Different types of small granule-containing cells and neurons in the guinea-pig adrenal medulla. An electron microscopic, histo- and biochemical study was carried out on the adrenal medulla of newborn and adult guinea-pigs giving special emphasis to small granule-containing (SGC) cells. Adrenaline (A) was the predominating catecholamine (CA) both in newborn (70-90% of total CA) and adult (85-90%) guinea-pig adrenals. In analogy to the biochemical findings electron microscopy revealed a high predominance of A cells, which contained large granular vesicles with an average diameter of 180 nm. Most noradrenaline (NA) storing cells showed granular vesicles of a considerably smaller average diameter (80 nm) and had a higher nuclear-cytoplasmic ratio. These cells were termed SGC-NA cells. NA cells with large granular vesicles (average diameter 170 nm) were extremely rare. Another type of SGC cells contained granular vesicles with cores of low to medium electron-density (SGC-NA-negative cells). Biochemical determinations made it unlikely that these cells contained predominantly dopamine (DA). SGC cells were scarcely innervated by cholinergic nerves. They formed processes, which were found both in the adrenal cortex and medulla contacting blood vessels including sinusoid capillaries, steroid producing cells of the reticularis and fasciculata zone and processes, which were interpreted to belong to medullary nerve cells. Two types of neurons were present in the guinea-pig adrenal medulla, one resembling the principal neurons in sympathetic ganglia, the other, which, principal neurons and SGC cells. In adrenomedullary grafts under the kidney capsule, which were studied three weeks after transplantation, "ordinary" A cells resembled SGC-NA negative cells with respect to their ultramorphology. Processes of transplanted principal neurons showed uptake of 5-hydroxydopamine and, hence, were considered to be adrenergic. Despite the lack of extrinsic nerves to the transplants, few principal neurons received cholinergic synapses, the origin of which is uncertain to date.


A scanning electron microscopic study of the luteo-follicular complex. III. Repair of ovulated follicle and the formation of the corpus luteum. As observed by SEM, the repair of an ovulated mammalian follicle is accompanied by a sequence of morphogenetic processes. In the initial phase, a mass of cells and coagulated fluids forms at the site of rupture. Shortly thereafter, connective cells, recruited from the adjacent and subjacent connective tissue stroma begin to proliferate and to migrate over this mass such that in the rabbit, the entire site of disruption is covered by a layer of connective cells by approximately 2 days following ovulation. Coincident with the migration of the connective tissue, superficial cells from undisturbed lateral and basal areas of an ovulated follicle also proliferate and begin to migrate over the newly established connective tissue matrix. By approximately 4 days following ovulation in the rabbit, the surface of an ovulated follicle is repopulated by elements of the superficial epithelium. The formation of the underlying corpus luteum (corpora lutea) involves characteristic morphological changes as granulosa cells transform into steroid secreting luteal cells. The luteal cells become organized into cords of cells which usually surround capillary vessels. When examined by SEM, the smooth-surfaced endoplasmic reticulum of the luteal cell is quite apparent and is observed to form a three-dimension network of anastomosing tubules which are continuous with the nuclear membrane. Variations in the appearance of the surface of the ovary which directly overlies corpora lutea were observed when the mouse, rat and rabbit were compared. The regression of corpora lutea involves the infiltration of the luteal mass by connective tissue and both degeneration and vacuolization of the luteal cells. The regressing corpus luteum is a honey-comb-like structure in which each space is occupied by a degenerating luteal cell.


Effect of castration and testosterone administration on the neuromuscular junction in the levator ani muscle of the rat. The ultrastructure of the neuromuscular junction (n.m.j.) of the androgen-sensitive levator ani muscle was studied in normal adult male rats, in 8-month-old rats castrated at the age of one month and in castrated rats treated with testosterone propionate (TP). Castration does not result in significant changes of the n.m.j. The density of synaptic vesicles and the postsynaptic junctional folds remain practically normal inspite of marked atrophy of the muscle. TP administration for 7 days results in marked changes in pre- and postsynaptic structures. There is slow progressive depletion of synaptic vesicles, appearance of cisternae and coated vesicles in axon terminals, and coalescence of coated vesicles with the plasma membrane. Coated vesicles are also found inside Schwann cells and among junctional folds. Dense core vesicles appear both in the axon terminals and in the postsynaptic area. Collateral sprouting of terminal axons with the formation of new immature junctions is observed. After 35 days of TP administration depletion of synaptic vesicles continues. Glycogen beta-particles, mostly freely dispersed, occasionally seen in axon terminals 7 days after TP administration, subsequently increase in number. In the endplate zone of the muscle fibre increased protein synthesis is indicated by a rapid increase in ribosomes and irregularly located myofilaments and myofibrils. The appearance of n.m.j. after testosterone administration resembles that described after nerve stimulation; the degree of change is however less pronounced.


The ultrastructure of the Herring bodies in rats subjected to different experimental conditions. A study of the morphology of Herring bodies of the posterior pituitary lobe of rats treated with colchicine and/or exposed to low temperatures has been performed. After treatment with colchicine (20 microgram in distilled water injected intracisternally) a predominance of Herring bodies with a large number of small "synaptic-like" vesicles surrounded by neurosecretory granules is found. Exposures to low temperature (4-6 degrees C) result in an increase in the neurosecretory material and the Herring bodies show many neurosecretory granules of different densities. After treatment with colchicine and subsequent exposure to low temperatures, the Herring bodies are characterized by having a great number of autophagic bodies which become more numerous as the length of the exposure is increased; later autophagic vacuoles and lamellar bodies become evident.


Cortisone induced alterations of costal cartilage in single and in parabiosed rats. Cortisone-treated Buffalo rats have been parabiosed with untreated controls of the same age. The optical and electron microscopy, including histochemistry, of costal cartilage of these rats has been compared with that in single cortisone treated rats, single controls, and control parabiosed with control rats, at 14 and 28 days after parabiosis. Single cortisone-treated rats, in comparison to controls, have shown the greatest alteration in cellular morphology and in the extracellular matrix both at 14 and at 28 days. Cortisone-treated parabiosed rats demonstrate a gradation of these alterations. Cellular alterations include enhancement of lipid and glycogen deposition concrurently with the presence of  numerous large cytoplasmic vacuoles containing beaded irregularly-shaped filaments, banded or unbanded collagen-like fibrils, and/or electron dense lamellar bodies. In the extracellular matrix, matrix vesicles, amianthoid fibers, randomly oriented unbanded fibrillar materials, and filament-like materials are most prominent in the single cortisone-treated rats and they are progressively less prominent in the cortisone-treated parabiosed rats, and in the parabiosed and single controls. Calcification of the extracellular matrix follows a similar pattern and is observed initially in pericellular halos of the single cortisone and in cortisone-treated rats parabiosed with controls. Histochemical techniques have shown that chondroitin sulfate is less demonstrable in the single cortisone and in the cortisone-treated parabiosed rats than it is in the single or parabiosed controls at 14 days but, at 28 days, all untreated or treated rats, single or parabiosed are basically comparable. Glycoproteins are prominent in the single cortisone-treated rats both at 14 and at 28 days and, at these same times, they are progressively less prominent in the cortisone-treated parabiosed rats and in the single or parabiosed controls. Many of the cortisone induced alterations in costal cartilage are suggestive of enhancement of the aging process.


The ultrastructure and histochemistry of the mesenteric connective tissue of the cockroach Periplaneta americana L. (Insecta, Dictyoptera). The thick layer of collagenous connective tissue around the mesenteron of the cockroach Periplaneta americana has been examined by light microscopy, electron microscopy and histochemical techniques. This sheath, connected to the mesenteric epithelium by a basal lamina and traversed by longitudinal and circular musculature, tracheae and nerves, contains numerous collagen fibrils, elastic fibers, fibroblasts and rare hemocytes. Development starts during the first instars with the production of connective tissue material by fibroblasts. Glycosaminoglycans have been observed with the use of various histochemical reactions, in particular alcian blue staining, metachromasia and enzymatic digestion. The reactions indicate the presence of hyaluronic acid in the various instars, the appearance and increase of sulphated components during development, and the presence of neutral glycoproteins.


A new method for the precise and complete correction of distortion on cineangiographic image: its effect on left ventricular measurements. Some of the problems associated with the precise evaluation of left ventricular volumes from angiographic images are due to the magnification and distortion of the pictures obtained from X-ray equipment. Magnification errors can be evaluated easily but the distortion (due to pincushion effect) are more difficult to measure. A mathematical method is proposed for a complete point-by-point correction of errors due to magnification and X-ray distortion. This method is applicable to most currently used angiographic equipment. Since more and more angiographic images are processed with local or remote digital computers, the applicability of the proposed technique becomes realistic. Results from this study show that a 5-30% overestimation of usual geometric ventricular parameters is encountered if no correction is made for distortion. End-diastolic measurements are more affected than end-systolic ones because of the greater degree of spherical distortion at the periphery of the picture area.


Angiographic anatomy of the left Kugel's artery. The left Kugel's artery is an uncommon but sometimes important collateral between the proximal circumflex artery and whatever artery supplies the crux of the heart. It passes anterior to the mitral valve ring, coursing in the interatrial septum, to anastomose usually with the atrioventricular node artery. The left Kugel's artery was prospectively identified on coronary arteriograms in 3.6% of patients with significant disease in dominant circumflex or right coronary arteries with associated collateral circulation. It was occasionally as large as 1 mm in diameter. The angiographic anatomy of this vessel is described.


Significance of collateral circulation in patients with left main coronary artery disease. Fifty patients with left main coronary artery disease were studied to evaluate the functional role of collateral circulation. The left main was narrowed 50-70% in 22 patients (group I), and more than 70% in 28 patients (group II). Significant disease in the other vessels was equally common in each group. There was no significant difference in the incidence of inter- and intracoronary collaterals in the two groups. Fifteen patients with no collaterals were compared with 35 patients with collaterals, and to a subset of 11 patients with very rich right-to-left collaterals, and there was no significant difference in historic or ECG evidence of old infarction, duration of angina, incidence of unstable angina, left ventricular end-diastolic pressure, cardiac index, ejection fraction, or segmental contraction abnormalities. We conclude that there is no evidence of protective effect of collateral vessels in patients with left main disease.


Submitral annular left ventricular aneurysm--unusual echocardiographic and angiographic features. A submitral annular left ventricular aneurysm in a Nigerian patient is described. The features consisted of an abnormal bulge on the left heart border with curvilinear calcification, an echo-free space behind the posterior left ventricular wall, and unusual angiographic appearance of the left circumflex coronary artery. The diagnosis was confirmed by pulmonary artery and left ventricular angiography. The patient underwent open heart surgery and a multiloculated submitral annular left ventricular aneurysm which was adherent to the pericardium was successfully resected.


