Evidence for amygdaloid projections to the contralateral hypothalamus and the ipsilateral midbrain in the rat. Light microscopic autoradiography was performed subsequent to injection of tritiated amino acids into various parts of the amygdaloid body of the rat. Evidence is provided for two hitherto unreported projections of the amygdala: from the medial amygdaloid nucleus to the contralateral premamillary nuclei and from the central amygdaloid nucleus to the mesencephalic central grey. The functional implications of these findings are discussed.


The spermatozoon of Oikopleura dioica Fol (Larvacea, Tunicata). The spermatozoon of Oikopleura dioica is about 30 micron long, with a spherical head, about 1 micron wide, a 3 micron long and 1 micron wide midpiece, and a 25 micron long tail with a tapered end piece. The head contains a nucleus with the chromatin volume limited to about 0.1 micron3. A small acrosome is found in an anterior inpocketing, and a flagellar basal body in a posterior inpocketing of the nucleus. The midpiece contains a single mitochondrion with the flagellar axoneme embedded in a grove along its medial surface. The flagellar axoneme has the typical 9 + 2 substructure, and the basal body the typical 9 + 0 substructure. A second centriole and special anchoring fibres are absent.


Ultrastructure and migration of screening pigments in the retina of Pieris rapae L. (Lepidoptera, Pieridae). The retinal morphology of the butterfly, Pieris rapae L., was investigated using light and electron microscopy with special emphasis on the morphology and distribution of its screening pigments. Pigment migration in pigment- and retinula cells was analysed after light-dark adaptation and after different selective chromatic adaptations. The primary pigment cells with white- to yellow-green pigments symmetrically surround the cone process and the distal half of the crystalline cone, whilst the six secondary pigment cells, around each ommatidium, contain dark brown pigment granules. The nine retinula cells in one ommatidium can be categorised into four types. Receptor cells 1-4, which have microvilli in the distal half of the ommatidium only, contain numerous dark brown pigment granules. On the basis of the pigment content and morphology of their pigment granules, two groups of cells, cells 1, 2 and cells 3, 4 can be distinguished. The four diagonally arranged cells (5-8), with rhabdomeric structures and pigments in the proximal half of the cells, contain small red pigment granules of irregular shape. The ninth cell, which has only a small number of microvilli, lacks pigment. Chromatic adaptation experiments in which the location of retinula cell pigment granules was used as a criterium reveal two UV-receptors (cells 1 and 2), two green receptors (cells 3 and 4) and four cells (5-8) containing the red screening pigment, with a yellow-green sensitivity.


Induction of paracrystalline arrays by vincristine in the synaptic formations of the teleost retina. After injection of 10 microliter 10(-3) M vincristine into the vitreous body of the eye of the rainbow trout (Salmo gairdneri, Richardson), paracrystalline arrays in cell bodies, cell processes and presynaptic formations of the retina cells were observed. The structures resemble the paracrystalline lattice identified by several authors as microtubular protein. The paracrystals in both microtubule-rich cell processes and in synaptic areas, which show only a few or no microtubules, appear to be equivalent. The synaptic paracrystals are suggested to arise from both soluble tubulin and synaptic vesicles, indicating a functional role of tubulin in synaptic transmission.


The influence of activation, removal or denervation of the pineal on the fine structure of the Leydig cell and seminal vesicle epithelium in golden hamsters. Fine structural changes of testicular interstitial cells of Leydig and secretory cells of seminal vesicles were studied in golden hamsters under different functional states of the pineal gland. Experiments were performed in the reproductive season (summer months). In the hamsters blinded for 8 weeks the testes and the seminal vesicles were markedly atrophic, and the Leydig cells and the secretory cells of seminal vesicles were extremely involuted. By contrast, both types of cells in the pinealectomized or superior cervical ganglionectomized hamsters exhibited cytological features suggestive of an enhanced secretory activity. This study shows that functional activity of Leydig cells as well as secretory cells of seminal vesicles in the hamster may be depressed or augmented by stimulating of inhibiting the pineal antigonadal function, respectively, without performing hypophysectomy or hormonal administration.


Culture of human pituitary prolactin and growth hormone cells. Fragments of pituitary tissue obtained from a total of 37 patients with either breast cancer, diabetic retinopathy, galactorrhea, or acromegaly were dissociated into single cell suspensions prior to cell culture. Release of human growth hormone (hGH) and human prolactin (hPRL) into the culture medium was measured by radioimmunoassay. During a 3-week culture period, prolactin cells released 9--13 times the intracellular levels of hPRL at the time of seeding, whereas hGH release from growth hormone cells was only 1--2 times that of their initial intracellular level during this same time. Both growth hormone and prolactin cells retained distinctive ultrastructural features during culture. The prolactin cells responded to TRH stimulation by elevated release of PRL into the medium. No evidence for mitotic division of prolactin cells in vitro was found.


Surface fine structure of the subfornical organ in the Japanese quail, Coturnix coturnix japonica. The surface ultrastructure of the subfornical organ (SFO) was investigated in the Japanese quail. The SFO consists of a body and a stalk. The body of the SFO can be divided into rostral and caudal parts. On the rostral part, each ependymal cell possesses a short central solitary cilium; clustered cilia are also occasionally seen. Microvilli are abundant. On the caudal part, cells with a solitary cilium are fewer in number, and clustered cilia are rarely found. Microvilli are not as abundant as on the rostral part. In addition, large bulbous protrusions, tufts of small protrusions, deep funnel-shaped hollows, small pinocytotic invaginations and possible cerebrospinal fluid-contacting axons are sporadically observed on the surface of various regions of the body. Each ependymal cell of the stalk has a wide apical surface. A central solitary cilium, microvilli and other structures are observed more rarely on the stalk than on the body, while clustered cilia are not seen on the stalk. These structures are compared with those of the mammalian SFO and further discussed in relation to the possible dipsogenic receptor function for angiotensin II.


Parenchymal fine structure of the subfornical organ in the Japanese quail, Coturnix coturnix japonica. The parenchyma of the subfornical organ (SFO) of the Japanese quail was studied by light and electron microscopy. The SFO consists of ependymal, intermediate, and basal (perimeningeal) layers. In the intermediate layer, neurons, glial cells, and their processes are found. Axons containing dense core granules approximately 80 nm in diameter are numerous, some of which make synaptic contact with the neuronal perikarya or dendrites. Synaptic vesicles in some axons contain a dense dot in the interior after treatment with 5-hydroxydopamine. The activity of the SFO, which is probably concerned with elicitation of drinking by angiotensin II, may be regulated at least partly by afferent monoaminergic axons. Capillaries with a non-fenestrated endothelium are occasionally found in the parenchyma. The basal layer is occupied by glial processes abutting on the digitating layer of perivascular connective tissue of meningeal vessels. The endothelium of these vessels is occasionally fenestrated. Trypan blue injected systemically accumulated in the SFO, but not in the deeper areas of the brain. The absence of a blood-brain barrier is suggested in the SFO.


Direct connections between mitochondria and catecholamine-storage vesicles demonstrated by high voltage electron microscopy in rat adrenal medulla. Tubular channels from mitochondria to catecholamine-storage vesicles have been demonstrated in thick sections of adrenal medullary tissue from hypoglycemia-stressed rats by the use of the high voltage electron microscope. The function of these connections is not presently known although they may serve as channels for the transport of materials such as high-energy nucleotides from one organelle to the other. The present study has examined only the adrenal medulla, but it should be considered that such connections may also exist in other neural cells and possibly other cells in which there is intracellular transport of ATP.


Structure of occluding junctions in ileal epithelial cells of suckling rats. Two kinds of occluding junctions are found between ileal epithelial cells of suckling rats: apical zonulae occludentes (ZO) and fasciae occludentes (FO) which are associated with the lateral plasma membranes of many epithelial cells. In unfixed preparations, glycerol treatment induces the further proliferation of extensive fasciae occludentes. Both kinds of junction have identical structural elements when visualized in freeze fracture replicas, although the arrangement of these elements differs. Zonulae occludentes consist of networks of branching and anastomosing linear ridges or rows of approximately 10 nm particles with 20--30 nm spaces between the rows which form narrow belt-like structures around the apical region of adjacent cells. Fasciae occludentes, on the other hand, consist of similar linear ridges or rows of particles but the junction strands are often discontinuous, open ended and only occasionally intersect with each other. Several different fracture planes through the plasma membrane in the region of the occluding junctions have been observed and these provide further evidence that two components, one from each membrane, fused at the level of the extracellular space, form the junction sealing element. Furthermore, we present evidence which indicates a staggered rather than an in-register arrangement of these two components.


Transient occurrence of chloride cells in the abdominal epidermis of the guppy, Poecilia reticulata Peters, adapted to sea water. Chloride cells are characterized by a great number of mitochondria and a uniquely proliferated agranular endoplasmic reticulum through which ions are transported to an apical pit for secretion to the environment. Chloride cells are extremely rare in the epidermis of the guppy, Poecilia reticulata Peters, adapted to fresh water. They are frequent in the epidermis of guppies adapted to sea water three weeks after adaptation, but have disappeared in fish that were kept in sea water for approximately one year. The fine structure of epidermal chloride cells and the influence of prolactin treatment on their number are described. The significance of these cells with respect to the possible roles of the epidermis and the pituitary hormone prolactin in teleost osmoregulation is discussed.


Cytological observations on the ventromedial hypothalamic nucleus. Two sorts of neurons are recognized in Golgi impregnations of the rat ventromedial hypothalamic nucleus (HVM). The two cell types, category I and II neurons, are differentiated on the basis of their somatic, dendritic, and axonal characteristics. Category  I neurons form most of the neuronal population and are located throughout HVM. The small number of category II neurons that have been studied occur in lateral HVM. Two varieties of neuronal profile, "common" and "uncommon cells", are seen in thin sections of HVM. The "uncommon cells", in comparison with the "common ones", appear to have a larger soma, a more electron-dense cytoplasmic matrix, an abundance of Nissl bodies, and a population of dense-cored vesicles (100--130 nm in diameter). Some of the somata and proximal dendrites of "common", but not "uncommon" cells, are wrapped in multiple layers of astrocytic processes. Although the correlation is tentative, it is argued that category I neurons correspond to "common cells" and category II, to "uncommon cells". One possible implication of this correspondence is discussed regarding neuronal alteration in response to change in the endocrinological environment of the brain.


