern diet group compared with the control. For serum lipoproteins fractionated by the Quick-Disc electrophoresis method, there was a significant decrease in the serum pre-beta lipoprotein fraction in all the rats of the Wheat-, Rice- and Miyazaki-pattern diet groups compared with the control. These observations provide firm support for the hypothesis that in rats given diets consisting of a low level of the Wheat-, Rice- and Miyazaki-pattern amino acid mixture, fatty liver may be produced mainly by an impaired transport of triacylglycerols from the liver.


A sero-epidemiological study of onchocerciasis with the indirect hemagglutination test.

A sero-epidemiological study of onchocerciasis was carried out with the IHA test in Guatemala. In the endemic area, 94.8% of 191 subjects with microfilariae, 82.1% of 172 cases with onchocercal nodules, and 22.5% of 236 cases without either microfilariae or nodules gave positive reactions in the IHA test. On the other hand, the positive rate was 3.4% and 2.0% in each of 2 nonendemic populations. A follow-up study of reactors in the IHA with neither microfilariae nor nodules showed that 11 (42%) of 26 cases were microfilaria positive 6--7 mo after the first examination. A clear correlation was found between the results of the IHA test and skin biopsy in surveyed communities. The age distribution curve of the IHA test was closely associated with skin biopsy although the former was higher. IHA titers rose in proportion to microfilarial density. A sex-related difference was evident in the IHA positive rate in the subjects from medium and low endemic areas, but little difference was shown in individuals from a higher endemic area.


Eimeria caprina sp. n. from the domestic goat, Capra hircus, from the USA.

The ellipsoidal to slightly ovoid oocysts of Eimeria caprina sp. n. from the domestic goat, Capra hircus, are 27--40 x 19.5--26 micrometers (mean 31.8 x 23.1 micrometers); their sporocysts are 13--17 x 7--10 micrometers (mean 15.3 x 8.5 micrometers). The oocyst wall is 1.7 micrometers thick, smooth, dark brown to brownish-yellow, and 2-layered. Micropyle, polar granule, and sporocyst residuum are present; micropylar cap and oocyst residuum are absent. These features distinguish the new species from other species in the genus which have a micropylar cap, or are smaller, or have a small micropyle with a small internal plug.


Home visiting: the part played by the "intermediary".

Home visits are usually initiated by someone other than the patient, whom we have called the ;intermediary'. It is seldom an objective decision on the part of the intermediary that a home visit is needed and is usually the result of his or her own anxieties. It is therefore important for the doctor consciously to aim at a diagnosis or formulation of the problems of the intermediary, and it may then become possible deliberately to help him or her with his needs and anxieties. If these are not considered, the strains or dissatisfaction of the intermediary which initiated the visit are likely to remain, and to influence both the way the doctor treats the patient and the doctor's sense of competence.


General practitioners and their staff.

We describe the practices and staff of a random sample of 158 doctors. There has been a significant increase in ancillary staff since the Doctors' Charter of 1966. The study showed that there was no economy in staff as the number of doctors in a practice increased. In the selection of receptionists, doctors preferred married women over 35 with children, and were in favour of good personal qualities rather than good academic attainments. However, a high standard of work is important as well as a good understanding of human behaviour.


'Red'flu': a study of an epidemic in a girls' boarding school in February 1978.

All the girls at a boarding school who presented with symptoms of influenza were interviewed and examined. Their symptoms and signs were recorded and related to age, date of last menstrual period, and previous influenza immunization.The age of the girls had no influence on the incidence of disease, nor did the number of girls in each form. However, the attack rate among those not immunized was 61 per cent and those not immunized 71 per cent. There was a highly significant variation in the incidence of influenza in relation to the menstrual cycle.


The self-assessment of confidence, by one vocational trainee.

A list of important topics in general practice was constructed and a trainee was asked to indicate his confidence about each topic on a one to five scale. Repeated use showed different confidence ratings by the same trainee, and an attempt was made to correlate factual knowledge by using a multiple choice questionnaire.Despite important limitations, which are described, this method may be useful in identifying suitable topics for teaching during the trainee year.