Late right ventricular failure after Mustard operation for transposition of the great arteries. The clinical course and postmortem findings of an unusual case of complete transposition of the great arteries, in which the Mustard operation was not performed until the patient was 12 years of age, are documented with serial catheterizations showing adequate repair. The clinical course of the patient was uneventful for 8 years after surgery, when right (systemic) ventricular failure occurred, resulting in the patient's death at 20 years of age. Late cardiac failure, the cause of death, is not a commonly described problem after successful surgical repair of transposition of the great arteries.


Congenital coronary arteriovenous fistula: a diagnostic challenge. This report describes a case of coronary arteriovenous (A-V) fistula involving both the right and the left coronary arteries. The venous component of the two circulations combines into a single channel that empties into the left ventricle just below the mitral valve.


Radionuclide scintigraphy in the evaluation of ventricular septal defect complicating acute myocardial infarction. A 67-year-old man with an autopsy-proven ventricular septal defect (VSD) following an acute inferoposterior myocardial infarction underwent bedside evaluation by radionuclide imaging techniques. The presence of the VSD was confirmed, and the size of the shunt determined by qualitative and quantitative radionuclide angiocardiography. In addition, radionuclide ventriculography permitted assessment of global as well as regional left ventricular function.


Mitral valve prolapse (MVP) and coronary artery ectasia. A case of isolated ectasia of the entire right coronary artery and mitral valve prolapse is reported. The patient presented with acute inferior myocardial infarction and progressive angina pectoris. The diagnosis of ectasia of the right coronary artery and mitral prolapse was established angiographically. During a two-year follow-up period, the patient has continued to have angina and has suffered a second inferior myocardial infarction. Subsequent angiographic reevaluation has failed to show occlusive coronary lesions. It is suggested that distal thromboembolism due to changes in the character of blood flow in the dilated vessel has been responsible for the two episodes of myocardial infarction and persistent angina pectoris.


Transseptal left heart catheterization in infants and children. Transseptal left heart catheterization was performed in 80 infants and children with various forms of congenital heart disease. The majority had left heart obstructive lesions. Forty percent were under 5 years of age and less than 20 kg in weight. Uncomplicated cardiac perforation occurred in two patients. The technique is described in detail with emphasis on measures which increase the safety of the procedure for the patient. We conclude that this is a useful technique and in selected patients may be the preferred approach to the left heart.


"Zero reference" during cardiac catheterization. Hemodynamic pressure measurements obtained during cardiac catheterization studies are calibrated to an arbitrary level where the pressure is considered zero. Despite the fundamental nature of the zero reference level, there is no standardized method for determination of "zero reference." A horizontal metal rod has been constructed to be level with the pressure transducer domes. With lateral radiographic viewing, the metal rod is adjusted to be level with the junction of the inferior vena cava and the right atrium. The technique is rapid, reliable, without risk and allows accurate comparison of hemodynamic data between patients and in the same patient longitudinally.


Regeneration of mirror symmetrical limbs in the axolotl. Mirrow symmetrical, double posterior limbs were produced by two types of operations on tailbud embryos of the axolotl, Ambystoma mexicanum. When the limbs had grown to maturity, they were amputated and a study was made of the structures which regenerated in their place. The majority of the regenerates were also reduplicated, although a significant minority showed other types of symmetry. Reduplicate symmetry was often conserved following several amputations, but the number of skeletal elements present in each regenerate tended to vary: both expansion and contraction were found, the changes always occurring in the center of the pattern. These results are discussed in terms of the possible mechanisms for pattern formation in the anteroposterior axis during regeneration. We suggest that control of normal regeneration is carried out by a monotonic gradient of positional information which is originally set up to control pattern formation during embryonic development.


Sodium butyrate inhibits histone deacetylation in cultured cells. Sodium butyrate in millimolar concentrations causes an accumulation of acetylated histone species in a variety of vertebrate cell lines. In all lines tested, butyrate caused hyperacetylation of H3 and H4, and in rat IRC8 cells, H2A and H2B were also affected. In Friend erythroleukemic cells, butyrate also induces the synthesis of a nonhistone chromosomal protein, IP25. butyrate does not affect the rate of histone acetylation in cell-free extracts of nuclei of Friend cells. Rather, this fatty acid inhibits histone deacetylation. Cell-free extracts of either control cells or butyrate-grown cells contain comparable levels of histone-deacetylating activity. This in vitro activity is inhibited by the addition of butyrate to the extracts. Thus butyrate appears to be an inhibitor of histone deacetylases both in vivo and in vitro.


The effect of sodium butyrate on histone modification. The hyperacetylation of histones due to treatment of cultured cells with sodium butyrate has been studied. The hyperacetylation is due to inhibition of histone deacetylase. Other short chain fatty acids including acetic, isobutyric and propionic acid also produce increased modification. Histone H4 already deposited on the chromosome can be rapidly acetylated to the extent of about 70%. That 80% of histone H4 is acetylated after a 24 hr exposure to butyrate is due to the fact that incoming H4 histone is 100% acetylated and does not return to the parental unmodified form in the presence of butyrate.


Sequence complexity of nuclear and polysomal RNA in leaves of the tobacco plant. The first measurements are reported of the sequence complexity of nuclear and polysomal RNA contained within the cells of a higher plant. Polysomal RNA from tobacco leaves was prepared by a procedure which minimized contamination with nuclear RNA. Hybridization of 3H-cDNA complementary to polysomal poly(A) RNA with an excess of tobacco DNA indicated that greater than 95% of the poly(A) mRNA was transcribed from single-copy sequences. RNA excess hybridization reactions with polysomal poly(A) RNA and 3H-cDNA revealed the presence of three abundance classes in the poly(A) mRNA. The best least-squares solution indicated that these classes comprise 9, 52, and 39% of the poly (A) mRNA and contain sequences present an average of 4500, 340 and 17 times per cell. Hybridization reactions containing an excess of nuclear or total polysomal RNA and 3H-single-copy DNA indicated the complexity of these RNA populations to be 1.19 X 10(8) nucleotides (nuclear) and 3.33 X 10(7) nucleotides (polysomal). Thus only 28% of the nuclear RNA sequence diversity (27,000 average-sized mRNA sequences) is respresented in leaf polysomes. These results suggest that there is a general similarity in the basic transcriptional processes of metaphytan and metazoan cells.


The transcription map of mouse mitochondrial DNA. Nine transcripts complementary to mouse L cell mitochondrial DNA have been detected, sized and mapped to restriction fragments using the method of Berk and Sharp (1977). RNA isolated from L cell mitochondria was hybridized to 32P-labeled, cloned L cell mitochondrial DNA restriction fragments in 70% formamide under conditions 5 degrees C above the melting temperature of the DNA-DNA duplex, but approximately 15 degrees C below the melting temperature of the RNA-DNA duplex. The heteroduplexed material was then treated with the single-strand-specific nuclease S1, whick cleaves the single-stranded DNA not protected by RNA-DNA duplex formation into oligonucleotides and leaves intact 32P-labeled, single-stranded DNA replicas complementary to the transcripts. The single-stranded DNA replicas were then resolved and sized by alkaline agarose gel electrophoresis. Hybridization to strand-separated, 32P-labeled L cell mitochondria DNA restriction fragments under the same conditions showed that all nine transcripts hybridized exclusively to the heavy strand (H strand) of restriction fragments isolated as the dense strand from alkaline CsCl gradients, indicating that all stable transcripts 300 bases or longer detected by this technique originate from genes on the H strand. The two most abundant transcripts homologous to mitochondrial DNA map adjacent to the origin of replication. This result is consistent with map positions assigned to the large and small mitochondrial ribosomal RNAs isolated from Xenopus laevis and HeLa cells. Six of the other seven transcripts map continuously in approximately 40% of the genome. Only one transcript of 950 bases maps in the first quadrant of the genome as defined by the origin and direction of mitochondrial DNA replication, and it does not lie within the D loop region. The genetic function of the remaining 75% of this region of the genome is yet to be determined.


The replication of kinetoplast DNA networks in Crithidia fasciculata. Kinetoplast DNA from the mitochondria of Crithidia is in the form of a two-dimensional network of thousands of minicircles each containing about 2.5 kb, and a small number of maxicircles each containing about 40 kb. Fractionation of kinetoplast DNA by equilibrium centrifugation in a CsCl-propidium diiodide gradient resolves it into three types of networks. Form I networks band at high density and contain minicircles which are covalently closed; form II networks band at low density and contain minicircles which are nicked or gapped; and replicating networks band at intermediate density and contain some minicircles of each type. Form I networks contain about 5000 minicircles; form II networks contain about 11,000; and replicating networks contain an intermediate number. When cells are pulse-labeled with 3H-thymidine, radioactivity in mitochondrial DNA is preferentially incorporated into replicating networks, but after a chase it appears first in form II networks and finally in form I. Examination of replicating networks by electron microscopy in the presence of ethidium bromide reveals that minicircles in the central region of the network are twisted and therefore covalently closed, whereas those in the peripheral region are not twisted and therefore must be nicked or gapped. The pulse-label is incorporated into the nicked or gapped minicircles of the replicating networks. These results indicate that replication of form I networks begins in peripheral minicircles and that progeny minicircles remain nicked or gapped. As replication proceeds, the size of the network increases, and the peripheral zone of nicked or gapped minicircles enlarges. Finally, when all minicircles have replicated, the network, now form II is double the size of form I and contains only nicked or gapped minicircles. The final step in replication presumably includes both the cleavage of the network into two form I species and the covalent closure of all the minicircles.


Kinetoplast RNA of Leishmania tarentolae. RNA has been isolated from highly purified kinetoplast-mitochondrial fractions of Leishmania tarentolae, and shown to consist of two major species that sediment at 9S and 12S in sucrose and also several additional low molecular weight species which were visualized by gel electrophoresis. The in vivo transcription of 9S and 12S RNAs was inhibited by ethidium bromide and rifampin, and was fairly insensitive to low actinomycin D and camptothecin. The 9S and 12S RNAs were isolated by acrylamide gel electrophoresis or by sedimentation in sucrose. Both RNAs contained approximately 80% A + U and did not contain long stretches of poly(A). The 9S and 12S RNAs were found to hybridize selectively to the maxicircle sequences of the kinetoplast DNA, implying that the maxicircle, and not the minicircle, represents the informational mitochondrial DNA in the kinetoplast.