Effects of hormone-releasing stimuli on the area of the perivascular space in the neural lobe of the rat. Neural lobes from rats subjected to neurohypophysial hormone-releasing stimuli were examined electron microscopically following fixation in 4% tannic acid in 2.5% glutaraldehyde. This fixation allowed the delineation of the perivascular space in the neural lobe tissue. Measurement of the area of the perivascular space showed that it was significantly increased in the rats subjected to vagal stimulation and intraarterial calcium ions compared to the control rats. The rats which had been subjected to haemorrhage as a hormone-releasing stimulus did not show any significant change in the area of the perivascular space. The significance of these findings in relation to hormone release is discussed.


Fine structure of the palisade zone of the rat median eminence as revealed by freeze-fracturing. Nerve terminals in the palisade zone of the rat median eminence were investigated with freeze-fracture electron microscopy. Fracture face P of the specific terminals showed two populations of intramembranous particles (IMP): a large and a small variety. The large IMP-s often formed small irregular clusters. In nerve terminals the total number of both populations of IMP-s was considerably less than that found on P membrane faces of ependymal feet. On fracture face E of the nerve terminals, the number of IMP-s was about a quarter of that seen on fracture face P. On both fracture faces of most terminals a few small round impressions (or elevations respectively) were found which may be interpreted as broken necks of either exo- or endocytotic vesicles. Neither gap nor tight junctions occurred at lateral membranes of the specific axon terminals. Similarly, no membrane specializations were observed with freeze-fracturing on membrane areas adjacent to the basement membrane. The findings are discussed in relation to a possible exocytosis mechanism of the hypothalamic releasing and inhibiting hormones.


The origin, structure and occurrence of corpora amylacea in the bovine mammary gland and in milk. Corpora amylacea were present in variable numbers in the mammary glands of 8 lactating cows but they were not observed in the glands of pregnant or virgin heifers. In primiparous cows they were relatively simple bodies composed of 2--3 concentric layers but in multiparous animals, more complex multilayered structures were also noted. Corpora amylacea were formed in alveoli lined with actively secreting cells. The initial stage of their formation involved the aggregation, fusion and compaction of casein micelles in the alveolar milk. The resulting structure acted as a nucleus for further growth which occurred by the addition of material of micellar origin to its surface. Bodies containing a large proportion of crystalline material were common. Contact of corpora amylacea with the alveolar epithelium often caused flattening or irreversible damage to the secretory cells. The corpora amylacea recovered from milk corresponded in appearance to various stages in their growth seen in the mammary gland. They were usually present in the colostrum of multiparous cows but never in that from primiparous animals. In only 1 cow was the colostrum and milk found to be consistently free of corpora amylacea.


Lymph node macrophages and reticulum cells in the immune response. I. The primary response to paratyphoid vaccine. The reactivity of rat lymph node macrophages and reticulum cells has been studied in relation to the course of the immune response. Special attention was paid to the interdigitating cells (IDC) in the paracortex and the dendritic reticulum cells (DRC) in the germinal centre. Different types of macrophage predominate in the three different phases of the immune response. In the induction phase exudate macrophages predominate. They transform their morphology depending on the micro-environment in which they settle. These cells may contain Birbeck granules. Our experiments suggest, that these granules are immunologically specific organelles, associated with the induction of a primary humoral response. In the proliferation phase transitional cells between newly arrived macrophages and IDC predominate in the paracortex and the number of IDC decreases. In the memory phase IDC repopulate this thymus dependent area, indicating that the characteristic morphology of these cells results from the immune response in the T cell environment. The DRC are specifically related to the germinal centre, the morphological transformation of this peculiar reticulum cell occurs under the influence of the germinal centre reaction. The germinal centre reaction originates by tingible body macrophages (TBM).


Ultrastructural analysis of the action of reserpine on the brain neuroendocrine system of the wax moth, Galleria mellonella L., Lepidoptera. This study concerns the influence of reserpine on the fine structure of peptidergic neurosecretory cells in the pars intercerebralis of Galleria mellonella, and of neurons containing smaller dense-cored vesicles (presumed to be aminergic) localized in the same area of the brain. The drug, administered in doses of 125 microgram and 250 microgram per g of insect body weight, reduces both the amount and the electron opacity of the dense-cored vesicles with a diameter of 60--80 nm in the neuronal perikarya as well as their terminals. Simultaneously, this treatment evokes an abnormal accumulation of secretory granules within the perikarya of peptidergic neurosecretory cells belonging to three types. This accumulation of secretory material is followed by some changes in the fine structure of these cells. One (fourth) type of neurosecretory cells seems to be insensitive to reserpine action. Participation of the aminergic system in the regulation of the secretory activity of some populations of peptidergic neurosecretory neurons of the insect brain is postulated.


Kinetics of functional and morphological changes during decoupling and recoupling induced by glycerol in isolated muscle fibres of the crayfish. The development of ultrastructural changes in the T-system of isolated muscle fibres of the crayfish by the glycerol procedure is described in correlation with the dissociation of excitation-contraction (E--C) coupling as well as with recoupling of the E--C link. The sequence of events in the process of disconnection of the tubules is as follows: dilation of the T-system tubules, disconnection of the constricted tubular segments from the surface membrane and from the T-system vesicle, disappearance of the lumen and its disintegration. The decoupled state is characterised by the presence of round vesicles uniformly distributed in the entire volume of the fibre. The volume of vesicles accounts well for the residual postglycerol volume increase (15%) of the muscle fibres. Functional and structural recovery can be induced by reapplication of glycerol to fibres decoupled and vesiculated with concentrations of glycerol less than or equal to 300mmol.l(-1) in crayfish saline. The restitution starts with the organisation of the material of the disintegrated connecting segment of the T-system tubule into small vesicles which coalesce to form the tubule from the vesicular site. At the same time the surface membrane is invaginated toward the vesicle, thus forming the tubule from the surface membrane site. Recovery starts already in the first minute after application of glycerol and is completed within approximately 15 min.


A dividing granule-containing cell in the pelvic ganglion of the guinea-pig. A dividing granule-containing cell is described in the pelvic ganglion of the guinea-pig two days after pelvic nerve section. This appears to be the first report of a dividing granule-containing cell in adult tissue.


Morphological changes in the esophageal epithelium of the eel, Anguilla japonica, during adaptation to seawater. The esophageal epithelium of the Japanese eel, Anguilla japonica, was studied by light and electron microscopy. In freshwater-adapted eels, longitudinal folds of the mucosal surface are simple in form and lined by a stratified epithelium composed of mucous cells, filament- and ribosome-rich cells. Mucous cells are numerous. The filament-rich cells form the outermost and the basal layers of the stratified epithelium and are scattered in the middle zone among the mucous cells. They are firmly bound to one another by many desmosomes and prominent interdigitations of plasma membrane. The distal free surface of the filament-rich cell has a fingerprint-like pattern of microridges. A small number of columnar cells occur at the apices of the folds. They are rich in mitochondria and their distal surfaces bear short microvilli. In seawater-adapted eels, irregularly meandering folds increase the surface area of the mucosa. The stratified epithelium is extensively replaced by a simple columnar epithelium free of mucous cells. The columnar cells resemble in many respects those found in freshwater-adapted eels. They are rich in mitochondria and their distal free surface were provided with short microvilli. However, prominent lateral intercellular spaces and elaborate interdigitations of cytoplasmic processes in the distal zone distinguish the former from the latter. Results are considered in connection with the changes in ion and water permeability of the epithelium after seawater adaptation.


Ultrastructure of capillaries in the conduction system of the heart in various mammals. The capillaries in the atrioventricular node and atriventricular bundle of the heart (mammals) were investigated by thin sections and freeze-etch replicas. Fenestrated endothelial cells were found to occur in all mammals examined in this study. These fenestrations (500 +/- 50 A diameter) are bridged by a diaphragm and occur in relatively small sieve areas. In contrast to the AV-node and AV-bundle, the non-specialized myocardium is supplied by continuous capillaries. The presence of the fenestrated capillary in the conduction system of the heart may serve to increase both permeability and passive transport capacity.


Endoneurial mast cells in peripheral nerves of the armadillo dermis. Electron microscopy of the dermis of the 9-banded armadillo, Dasypus novemcinctus, reveals an intimate relationship of mast cells to peripheral nerves. Mast cells are routinely found in the dermis in close proximity to nerves, and mast cells are also present within the perineurial cell sheath in the endoneurial space proper. These cells contain electron opaque granules and exhibit numerous surface folds as is characteristic of other species. The degranulation of the mast cells by injection of 0.5% aqueous trypan blue reveals sequential exocytosis of granules, similar to that described for the rat. Degranulation occurs in most dermal and endoneurial mast cells following a single intradermal injection of trypan blue. A small number of mast cells partially degranulate and a few exhibit no degranulation, suggesting a heterogeneous population. It is also noted that mitochondrial morphology differs in degranulating cells, in that they become swollen and vacuolated. The results of this study are discussed in light of previous results reported for other species, mainly at the light microscope level.


The structural organization of DNA packaged within the heads of T4 wild-type, isometric and giant bacteriophages. We present electron microscopic and X-ray diffraction evidence concerning the structural organization of condensed DNA within a series of T4 bacteriophage with the following head morphologies: prolate (wild-type), isometric and giant (with greatly increased axial ratio). In all cases, the DNA helix segments are locally parallel and 27 A apart. For the giant particles, we show that the DNA forms a large coil whose axis is perpendicular to the axis of the phage tail. This evidence, combined with previous results from a series of isometric bacteriophages (Earnshaw and Harrison, 1977), leads to a model for the organization of condensed DNA that may apply to most dsDNA-containing bacteriophages.