The accessibility of primary care to urban patients: a geographical analysis.

Techniques of analysis now used in geography can be applied to general practice. They show that the locations of general practitioners' premises are not randomly distributed and are not related to the sites of populations or to patients in greatest need of medical services. The findings suggest a concentration of surgeries in middle class areas and lack of surgeries in areas of low social class.


The prevalence of epigastric bruit.

A study was undertaken to determine the prevalence of an epigastric bruit in a family practice population.Auscultation of the abdomen in the supine position was carried out on 567 consecutive unselected patients. A systolic bruit, localized to the epigastrium, was heard in 80 patients, none of whom had symptoms of intra-abdominal vascular disorder.The results are tabulated by age and sex, and show statistically significant differences in prevalence between males and females of similar ages, and also between different age groups of the same sex. The highest prevalence is in females aged between 10 and 34 years and no significant difference is found between pregnant and non-pregnant females of comparable age.


Group discussions with doctors' wives.

Group discussions with the wives of general practitioners and vocational trainees have been held in the Boston vocational training scheme. Many problems such as being tied to the telephone and the registration of doctors' families with other general practitioners were discussed.A consensus view emerged on several topics and I believe that further discussions with wives on subjects of this kind would be valuable.


Prodromal itching in asthma.

In a survey of 50 young asthmatic patients it was found that 35 (70 per cent) recognized a sensation of itching of the lower face, or anterior neck, in association with their asthmatic attacks. Sixteen (32 per cent) said that the itching closely preceded subjective wheezing, and 15 (30 per cent) said that it occurred at the onset of wheezing. Recognition of this early sign may be of use in the prompt treatment of an asthmatic attack.


Family practice in the United States of America: the first 10 years.

Family medicine was officially recognized as an independent discipline and as the twentieth specialty in the USA in 1969, when the American Board of Family Practice was established.The main achievements of the first 10 years has been the establishment of departments of family practice in two thirds of the medical schools and the growth of graduate training has advanced rapidly from a total of 290 residents in 1970 to over 6,000 in 1978. Developments in group practice, team work, and medical records have been considerable and research is expanding.In 1976, the 50-year trend of falling numbers of family physicians in the USA was reversed for the first time and excellent progress is also being made in countering the geographical maldistribution of physicians.The challenges for the future of family practice are different from those in the past and are discussed.


French general practice.

Five British general practitioners spent a week with French colleagues in various parts of France. We report results of the observations which we agreed in advance to make about some aspects of clinical practice in France.


Anglo-Australian exchange in general practice.

An exchange visit was arranged between the two partners in our practice in Chagford, Devon and two partners in a practice in Canberra, Australia.This enabled us to experience a completely different type of practice and financial system with the opportunity of working closely with other colleagues, while maintaining continuity of treatment for our own patients, since only one partner was exchanged at a time.We also benefited from seeing new patients with new diseases, and felt that our patients probably benefited from seeing a new doctor. We returned to the practice refreshed and stimulated and our patients appeared to have been similarly stimulated by the impact of a different culture.


Night calls in a group practice.

Night calls attended in 1977 by four doctors in a group practice were analysed and divided into categories of urgency.Slightly over half the night calls were genuine emergencies, and only four per cent were totally unnecessary.


Killing, letting die and euthanasia.

Medical ethicists debate whether or not the moral assessment of cases of euthanasia should depend on whether the patient is 'killed' or 'allowed to die'. The usual presupposition is that a clear distinction between killing and letting die can be drawn so that this substantive question is not begged. I contend that the categorisation of cases of instances of killing rather than as instances of letting die depends in part on a prior moral assessment of the case. Hence is it trivially rather than substantively true that the distinction has moral significance. But even if a morally neutral (ie non-question begging) distinction could be drawn, its application to the euthanasia controversy is problematic. I illustrate the difficulties of employing this distinction to reach moral conclusions by critically discussing Philippa Foot's recent treatment of euthanasia. I conclude that even if an act of euthanasia is an instance of killing, and there exists a prima facie moral duty not to kill, and no more stringent duty overrides this duty, one still cannot determine such an act to be morally impermissible.