Induction of the Drosophila heat shock response in isolated polytene nuclei. When Drosophila salivary glands are disrupted in cytoplasm from heat-shocked Drosophila Kc cells in culture, puffs are induced in the polytene nuclei at the heat shock loci. The in vitro response increases with time of incubation, reaching in vivo levels after 2 hr, and is sensitive to alpha-amanitin. Cytoplasm from control Kc cells induces poorly, but prolonged exposure to air or the addition of hydrogen peroxide stimulates induction to the levels observed with cytoplasm from heat-shocked Kc cells. Autoradiography confirms that the heat shock loci induced to puff in vitro are the most actively transcribing sites in the genome.


The synthesis and properties of T25 blycoprotein in Thy-1-negative mutant lymphoma cells. The synthesis and properties of T25 glycoprotein which bears the serological specificity Thy-1 have been studied in mutants of cultured mouse lymphoma cells that do not express Thy-1 on their surface. Five complementation classes of mutant cells were previously characterized by somatic genetic analysis. Synthesis of abnormal T25 glycoproteins was detected in four classes of mutants. Each of these aberrant products was degraded move rapidly than T25 glycoprotein of wild-type cells. Defects in the oligosaccharide units of T25 glycoprotein were demonstrated in three classes of mutants. In one of these mutant classes, evidence for a general defect in glycosylation of cell surface glycoproteins was obtained. These data indicate that normal glycosylation of T25 glycoprotein is probably essential for the molecule to be incorporated into the plasma membrane and expressed on the cell surface.


The effect of inhibitors of protein synthesis on the reexpression of surface immunoglobulin following antigenic modulation. P3, a cell line derived from the plasmacytoma MOPC-21 secretes IgG1 (K) and is sensitive to complement (C')-mediated lysis by antibody directed against gamma1 or K. Sensitivity is attributed to the presence of immunoglobulin molecules on the surface membrane, designed Ig-mem. This sensitivity is abolished by antigenic modulation of Ig-mem. Modulated cells, when incubated in the absence of antibody, recover sensitivity to lysis in 4 hr. By measuring the rate of recovery, it has been possible to study the effects of various drugs on the reexpression of Ig-mem. Treatment of modulated cells with cycloheximide (Cx), pactamycin Pc), anisomycin (An), homoharringtonine (Ha) or sparsomycin (Sm), each a specific inhibitor of a different step in protein synthesis, produces a significant reduction in the rate of recovery. Paradoxically, puromycin (Pm), also a specific inhibitor of protein synthesis, does not reduce the rate of recovery. Studies were performed using Pm together with each of the other drugs to gain an understanding of the relationship between protein synthesis and recovery from modulation. Based upon these studies, we conclude that continued operation of the initiation cycle of protein synthesis is required for reexpression of Ig-mem in the absence of de novo protein formation.


Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis. Antibody-secreting hybrid cells have been derived from a fusion between mouse myeloma cells and spleen cells from a mouse immunized with membrane from human tonsil lymphocyte preparations. Hybrids secreting antibodies to cell surface antigens were detected by assaying culture supernatants for antibody binding to human tonsil cells. Six different antibodies (called W6/1, /28, /32, /34, /45 and /46 were analyzed. These were either against antigens of wide tissue distribution (W6/32, /34, and /46) or mainly on erythrocytes (W6/1 and W6/28). One of the anti-erythrocyte antibodies (W6/1) detected a polymorphic antigen, since blood group A1 and A2 erythrocytes were labeled while B and O were not. Antibodies W6/34, /45 and /46 were all against antigens which were mapped to the short arm of chromosome 11 by segregation analysis of mouse-human hybrids. Immunoprecipitation studies suggest that W6/45 antigen may be a protein of 16,000 dalton, apparent molecular weight, while W6/34 and /46 antigens could not be detected by this technique. Antibody W6/32 is against a determinant common to most, if not all, of the 43,000 dalton molecular weight chains of HLA-A, B and C antigens. This was established by somatic cell genetic techniques and by immunoprecipitation analysis. Tonsil leucocytes bound 370,000 W6/32 antibody molecules per cell at saturation. The hybrid myelomas W6/32 and W6/34 have been cloned, and both secrete an IgG2 antibody. W6/32 cells were grown in mice, and the serum of the tumor-bearing animals contained greater than 10 mg/ml of monoclonal antibody. The experiments established the usefulness of the bybrid myeloma technique in preparing monospecific antibodies against human cell surface antigens. In particular, this study highlights the possibilities not only of obtaining reagents for somatic cell genetics, but also of obtaining mouse antibodies detecting human antigenic polymorphisms.


Length and sequence heterogeneity of the histone gene repeat unit of the sea urchin, S. purpuratus. Histone gene repeats in S. purpuratus are shown to be of variable length and sequence. Two recombinant plasmids containing the full-length 6.3 kb histone repeat unit are found to differer in length at two sites in the repeating structure and in the occurrence of two restriction enzyme recognition sites. Variation in repeat length is also demonstrated in the unfractionated DNA of five sea urchins and in a sample of DNA enriched for histone gene sequences by density gradient methods. The repeats in each individual are of a very limited number of major classes, which may differ from one another in overall length or in distribution and presence of particular restriction enzyme sites. Variations are found to occur at many regions of the repeat; some have been mapped specifically to spacer regions. Repeats may differ dramatically from individual to individual since there is no one type of repeat class common to all, although the absolute length differences of the repeats that are found are small.


Characterization of beta-globin mRNA in the beta0 thalassemias. A number of cases of beta0 thalassemia have been examined for the presence or absence of beta-globin mRNA. Total RNA extracted from peripheral blood was hybridized to purified complementary DNA specific for beta-globin mRNA, and to beta-cDNA probes specific for the 5' and 3' noncoding regions of beta-globin mRNA. Three clear-cut categories of beta0 thalassemia were identified. The first type had no detectable beta-globin mRNA. A second typed had beta-globin mRNA sequences which hybridized incompletely to the cDNA probes and probably represented mRNAs with grossly altered structures. A third type appeared to have essentially intact, though untranslatable, beta-globin mRNA. Depurination products from 5' and 3' beta-cDNAs synthesized from this latter mRNA were identical to those from normal beta-globin mRNA, but the relative yields were different, suggesting a possible defect near the initiation codon.


Variability in the amount of beta-globin mRNA in beta0 thalassemia. Globin mRNA isolated from a number of beta0 thalassemia patients of different ethnic origins was analyzed by RNA-cDNA hybridization and, in two cases, by fingerprint analysis of 125I-labeled mRNA. Quantitation of the relative amounts of alpha- and beta-mRNA by hybridization to purified alpha-and beta-cDNA revealed that in approximately half the cases, there was less than 1% as much beta-mRNA as alpha-mRNA. In the rest of the cases, low levels of beta-like mRNA were detected in amounts 4-12% as abundant as alpha-mRNA. There was variability in the yield of beta-like mRNA in patients of the same racial group, in the same patient at different times and in similarly affected siblings: beta-mRNA was virtually absent in some samples, whereas low but significant levels were found in other samples. In one patient, beta-like mRNA was not detected in peripheral blood RNA, but was present in the RNA of bone marrow cells. In one case, the thermal stability of the beta0 thalassemia mRNA-beta-cDNA hybrid was measured and found to be slightly lower than that of the authentic beta-mRNA-beta-cDNA hybrid. In none of the cases tested was there synthesis of beta-globin chains directed by beta0 thalassemia mRNA in a cell-free protein-synthesizing system, even when beta-like mRNA was detected in the sample by hybridization assays. mRNA from two patients was labeled in vitro with 125I, digested with T1 RNAase and fractionated in two dimensions. Analysis of the resulting fingerprints revealed the presence of prominent alpha chain-specific oligonucleotides without detectable beta chain-specific oligonucleotides, and thereby confirmed the results of hybridization assays showing absent or very low levels of beta-mRNA in the same RNA samples. Our results support the concept that beta0 thalassemia is heterogeneous in its molecular basis even within the same racial group: in some patients, it is associated with absent beta globin mRNA, whereas in other patients, it is associated with low but significant levels of nonfunctional beta or beta-like globin mRNA. The variable amounts of beta-like mRNA detected in different samples from the same patient, and in patients with the same genotype, indicate that as yet undefined factors can influence the yield of beta-like mRNA observed in beta0 thalassemia.


Histone gene transcripts in the cleavage and mesenchyme blastula embryo of the sea urchin, S. purpuratus. Two distinct populations of histone gene transcripts have been identified in the sea urchin embryo. Both late cleavage and mesenchyme blastula stages contain histone transcripts which hybridize to a full-length histone repeat recombinant DNA, pCO1. The histone RNAs of the two stages, however, are dissimilar in sequence. While the transcripts of the cleavage embryo form well matched hybrids with the plasmid DNA which are relatively resistant to RNAase, the hybrids containing the mesenchyme blastula transcripts melt some 10 degrees C lower and are twice as sensitive to RNAase. Hybridization of the two RNA samples to the Hha I fragments of the histone DNA, or to segments of the histone repeat subcloned in other plasmids, shows that many regions scattered along the repeat are complementary to widely diverged transcripts in the mesenchyme blastula RNA. The two RNA populations consist predominantly of polysomal RNA sequences and are most probably mRNAs for the five histones. The mesenchyme blastula RNA sequences in both S. purpuratus and L. pictus form hybrids with pCO1 DNA that are less stable than those containing L. pictus cleavage RNA, indicating the wide divergence of the two histone RNA populations. The bulk of the histone genes in S. purpuratus appear to be of the type coding for the early mRNAs. Only a small percentage of the several hundred gene copies are candidates for the type coding for the late mRNAs. The melting characteristics of the hybrids and the sensitivity of RNAase provide an assay for the late embryonic histone genes. Of the total RNA labeled during a 10 min pulse in the cleavage embryo, histone transcripts represent approximately 9.7 and 6.5% of the radioactivity in S. purpuratus and L. pictus, respectively. These values fall to 0.57 and 1.4%, respectively, at the mesenchyme blastula stage. Although histone genes are transcribed during these two periods, the type of gene which is active is switched at some point prior to the mesenchyme blastula stage.


A change in the stability of globin mRNA during the induction of murine erythroleukemia cells. The stability of globin mRNA in murine erythroleukemia cells (Friend cells) before and during DMSO-induced differentiation was investigated. Cells were exposed to 3H-uridine for 2 hr and then transferred to medium without the radioactive precursor. The loss of radioactivity in total RNA, poly(A)-containing RNA and globin mRNA was followed. The globin mRNA was isolated using a highly specific globin cDNA column. In uninduced cells and cells early in differentiation, the globin mRNA decays with a half-life of less than 50 hr. After 4 days of induction, the globin mRNA decays with a half-life of 17 hr, demonstrating a change in stability during the induction process. Although the stability of globin mRNA changes during induction, this is not true for total poly(A)-containing RNA. At all times of induction, the poly(A)-containing RNA decays as two populations, one with a half-life of 6 hr and the other with a half-life of 36 hr. The half-life of the rRNA also remains unchanged during differentiation.