Patterns of integration of viral DNA sequences in the genomes of adenovirus type 12-transformed hamster cells. The patterns of integration of the viral genome have been analyzed in four hamster cell lines transformed by adenovirus type 12 (Ad12). It has previously been shown that in each of the cell lines HA12/7, T637, A2497-2 and A2497-3, the viral genome persists in multiple copies, and that different parts of the viral DNA are represented non-stoichiometrically (Fanning and Doerfler, 1976). All four cell lines are oncogenic when injected into hamsters. The DNA from each of the cell lines was extracted and cleaved in different experiments with restriction endonucleases Bam HI, Bgl II, Eco RI, Hind III, Hpa II or Sma I. The DNA fragments were separated on 1% agarose slab gels and transferred to nitrocellulose filters by the Southern technique. Ad12 DNA sequences were detected by hybridization to Ad12 DNA, which was 32 P-labeled by nick translation, and by subsequent autoradiography. In some experiments, the 32P-labeled Eco RI restriction endonuclease fragments of Ad12 DNA were used to investigate the distribution of specific segments of the viral genome in the cellular DNA. For each cell line, a distinct and specific pattern of integrated viral DNA sequences is observed for each of the restriction endonucleases used. Moreover, viral sequences complementary to the isolated Eco RI restriction endonuclease fragments are also distributed in patterns specific for each cell line. There are striking differences in integration patterns among the four different lines; there are also similarities. Because the organization of cellular genes in virus-transformed as compared to normal cells has not yet been determined, conclusions about the existence or absence of specific integration sites for adenovirus DNA appear premature. Analysis of the integration patterns of Ad12 DNA in the four hamster lines investigated reveals that some of the viral DNA molecules are fragmented prior to or during integration. Analysis with specific restriction endonuclease fragments demonstrates that the Eco RI B, D and E fragments, comprising a contiguous segment from 0.17-0.62 fractional length units of the viral DNA, remain intact during integration in a portion of the viral DNA molecules. Although each cell line carries multiple copies of Ad12 DNA, the viral DNA sequences are concentrated in a small number of distinct size classes of fragments. This finding is compatible with, but does not prove, the notion that at least a portion of the viral DNA sequences, is integrated into repetitive sequences, or else that the integrated viral sequences have been amplified after integration. In the three cell lines which were tested, the integration pattern is stable over many generations, with continuous passage-twice weekly-of cells for 6-7 months. In the three cell lines which were examined, the integration pattern is identical in a number of randomly isolated clones. Hence it can be concluded that the patterns of integration are identical among all cells in a population of a given line of transformed cells.


DNAase I, DNAase II and staphylococcal nuclease cut at different, yet symmetrically located, sites in the nucleosome core. We have determined the relative location of pancreatic DNAase (DNAase I), spleen acid DNAase (DNAase II) and staphylococcal nuclease cleavage sites in the nucleosome core. Each of these three enzymes cleaves the DNA of chromatin at 10.n nucleotide intervals (n integer); this specificity presumably reflects the internal structure of the nucleosome. We have already reported that DNAase I cleaves nucleosomal DNA so that nearest adjacent cuts on opposite strands are staggered by 2 nucleotides, 3' end extending (Sollner-Webb and Felsenfeld, 1977). Here we show that the nearest cuts made by DNAase II in nucleosomal DNA are staggered by 4 nucleotides, 3' end extending, while cuts made by staphylococcal nuclease have a stagger of 2 nucleotides, 5' end extending. The cutting sites of the three enzymes thus do not coincide. Each pair of staggered cuts, however, is symmetrically located about a common axis-that is, the "dyad axes" that bisect nearest pairs of cutting sites coincide for all three enzymes. This result is consistent with the presence of a true dyad axis in the nucleosome core. Our results support the conclusion that a structural feature of the nucleosome, having a 10 nucleotide periodicity, is the common recognition site for all three nucleases. The position of the cut is determined, however, by the individual characteristics of each enzyme. Sites potentially available to nuclease cleavage span a region of 4 nucleotides out of this 10 nucleotide repeat, and a large fraction of these sites are actually cut. Thus much of the nucleosomal DNA must in some sense be accessible to the environment.


Electron microscopy and restriction enzyme mapping reveal additional intervening sequences in the chicken ovalbumin split gene. The Eco RI fragment "b" of chicken DNA (Breathnach, Mandel and Chambon, 1977), which contains the sequences coding for the 5' quarter of ovalbumin mRNA (ov mRNA), has been isolated by molecular cloning using a "shotgun" approach. Electron microscopy and restriction enzyme analysis have revealed that the sequences coding for the 5' quarter (approximately 500 nucleotides) of ov mRNA are split into four regions separated by three intervening sequences. The cloning procedure seems to be reliable, since the restriction enzyme pattern of the cloned Econ RI fragment "b" is similar to that of the corresponding chromosomal DNA fragment. There is no evidence supporting the existence of a 150-200 nucleotide long sequence at the 5' end of the ov mRNA similar to the "leader" sequences found at the 5' end of some adenovirus and SV40 mRNAs.


Organization of coding and intervening sequences in the chicken ovalbumin split gene. The interruptions in the chicken ovalbumin gene which were reported previously (Breathnach, Mandel and Chambon, 1977) are shown to be due to the presence of intervening sequences which separate the messenger-coding sequences. We present evidence for an additional interruption of the gene, which, together with those reported earlier and by Garapin et al. (1978b), make a total of six intervening sequences. All of these intervening sequences are located in the DNA region that corresponds to the part of the ov mRNA which codes for amino acids. The seven coding fragments of the split ovalbumin gene are arranged in the same order and relative orientation as in the ovalbumin double-stranded cDNA. All the sequences coding for ov mRNA are contained in a chromosomal DNA region of 6000 bp, which is more than 3 times longer than ov mRNA. The general organization of the ovalbumin split gene is discussed.


Genes and spacers of cloned sea urchin histone DNA analyzed by sequencing. A cloned histone gene cluster of the highly reiterated type from the sea urchin Psammechinus miliaris was analyzed by DNA sequencing. More than half of the 6 kb repeat was sequenced, including coding regions of all five histones, some prelude and trailing sequences lying adjacent to the structural gense, and segments of the AT-rich spacer DNA. The gene cluster does not code for gonad-specific histone variants but may instead be active in early sea urchin development, as indicated by comparison to reference histones. The encoded histones seem not to be derived from longer precursor proteins, not is there any evidence for insert sequences within the coding regions. Sequence similarities exist among the putative ribosome-binding sites adjacent to the initiator codons of individual genes. The AT-rich spacer segments between the genes differ from each other, are made up from relatively simple nucleotide arrangements, but are not repetitious, and apparently do not code for additional large proteins.


The precursor of mouse beta-globin messenger RNA contains two intervening RNA sequences. We have investigated the locations of the poly(A), the mRNA-specific sequences and the RNA sequences that are eventually cleaved from the 1860 nucleotide precursor of mouse beta-globin mRNA. Biochemical and electron microscopic data demonstrate that there are two intervening RNA moieties in the precursor which separate the beta-globin mRNA sequences into three portions containing 480, 205 and 155 nucleotides. One of the two intervening RNA moieties contains 780 nucleotides. The size of the smaller intervening RNA has not been determined precisely, but it is 125 nucleotides or less. The largest mRNA-specific fragment is derived from the 3' terminus of the precursor, and contains the 3' terminal poly (A) and 330 mRNA-specific transcribed nucleotides. At least one, and probably both, intervening RNAs occur within the coding portion of the mRNA sequences. The larger of the intervening RNAs is located next to the 480 nucleotide mRNA-specific fragment, and the smaller intervening RNA is located between the 205 and 155 nucleotide mRNA-specific fragments. These experiments are consistent with the notion that the intervening sequences in the DNA of mouse beta-globin genes are transcribed into the mRNA precursor and are excised from the RNA by post-transcriptional events.


Structure of the adenovirus 2 early mRNAs. We have defined the structure of adenovirus 2 (Ad2) cytoplasmic RNAs produced during the early phase of infection. Hybrids between cytoplasmic RNA and DNA restriction fragments of the viral genome were digested with endonuclease S1 or exonuclease VII, and the products were analyzed by gel electrophoresis. Seven abundant cytoplasmic RNAs (assumed to be mRNAs) were identified, and all have a spliced structure. Different mRNAs produced from a single transcriptional unit contain extensively overlapping sequences, and differ from each other by the pattern in which genome sequences are spliced together. The structures of the early Ad2 mRNAs are consistent with a model for mRNA biosynthesis in which an initial transcript is processed into a mature mRNA by "splicing out" internal sequences. The pattern of spliced mRNAs produced from the early region responsible for the transforming activity of Ad2 resembles the splicing pattern of the oncogenic early mRNAs of simian virus 40 (SV40). This fact, in conjunction with recent DNA sequencing results, leads us to suggest that, like the SV40 tumor antigens, the polypeptides encoded by these Ad2 mRNAs have an identical amino acid sequence at their N terminal ends, but have different C terminal sequences.


Sequence organization of a cloned tDNA met fragment from Xenopus laevis. 3.18 kb fragments of X. laevis DNA coding for tRNA 1 met have been inserted into a lambda vector via Hind III termini and cloned in E. coli. The organization of one cloned fragment has been analyzed by restriction endonuclease digestion and RNA-DNA hybridization. From the distribution of sites for three enzymes, this fragment appears to be typical of the majority of X. laevis tandem tDNA 1 met repeat units. Evidence is presented to suggest that it contains two genes coding for tRNA 1 met and at least one gene coding for a second as yet unidentified 4S RNA species. The two tRNA 1 met genes are located on the same DNA strand 0.96 and 1.38 kb from one end of the repeat unit. A detailed restriction map for 19 enzymes reveals that the spacers between these genes are not identical, and it provides no indication of short repetitive sequence elements within the spacers.


Evidence for functional hemizygosity at the Emtr locus in CHO cells through segregation analysis. The hypothesis of functional hemizygostiy at the emetine-resistant (Emtr, a non-X-linked recessive marker) locus in Chinese hamster ovary (CHO) cells has been examined by segregation analysis. The frequencies and the rates of segregation of the Emtr and Thgr (thioguanine-resistant, an X-linked recessive mutation) markers were determined from hybrids constructed between an Emtr-Thgr CHO cell line and various other Chinese hamster lines (V79, M3-1, CHO, GM7S, CHW and CHL). Thgr segregants were obtained at similar frequencies (10(-2)-10(-3)) from all the hybrids. The frequency of segregation of the Emtr marker, however, was similar to that of Thgr only in the CHO x CHO hybrids and was much lower (10(-4)-10(-6)) than the CHO x other Chinese hamster hybrids. Similar results were obtained when the segregation rates for the two markers from various hybrids were determined. These results are consistent with the hypothesis that in CHO cells, the gene responsible for Emtr is present in a single (functional) copy, whereas two copies of this gene are present in other Chinese hamster lines examined.