Functional defects of RNA-negative temperature-sensitive mutants of Sindbis and Semliki Forest viruses.

Defects in RNA and protein synthesis of seven Sindbis virus and seven Semliki Forest virus RNA-negative, temperature-sensitive mutants were studied after shift to the restrictive temperature (39 degrees C) in the middle of the growth cycle. Only one of the mutants, Ts-6 of Sindbis virus, a representative of complementation group F, was clearly unable to continue RNA synthesis at 39 degrees C, apparently due to temperature-sensitive polymerase. The defect was reversible and affected the synthesis of both 42S and 26S RNA equally, suggesting that the same polymerase component(s) is required for the synthesis of both RNA species. One of the three Sindbis virus mutants of complementation group A, Ts-4, and one RNA +/- mutant of Semliki Forest virus, ts-10, showed a polymerase defect even at the permissive temperature. Seven of the 14 RNA-negative mutants showed a preferential reduction in 26S RNA synthesis. The 26S RNA-defective mutants of Sindbis virus were from two different complementation groups, A and G, indicating that functions of two viral nonstructural proteins ("A" and "G") are required in the regulation of the synthesis of 26S RNA. Since the synthesis of 42S RNA continued, these functions of proteins A and G are not needed for the polymerization of RNA late in infection. The RNA-negative phenotype of 26S RNA-deficient mutants implies that proteins regulating the synthesis of this subgenomic RNA must have another function vital for RNA synthesis early in infection or in the assembly of functional polymerase. Several of the mutants having a specific defect in the synthesis of 26S RNA showed an accumulation of a large nonstructural precursor protein with a molecular weight of about 200,000. One even larger protein was demonstrated in both Semliki Forest virus- and Sindbis virus-infected cells which probably represents the entire nonstructural polyprotein.


Structural and biological properties of mycoplasmavirus MVL3: an unusual virus-procaryote interaction.

The kinetics of adsorption and growth of mycoplasmavirus MVL3 in Acholeplasma laidlawii 1305/68 host cells have been studied with one-step growth, premature lysis, and single-burst experiments. The virus was found to kill infected host cells. Virus release starts 90 min after infection and continues for about 10 to 15 h. Hence, virus production is unlike the classical lytic bacteriophages and instead resembles nonlytic cytocidal animal viruses. Structural details of the virus are described, and the molecular weight of the viral linear DNA has beenfound to be 26 x 10(6).


Mappings of adenovirus type 7 cytoplasmic RNA species synthesized early in lytically infected cells and synthesized in transformed cells.

Early virus-specific RNA synthesized in KB cells infected with adenovirus type 7 and virus-specific RNA synthesized in rat embryo cells (71JY1-2) transformed by the adenovirus type 7 HindIII-I.J fragment (left-hand 8.1% of the viral genome) have been mapped on the viral genome. About 25% of the viral genome, four discrete regions, two on each strand of the viral genome, are expressed as "early" mRNA. Almost similar regions in the left-hand 8.1% of the viral genome are transcribed both in KB cells at early times after infection and in 71JY1-2 cells.


Hemodynamic effects of sublingual nifedipine in congestive heart failure.

Hemodynamic response of nifedipine in 11 patients with congestive heart failure was examined. Thirty minutes after sublingual administration of nifedipine, CI and SVI were increased together with a decrease in PAEDP and TSVRI. The increase in CI was proportional to the decrease in TSVRI. Twenty-four hours after continuous administration of nifedipine, however, six of 11 patients did not show the improvement of left ventricular function. These results indicate that sublingual administration of nifedipine is effective in the short-term therapy of the patients with a markedly decreased cardiac output and an increased systemic vascular resistance.


Electrophysiologic and pathologic correlations in six cases of atrioventricular block.