Studies on intercellular LETS glycoprotein matrices. Intercellular matrices secreted by chick embryo fibroblasts in culture were studied by scanning electron microscopy. Cell-cell contact is a prerequisite for the expression of such matrices. The smallest fiber detected by transmission electron microscopy is 5--10 nm in diameter. These matrix fibers tend to cluster to form bundles. Immunofluorescence and immunoferritin procedures reveal that LETS protein is one of the components of the matrices. The matrices are isolated from other cellular organelles by detergent treatment. More than 90% of the proteins in cell-free matrices are LETS protein, suggesting that the matrices are probably made of only one component--LETS protein. Since the solubilization of matrices requires beta-mercaptoethanol, LETS protein matrices may be the first known polymer system in nature to use disulfide linkage as an intermolecular polymerization vehicle. Collagen does not appear to be involved in such matrices. The LETS protein matrix supports the morphological conversion of rounded cells into spindle-shaped, and also promotes myoblast fusion. It does not, however, exert an effect upon cell growth, the rate of glucose uptake or protease production.


Synthesis of tropomyosin in myogenic cultures and in RNA-directed cell-free systems: qualitative changes in the polypeptides. The synthesis of polypeptides with the properties of alpha and beta tropomyosin was investigated in differentiating cultures of a myogenic cell line and in a wheat germ cell-free system directed by purified RNA extracted at different stages of differentiation. The polypeptides co-migrate with tropomyosin in isoelectric focusing and SDS two-dimensional gel electrophoresis and SDS-urea/SDS two-dimensional gels. Like authentic tropomyosin, these polypeptides change their mobility greatly in the presence of urea and do not become labeled with proline. The beta tropomyosin synthesized in the intact cells and in the cell-free system can be separated by isoelectric focusing into at least two components. One component (designated beta1) is present in a small amount at all developmental stages examined, and a more basic component (beta2) is specific for differentiated cultures. The synthesis of beta2 in the intact cells and the capacity of purified RNA to direct its synthesis in a cell-free system become detectable and increase greatly during the period of fusion of the mononucleated cells into multinucleated fibers. The results suggest that the beta1 and beta2 tropomyosins are coded for by different genes.


Isolation and characterization of amphotericin B-resistant cell lines in Chinese hamster cells. Amphotericin B is a polyene macrolide antibiotic which interacts specifically with sterols in mammalian cell membranes. Amphotericin B-resistant (AMBr) lines of stable phenotype have been isolated from cultured Chinese hamster (V79) cells. Three AMBr clones (AMBr-1, -2 and -3) isolated independently after treatment with nitrosoguanidine were resistant to greater than or equal to microgram/ml of the antibiotic, while DNA synthesis as well as the colony-forming ability of the parental V79 cells was blocked by greater than 80% of control in the presence of 20--50 microgram/ml amphotericin B. The AMBr cell line also exhibited increased resistance to other polyene macrolide antibiotics such as nystatin and pentamycin. Other agents, however, such as cytosine arabinoside or ricin, blocked DNA synthesis in AMBr cells to the same extent as in V79 cells. The amphotericin B resistance phenotype was stably retained even after AMBr cells were cultured in the absence of the drug for over 200 generations. The control of free cholesterol or its esters was significantly decreased in all three resistant clones. Furthermore, cholesterol synthesis from acetate as well as mevalonate was partly defective in AMBr cells, compared with that in V79 cells.


The adhesive interaction between polymorphonuclear leukocytes and endothelial cells in vitro. The results of this investigation indicate an adhesive specificity between PMN and cultured endothelial cells. This was monitored by the mono-layer collection assay and by direct cell counts using the scanning electron microscope. Both techniques showed that significantly more PMN attached to endothelial cells than to a variety of other cell types. The interaction can be modulated by divalent cations and neuraminidase, implying a role for surface charge. In the presence of chemotactic agents, the number of PMN adhering to endothelial cells increases. This system presents a good model for studying the process of PMN margination which occurs in vivo during the acute inflammatory response.


Effects of LETS glycoprotein on cell motility. Addition of LETS glycoprotein to normal or transformed cells produces increased migration of the cells, as determined by formation of phagokinetic tracks on gold particle-coated coverslips. These tracks arise by a combination of phagocytosis of the gold particles and cellular migration. Increased motility is also evident on plastic in the absence of gold particles. The added LETS protein attaches to the cells in a fibrillar network, and binding is greater to normal than to transformed cells. The effects of LETS protein on migration are consistent with its effects on cell adhesion, morphology and cytoskeleton, and have potential implications for the determination of cellular migration in vivo.


Developmental relationships between vitelline and intra-embryonic haemopoiesis studied in avian 'yolk sac chimaeras'. Using the quail-chick marker technique, cells have been traced in the haemopoietic organs of 149 developing chimaeras composed of a quail embryo with a chick yolk sac. The existence of intraembryonic stem cell relaying yolk sac stem cells, previously demonstrated with this system, is confirmed. Thymus, bursa of Fabricius and bone marrow are preferentially populated by intraembryonic stem cells. The spleen shows a transitory phase of colonization by chick yolk sac stem cells at 11--12 days of incubation. From 7 days onwards the yolk sac receives quail stem cell emigrating from the embryo.


On the occurence and composition of dense particles in mitochondria in ultrathin frozen dry sections. Clusters of dense particles were observed irregularly in mitochondrial profiles of shock-frozen brown adipose tissue and liver when cryosectioned at a chamber temperature of -80 degrees C and not if sectioning was performed at -110 degrees C. The particles are rich in Ca and P (Ca/P =0.5); they are therefore not any form of tricalcium orthophosphate (e.g. hydroxyapatite). They apparently form by electrolyte diffusion in the section and therefore cannot be considered as representing the in vivo pool of mitochondrial calcium.


Unidirectional motility occurring in association with the axopodial membrane of Echinosphaerium nucleofilum. A system of rapid, unidirectional motility associated with surface of Echinosphaerium axopodia is demonstrated using polystyrene micropheres as exogenous markers.


Stimulation of cell proliferation by histamine H2 receptors in dimethylhdrazine-induced adenocarcinomata. Cell proliferation in dimethylhydrazine-induced colonic carcinomata was stimulated by histamine and by the histamine H2 receptor agonist dimaprit and inhibited by the histamine H2 receptor antagonists Metiamide and Cimetidine but not by the histamine H1 receptor antagonist Mepyramine. In contrast histamine had no effect on colonic crypt cell proliferation in normal or dimethylhydrazine-treated rats.


Delphinidin- and cyanidin-chlorides, promotors of cell aggregation in vitro. Delphinidin-chloride and Cyanidinchloride cause cells in cell culture to form aggregates instead of the usual monolayers, without impairing viability or proliferation. The process is reversible.


Characterization of globin mRNA from Xenopus laevis. A 9S polyadenylated mRNA was isolated from 'xenopus laevis reticulocytes and purified by sedimentation and oligo-dT-cellulose chromatography. The size of this RNA estimated in denaturing agarose gels was 2.2 x 10(5) daltons. 9S polyadenylated mRNA hydridized to total DNA with a Cot1/2 of 3.8 x 10(3).


3' deoxycytidine, like hydroxyurea, inhibits DNA synthesis without preventing the initiation of the cell cycle. 3' Deoxycytidine, the cytidine analogue of cordycepin and selective inhibitor of pre-ribosomal RNA in HeLa cells, has been found to be a reversible inhibitor of DNA replication in RNA accumulation. Like other inhibitors of DNA replication such as hydroxyurea, it does not prevent serum-stimulated quiescent 3T3 cells from undergoing the random transition which is rate-limiting for entry into S phase.


RNA metabolism and poly(A) distribution in mouse liver following administration of dimethylnitrosamine. Dimethylnitrosamine (DMNA) strongly inhibited RNA synthesis in mouse liver under conditions when the nucleotide pattern, rate of nucleotide synthesis and phosphorylation ratio were unaffected. (An unidentified, probably non-nucleotide, component in the acid-soluble liver fraction was selectively reduced.) The inhibition of RNA synthesis was associated with a decrease in the RNA polymerase activity of isolated liver nuclei, well established already 45 min after DMNA administration. The reduced activity included both Mg2+- and Mn2+/(NH4)2SO4-stimulated polymerase functions. The inhibition in vivo involved the whole complement of RNA, including poly (A)-containing RNA and isolated poly(A) sequences. The transfer of labelled RNA from the nucleus to the cytoplasm was not impaired. There was no detachment of poly(A)-containing RNA from the microsomes, and the proportion of tightly membrane-bound microsomal RNA and poly(A) sequences was not reduced as determined by use of a flotation technique. No breakage or shortening of the poly(A) chains was indicated by sedimentation analysis.


Sterol synthesis. Chemical synthesis, structure determination and metabolism of 14 alpha-methyl-5 alpha-cholest-7-en-3 beta, 15 beta-diol and 14 alpha-methyl-5 alpha-cholest-7-en-3 beta, 15 alpha-diol. 14alpha-Methyl-5alpha-cholest-7-en-3beta,15beta-diol and 14alpha-methyl-5alpha-cholest-7-en-3beta,15alpha-diol have been prepared by chemical synthesis. Unequivocal establishment of these structures was based upon X-ray crystallographic analysis of 3beta-p-bromobenzoyloxy-14alpha-methyl-5alpha-cholest-7-en-15beta-ol and was supported by other spectroscopic data. Spectroscopic data were presented for the following compounds prepared in this study: 3beta-benzoyloxy-5alpha-cholest-8(14)-en-15-one, 3beta-benzoyloxy-14alpha-methyl-5alpha-cholest-7-en-15-one, 3beta-benzoyloxy-14alpha-methyl-5alpha-cholest-7-en-15beta-ol, 14alpha-methyl-5alpha-cholest-7-en-3beta, 15beta-diol, 14alpha-methyl-5alpha-cholest-7-en-3beta, 15alpha-cholest-7-en-15beta-diol, 14alpha-methyl-5alpha-cholest-7-en-3beta, 15alpha-diol, 3beta, 15alpha-bis-benzoyloxy-14alpha-methyl-5alpha-cholest-7-ene, 3beta-p-bromobenzoyloxy-14alpha-methyl-5alpha-cholest-7-en-15beta-ol and 3beta, 15beta-bis-p-bromobenzoyloxy-14alpha-methyl-5alpha-cholest-7-ene. Studies of the metabolism of [16-3H]-14alpha-methyl-5alpha-cholest-7-en-3beta, 15beta-diol and [16-3H]-14 alpha-methyl-5 alpha-cholest-7-en-3beta, 15alpha-diol in liver homogenates of female rats indicated that only the 3beta, 15beta-diol was convertible to cholesterol.