Radioimmunoassay for tubulin: a quantitative comparison of the tubulin content of different established tissue culture cells and tissues. A quantitative estimate of the cellular tubulin concentration can be obtained by the use of a radioimmunoassay based upon the competition between tubulin in cell extracts and a known amount of radioactively labeled homogeneous tubulin during binding to a limited amount of anti-tubulin antibodies. This assay shows that a variety of widely used tissue culture cells (mouse L cells, mouse 3T3 cells, chick embryo fibroblasts) have a tubulin content which corresponds to approximately 2.5--3.3% of their total protein. Transformation of mouse 3T3 cells by the DNA virus SV40, and of chick embryo cells by the RNA Rous sarcoma virus, does not change the intracellular tubulin concentration. Transformed cells of brain origin, such as some glia tumor cell lines and some neuroblastoma cell lines, have a much lower tubulin content than does normal brain tissue. The intracellular concentration of tubulin in mouse 3T3 cells is discussed in relation to the number of microtubules detected during interphase by immunofluorescence microscopy. These results are also discussed in view of a mechanism of microtubule elongation in vivo driven by self-assembly.


Cell surface action of thrombin is sufficient to initiate division of chick cells. Thrombin covalently linked to carboxylate-modified polystyrene beads initiated division of quiescent chick embryo (CE) cells either in medium containing low levels of serum or in serum-free medium. Release of thrombin was monitored by measuring acid-precipitable radioactivity released from 125I-thrombin beads into the medium during incubation with cells. Even if all of the acid-precipitable material released from the beads were active thrombin, it was not sufficient to account for any of the observed cell division, and was 10-30 fold less than the amount necessary to produce the increase in cell number caused by the thrombin beads. Two other kinds of experiments also showed that material released into the medium did not account for the observed initiation of cell division. First, medium taken from cultures incubated with thrombin beads did not initiate cell division when added to new quiescent cultures. Second, in coverslip experiments where populations of cells with an without thrombin feads shared the same medium, only bead-contacted cells divided. Several results suggested that the material which was released from the thrombin beads resulted from cell-associated proteolysis rather than from "leakage" of intact thrombin from the beads. For example, after incubating 125I-thrombin beads with or without CE cells, we were unable to detect any intact thrombin released into the medium. In addition, most of the material released from the beads was acid-soluble and was only released in the presence of CE cells. A few thrombin beads were endocytosed by CE cells, but they were surrounded by an intact plasma membrane. Thus they did not directly interact with the cytoplasm. The close association of many of the beads with the cell surface and the presence of a few beads in endocytic vesicles made it important to consider the possibility that thrombin might be released from the beads directly into the cells. This possibility was explored using ultrastructural (EM) autoradiography. With this technique (where one grain represented 700--900 thrombin molecules), we found that beads inside the cells had approximately the same number of grains as beads not in contact with cells. This suggested that little, if any, additional radioactive material had been released from the beads which were in contact with the cells. In addition, we were unable to detect any grains in the cytoplasm which could be attributed to released thrombin, even using an amount of 125I-thrombin beads which was 8 fold greater than the amount which produced maximal cell division. Taken together, these results provide direct evidence that thrombin action at the cell surface is sufficient to initiate division of CE cells.


Interferon-associated, dsRNA-dependent enzyme activities in a mutant 3T6 cell engaged in the semiconstitutive synthesis of interferon. Cytoplasmic extracts of untreated cultures of a virus-resistant mutant of mouse 3T6 cells, designated 3T6-VrB2, contain two double-stranded, RNA-activated enzyme activities associated with interferon action. These are the synthesis of a low molecular weight oligonucleotide inhibitor of cell-free protein synthesis from ATP, and the phosphorylation of a 67,000 dalton polypeptide by transfer of the gamma phosphate of ATP. Basal levels of both enzyme activities are detectable in extracts of untreated parental 3T6 cells, and are greatly enhanced upon interferon pretreatment. A procedure was developed, using a nonionic detergent to effect cell lysis, which allowed the analysis of the protein kinase activity from as few as 2 x 10(7) cells. Using this procedure, direct proportionalities were demonstrated between the concentration of interferon to which 3T6 cells were exposed, and both the level of protein kinase activity and the magnitude of the antiviral state were established in these cells. Furthermore, untreated cultures of 3T6-VrB2 exhibited both an antiviral state and an intracellular protein kinase activity equal to that of cultures of the parental 3T6 cells pretreated with a single concentration of mouse interferon.


Characteristics of individual repetitive sequence families in the sea urchin genome studied with cloned repeats. Cloned repetitive sequences from the S. purpuratus genome a few hundred to approximately 1000 nucleotides long were used to investigate the characteristics of individual repetitive sequence families. They wer terminally labeled by the kinase procedure and reacted with sheared S. purpuratus DNA. Repetition frequencies were measured for 26 individual families and were found to vary from a few to several thousand copies per genome. Estimates of sequence divergence were made for 18 cloned repeat families by measuring thermal stability of the heteroduplexes formed between the genomic DNA and the cloned fragments, compared with that of the renatured cloned fragments. The difference was less than 4 degrees C for three of the 18 families, and less than 10 degress C for 13 of the 18 families. These 13 repetitive sequence families lack any detectable highly divergent sequence relatives, and the results reported are shown not to change when the renaturation criterion is lowered below 55 degrees C in 0.18 M Na+. Five of the 18 cloned families displayed greater sequence divergence. The average sequence divergence of the total short repetitive sequence fraction of S. purpuratus DNA was found to match closely the average of the divergences of the cloned repeat sequences.


[Aortic valve stenosis with calcification]. A group of 52 patients was studied with a view to defining the characteristic features of aortic valve stenosis with calcification. There is no uniform aetiopathogenesis to explain the condition. 3 basic types could be distinguished--post-rheumatic, senile, and due to congenital malformation of the valve; all were represented about equally.


[The proportion of potassium and calcium in the vitreous body in relation to the time of death]. The method of atomic absorption spectrophotometry was employed to estimate the proportion of potassium and calcium in vitreous humour in 47 deceased persons. The amount of both was found to increase in linear proportion to the time elapsed from death. The correlation can be expressed in the form of equations. The simultaneous estimation of the proportion of potassium and calcium in vitreous humour enables a more precise ascertainment of the time of death than the estimation of merely one of the two elements. In persons who died of brain injuries, strangulation or who had suffered from conditions associated with metabolic breakdown, the time of death should not be determined according to the amount of calcium in vitreous humour; in such cases only the proportion of potassium should be exploited.


[Nature of the pigment in multilayer neurofibromas]. Two cases of storiform neurofibroma were examined electronmicroscopically. Their pigment was identified as melanin both by histochemical methods and by the presence of melanosomes. In one case moreover another pigment of lysosomal location was found.


[Cellular irregularities in nasal polyps]. 50 patients of different ages were found to have irregular multinuclear cells lodged in their inflammatory hyperplastic nasal mucosa. In only isolated cases were they numerous enough to give the tissue a sarcomatoid appearance. Most of those could clearly be described as proliferating endothelial cells. Mechanical irritation seemed to be the obvious explanation of their pathogenetic relationships.


[Unusual trabeculra material in the fine structure of intercellular matter in pulmonary hamartoma]. Numerous fibrous structures stainable with resorcin--fuchsin were found by light microscopy in five pulmonary hamartomas. Their electronoptic correlates were granular dense trabecular formations reminiscent of oxytalan fibres whereas trabeculae typical of elastica were rare. The trabecular material orientation according to collagenous fibrils as well as its occasional transverse striation suggest a special form of collagen to be involved.


[Adenomatoid hamartoma of the lung].

[Adenomatoid hamartoma of the lung]. The right lung of a stillborn premature infant was found to have a growth occupying most of the pleural cavity and displacing the mediastinum. It was predominantly solid with minor cysts and with structure reminiscent of a clusters of primitive alveoli and bronchioles, or else it contained major gaps with papillomatous proliferating cylindrical cells.


[Intrapulmonary thymoma].

[Intrapulmonary thymoma]. A 53-year old woman had for 14 years been under clinical observation for a right-sided intrapulmonary thymoma. As a terminal development, metastases appeared in the liver. The tumour was not classified until necropsy, and that with the assistance of electronoptic proof of the epithelial squamous appearance of large tumour cells.


[Echinococcal cysts in the liver].

[Echinococcal cysts in the liver]. A report on 2 cases of hydatid cysts in the liver accidentally detected in an autopsy and on one case found in a biopsy. A brief list is given of the macro- and microscopic findings in correlation with clinical tests.


[Dysontogenic cyst of the tongue lined with heterotopic gastrointestinal mucosa]. A dysontogenetic cyst in the edge of the tongue in 1-year old boy was found lined with gastric, duodenal, and intestinal mucosa with its own lamina muscularis mucosae, with a thin layer of submucous connective tissue encircled by bundles of smooths muscle imprecisely separated from the striated lingual muscle.


[Immunohistologic demonstration of hepatitis B antigen]. A description is presented of the results of immunohistological reaction to HBAg, using home-prepared sera, in bioptical material routinely processed in the department by standard methods. The results are essentially in agreement with literary data. One conspicuous feature was the substantially reduced quantity of positive elements in comparison with the results of staining with aldehydefuchsine and orceine.


[Immunoglobulins in sudden and violent death]. In the blood serum of 65 subjects (33 sudden deaths and 32 deaths due to injury) immunoglobulins IgG, IgM, IgA and IgD were examined using the method of simple radial diffusion. The average values in the whole series of infants, children and adults were as follows: IgG 104, IgM 147, IgA 91, IgD 37 international units per 1 ml. In sudden deaths the immunoglobulin levels were found to be higher.