1) A prolonged A-H interval suggested A-V nodal involvement. 2) A prolonged duration of the His potential suggested moderate His bundle involvement. 3) Complete block distal to H appeared to reflect total disruption of both bundle branches. 4) The lesion at the penetrating portion of the His bundle could be responsible for A-H block. 5) A-H block occurred in a case of cellular infiltration in the A-V node and the His bundle, in which bilateral bundle branch showed severe fibrosis. 6) A combination of right bundle branch block, marked left axis deviation and H-V prolongation suggested trifascicular disease. 7) In case 6, there was a severe pathologic lesion at the origin of the left bundle branch, yet left bundle branch block was not indicated electrocardiographically. This study revealed a close correlation between electrophysiologic and pathologic findings in 4 out of 6 cases.


Ectopic aortic origin of the right pulmonary artery in tetralogy of Fallot.

A case is described wherein Fallot's tetralogy was associated with aortic origin of the right pulmonary artery. The left pulmonary artery was the continuation of the pulmonary trunk. Although a number of examples of Fallot's tetralogy associated with aortic origin of the left pulmonary artery are known, combination of Fallot's tetralogy and aortic origin of the right pulmonary artery is rare and has been described in only two cases previously.


Echocardiographic features of Uhl's anomaly. A case report.

M-mode echocardiography on a fifty-eight year old female with Uhl's anomaly showed several characteristic findings, which were considered to be useful in differentiating Uhl's anomaly from Ebstein's. Those findings were normal tricuspid diastolic closing velocity, the relatively early tricuspid valve opening, easy visualization of the tricuspid valve in usual position and the unusual mid-diastolic pulmonic valve full opening. The premature opening of the pulmonic valve correlated with the right atrial and ventricular mid-diastolic pressure exceeding that of the pulmonary artery.


A lectin-like substance from bovine spleen.

A lectin-like substance was isolated and purified from bovine spleen. This protein was named bovine spleen binding protein (BSBP). BSBP has binding activity for sialidase-treated bovine erythrocytes but not for normal erythrocytes. It specifically recognizes terminal galactose residues of sugar moieties of the bovine erythrocyte cell surface. BSBP shows an apparent molecular weight of 240,000. It is an acidic protein having a pI value of 4.8 and containing 13% carbohydrates. It has mitogenic activity against human peripheral lymphocytes. The physiological role of BSBP has not been clarified yet, but it may play a significant role in the clearance of aged erythrocytes which have a decreased amount of sialic acid on the cell surface.


Detection of IgG antibody to measles virus by radioimmunoassay technique.

A highly sensitive procedure of solid-phase radioimmunoassay (RIA) was developed for the detection of measles IgG antibody. HeLa cells persistently infected with measles virus were used as a solid-phase antigen. This technique was applied to the detection of measles IgG antibody in patients with subacute sclerosing panencephalitis (SSPE) and multiple sclerosis. Normal subjects having experienced natural measles or measles vaccination and patients with various neurological diseases of non-virus nature were also examined as control groups. Measles antibody was detected at high titers in both the sera and cerebrospinal fluid of SSPE patients. Moreover, RIA/HI ratios of SSPE patients were significantly higher than those of normal subjects, suggesting the presence in the formers of antibodies to nucleocapsids at high titers as well as to viral envelopes. On the other hand, no significant difference was found in both RIA and HI titers between the sera of multiple sclerosis and those of various neurological diseases.


Induction of pupal diapause and photoperiodic sensitivity during early development of Sarcophaga peregrina larvae.

Effects of long day and short day treatments during the embryonic and larval stages on induction of pupal diapause were studied on a diapausing race of Sarcophaga peregrina. Two long day (15L 9D) cycles during 2 days before or after the larviposition completely stopped the induction of pupal diapause on larvae which grow in short day condition before and after the long day treatment. The sensitivity appeared to decrease during the early stage of the third instar and to increase again to some extent in the prepupal stage.


A putative role for cell-cell epithelial contacts in lactose secretion.