Alterations in brain and muscle oxygenation during hypovolemia and replacement with plasma substitutes in rats. A study was conducted on the effects of mild hemorrhage-induced hypovolemia and subsequent replacement with lactated Ringer's dextran or hemoglobin solution in 52 pentobarbital-anesthesized rats. Blood loss of 1.25% of body weight had no significant effect on the blood flow or relative tissue PO2 of the white matter of the cerebral cortex. Blood flow and relative tissue PO2 of the biceps brachii muscle were lowered during hypovolemia. Volume replacement with any of the three plasma substitutes had no effect on blood flow or relative tissue PO2 of cortical white matter. They all caused partial restoration of blood flow and relative tissue PO2 in the biceps brachii muscle. Full return in the muscle was not achieved, nor were there any significant differences between the treatment groups.


Comparison of the cytotoxic actions of hypoxia and endotoxin in the perfused cat liver. The isolated cat liver perfused at a constant flow with Krebs-Henseleit solution containing low-molecular-weight dextran was employed to ascertain the direct effects of hypoxia or endotoxin on hepatic integrity. Hypoxia resulted in large increases in circulating lactate dehydrogenase (LDH) activity and in amino-nitrogen concentration, whereas endotoxin at a dose of 0.75 microgram/gm liver wet weight resulted in only small changes in these variables after 150 minutes of perfusion. Perfusion pressure and perfusate pH did not change significantly in response to either intervention. Both hypoxia and endotoxin significantly compromised lysosomal stability as evidenced by large increases in circulating levels of cathepsin D, large increases in the nonsedimentable fraction of tissue cathepsin D (ie, increased percentage of free activity), and changes in the ultrastructural appearance of liver lysosomes associated with enhanced fragility (eg, swelling, increased vacuolization). Both interventions also significantly impaired phagocytosis by reticuloendothelial cells within the liver. However, neither intervention altered BSP clearance, indicative of a lack of effect on parenchymal cell clearance. These findings indicate that both endotoxin and hypoxia induce direct cellular damage within the liver; however, endotoxin exerted a more selective action on lysosomes, whereas hypoxia produced more of a diffuse cytotoxic effect.


Increased template activity of chromatin isolated from shocked liver. Hemorrhagic shock causes an increase in the template activity of chromatin isolated from rat liver, as measured with E. coli RNA polymerase. The increase in template activity is directly related to the time of shock: The longer the shock period, the greater the increase in template activity. Reinfusion reduces the increased template activity. The increased template activity of the chromatin during hemorrhagic shock may be indicative of nuclear changes which contribute to the inability of the cell to recover from severe shock.


Behaviour therapy in clinical child psychiatry: a preliminary report. This paper reports on the practice of behaviour therapy with children and families in a clinical setting. The behavioural treatment of 33 cases is discussed. The cases are characterized by the range of problems treated and the variety of behavioural techniques used. Behaviour therapy is shown to be an approach that can be used effectively in a variety of different settings and alongside, or together with treatments based on different theoretical orientations.


Demonstration of nonperfused myocardium in late hemorrhagic shock. Studies were performed to determine whether or not regions of left ventricle become nonperfused in the normovolemic stages of hemorrhagic shock. Using the endothelial fluorescent stain thioflavin S as a tracer, nonperfused myocardium was assessed in six control dogs, eight dogs in early normovolemic shock, and 15 dogs in late normovolemic shock. Nonperfused myocardium was not demonstrable in any of the control animals and in only one of the early shock animals. However, in nine of the late shock animals, nonperfused myocardium was clearly evident, particularly in the anterior and posterior papillary muscles. The demonstration of nonperfused myocardium in late shock is compatible with the concept of a progressively developing edema with a consequent interruption of blood flow.


Profile: the National Association for Gifted Children. Until the foundation of NAGC in 1966, the general impression was that, of all sections of the child population, gifted children were among the best endowed and therefore the least worthy of our compassion and concern. Work with gifted children over the past 10 years has led us to reconsider the situation. Certainly the majority of them are successful, well-balanced and reach adult life with a large measure of happiness, but a sizeable minority unfortunately do not do so. For these children, giftedness itself brings problems, sometimes referred to as "the handicap of brilliance".


Enhancement of platelet aggregation by tranylcypromine in mouse cerebral microvessels. Tranylcypromine, given intraperitoneally at doses greater than or equal to 10 mg/kg, enhanced platelet aggregation in the arterioles on the cerebral surface in mice. Tranylcypromine inhibits prostacyclin synthesis in vitro. Iproniazid, which inhibits monoamine oxidase but not prostacyclin synthesis, failed to enhance platelet aggregation. The failure of iproniazid to enhance aggregation in this study rules out an effect on monoamine oxidase as the cause of tranylcypromine's action. That iproniazid inhibited aggregation indicates it has an effect opposite to that of tranylcypromine. Imidazole, a drug known to inhibit synthesis of both prostacyclin and thromboxane, failed to affect platelet aggregation. All of our data are compatible with the hypothesis that prostacyclin is an inhibitor of platelet aggregation. This hypothesis has been based largely on in vitro data, to which we now add in vivo support.


Pressure-flow characteristics of coronary stenoses in unsedated dogs at rest and during coronary vasodilation. The pressure-flow characteristics of 100 left circumflex stenoses in 10 chronically instrumented unsedated dogs were studied under resting conditions and during pharmacological coronary vasodilation. At rest, the pressure loss (deltaP) due to a stenosis and arterial flow velocity (V) were related by the equation, deltaP = FV + SV2, where F is the coefficient of pressure loss due to viscous friction in the stenotic segment and S is the coefficient of pressure loss due to flow separation at the diverging end of the stenosis. The linear term due to viscous friction accounted for 65% and the nonlinear term due to flow separation accounted for 35% of the total pressure loss at resting coronary flow. At peak coronary flow after coronary vasodilation, the pressure loss due to viscous friction accounted for 33% and pressure loss due to flow separation accounted for 67% of the total pressure loss. The pressure gradient-velocity relationship at high flows was characterized by the same general equation but with proportionately larger values of the coefficient S and therefore greater pressure loss associated with flow separation than predicted by the resting gradient-velocity relationship. The pressure loss predicted for high coronary flow velocities on the basis of the gradient-velocity equation at rest was only 64% of the actual experimentally observed pressure gradient at peak coronary flow. The augmented separation loss following coronary vasodilation probably was due to dilation of the epicardial artery adjacent to the fixed stenotic segment which caused more severe relative percent narrowing and a larger divergence angle at the distal end of the stenosis, the primary geometric determinants of separation losses.


Blood pressure and vascular resistance in genetically hypertensive rats treated at birth with 6-hydroxydopamine. Genetically hypertensive (GH) rats of the New Zealand strain and normotensive (N) rats were sympathectomized from birth with 6-hydroxydopamine (100 mg/kg,s.c, on alternate days, seven treatments). In adult treated rats from each strain (GHTr and NTr), blood pressure was lower than normal. Functional tests and electron microscopy showed that denervation was virtually complete in mesenteric and hindlimb arteries; the innervation of the renal artery was little affected. Ganglionic blockade still caused a large fall in blood pressure in treated rats. Vascular resistance was higher in blood-perfused hindlimbs and tails of GH rats than in those of N rats; in contrast, resistance was similar in limbs and tails of GHTr and NTr rats and was greater than that found in untreated N rats. Saline-perfused limb vessels had neither neurogenic nor myogenic tone and resistance was higher in GH limbs (whether these were from treated rats or not) than in untreated N limbs. In saline-perfused NTr limbs, there was a paradoxical structural adaptation (probably luminal narrowing) of the hindlimb blood vessels and resistance was higher than in untreated N rats. The resistance of saline-perfused GH and GHTr limbs was similar. A high peripheral resistance appears to be the main mechanism sustaining genetic hypertension, and the integrity of the vasomotor sympathetic nerves is necessary for the development of this form of experimental hypertension.


Simulation studies of the electrocardiogram. I. The normal heart. A digital computer model is presented for the simulation of the body surface electrocardiogram (ECG) during ventricular activation and recovery. The ventricles of the heart are represented in detail by a three-dimensional array of approximately 4000 points which is subdivided into 23 regions. Excitation sequence and cellular action potential data taken from the literature are used to determine the spatial distribution of intracellular potentials at each instant of time during a simulated cardiac cycle. The moment of the single dipole representing each region is determined by summing the spatial gradient of the intracellular potential distribution throughout the region. The resulting set of 23 dipoles is then used to calculate the potentials on the surface of a bounded homogeneous volume conductor with the shape of an adult torso. Simulated isopotential surface maps during both activation and recovery are in good agreement with data for humans reported in the literature.


Simulation studies of the electrocardiogram. II. Ischemia and infarction. Experimental studies of the myocardial action potentials following coronary artery occlusion have shown that the resulting regional ischemia is reflected by characteristics changes in the  shapes of the action potentials in the ischemic region. The principal changes are decreases in the magnitude of the resting potential and in the action potential duration. Action potentials with prolonged durations have been observed in the infarcted regions of experimental animals after the development of inverted T waves in the surface electrocardiogram (ECG). We use such abnormal action potentials in our digital computer model to study the effects of acute myocardial ischemia and infarction on the surface ECG. The heart is represented in sufficient detail to allow variations in the location and size of the ischemic injury and in the distribution of the severity of injury within the injured region. The evolution of acute infarctions is simulated by progressively modifying the abnormal action potentials assigned to the injured region. Calculated standard 12 lead ECGs and torso isopotential surface maps for simulated acute ischemia and infarction are in good agreement with patient data reported in the literature. Typical simulations include anterior and inferior transmural ischemia and infarction and anterior subendocardial ischemia. The model is used to examine relationships between torso surface potentials during ventricular activation and recovery and the site and size of the ischemic injury.


Local cerebral blood volume response to carbon dioxide in man. We used an emission tomographic brain scanner to investigate the relationship between local cerebral blood volume (LCBV) and arterial blood carbon dioxide tension (PaCO2) in normal awake man. Measurements were made separately in three dimensions in various regions of grey and white matter, and the resting LCBV as well as the difference in sensitivity among these regions were compared. Over the range of PaCO2 studied (20-42 torr), the response of both the grey matter and the white matter to carbon dioxide was linear. The LCBV sensitivity of the grey matter to changes in PaCO2 was 0.053 ml/100 g per torr PaCO2 and in the white matter this sensitivity was 0.046 ml/100 g per torr PaCO2. These sensitivities were found not to be significantly different, yielding a slope of 0.049 ml/100 g per torr PaCO2 for the LCBV-PaCO2 curve for the entire brain. This is in excellent agreement with the other data for the whole brain. The resting cerebral blood volume of the grey matter at a PaCO2 of 34.4 torr, which was the average resting arterial carbon dioxide tension of the subjects, was 5.0 ml/100 g and was significantly higher than for white matter, which was 3.6 ml/100 g. At the local level, the cerebral blood volume of the frontal cortex is significantly less than that of the thalamus, whereas the frontoparietal cortex in the region of the sylvian fissure has a local cerebral blood volume significantly greater than that of the thalamus.