Glucuronidation of benzo[a]pyrene in hamster embryo cells. The formation of water-soluble metabolites of tritium-labeled benzo[a]pyrene (BP) by cultured hamster embryo cells was studied. The ratio of the radioactivity in the aqueous phase to that in the organic phase increased with the incubation period. After incubation for 48 h with 3.75 nmol/ml of [3H] BP in the medium more than 90% of the 3H-radioactivity was found in the aqueous phase, whereas with 10-fold more BP about half the radioactivity remained in the organic phase. The main metabolites extracted from the medium at 37.5 nmol/ml BP with ethyl acetate by high pressure liquid chromatography (HPLC) were 9,10-diol and 7,8-diol; but after treatment of the medium with beta-glucuronidase the main oxygenated metabolites were phenols, the amount of 9-OH BP being more than that of 3-OH BP. beta-Glucuronidase also released 9,10-diol and 7,8-diol, but most of these diols were in the free form in the medium. The medium from cells treated with 3.75 nmol/ml BP has a quantitatively different profile, and most of the radioactivity obtained by extraction with organic solvent and digestion with beta-glucuronidase was eluted in the regions of phenols. These results show that in hamster embryo cells BP is mainly metabolised to conjugates of phenols with glucuronic acid.


Pyruvate metabolism after in vivo exposure to oral arsenic. This study investigated altered pyruvate metabolism after prolonged oral arsenic exposure. Male rats were given access to deionized drinking water containing 0, 40 or 85 ppm sodium arsenate (As5+) for 3 weeks. Respiration studies with mitochondria isolated from treated animals indicated decreased state 3 respiration (with ADP) and decreased respiratory control ratios (RCR) for pyruvate/malate-mediated respiration, but not for succinate-mediated respiration, as compared to control respiration values. In addition, pyruvate dehydrogenase activity was measured, in both liver and intestine, before and after Mg-activation in vitro. After 3 weeks, the effects of arsenic at the highest dose level were pronounced on the basal pyruvate dehydrogenase activity (before activation) as well as the total pyruvate dehydrogenase (after activation). The inhibition of pyruvate dehydrogenase activity both before and after Mg-activation suggests an arsenic effect on mitochondrial pyruvate metabolism which, in part, involves inhibition of pyruvate decarboxylase. Evidence is also presented which may indicate an arsenic effect on the kinase and/or phosphatase which regulate pyruvate dehydrogenase activity.


Binding of benzo[a]pyrene at the 1,3,6 positions to nucleic acids in vivo on mouse skin and in vitro with rat liver microsomes and nuclei. Loss of tritium from specific positions in [3H,14C] aromatic hydrocarbons can elucidate their binding site(s) to DNA and RNA and indicate the mechanism of activation. Studies of tritium loss from [6-3H,14C]benzo[a]pyrene (B[a]P), [1,3-3H,14C]B[a]P, [1,3,6-3H,14C]B[a]P, [6,7-3H,14C]B[a]P, and [7-3H,14C]B[a]P were conducted in vitro using liver nuclei and microsomes from 3-methylcholanthrene-induced Sprague-Dawley rats and in vivo on the skin of Charles River CD-1 mice. The relative loss of tritium from [3H, 14C]B[a]P was measured after binding to skin DNA and RNA, to nuclear DNA, and to native and denatured calf thymus and rat liver DNA's and poly(G) by microsomal activation. In skin, nuclei, and microsomes plus native DNA, virtually all B[a]P binding occurred at positions 1,3 and 6; while with microsomes plus denatured DNA or poly(G), B[a]P showed no binding at the 6 position and a small amount at the 1 and 3 positions. In vivo and with nuclei, binding at the 6 position predominated. Little loss of tritium from the 7 position was seen; this was expected because binding at this position is not thought to occur. This confirms the interpretation of loss of tritium as an indication of binding at a given position. These results demonstrate that the use of microsomes to activate B[a]P is not a valid model system for delineating the in vivo mechanism of B[a]P activation, and support previous evidence for one-electron oxidation as the mechanism of activation of hydrocarbons in binding to nucleic acids.


Carcinogenicity and metabolic profiles of 6-substituted benzo[a]pyrene derivatives on mouse skin. The ability was tested of appropriate substituents of benzo[a]pyrene (BP) at C-6 to decrease or suppress the carcinogenic activity for these BP derivatives relative to the parent compound. 8-week-old female Swiss mice in 9 groups of 30 were treated on the back with 0.2 mumol of compound in acetone 4 times weekly for 20 weeks. The following compounds were administered: BP, 6-methylbenzo[a]pyrene (BP-6-CH3), 6-hydroxymethylbenzo[a]pyrene (BP-6-CH2OH), benzo[a]pyrene-6-carboxaldehyde (BP-6-CHO), benzo[a]pyrene-6-carboxylic acid, 6-methoxybenzo[a]pyrene, 6-acetoxybenzo[a]pyrene, 6-bromobenzo[a]pyrene, and 6-iodobenzo[a]pyrene. Two additional groups received BP or BP-6-CH3 twice weekly for 20 weeks at a total dose 25% of that above. In addition, the metabolism of selected 6-substituted BP derivatives was studied, using mouse skin homogenates in vitro and mouse skin in vivo. Only four compounds were carcinogenic; the order of potency was BP greater than BP-6-CH3 greater than BP-6-CH2OH and BP-6-CHO. The difference in carcinogenicity between BP-6-CH2OH and BP-6-CHO could not be assessed by this experiment. In a further tumorigenesis experiment the carcinogenicity of BP-6-CH2OH was compared to that of BP-6 CHO, BP-6-CH3 and 6-hydroxymethylbenzo[a]pyrere sulfate ester (BP-6-CH2OSO3Na) on mouse skin. 9-week-old female Swiss mice in groups of 28 were treated at three dose levels with 0.8, 0.2 and 0.05 mumol of compounds in dioxane--dimethyl sulfoxide (75 : 25) twice weekly for 40 weeks. After 40 experimental weeks BP-6-CH2OSO3Na proved to be a more potent carcinogen than BP-6-CH2OH, which, in turn was more active than BP-6-CHO. The greater carcinogenicity of BP-6-CH3 relative to BP-6-CH2OH and BP-6-CHO is confirmed, suggesting that BP-6-CH2OH is not a proximate carcinogenic metabolite for BP-6-CH3. Since BP-6-CHO is a weaker carcinogen than BP-6-CH2OH and is efficiently reduced metabolically to BP-6-CH2OH, the latter compound may be a common proximal carcinogenic metabolite. The stronger potency of BP-6-CH2OSO3Na, compared to its alcohol, suggests that an ester of BP-6-CH2OH might be the ultimate alkylating compound reacting with cellular nucleophiles.


Carcinogenicity and metabolic profiles of 3-methylcholanthrene oxygenated derivatives at the 1 and 2 positions. Trapping of 3-methylcholanthrene (MC) radical cation by nucleophilic compounds occurs specifically at the 1-carbon atom. With the purpose of providing more evidence for the hypothesis that the critical mechanism of activation of MC is one-electron oxidation, the carcinogenicity of MC was compared to that of 1-hydroxy-3-methylcholanthrene (MC-1-OH), 3-methylcholanthrene-1-one (MC-1-one), 2-hydroxy-3-methylcholanthrene (MC-2-OH), 3-methylcholanthrene-2-one (MC-2-one) and 3-methylcholanthrylene (MCL) by repeated application on mouse skin. Seven-week-old female Swiss mice in 6 groups of 30 were treated on the back with 0.2 mumol of compound in acetone twice weekly for 20 weeks. In addition, the metabolism of MC and its derivatives was studied using mouse skin homogenates. The compounds tested were classified according to carcinogenicity in 4 groups: MC and MC-2-OH, the strongest carcinogens; MC-2-one and MCL, weaker than MC and MC-2-OH; MC-1-OH, the weakest carcinogen; and MC-1-one, non-carcinogenic. These results support the hypothesis that one-electron oxidation for MC, MC-2-OH and MC-1-one might be the critical mechanism of carcinogenic activation, with C-1 the binding site to cellular nucleophiles. The carcinogenic effect of MC-1-OH is speculated to be the formation of an ester bearing a good leaving group, which might be the ultimate alkylating compound in the in vivo reaction. The lack of carcinogenic activity for MC-1-one may be attributed to absence of nucleophilic trapping at C-1 via the radical cation pathway as well as the inability of mouse skin to reduce MC-1-one to the carcinogenic MC-1-OH.


Microsomal metabolism of N-nitrosodi-n-propylamine: formation of products resulting from alpha- and beta-oxidation. We have identified propionaldehyde, n-propranolo, isopropanol and N-nitroso-2-hydroxy-propylpropylamine following incubation of N-nitrosodi-n-propylamine with a microsomal fraction from rat liver. Based on the yields of the various products, we have shown that beta-oxidation occurs at about 15% of the level of alpha-oxidation, beta- as well as alpha-oxidation was shown to be carried out by the microsomal mixed function oxidase system. N-nitroso-2-hydroxy-propylpropylamine is further oxidized by the microsomal preparation to yield N-nitroso-2-oxopropylpropylamine.


Membrane lipid modifications: biosynthesis and identification of phosphatidyl-N-methyl-N-isopropylethanolamine in rat liver microsomes. In previous studies on the modification of polar head groups of membrane phospholipids with the unnatural base analog, N-isopropylethanolamine, we reported an unidentified phospholipid in addition to phosphatidyl-N-isopropylethanolamine in the various membrane fractions of rat liver. The structure of this phospholipid has now been identified as phosphatidyl-N-methyl-N-isopropylethanolamine by nuclear magnetic resonance spectroscopy, and by chromatographic and enzymic analysis. In addition, we found that when rats were injected intraperitoneally with the N-methyl-N-isopropylethanolamine, 19% of the liver microsomal phospholipid was phosphatidyl-N-methyl-N-isopropylethanolamine.