Rates of synthesis and secretory release and the maximal requisite intracellular transit time (Tsec) for lactose were measured in vitro for three preparations of lactating guinea pig mammary tissue: tissue slices, mammary epithelial cell (MEC) acini, and mono-dispersed MEC. The Tsec values for tissue slice and acini preparations were similar, lactose required approximately 16 min to pass from its site of synthesis (Golgi) to the extracellular medium. Dispersal of mammary tissue into single cells by collagenase disruption of all cell-cell junctional complexes increased the Tsec value to approximately 25 min but did not alter kinetics of lactose synthesis and secretory release. These data suggest a possible involvement of cell-cell junctional contacts in intracellular transport of lactose.


Seasonal variation of plasma glucagon concentrations in men.

In an attempt to understand a role of glucagon in seasonal acclimatization in men, measurements of plasma glucagon, blood free fatty acids (FFA), blood glucose, blood ketone body (beta-hydroxybutyrate) and hematocrit were made in 13 male and 8 female college staff members, aged 20 to 42, once a month for one year. Blood samples were obtained at 4:00 to 5:00 p.m, between meals. Average monthly temperatures during the study were as follows; Jan. -7.8, Feb. -5.6, March -3.7, Apr. 4.6, May 11.4, June 18.3, July 23.9, Aug. 22.0, Sept. 14.7, Oct. 7.5, Nov. 1.4, Dec. -4.2 (degrees C). Plasma glucagon, blood FFA and blood ketone body exhibited significant monthly variation in both sexes. Plasma glucagon as well as blood FFA level was significantly higher in winter (Dec., Jan., Feb.) than in summer (June, July, Aug), whereas blood ketone body level was lower in winter than in summer. Plasma glucagon level was significantly lower in female than in male subjects. A significant positive correlation was observed between plasma glucagon and blood FFA levels throughout the year. Seasonal variations of blood glucose and hematocrit were not observed. These results suggest that seasonal variation in glucagon secretion is associated with seasonal changes in ambient temperatures as one of the strategies for climatic acclimatization through regulation of lipid metabolism.


Excitation of squid giant axons in hypotonic and hypertonic solutions.

Excitability of intracellularly perfused squid giant axons was maintained in hypotonic solutions (down to 300 mOSM) and in hypertonic solutions (up to about 10 OSM), when osmolalities of internal and external solutions were adjusted to be equal with glycerol, glucose, or sucrose. Molar concentrations of ions were kept constant during one series of experiments. The resting potential and the amplitude of the action potential did not change in both hypotonic and hypertonic solutions. With reduction of osmolality, the duration of action potential decreased and the maximum rate of rise and conduction velocity increased. By raising osmolality, the duration was prolonged and the maximum rate of rise and the conduction velocity decreased. Effects of osmolality change were almost reversible. However, these effects were not directly related to the osmolality change but seemed to be related to the viscosity change of the solutions. When the osmolality of external solution was raised with NaCl (up to 2.6 M NaCl), the overshoot increased in porportion to the logarithm of the NaCl concentration. The slope of increase was about 50 mV/decade. However, the resting potential showed little change. With increase of the NaCl concentration, the duration of the action potential increased.


Thermogenic responses of brown adipocytes to noradrenaline and glucagon in heat-acclimated and cold-acclimated rats.

Heat production of isolated brown-fat cells by addition of noradrenaline and glucagon was measured in warm-acclimated control, cold-acclimated and heat-acclimated rats by use of a twin-type conduction microcalorimeter. Noradrenaline and glucagon induced maximum heat production per 10(6) cells in dose of 1 microgram/ml. Heat produced by maximum thermogenic response to glucagon was twice as much as that to noradrenaline. Thermogenic response to noradrenaline was markedly increased in cold-acclimated brown adipocytes, while it was reduced in heat-acclimated ones. Thermogenic response to glucagon was significantly reduced in heat-acclimated brown adipocytes, while it was not affected in cold-acclimated brown adipocytes.


Denervated muscle: rate of propagation and effects of manganese ions and glycerol on the fibrillatory activity of frog semitendinosus.