On the interpretation of the exercise test. The widespread popularization of stress testing and the extensive application of its results in a variety of clinical settings make it imperative that a better method than the presently used simplistic "positive" or "negative" interpretation be devised. As a step toward a comprehensive diagnostic and prognostic index based on stress tests, a simple grading system of electrocardiographic changes, together with recognition of clinical modifiers, is suggested.


Early intraaortic balloon pumping for anterior myocardial infarction without shock. Eleven patients with anterior myocardial infarction less than six hours old underwent intraaortic balloon pumping (IABP) in an attempt to control injury. Direct hemodynamic measurements excluded shock. Response to therapy was judged individually by comparison to a one-hour pretreatment period. There was no parallel control group. Five patients responded with an 84% fall in ST elevation in one hour, with preservation of precordial R waves and good ventricular function. In contrast, six patients responded poorly, with a 40% fall in ST elevation in one hour, Q wave development and poorer residual left ventricular function. Coronary angiography demonstrated a significant correlation between response and presence or absence of complete left anterior descending coronary artery occlusion. Early use of IABP in anterior myocardial infarction may interrupt injury, though in this series only in cases with residual left anterior descending patency.


The effect of ischemia on thallium-201 clearance from the myocardium. To determine the effect of ischemia on myocardial clearance of thallium-201 (201Tl), we studied 12 dogs with ischemia produced after the injection of Tl. Tl was given I.V. 10 minutes before left anterior descending (LAD) coronary artery ligation. 85Sr-microspheres (MS) were administered 5 minutes later, and control biopsies were obtained from the myocardium. The LAD was tied and repeat biopsies obtained from the ischemic zone (IZ) and normal zone (NZ) 15 minures and 2 hours later. 46Sc-MS were given just before the final giopsy. Tl activity in the IZ was not significantly different from that in the NZ either before LAD occlusion or 15 minutes and 2 hours later. Tl clearance at the end of 2 hours was not significantly different (27 +/- 5% vs 28 +/- 5%, IZ vs NZ respectively) between the two zones. The half-time of Tl clearance from both the IZ and NZ was calculated at 4.5 hours (consistent with previously reported normal values). This occurred despite a decrease in regional myocardial blood flow to 24 +/- 6% of control (P less than 0.01) in the IZ  and an increase to 47 +/- 14% of control (P less than 0.01) in the NZ during the study. We conclude that myocardial ischemia does not alter the normal rate of Tl clearance from the myocardium.


Long-term preservation of ischemic myocardium in the dog by hyaluronidase. The administration of hyaluronidase is a promising intervention to protect the ischemic myocardium in man, but evidence of the extent to which it may reduce the ultimate size of an infarct is not well-defined. Hence, open chest, anesthetized dogs were randomized into 10 control dogs which received saline and eight treated dogs which received three doses of hyaluronidase (500 NF units/kg I.V.) at 15 minutes, 2 hours and 24 hours after occlusion of the left anterior descending coronary artery (CAO). Regional myocardial blood flow (RMBF) assessed by the microsphere technique was measured 12 minutes after CAO. The chest was then closed and the dogs were allowed to recover. Twenty-one days after CAO, the hearts were excised, divided into 1 cm thick slices and incubated in triphenyl tetrazolium chloride. Infarct size was then determined by planimetry. The left ventricular myocardium was divided into multiple samples for RMBF analysis. In control dogs 23.2 +/- 2% of the left ventricle was infarcted, compared to only 9 +/- 2.8% (P less than 0.001) in hyaluronidase-treated dogs. RMBF in noninfarcted myocardium directly adjacent to the infarct was similar to that in the normal zone remote from the infarct in the control dogs; however, in the hyaluronidase-treated dogs, blood flow in the myocardium adjacent to the infarct was significantly reduced to 68% of normal (P less than 0.01) in the outer myocardial wall and to 86% of normal (P less than 0.02) in the inner myocardial wall, which indicates that this tissue, at least in some part, was in jeopardy, but was salvaged by hyaluronidase. Epicardial electrocardiographic data showed that three weeks after CAO, Q waves were less frequent and smaller in hyaluronidase compared to untreated dogs. Preservation of the frequency and magnitude of R waves was greater in the hyaluronidase-treated group at three weeks. We conclude that hyaluronidase resulted in long-term preservation of the ischemic myocardium.


Myocardial infarct quantification in the dog by single photon emission computed tomography. Radionuclide techniques for sizing acute myocardial infarction have been hampered by the intrinsic limitations of the scintillation camera. Emission computed tomography can overcome these limitations. Single photon emission computed tomograms of the distribution of technetium-99m pyrophosphate in acute anterior and posterior infarcts were obtained in 16 dogs after death. Tomograms were also obtained in 10 dogs during life without gating. The size of the infarcts was determined by staining gross sections of the heart with nitro blue tetrazolium, dissecting out the infarcted tissue and weighing it. Infarct sizes were determined from the tomographic images and compared with the measured infarct sizes. Good images showing the location and three-dimensional extent of the infarcts were obtained in all dogs. The measured and calculated infarct sizes correlated well (r = 0.85). Comparison of the calculated sizes in the living (non-gated) and dead ("physiologically" gated) animals showed reasonable agreement (r = 0.87). Single photon emission computed tomography is a feasible and useful technique for localizing and sizing acute myocardial infarctions.


Pressure-flow characteristics of the coronary collateral circulation during cardiopulmonary bypass. Effects of ventricualr fibrillation. Even though ventricular fibrillation is used frequently during cardiopulmonary bypass (CPB), the effects of fibrillation on myocardial regions supplied by collateral vessels have not been determined. To study these effects, nine dogs with left ventricles (ameroid model) consisting of a region of myocardium supplied by collateral vessels (CR) and a region supplied by normal coronary arteries (NR) were subjected to normothermic CPB at two perfusion pressures. In both the empty beating heart (EBH) and empty fibrillating heart (EFH) regional myocardial flow was determined by tracer microspheres. Retrograde coronary pressure was measured via cannulation of the circumflex artery distal to the ameroid induced occlusion. When perfusion pressure was maintained at 80 mm Hg, retrograde coronary pressure was similar in the EBH (46 +/- 4 mm Hg) and in the EFH (48 +/- 3 mm Hg). During fibrillation subendocardial flow in the CR was unchanged, while flow in the NR increased (P less than 0.02). In addition, the endo/epi was greater in the NR than in the CR (P less than 0.01), a difference which did not exist in the EBH. The flow response to fibrillation in the CR could be produced in the NR by reducing the perfusion pressure to 50 mm Hg. These data suggest that during CPB, fibrillation exaggerates existing subendocardial perfusion deficits in collateral regions and the impaired flow response appears to be related to a low regional intravascular pressure.


Severe aortic stenosis with impaired left ventricular function and clinical heart failure: results of valve replacement. Nineteen patients, aged 58-80 years, with severe isolated aortic valve stenosis, severely reduced ejection fraction and clinical heart failure underwent aortic valve replacement between January 1970 and April 1977. Ten had concomitant coronary artery disease (all underwent additional coronary bypass surgery), 17 had angina pectoris and four had syncope. Aortic valve area index was 0.32 +/- 0.03 cm2/m2 (mean +/- SEM); left ventricular (LV) end-diastolic volume index was 117 +/- 9 ml/m2 and LV ejection fraction was 0.37 +/- 0.02. There were four operative deaths and one late death. The follow-up time ranged from six to 74 months (38 +/- 6 months). Actuarially determined three-year survival is 74 +/- 10%; the expected five-year survival is the same. One patient had a serious cerebrovascular accident. Of the remaining survivors, seven were initially Functional Class IV and six  Class III; currently, six are Class I and seven Class II (New York Heart Association classifications). The cardiothoracic ratio has decreased from 0.54 +/- 0.03 to 0.49 +/- 0.03. Repeat hemodynamic evaluation has been performed in 10 patients, 22 +/- 6 months after surgery. In these 10 patients, the aortic valve gradient decreased from 55 +/- 7 11 +/- 1.3 mm Hg; LV end-diastolic pressure from 22 +/- 2.4 to 9 +/- 1.9 mm Hg; LV end-diastolic volume index from 119 +/- 16 ml/m2 to 107 +/- 11 ml/m2. LV ejection fraction has increased dramatically from 0.34 +/- 0.03 to 0.63 +/- 0.05 and mean velocity of circumferential fiber shortening from 0.57 +/- 0.08 to 1.3 +/- 0.18 circ/sec. The encouraging long-term survival, improved functional class and the marked improvement in left ventricular function that occurred in our patients indicate that all patients with severe aortic stenosis in clinical heart failure should be offered aortic valve replacement.


Cardiac tamponade: hemodynamic observations in man. Hemodynamic studies were performed before and after pericardiocentesis in 19 patients with pericardial effusion. Right atrial pressure decreases significantly, from 16 +/- 4 mm Hg (mean +/- SD) to 7 +/- 5 mm Hg in 14 patients with cardiac tamponade. This change was accompanied by significant increases in cardiac output (3.87 +/- 1.77 to 7 +/- 2.2 l/min) and inspiratory systemic arterial pulse pressure (45 +/- 29 to 81 +/- 23 mm Hg). The remaining five patients did not demonstrate cardiac tamponade, as evidenced by lack of significant change in these hemodynamic parameters. In all patients with tamponade, right ventricular end-diastolic pressure (RVEDP) was elevated and equal to pericardial pressure; equilibration was uniformly absent in patients without tamponade. During gradual fluid withdrawal in the tamponade group, significant hemodynamic improvement was largely confined to the period when right ventricular filling pressure remained equilibrated with pericardial pressure. In 10 patients with tamponade and pulsus paradoxus, pulmonary arterial wedge pressure (PAW) was equal to pericardial pressure except during early inspiration and expiration when it was transiently less and greater, respectively; however, inspiratory right atrial pressure never fell below pericardial pressure. In these 10 patients, PAW decreased significantly following pericardiocentesis (P less than 0.001). In the remaining four patients with tamponade but without pulsus paradoxus, all of whom had chronic renal failure, PAW was consistently higher than pericardial pressure or RVEDP and did not decrease after pericardiocentesis. These data tend to confirm the hypothesis that in patients with tamponade, the venous pressure required to maintain any given cardiac volume is determined by pericardial rather than ventricular compliance. When pericardial compliance determines diastolic pressure in both ventricles, relative filling of the ventricles will be competitive and determined by their respective venous pressures (pulmonary vs systemic), which vary with respiration and alternately favor right and left ventricular filling. This results in pulsus paradoxus. However, if pulmonary arterial wedge pressure is markedly elevated before the onset of tamponade, as in patients with chronic renal failure, then pericardial compliance may only determine right ventricular filling pressure. In such cases, pulsus paradoxus may be absent.