Cross-linking of membrane phospholipid and protein using putative monofunctional imidoesters. The reaction of methyl acetimidate or isethionyl acetimidate with mitoplasts at pH 8.5 yields two derivatives of phosphatidylethanolamine. These derivatives are shown to be the mono-amidine derivative and the bis-derivative of phosphatidylethanolamine. The bis-derivative represents one phosphatidylethanolamine cross-linked to another phosphatidylethanolamine. Similar derivatives are formed by the reaction of dipalmitoyl phosphatidylethanolamine with these imidoesters in organic solution with the exception that much more monoderivative is produced. Methyl picolinimidate reacts with phosphatidylethanolamine of mitoplasts to form primarily the mono-derivative. The bis-derivative was not detected. The reaction of bovine rod outer segment discs with methyl acetimidate causes cross-linking of 30% of the membrane rhodospin as dimers. Putative monofunctional imidoesters cause considerable cross-linking of both phospholipids and proteins in cell membranes. Cross-linking can be minimized at pH 9.0.


[Bronchogenic cyst of the lung and pulmonary tuberculosis. Presentation of a case]. The paper considers a clinical case of multiple bronchogenic cysts associated with a pneumonitic process of TB character, operated successfully by means of an upper right and middle bilobectomy. No similar case was found in literature. The malformations were of congenital origin, characterised histologically by a cavity, the walls of which contain elements proper to the bronchus: cartilage and ciliate columnar epithelium. The squamous metaplasia is rare, and the neoplastic transformation very rare. From the clinical point of view a distinction is made between asymptomatic cysts, in which the clinical picture is dominated by compressive phenomena affecting the surrounding structures, or sometimes by infection due to supervening bacterial, mycotic or--as in our case--tubercular contamination. The therapeutic indication is exclusively surgical, with methods of exeresis varying according to the extent and location of the morbid process.


[Physiopathology of the gastrectomized patient]. The Authors re-examined 50 cases of various ages and both sexes subjected to total gastrectomy for neoplasia, without any other chemotherapic or radiant collateral treatmente, and analyse the sequelae following total removal of the stomach. On the basis of the data collected, they conclude on the importance of an adequate reconstructive technique, in particular stressing the utility of preservation of the duodenal transit and the need for suitable dietetic-pharmacological conduct in the years following the operation.


[Gardner's syndrome in familial popyposis: clinical contribution and etiopathogenetic, therapeutic and prognostic considerations]. Starting from one case of Gardner's syndrome of their own observation, and in light of Watne's results, the authors argue that in the syndrome just named--and in familial polyposis in a broader sense--genetic factors do not play a predominant role; rather, polyps seem to result from the prolonged action of noxious agents. At any rate, said action can be neutralized, and possibly prevented either with a surgical modification of the distal gut or by treatment with a chemical substance identified as ascorbic acid. In view of these considerations, the authors recommend preventive medical therapy.


[Acute pancreatitis in surgery of the common bile duct]. After commeting on the importance of acute postoperative pancreatiti (APP) as a complication of surgery involving the main biliary passages, the authors give a detailed account of the 5 cases of this complication seen in their own clinical material over the last 5 years. In their summation, they identify the presence of pathological conditions of the papilla and the manipulation of the terminal segment of the choledochus as factors conducive to APP in the short-term postoperative period. In their discussion of therapeutic guidelines in the event of APP they call attention to the need for a careful assessment of pancreatic function before surgery.


[Primary carcinoid of the testis. Clinical case]. A very rare case of carcinoid of the testicle combined with teratoma is illustrated. In fact the literature of the last twenty years reports only eleven cases of carcinoid of the testicle, of which only four combined with teratoma. It is stressed that it is possible that these neoplasias arise on already existing teratomas which may be very small and hence not always evidenced.


The effects of intra-arterial and intraportal injections of vasopressin on the simultaneously perfused hepatic arterial and portal venous vascular beds of the dog. The hepatic arterial and hepatic portal venous vascular beds of the chloralose-urethane anesthetized dog were perfused simultaneously in situ. Vasopressin (10 mU = 1 unit) was injected in graded increasing doses into the hepatic artery and into the portal vein. Both intra-arterial and intraportal vasopressin elicited both hepatic arterial vasoconstriction and hepatic venous dilation; the delay in onset of both hepatic vascular effects was significantly shorter than that for any succeeding systemic effects (a rise in systemic arterial pressure and fall in heart rate), showing that they were not attributable to recirculation or to arterial baroreceptor reflexes. Injections of vasopressin into the inferior vena cava at the level of the hepatic veins consistently produced smaller hepatic vascular effects than either intra-arterial or intraportal injections of the same doses. The results are discussed in the context of the therapeutic role of vasopressin in controlling gastrointestinal bleeding and portal hypertension.


Subpressor angiotensin infusion, renal sodium handling, and salt-induced hypertension in the dog. We studied the combined effect of subpressor amounts of angiotensin and long-term sodium chloride infusion on arterial pressure in 16 dogs for periods of 2--8 weeks. In dogs receiving 3.5 liters of isotonic NaCl daily, but no angiotensin, the arterial pressure increased an average of only 3 mm Hg. When angiotensin was infused continuously at a rate of 5 ng/kg per min (a rate too small to cause an observable immediate increase in pressure, subsequent infusion of 3.5 liters of saline daily then increased the pressure by 39 mm Hg. The urinary output of sodium increased to the same extent in both instances, that is, there was no extra sodium loss because of the elevated pressure. This suggests that the angiotensin significantly blocked the normal "pressure natriuresis" usually seen with such large increases in pressure. However, the plasma aldosterone levels during angiotensin infusion were not found to be different from those in the absence of angiotensin. Therefore, we have suggested that the tendency of the kidneys to retain sodium under the influence of angiotensin was probably caused mainly by a direct effect of angiotensin on the kidney itself. Such a direct renal sodium-retaining effect also could be a contributing factor in the marked hypertension that results from salt administration in the presence of small amounts of angiotensin.


Preferential distribution of inhibitory cardiac receptors with vagal afferents to the inferoposterior wall of the left ventricle activated during coronary occlusion in the dog. The purpose of this study was to determine the relative magnitudes of the reflex effects mediated by cardiac receptors during anterior as opposed to inferoposterior ischemia of the left ventricle of the dog. Cessation of perfusion (coronary "occlusion") of the circumflex coronary artery (Cx) in 29 chloralose-anesthetized dogs with common carotids ligated (group I) resulted in significant bradycardia and hypotension, but in no significant change in perfusion pressure in the gracilis muscle perfused at constant flow. Occlusion of the left anterior descending coronary artery (LAD) produced less hypotension, no change in heart rate, and vasoconstriction in the gracilis. After vagotomy and aortic nerve section, no significant change in heart rate or gracilis perfusion pressure was observed during lad or Cx occlusion, and the blood pressure responses to LAD and Cx occlusion were not different. In nine dogs with sinoaortic denervation (group II), brief Cx occlusion resulted in bradycardia, hypotension, and vasodilation in the gracilis muscle. LAD occlusion in group II dogs caused less hypotension and no change in heart rate or gracilis perfusion pressure. After vagotomy, the bradycardia and vasodilation resulting from Cx occlusion were abolished and the blood pressure responses to LAD and Cx occlusion were not different. The weights of left ventricle perfused by each occluded vessel were not different. These data show that left ventricular receptors with vagal afferents which are activated during coronary occlusion and which mediate cardioinhibitory and vasodepressor responses are located mainly in the inferoposterior left ventricle of the dog heart.


Renal chemoreceptors in the rat.

Renal chemoreceptors in the rat. There are afferent nerve fibers responsive to alterations of the kidney's chemical environment in the renal nerves of the rat. In anesthetized, artificially ventilated, male Sprague-Dawley rats, single unit recordings were prepared by dissection of the centrally cut nerves of the right kidney. The stimuli used included occlusion of the renal artery, systemic asphyxia, changes in renal arterial and venous pressures, changes in ureteral pressure, and cyanide infusion. We found a population of sensory nerve fibers whose endings are activated only during markedly impaired renal blood flow (produced by clamping the renal artery, severe hypotension below 40 mm Hg, and prolonged occlusion of the renal vein), and during systemic asphyxia. The same units are not responsive to increases and decreases in systemic arterial pressure (range: 40--190 mm Hg), to ureteral pressure (range: 0--50 mm Hg), or to changes in renal venous pressure. None of the 40 single units studied was spontaneously active; their pattern of activation during renal ischemia always was characterized by trains of impulses. These sensory units have functional properties distinctly different from those of known renal mechanoreceptors. They appear to be a homogeneous group of sensory elements, and we have termed them renal ("R") chemoreceptors. Evidence also is presented which is consistent with the concept that a chemical substance released by or accumulated within the kidney might be the agent activating these chemoreceptors during renal ischemia.


Increased renal vascular reactivity to angiotensin II but not to nerve stimulation or exogenous norepinephrine in renal hypertensive rats. We isolated and perfused both the "clipped" and "contralateral" kidneys from Goldblatt renal hypertensive and sham-operated control rats, 1--104 days postoperatively. Responses to renal nerve stimulation were depressed in clipped kidneys from hypertensive rats (1 day postoperative), and these kidneys were supersensitive to exogenous norepinephrine (1--31 day) when compared with the contralateral organ of the same animal. Similar alterations were found between clipped and contralateral kidneys from sham-operated control rats. There was no difference in responses to renal nerve stimulation of norepinephrine between clipped kidneys from hypertensive and control rats, but clipped kidneys from hypertensive rats were supersensitive to angiotensin II (17 and 31 days). Comparison of contralateral kidneys from hypertensive and control rats revealed no change in norepinephrine sensitivity or in responses to renal nerve stimulation, but there was a reduction in the slope of the dose-response curve to norepinephrine and of the maximal effect of the catecholamine (104 days) and a pronounced supersensitivity to angiotensin II (17--104 days) in the hypertensive rats. These results indicate that (1) renal nerve function and norepinephrine sensitivity of the isolated renal vasculature are unchanged in renal hypertension, but clipping partially denervates the kidney causing depressed nerve function and unilateral norepinephrine supersensitivity, unrelated to hypertension; (2) the prolonged high pressure load on the contralateral kidney may impair the function of the vascular smooth muscle; and (3) bilateral supersensitivity to angiotensin II is associated with hypertension but is not solely a consequence of the high pressure.