Denervated frog's semitendinosus muscles were studied. Fibrillation potentials generate propagated spikes which are conducted at a rate similar to that calculated for innervated muscles. The twitch tension per unit area was similar in denervated and innervated single fibres; the average value corresponded with values reported in the literature for fibres stimulated at 125/sec. Manganese ions (1-3 mM), selectively inhibit fibrillation potentials and have no appreciable effect upon evoked spikes. Mn2+ also inhibits (40-50%) the twitch responses and abolishes the resting tension of muscle fibres. The latter effects were observed in denervated as well as in innervated preparations when the fibres studied belonged to fasicles located at the site of entrance of the nerve. Disruption of the transverse tubular system by glycerol treatment does not interfere with the generation of fibrillation potentials, although the effects on twitching are similar to those reported in the literature. A possible mechanism is proposed to explain the generation of fibrillation potentials in frog denervated muscles.


Motor nuclear representation of masticatory muscles in the rat.

Representation of the masticatory muscles within the motor trigeminal nucleus was studied in rats by the horseradish peroxidase (HRP) method and the antidromic field potential method. The motor trigeminal nucleus of the rat could be divided cytoarchitecturally into a dorsolateral and  a ventromedial division. Within the dorsolateral division, the temporal muscle was represented dorsomedially, the masseter muscle dorsolaterally and laterally, and the lateral and medial pterygoid muscles ventrolaterally. Within the ventromedial division, the anterior digastric muscle was represented dorsomedially and the mylohyoid muscle ventrolaterally. Distribution of antidromic field potentials evoked by stimulation of the mylohyoid and masseteric nerves coincided with the results from the HRP investigation.


Neurogenic constriction of the superior mesenteric and femoral veins during systemic blood pressure oscillation in rabbits.

Behavior of the mesenteric and femoral veins was studied in rabbits during an arterial pressure oscillation elicited by a method that we call the "side pressure exertion experiment." A segment of the mesenteric or femoral vein with intact innervation was vascularly isolated and was perfused under an isovolumetric condition. The force of the isovolumetric constriction of the segments was recorded in terms of the intrasegmental pressure duirng arterial pressure oscillation. The intrasegmental pressure of the superior mesenteric vein indicated an alternation of marked rise and fall during the oscillation. On the other hand, the response of the femoral segment during oscillation was less phasic but was better maintained at the higher range of initial pressures than that of the superior mesenteric segment. Simultaneously with the maximum rise of systemic arterial pressure, maximum elevation by 6.1, 6.1 and 6.4 mmHg was obtained in the mesenteric vein segment at initial intrasegmental pressures of 6, 10 and 14 mmHg, respectively. In the same circumstances, the femoral vein segment indicated a maximum rise of 8.1 to 9.3 mmHg at initial pressures of 10, 18, 26 and 34 mmHg. It can be concluded that there  are marked differences between the mesenteric and femoral segments in behavior during systemic arterial oscillation and in the range of bearable intrasegmental pressure. The characteristics of the two veins might be related to the difference of specific circumstances in which the respective veins send blood back to the heart.


Factors affecting frequency dependence of resistance observed in healthy lungs.

The frequency dependence of effective resistance of the respiratory system was studied in seated and supine healthy subjects by use of a forced oscillation technique for the purpose of estimating the distribution of regional ventilation. Effective resistance determined at respective FRC levels of both these body postures fell with an increase in the breathing frequency, although the frequency dependence of resistance delta R/delta f was more dominant in the supine posture than when seated. The delta R/delta f increased to an appreciable  extent by reducing the lung volume in the seated posture, while in the supine position the value  was relatively insensitive to the volume change. The chest wall vibration measured during the forced oscillation was found to be larger in amplitude in the basilar than in the apical regions, and the regional difference became less dominant by postural change from seated to supine. Transthoracic electrical impedance change measured during voluntary ventilations with different frequencies gave a slight frequency dependence when measured in the basilar region either in the seated or supine posture. The above results lead to the conclusion that an inhomogeneous distribution of time constants independent of gravity appears between the apex and the base, and the gravity-independent distribution of time constants, either interlobular or intralobular, would be a prime determinant of the frequency-dependent resistance.