Beneficial effects of hydralazine in severe mitral regurgitation. The severity of mitral regurgitation is, in part, determined by aortic impedance to left ventricular outflow. Sodium nitroprusside acutely decreases regurgitant flow, but the importance of its dual vasodilating effects, the lowering of peripheral vascular resistance and increasing of venous capacitance, is unclear. We studied the hemodynamic response to intravenous hydralazine, which selectively acts on the arteriolar resistance bed, in 10 patients with severe mitral regurgitation. Hydralazine produced a 50% increase in forward stroke volume (22 +/- 2 to 33 +/- 3 ml/m2, P less than 0.001) and a 33% reduction in regurgitant stroke volume (40 +/- 6 to 27 +/- 6 ml/m2, P less than 0.001), with a resultant fall in pulmonary capillary wedge v wave and mean pressures. Unlike nitroprusside, it did not alter left ventricular end-diastolic volume or pressure. Oral hydralazine maintained this hemodynamic improvement for at least 48 hours and, in three patients, provided more sustained clinical improvement. We conclude that hydralazine, by virtue of its selective lowering of aortic impedance, reduces the amount of mitral regurgitation and thus may be a useful mode of interim or chronic therapy in selected patients.


Contrasting rates of reversal of digoxin toxicity by digoxin-specific IgG and Fab fragments. Both heterologous IgG and Fab fragments of appropriate affinity and specificity have been shown capable of reversing advanced cardiac glycoside toxicity. Fab fragments are more rapidly excreted and theoretically have a smaller risk of unwanted immunologic effects, but relative rates of toxicity reversal have not been established. Rates of reversal of advanced digoxin toxicity by digoxin-specific IgG and Fab fragments were therefore compared in a dog model of advanced digoxin intoxication. Initial studies confirmed more rapid distribution of sheep Fab fragments (M.W. 50,000) than of the parent IgG molecule (M.W. 150,000) after intravenous injection. Twenty-five pentobarbital-anesthetized dogs were given 0.3 mg/kg digoxin intravenously, resulting in rapid onset of ventricular tachycardia in all animals. Eight dogs subsequently given nonspecific IgG or Fab died in asystole or ventricular fibrillation an average of 55 minutes after digoxin administration. Ten of 11 dogs given 1.33 moles of binding sites per mole of digoxin as intact IgG returned to sinus rhythm at a mean time of 85 minutes after antibody infusion. In contrast, six of six dogs given an equivalent dose of specific Fab fragments returned to sinus rhythm in a significantly shorter mean time of 36 minutes (P less than 0.01). Variability of time to arrhythmia reversion was less in Fab-treated dogs. These data demonstrate a decisive advantage of specific Fab fragments over intact IgG for potential clinical use in advanced, life-threatening digoxin intoxication.


The effect of digitalis on refractoriness of the intact canine His-Purkinje system. The effect of therapeutic doses of digitalis on functional (F), relative (R) and effective (E) refractory periods (RP) of the His-Purkinje system (HPS) was studied in 12 open-chested, innervated adult mongrel dogs (10-20 kg) during control and 15, 30 and 45-60 min after 0.016 mg/kg of intravenous ouabain. To determine the stability of the preparation and to assess time-dependent changes in His-Purkinje refractoriness, another six dogs (Group II) had similar studies, but without drug administration. In all dogs, the His bundle was paced by using the plunge wire technique at a predetermined cycle length (CL) and a premature stimulus (S2) to the His bundle was introduced at decreasing S1 S2 intervals. Following ouabain, in Group I dogs, at the longest Cls tested  (458 +/- 125 msec; +/- SD) there was significant increase in the FRP (+4.34%; P less than 0.05), RRP (S2 V2 (+6.57%, P less than 0.05), RRP (Ab) (+6%, P less than 0.05) and ERP (52%, P less than 0.05) of the HPS. These significant changes were generally observed 30 minutes after drug administration. Changes in RPs were of greater magnitude at longer CLs (greater than 400 msec), but insignificant at shorter CLs (less than 400 msec). The H-V interval during sinus rhythm and the S1 V1 interval during His bundle pacing at all CLs did not change after ouabain. In Group II dogs there were no significant change in His-Purkinje refractoriness over 60 minutes. These findings suggest that therapeutic doses of digitalis 1) tend to increase refractoriness within the HPS to a very small degree, 2) have no appreciable effect on His-Purkinje conduction, and 3) affect CL-dependent changes in refractoriness. The His bundle extrastimulus method is useful in studying the HPS in the intact heart.


Reentry within the His-Purkinje system. Elucidation of reentrant circuit using right bundle branch and His bundle recordings. Routes of impulse propagation during reentry in the His-Purkinje system (Re-HPS) in 13 patients were studied using the right ventricular extra stimulus (V2) method and right bundle branch (RBB) recordings in addition to the conventional His (H) bundle recordings. The H-V and RB-V intervals during sinus rhythm averaged 44.2 and 27.6 msec respectively, with the average antegrade H-RB interval 16.6 msec. All 13 patients demonstrated Re-HPS in the form of V3 showing a left bundle branch block (LBBB) pattern, and two of the 13 patients also manifested V3 with a right bundle branch block (RBBB) pattern. During Re-HPS showing a LBBB pattern, the retrograde activation of the His bundle (H2) in response to V2 occurred via the left bundle branch (LBB), as indicated by inscription of retrograde H2 prior to RB2 in nine of 13 cases (average H2-RB2 = 9.4 msec), and simultaneous incription of retrograde H2 and RB2 in the remaining four. When V3 showed a RBBB pattern the retrograde RB2 preceded H2, suggesting H2 activation via the RBB. These findings suggest that circuit of Re-HPS incorporates both bundle branches and the bundle of His. The H-RB recordings were also useful in distinguishing sustained Re-HPS from atrioventricular nodal reentry when in both instances the QRS complex showed a LBBB pattern.


Clinical and electrophysiologic findings in patients with paroxysmal slowing of the sinus rate and apparent Mobitz type II atrioventricular block. Over five years, 13 patients with episodic apparent type II atrioventricular (AV) block associated with sinus slowing were seen. This phenomenon occurred only transiently during an acute illness in eight patients (group I) but recurred chronically in five (groupII). For the group as a whole, the mean spontaneous cycle length was 42% longer during the period of AV block compared with periods of 1:1 AV conduction (800 +/- 116 msec to 1138 +/- 489 msec) (P less than 0.05). Electrophysiologic studies in four group I patients showed no abnormalities, whereas abnormalities in AV nodal conduction and refractoriness or provocation of intranodal Mobitz type II AV block (during carotid massage) were observed in three patients in group II and were totally abolished by atropine. In group I patients, apparent type II AV block was self-limited. In the chronic group, recurrent symptoms required insertion of permanent pacemakers in two patients. Simultaneous type II block and sinus slowing appeared to be related to the effects of increased vagal tone on both nodal structures. Intracardiac pacing is not indicated for patients with transient episodes associated with an acute illness, but may be required for symptomatic patients with recurrent episodes.


Dual echocardiographic determination of atrial contraction sequence in atrial flutter and other related atrial arrhythmias. We have applied the new technique of dual echocardiography to determine the sequence of atrial contraction as reflected in the simultaneously recorded movements of the tricuspid and mitral valves. The study group included 29 normal subjects and 23 patients with either atrial flutter, coarse atrial fibrillation or atrial tachycardia with block. In normal individuals, right atrial contraction preceded left atrial contraction, with an average interatrial contraction time of 17 +/- 8 msec. In contrast, the atrial contraction sequence was reversed in atrial flutter, with left preceding right atrial contraction and a prolonged interatrial contraction time of 82 +/- 20 msec. In two patients with atrial tachycardia with block, atrial contraction was either simultaneous or left preceded right atrial contraction by a brief interval. The sequence of atrial excitation, as determined by electrode catheter recordings from the right and left atria in one patient with atrial flutter and one patient with normal sinus rhythm, was the same as the contraction sequence. Left atrial pacing reversed both excitation and contraction sequences. After cardioversion of three patients from atrial flutter to normal sinus rhythm, interatrial contraction time was shortened but remained longer than in normal subjects, suggesting an interatrial conduction disturbance in patients with atrial flutter. In coarse atrial fibrillation, the contraction sequence varied. Significant motion of both mitral and tricuspid valves coincident with fibrillary waves occurred frequently, especially when the fibrillary waves were coarse and regular. Dual echocardiography permits the noninvasive determination of the sequence of atrial contraction and excitation, and may be useful in studying the characteristics of atrial arrhythmias.


Diagnosis of ventricular septal defect by pulsed Doppler echocardiography. Sensitivity, specificity and limitations. The M-mode echocardiographic findings of ventricular septal defect (VSD) are nonspecific. A specific pulsed Doppler echocardiographic (PDE) diagnosis of VSD can be made by following the turbulent VSD jet through the septum. To assess the sensitivity, specificity and limitations of PDE diagnosis of VSD, 105 children undergoing cardiac catheterization were examined by PDE. These children had a variety of cardiac defects, and a PDE diagnosis of VSD was made in 46/51 (90%) who had VSD proven at catheterization. There was one false positive PDE diagnosis of VSD, for a specificity of 98%. Factors influencing the ability to diagnose VSD by PDE include the location of the defect, level of pulmonary vascular resistance and direction of blood flow through the VSD. The presence of additional defects did not interfere with PDE diagnosis of VSD. The PDE detection of additional defects may identify situations where M-mode echocardiographic estimation of dimensions may not be indicative of the size of VSD shunt.


Two-dimensional echocardiographic assessment of Ebstein's anomaly. Nine patients with Ebstein's anomaly of the tricuspid valve were studied by two-dimensional echocardiography, using the standard long and short axis views as well as the apex four chamber view. With this latter view, the displacement of the tricuspid valve into the right ventricle was clearly seen in all nine cases of Ebstein's anomaly and was not noted in a control population. The severity of the tricuspid displacement was assessed by comparing the position of the mitral and tricuspid valves relative to the cardiac apex. The apex four chamber view allowed visualization of the atrioventricular (AV) ring simultaneously with the displaced tricuspid valve, and therefore the size of the "atrialized" right ventricle, true right ventricle and right atrium could be determined. These dimensions compared favorably with angiography.