The effect of changing interpulse intervals on the negative chronotropic response to repetitive bursts of vagal stimuli in the dog. The chronotropic responses to repetitive bursts of vagal stimulation were determined in open-chest, anesthetized dogs. Either 5 or 10 electrical pulses were included in each stimulus burst, and the interpulse interval (deltat) was varied over the range of 5 to 150 msec. As the frequency of the stimulus bursts was progressively changed, the sinoatrial (SA) nodal pacemaker cells became synchronized with the repetitive bursts of stimuli over a certain range of burst frequencies. The magnitude of this frequency range varied with deltat. For 5 and 10 pulses/burst, the values of deltat that produced the greatest magnitude of this frequency range were 30.2 and 24.3 msec, respectively. Also, over the range of values of deltat from 5 to 50 msec, the magnitude of the negative chronotropic effect of the vagal stimulus burst varied directly with deltat. It is likely that, as the interpulse interval is increased within the range of values, either more acetylcholine is released from the vagal nerve endings per pulse or there is less saturation of the receptors on the pacemaker cell membranes during each burst.


Contribution of systemic venous hypertension to the development of pulmonary edema in dogs. Systemic venous hypertension (SVH) is a frequent finding in pulmonary edema. To study the possible contributory or even causal role of SVH in pulmonary edema, a dog model was developed in which balloon catheters were placed in the left and right atria. Inflation of the left atrial balloon produced a tendency to pulmonary edema by causing pulmonary venous hypertension (PVH) (pulmonary artery wedge pressure of 20 mmHg). Inflation of the right atrial balloon produced SVH (central venous pressure of 15 mm Hg). After 2 hours, dogs with SVH with or without PVH demonstrated a greater amount of lung fluid accumulation (P less than 0.01) compared to controls or PVH alone. There was no significant difference in lung water in SVH dogs with or without PVH. Pulmonary blood flow was not significantly different between the experimental groups, each of which was less than control. Impairment of pulmonary lymphatic flow is one possible mechanism producing the worsening edema; however, bronchial venous hypertension or neurogenic reflexes cannot be excluded. We conclude that the contribution of systemic venous hypertension to the development of pulmonary edema may have therapeutic implications.


Experimental evaluation of streamline patterns and separated flows in a series of branching vessels with implications for atherosclerosis and thrombosis. Flow conditions in four models representing the aortic bifurcation, iliac bifuraction, and a renal artery branch were investigated at volumetric flow rates corresponding to Reynolds numbers from 1000 to 4000 over the complete range of flow division between daughter vessels. Qualitative flow streamline patterns and quantitative definition of those flow conditions leading to disturbed flow (flow separation ) were determined primarily at steady flow with a limited set of pulsatie experiments. Under conditions of no flow separation, common characteristic streamline patterns not parallel to the center lines of parent or daughter tubes were found for all models. These effects were accentuated with increasing Reynolds number. Flow separation was inducible through alteration of flow division between daughter vessels or by an increase in flow rate. Each of the four models had distinct combinations of flow division ratio and flow rate which gave: (1) no flow separation, (2) flow separation at the outside of the right daughter tube, and (3) flow separation at the outside of the left daughter tube. Models representing the renal artery also had regions of simultaneous left- and righthand separation on the outside of their daughter tubes. The separated flows observed here displayed streamlines forming an open vortex with flows entering and leaving. These regions, which occur only at distinct combinations of flow rate and flow division, may be key centers where platelet aggregates may form, release constituents, and cause vessel injury.


Sites of action and active forms of lidocaine and some derivatives on cardiac Purkinje fibers. We have compared the effects of lidocaine (L) to those of several derivatives in an attempt to determine the sites of action and active forms of the molecules. We studied cardiac Purkinje fibers with intracellular microelectrodes, and drugs were administered by superfusion or by iontophoretic intracellular injection. We compared to L the effects of QX-314 and QX-572, two quaternary derivatives, and also compound 6603, a tertiary analogue with a pKa of 9.81. When administered by superfusion in concentrations of 10(-5) and 10(-4) M, all four agents exerted qualitatively similar effects on the transmembrane action potential. The rate of onset of action of the quaternary derivatives was considerably slower than that of L and their actions were not reversed by superfusion for 1 hour with drug-free solution. Comparison of effects of L and QX-314 after intracellular injection showed that attenuation of Vmax, and thus of the fast inward current, results from interaction of the charged form acting form acting from the inner surface of the sarcolemma. After intracellular injection, QX-314 diffused readily along the long axis of fiber bundles, and it was thus possible to expose the inner surface of a large number of contiguous cells to drug. Comparison of results of extracellular and intracellular application suggests that effects on the voltate during phase 2 result from the charged form acting from the inner surface, but effects on total action potential duration result only from drug acting from the outer surface of the sarcolemma. Unlike L, the three derivatives did not decrease the slope of normal phase 4 depolarization. The results may help relate antiarrhythmic action to specific effects on transmembrane potentials.


Valve replacement in patients with active infective endocarditis. Eleven of 138 patients with infective endocarditis (IE) who underwent cardiac valve replacement for IE during a 12 1/2-year period had active IE. Eight of the 11 (all with aortic IE) had positive blood cultures within 48 hours preoperatively; six of the eight had positive Gram stains and cultures of the excised cardiac tissue. All 11 patients had Class IV cardiac functional disability (New York Heart Association classification) at the time of surgery. Staphylococci (three patients with Staphylococcus aureus and one with S. epidermidis) were the most frequent isolates. Three patients died; two of these three deaths occurred in patients who had a sudden onset preoperatively of severe aortic regurgitation and heart failure. In one patient (S. epidermidis infection) prosthetic valve endocarditis developed. Cardiac valve replacement may be performed successfully in patients with active IE even when blood cultures are positive in the immediate perioperative period. The hemodynamic status of patients with IE should be the determining factor in the timing of cardiac valve replacement, rather than the activity of the infection or the length of preoperative antimicrobial therapy. A radical surgical procedure may be necessary in patients with myocardial or aortic abscesses in whom conventional aortic valve replacement is not possible.


Treatment of infective endocarditis: a 10-year comparative analysis. The results of surgical and non-surgical treatment of active infective endocarditis in 182 patients over a 10-year period were analyzed. Heart failure, annular and myocardial abscesses, heart block, and coronary embolism, seen most frequently with staphylococcal and fungal endocarditis, were the primary causes of death in both native valve endocarditis (NVE) and prosthetic valve endocarditis (PVE). In NVE, surgery significantly improved the survival in patients with moderate or severe heart failure (P less than 0.05) and in all patients with staphylococcal endocarditis (P less than 0.03). In PVE, surgery significantly influenced survival in patients with moderate or severe heart failure (P less than 0.05) and in the entire group with late PVE (P less than 0.01). Early surgery is recommended for patients with native valve endocarditis and moderate or severe heart failure; those patients with staphylococcal NVE, regardless of hemodynamic state, should undergo early valve replacement. Early surgery is recommended for PVE patients with moderate or severe heart failure. We also recommend early valve replacement for early and late staphylococcal PVE.


Clinical use of blade atrial septostomy. A cardiac catheter enclosing an extensible blade was used to enlarge the interatrial opening in seven patients. Two patients with transposition of the great arteries who had balloon atrial septostomy as newborns subsequently presented with clinical evidence of a restrictive interatrial opening at 1 and 4 months of age. Cardiac catheterization confirmed restenosis of the interatrial opening and inadequate intracardiac mixing. After blade atrial septostomy the systemic arterial oxygen saturation increased by 20% and 30%, respectively. Five patients with mitral atresia complex, ages 2 months-9 1/2 years, had a restrictive interatrial communication and severe pulmonary venous hypertension (mean left atrial pressures ranged from 20-38 mm Hg). Following blade atrial septostomy, the pressure gradient between the atria was almost completely abolished and prompt clinical improvement was observed in each patient. All patients tolerated the procedure without complications. Blade atrial septostomy was a safe, effective procedure for enlarging the interatrial communication in this limited series of patients with an interatrial septum too thick to permit adequate rupture by conventional balloon atrial septostomy.


Association between coronary heart disease and the C3F-gene in essential hypertension. The occurrence of the C3F allele was investigated in the following three groups: 69 consecutive referred patients with untreated essential hypertension, including borderline hypertension; 70 patients with established and treated essential hypertension, already attending the same outpatient clinic, and 62 age- and sex-matched normotensive healthy subjects without clinical signs of atherosclerosis or familial predisposition to hypertension. In the three groups the C3F allele was found in 38.2%, 29% and 20%, respectively. Among the treated hypertensive patients with C3F gene, 40% had coronary heart disease (CHD) compared to 6.1% among the C3F negative (P less than 0.005), and the relative risk of CHD among the treated hypertensive patients with this allele was found to be 10.2 (P less than 0.002). The C3F gene was present in 72.7% of the treated patients with CHD. In the untreated patients the occurrence of CHD was low, and no differences between C3F positive and negative patients could be demonstrated. No association of the C3F allele with familial predisposition to hypertension was found. This study provides further evidence of a positive association of the C3F allele with atherosclerosis, and it is concluded that this allele in a hypertensive patient might accelerate the atherosclerotic process, with subsequent premature development of vascular complications.


Tracking of blood lipids and blood pressures in school age children: the Muscatine study. In four cross sectional school screens, the Muscatine Study has sampled 8,909 school children; 820 have been studied repeatedly over a six-year period. Tracking of measurements described by the relationship between repeated observations and the relationship between peer rank orderings over the six-year period has been studied. For height and weight, correlations between observations six years apart were approximately 0.74 and about 60% of children initially in the upper quintile were there again six years later. Six-year correlations were 0.65 for skinfold and 0.61 for cholesterol. Four-year correlation for fasting triglyceride was 0.40. Six-year correlations were 0.30 for casual systolic blood pressure and 0.18 for diastolic blood pressure. Peer rank orderings for both blood pressures were highly variable. Height and weight track well, and thus routine measurement of these variables are useful in identifying children with growth perturbing disorders. Cholesterol and, to a lesser degree, triglycerides also track, and a significant proportion of children with initially high values demonstrated consistently high values throughout the study period. Blood pressures do not track as well; consistently high blood pressures are unusual, thus indicating the need for repeated blood pressure measurements to identify children with persistent elevated levels. The future significance of transient blood pressure elevations has yet to be established.