Significance of skin pressure in body heat balance.

It has been demonstrated by Takagi and his colleagues that pressure on a specified area of the body surface causes depression of sweating in a certain body division and changes in the relative sweat rates between body divisions. Furthermore, skin pressure has been assumed to suppress  the central thermoregulatory activity, thus bringing about a rise or fall in body temperature in a hot or cool environment, respectively. We examined the effect of skin pressure applied to the bilateral subaxillary regions on body heat balance by means of continuous recordings of evaporative weight loss (total sweat rate), local sweat rates at various areas and rectal and skin temperatures and measurements of metabolic rate. Most experiments were carried out at a room temperature of 36 degrees C with 40% rh and a few were done at 27 degrees C in the absence of thermal sweating. Various strengths of pressure up to 5 kg/50 cm2 were employed. It  was observed that the total sweat rate was either unchanged, decreased or occasionally even increased. There  was an apparent tendency that the stronger the pressure was, the more depressed was the total sweating. A weaker pressure, on the other hand, often caused facilitation of total sweating. Changes in rectal and mean body temperatures and in metabolic rate were minimal in the majority of cases, and bore no relationship to the changes in the total sweat rate. These results offer no evidence that skin pressure of up to 5 kg/50 cm2 affects human central thermoregulatory activity but suggest that it may exert a sweat-inhibitory effect, primarily through the interaction of sudomotor impulses somewhere along the efferent pathways, possibly at the spinal segmental level.


High potassium-induced, calcium-dependent monoamine release from brain slices of the newborn rat.

To determine the functional significance of monoamines taken up into the newborn rat brain, a high K+-induced, Ca2+-dependent release of noradrenaline (NA), dopamine (DA) or serotonin (5-HT) from brain slices of the newborn rat was investigated and compared with that of the adult animal. Depolarization by increasing the potassium concentration in the medium induced the release of L-[3H]NA,[3H]DA or [3H]5-HT from brain slices prelabelled with the radioactive monoamines in the 2-day- or 3-day-old rat as well as in the adult. The release of three [3H]amines was markedly inhibited by perfusing with Ca2+- free medium in the presence of EGTA (2.0 mM) at both newborn and adult stages, although the inhibitory potency of Ca2+- deficiency in the newborn preparation was lower than in the adult. The degree of the release against [3H]amines initially taken up in the newborn was of the same order of magnitude as those in the adult. It is suggested that NA, DA and 5-HT are stored in a functionally releasable pool of the nerve terminals of the central monoamine neurons in the rat and that these compounds act as neurotransmitters at birth and at the adult stage.


Interaction between aspirin and prostaglandins in the isolated guinea-pig tracheal muscle.

The effects of PGE2 and PGF2 alpha on the tonus of isolated guinea-pig tracheal chain were investigated and compared with those of histamine and acetylcholine. PGE2 reduced tonus in normal resting state, but elevated tracheal tonus reduced by aspirin. Such PGE2-induced contractions did not exceed the initial resting tonus, and the magnitude and duration of the contractions progressively diminished with increase of PGE2 concentrations. Aspirin produced neither relaxation nor contraction in the presence of a low dose of PGE2. Unlike PGE2, PGF2 alpha produced a dose-related contraction in the normal tracheal chain, and the contractile response to PGF2 alpha was markedly potentiated by aspirin. In the presence of PGF2 alpha, aspirin no longer produced tracheal relaxation but produced a dose-related contraction. The contractile effect of histamine but not of acetylcholine was also potentiated by aspirin, but there was a slight difference between PGF2 alpha and histamine in that the potentiation of action of PGF2 alpha by aspirin was more easily diminished by PGE2. These results suggest that PGE2 plays an important role in the maintenance of the resting tonus of the isolated guinea-pig tracheal chain, and in large doses it also acts as a tracheal relaxant and attenuates the tracheal responses to PGF2 alpha and histamine.