Real-time cross-sectional echocardiographic imaging and measurement of the patent ductus arteriosus in infants and children. While echocardiography has been used to noninvasively document indirect effects on the heart of left-to-right shunting through a patent ductus arteriosus, no noninvasive technique has been developed to image the duct itself. In this study, 35 sequential studies were performed on 28 patients with a mechanical sector scanner to image the distal pulmonary artery and its bifurcation by scanning along the axis of the right ventricular outflow tract. Cross-sectional imaging, just superior to the take-off of the right pulmonary artery, provided visualization of the patent ductus as a distal continuation of the pulmonary artery connecting to the descending aorta. Ductal visualization by cross-sectional echo was validated by saline echo contrast observations of right-to-left and left-to-right shunting through the duct in 14 patients, by surgical observations in 11, angiographic observations in 13 and autopsy observations in three. Angiographic size of the ductus arteriosus, whether constricted or widely patent, tortuous or straight, was predicted correctly and echo/angiographic correlations for smallest inner ductal dimension were excellent (r = 0.97). This study provides a method for and validates the direct imaging of the ductus arteriosus and suggests that cross-sectional echocardiography can accurately predict ductal contour and quantitative ductal cross-sectional size noninvasively.


Models of congenital heart disease in fetal lambs. Intracardiac flow patterns were chronically altered by partially obstructing left ventricular (LV) inflow or outflow in midgestational fetal lambs. Physiological measurements of the fetal circulation were made serially through indwelling catheters and the use of radioactive microspheres. With LV inflow obstruction, mean LV output (LVO) decreased to 30% of control (P less than 0.01). Within seven days, the LV/right ventricular (RV) weight ratio decreased to 70% of control (P less than 0.01), and the mean LV/RV chamber volume decreased to less than one-half of control (P less than 0.001), simulating an early form of the hypoplastic left heart syndrome. With LV outflow obstruction, mean LVO decreased to 64% of control (P less than 0.05). Mean LV/RV wall thickness doubled (P less than 0.0001) and mean LF/RV chamber volume decreased to less than one-half of control (P less than 0.0001). Within four to ten days after increasing LV afterload, a large increase in LV mass occurred, which was demonstrated by morphometric analysis to be due to hyperplasia of ventricular myocytes. LV chamber volume decreased somewhat, simulating moderately severe congenital aortic stenosis. Over the long term (30--36 days), the mean LV/RV weight ratio decreased and the LV chamber was nearly obliterated, simulating very severe congenital aortic stenosis. The results suggest that by varying preload and afterload in both ventricles of the fetus, various forms of congenital heart disease may be simulated.


Effects of dietary and pharmacologic alteration of serum lipids on platelet survival time. Platelet survival time (SURV) (autologous labelling with 51Chromium) was shortened (3 +/- 0.03 days; average t 1/2 +/- SEM; normal t 1/2 3.7 +/- 0.03 days) in 88 out of 128 (69%) men with coronary disease. In 35 out of 47 men with hyperprebetalipoproteinemia, SURV was shortened (3 +/- 0.09 days) (74%). Of 30 men with hyperbetalipoproteinemia, SURV was shortened (2.5 +/- 0.10 days) in 26 (87%). Of 51 men without hyperlipoproteinemia, SURV was normal (3.3 +/- 0.10 days) in 24 (47%). Dietary alteration of serum triglyceride was undertaken in 12 men with hyperprebetalipoproteinemia, and in eight a decrease of triglyceride of more than 75 mg% was achieved (324 +/- 21-219 +/- 18 mg%; P less than 0.001) with an increase of SURV (2.2 +/- 0.11-2.8 +/- 0.13 days; P less than 0.001). In four, serum triglyceride increased by more than 75 mg% (279 +/- 14-451 +/- 28 mg%) and SURV decreased (2.7 +/- 0.16-2.3 +/- 0.21 days). Cholestyramine (16 g g.d.) and diet decreased serum cholesterol (348 +/- 7.6-319 +/- 6.3 mg%; P less than 0.001) in 15 men with hyperbetalipoproteinemia and SURV increased (2.3 +/- 0.08-2.7 +/- 0.07 days; P less than 0.001). Results suggest that SURV is shortened in men with coronary disease, particularly in those with hyperlipoproteinemia, and that alteration of triglyceride and cholesterol are associated with alteration of SURV.


Cardiovascular responses to dynamic and static effort soon after myocardial infarction. Application to occupational work assessment. Static and dynamic work involving the arms and the legs was performed by 40 men seven weeks after myocardial infarction. Leg ergometry produced a significantly higher peak work load, systolic blood pressure (BPs), heart rate (HR), and HR X BPs X 10(-2) product (DP) than did arm ergometry: 842 +/- 178 vs 546 +/- 135 kg-m/min, 176 +/- 24 vs 154 +/- 19 mm Hg and 256 +/- 54 vs 219 +/- 48 (SD). Peak heart rates were 145 and 142. Endpoints were primarily muscular and generalized fatigue and dyspnea. Ischemic abnormalities and ventricular ectopy were more frequent with leg ergometry. Sustained forearm lifting elicited higher HR, PBs and DP responses than sustained handgrip contraction: 95 +/- 16 vs 91 +/- 16 beats/min, 162 +/- 18 vs 152 +/- 17 mm Hg and 154 +/- 33 vs 139 +/- 33 (SD). Ischemic ST segment depression and significant ventricuar arrhythmias were infrequent with static effort. Dynamic leg testing is superior to dynamic or static arm testing in assessing the capacity of patients to perform physical work tasks after myocardial infarction.


Value and safety of diet modification to control hyperlipidemia in childhood and adolescence. A statement for physicians. Ad Hoc Committee of the Steering Committee for Medical and Community Program of the American Heart Association. Hyperlipidemia clearly is a risk factor for atherosclerotic disease in adults and very likely contributes to atherogenesis in children. The American Heart Association recommends that children with elevated plasma cholesterol or triglyceride be placed on an appropriate diet in order to reduce their risk of hyperlipidemia and possibly to reduce their risk of atherosclerotic disease when they become adults. Although the evidence does not yet support the recommendation that cholesterol and saturated fat should be reduced in the diet of all children, the public should be advised that such modification appears safe and very likely to be beneficial. Measuring plasma lipids of all children in order to prevent atherosclerotic disease probably is not cost effective. However, children of high risk families should be examined for hyperlipidemia.


Computer analysis of allergic histories taken by questionnaire. We report the development of a detailed allergy questionnaire designed so that the answers can be typed into a mini-computer. Computer programmes have been written to file and to retrieve these answers, and to print a summary which is then 'weighed', so as to produce a clinical atopy score which is then further broken down into indoor and outdoor scores. In a pilot survey of ninety patients, their clinical atopy scores and immunoglobulin E (IgE) profiles have been compared, and additional computer programmes have been written which will (a) assess if a more detailed IgE investigations is warranted, and (b) suggest which allergens, if any, are likely to be responsible for the patients' symptoms.


RAST atopy screen.

RAST atopy screen. A single in vitro screening test for atopic allergy is described. It uses the radioallergosorbent test (RAST) principle, and has been applied to screening sera from U.K. patients with possible atopic symptoms. When compared with the Phadebas RAST, the test sensitivity was 97% and the specificity 100%, and these results suggest that it is useful in any preliminary allergy investigation.


Serum IgE in atopic dermatitis: relationship to severity of cutaneous involvement and course of disease as well as coexistence of atopic respiratory diseases. Serum IgE concentrations were determined according to the radioimmunosorbent technique (RIST, Phadebas) on 116 adult patients with atopic dermatitis of varying severity and activity. Geometric mean IgE levels of patients with atopic dermatitis were significantly higher compared with the mean level of ninety-three non-atopic adult subjects without parasitic infestation. Severity of the atopic dermatitis was highly correlated to the levels of serum IgE. Severe chronic cases with ever-recurrent exacerbations show the most extreme values. In the moderate forms of atopic dermatitis, coexistent bronchial asthma causes a greater increase in the IgE values. Among the mild or abortive forms higher IgE levels were found in cases with allergic rhinitis than in the cases with 'pure' atopic dermatitis. Other findings in connection with IgE in atopic dermatitis are summarized. The pathogenetic significance of IgE in the cutaneous changes is briefly discussed.


Hymenoptera sting hypersensitivy: IgE, IgG and haemagglutinating antibodies to bee venom constituents in relation to exposure and clinical reaction to bee stings. High levels of IgE antibodies (Ab) to bee venom constituents were mainly found in patients with bee sting hypersensitivity, whereas the sera of bee keepers usually contained high levels of IgG Ab and haemagglutinins, representing blocking antibody activity. In bee keepers there was a positive correlation between the degree of severity of an adverse reaction to bee stings and IgE Ab, a negative one between the severity of reaction and IgG Ab and haemagglutinins, a positive one between the number of stings per season and IgG Ab and haemagglutinins, and negative one between the number of stings and IgE Ab. This suggests a mutual dependence of the production of IgE Ab and blocking Ab in a regularly exposed population. No such correlation was found in bee sting-allergic patients, indicating that other factors influence the severity of reaction in this only sporadically exposed group. The values of IgG Ab and haemagglutinins correlated well with each other, with IgG Ab giving a better correlation with severity of reaction and number of stings than the haemagglutinins.


Asthma, allergy and bronchial hyper-reactivity to histamine in patients with bronchiectasis. An allergy examination was made on forty-eight consecutive bronchiectasis patients, with a mean age of 37 years. Eleven had confirmed asthma and another nine, making twenty in all (42%), had distinct allergic diathesis. Allergy to a specific allergen was established in five cases. 50% of the patients in the total series had bronchial histamine hyper-reactivity, eleven without asthma or distinct allergic diathesis. The results corroborate views previously reported of the role of asthmatic symptoms and allergic diathesis as aetiological factors of bronchiectasis.


The effect of ICI 74,917 on asthma and bronchial provocation tests. A study was made of the protective action of the anti-allergic drug ICI 74,917, a phenanthroline derivative, in specific allergen provocation tests and in combating clinical symptoms in thirteen asthmatic out-patients. It was found that ICI 74,917 was able to diminish allergen-induced bronchoconstriction significantly (P less than 0.01) during both the placebo period and during chronic treatment with ICI 74,917 for 4 weeks. The results obtained during these two periods did not differ significantly. There was not, however, any statistically significant difference in the symptom scores or PEF rates during the treatment with ICI 74,917 or placebo for 1 month. This may indicate that although ICI 74,197 is able to prevent bronchoconstriction in an allergen provocation test, it failed to improve asthma clinically. No signs of the phenomenon of tachyphylaxis were observed in the present study. The Student's t-test was used in the statistical analysis.