Type A behavior pattern and coronary atherosclerosis. Previous research has demonstrated an increased rate of clinical coronary heart disease (CHD) events among people who exhibit a "coronary prone" (Type A) behavior pattern. This study was undertaken to determine whether the association between behavior pattern Type A and CHD might be extended beyond clinical CHD events to include also the coronary atherosclerotic process. In addition to usual clinical evaluation, 156 consecutive patients referred for diagnostic coronary angiography were independently assessed on the basis of a structured interview and assigned a rating of Type A, Type B, or Type X (indeterminate). Traditional physiologic factors--age, sex, cholesterol and cigarette smoking--were found to correlate with atherosclerotic disease. Type A patients were found in increasing proportions among groups of patients with coronary occlusions of moderate to severe degree compared with patients with only mild occlusions. This increasing proportion of Type A patients with increasing disease severity remained significant, even when age, sex, blood pressure, serum cholesterol level and cigarette smoking history were all simultaneously covaried. These findings suggest that, independently of traditional risk factors, behavior pattern Type A may contribute to the risk of clinical CHD events via effects on the atherosclerotic process.


Beneficial actions of N-dimethyl propranolol on myocardial oxygen balance and transmural perfusion gradients distal to a severe coronary artery stenosis in the canine heart. The purpose of the present study was to compare the effects of N-dimethyl propranolol (DMP), the quaternary derivative of propranolol, and propranolol on the transmural distribution (endo/epi) of coronary blood flow in normal and ischemic regions of the myocardium. The distribution of blood flow between subendocardium and subepicardium of a nonischemic region and one distal to a severe left circumflex coronary artery stenosis was determined by use of tracer microspheres (15 microgram) in intact dog hearts. DMP (1,5 and 10 mg/kg I.V.) produced a small dose-related increase in endo/epi of the nonischemic region (1.15 +/- 0.04--1.24 +/- 0.05), whereas a larger increase was observed in the ischemic region (0.61 +/- 0.08--1.09 +/- 0.10). DMP also produced a significant increase in ischemic subendocardial blood flow (0.59 +/- 0.12--0.76 +/- 0.11 ml/min/g). Similarly, propranolol (0.5 and 1.0 mg/kg I.V.) produced a small increase in endo/epi of the nonischemic region (1.18 +/- 0.08--1.30 +/- 0.07) and a larger increase in the ischemic region (0.72 +/- 0.17--1.18 +/- 0.09). However, propranolol did not increase ischemic subendocardial blood flow. It is concluded that DMP may be an alternative to propranolol in certain types of acute myocardial ischemia when beta-adrenergic blockade is undesirable.


Electrophysiologic effects of partial coronary flow reduction in the exposed canine heart. Effects of ischemia and ischemic-induced regional hypothermia on refractoriness and conduction delay. In the open-chested dog, coronary flow reduction results in a decrese of regional myocardial temperature (T). We assessed the contribution of T decrease to changes in refractoriness and conduction delay attributed to ischemia. The independent effect of regional hypothermia on effective refractory period (ERP) was a linear function of the temperature (ERP = -b T +a) with a -r = 0.97 0.02 in 11 dogs. The effect on conduction time of a ventricular premature beat was a linear function of the dog T at both endocardium (-r = 0.95 +/- 0.02) and epicardium (-r = 0.96 +/- 0.01). A 75% reduction in coronary flow resulted in a mean T decrease of 1.0 +/- 0.3 degrees C. The T decrease was sufficient to mask the effects of ischemia on shortening of the ERP. Furthermore, the conduction delay of ventricular premature beats during 75% coronary flow reduction could be accounted for by the decrease in T alone in five of seven dogs. We conclude that changes in refractoriness and conduction during acute coronary flow reduction in the open-chested dog are due to the composite effects of ischemia and the decrease in regional temperature. The open-chested model may have important limitations in understanding the electrophysiologic effects of acute coronary insufficiency. However, it may have important applications in defining the electrophysiologic environment at the time of coronary artery surgery.


The clinical significance of bundle branch block complicating acute myocardial infarction. 1. Clinical characteristics, hospital mortality, and one-year follow-up. To provide an understanding of the clinical characteristics of patients with acute myocardial infarction (MI) and bundle branch block, experience from five centers was accumulated. Patients in whom bundle branch block first appeared after the onset of cardiogenic shock were excluded. In 432 patients, the most common types of block were left (38%) and right with left anterior fascicular block (34%). In 42% of the patients, bundle branch block was new. Progression to high degree (second or third degree) atrioventricular (AV) block via a Type II pattern occurred in 22% of the patients. Hospital and first year follow-up mortality rates were 28% and 28%, respectively. Only 46% of the patients developed pulmonary edema or shock (Killip Class III or IV), and hospital mortality was related to the amount of heart failure (8%, 7%, 27%, 83% for Killip Classes I-IV, respectively). Patients with progression to second degree or third degree AV block via a Type II pattern had increased hospital mortality compared with patients without this complication (47% vs 23%, P less than 0.001). In the absence of pulmonary edema or shock, patients with Type II second degree or third degree AV block still had a higher mortality rate than patients without advanced AV block (31% vs 2%, P less than 0.005), with nearly all the deaths due to abrupt development of AV block. Thus, in many patients MI with bundle branch block is associated with severe heart failure. However, this was not true for a majority of the patients, in whom therapy aimed at preventing morbidity and mortality due to the bradyarrhythmia of advanced AV block might be beneficial.


The clinical significance of bundle branch block complicating acute myocardial infarction. 2. Indications for temporary and permanent pacemaker insertion. The indication for prophylactic temporary and permanent pacing during acute myocardial infarction (MI) complicated by bundle branch block is high risk of progression via a Type II pattern to second or third degree (high degree) AV block during hospitalization or follow-up. In this study, determinants of high degree AV block during hospitalization and sudden death or recurrent high degree block during the first year of follow-up were examined in 432 patients with MI and bundle branch block. Timing of onset of bundle branch block, the involved fascicles, and the PR interval were examined as determinants of risk of progression to high degree AV block during MI. At highest risk were 186 patients with blocks involving the right bundle and at least one fascicle of the left bundle which were not documented on prior electrocardiograms. Risk was similar with (38%) or without (31%) accompanying first degree AV block. Patients with transient high degree AV block during MI had a 28% incidence of sudden death or recurrent high degree block during the first year of follow-up. Patients not continuously paced had a higher incidence of sudden death or recurrent high degree block than patients continuously paced (65% vs 10%, P less than 0.001). Sudden death during follow-up also occurred in 13% of patients without high degree block during MI. A subgroup with 1) documented prior MI, 2) anterior or indeterminant acute MI, and 3) no symptoms of cardiac failure had a 35% risk of sudden death. The role of permanent pacing in this group is unknown. Thus, patients at high risk of high degree AV block should receive prophylactic temporary pacing. Patients who survive high degree block with MI should receive temporary and then permanent pacing. Patients without high degree AV block during MI who nervertheless have a high risk of sudden death may benefit from permanent pacing.


A new method for measurement of sinoatrial conduction time. This study describes a new method (NM) for estimation of sinoatrial conduction time (SACT), which utilizes constant atrial pacing (AP) instead of the premature atrial beats (PABs) used in the method reported in 1973 by Strauss et al. The SACTs were obtained by both methods in 20 patients. The SACT by the Strauss method (SM) was calculated as A2A3 minus A1A1. The NM consists of high right AP for a train of eight consecutive beats at rates less than or equal to 10 beats/min faster than the sinus rhythm. The interval between the last paced atrial electrogram (Ap) and the first escape atrial electrogram (A) of sinus origin (Ap-A) was measured along with several post pacing sinus cycles. The SACT by the NM was calculated as follows: SACT = Ap-A minus A1A1. The effect of AP at higher rates was also analyzed. In two patients, the SACT with the SM could not be defined, as all the A2A3 intervals were fully compensatory; with the NM the SACT was 217 and 320 msec. In the remaining 18 patients the SACT was obtainable by both methods. With SM, the SACT ranged 105--452 msec (mean 219 +/- 102 SD) and with the NM it was 85--492 msec (mean 201 +/- 112 SD), and the difference was statistically significant (P = 0.0162). The coefficient of correlation between the two methods was r = 0.97. During AP at faster rates, a rate related increment in Ap-A intervals and also post pacing sinus cycles was noted. This study describes a new and simple method for measurement of SACT in man.


Left ventricular volume during maximal supine exercise: a study using metallic epicardial markers. Changes in left ventricular (LV) volumes and ejection fraction between rest and maximal supine exercise were evaluated in 11 patients who had had four epicardial markers placed during coronary artery surgery. After calibrating marker distances with respect to volume (r = 0.92--0.99) over one cardiac cycle for each patient, regression equations were used to compute LV volume from marker measurements for beats before and during exercise. The response of the left ventricle to exercise and the extent of revascularization could not be predicted from resting LV volume or ejection fraction. Ten patients had normal resting end-diastolic volumes and eight had normal resting ejection fractions. With exercise, three had a rise in end-diastolic volume and four had a fall in ejection fraction. Graft patency was greater in the group with an unchanged or increased ejection fraction (86 vs 50%, P less than 0.05). Epicardial clip motion can be used to determine LV volumes and ejection fraction during supine maximal exercise in man. The revascularized ventricle with normal or nearly normal performance in studies done at rest responds by decreasing end-diastolic and end-systolic volume and by increasing the ejection fraction. Increases in volumes or decreases in ejection fraction reflect old myocardial damage from infarction, fibrosis or ischemia from incomplete revascularization.


Correlation of angiographic and autopsy left ventricular dimensions in children. To evaluate controlled pressure coronary artery perfusion fixation of hearts as a means of obtaining meaningful cardiac measurements at autopsy, left ventricular (LV) autopsy measurements were correlated with in vivo end-diastolic (D) and end-systolic (S) angiographic (angio) values from biplane cineangiograms in the same patients. Mitral (MV) and aortic valve (AV) circumferences, LV equatorial circumference (Circ), spatial inflow and outflow lengths, wall thickness (W) and LV volume (Vol) were measured in 34 children with congenital heart disease. All dimensions showed significant correlation of angiographic and autopsy data from which linear regression equations were derived. The r values obtained were: MV 0.74; AV 0.85; D Circ 0.83; S Circ 0.82; D inflow 0.92; S inflow 0.90; D and S outflow 0.96; W 0.78; D Vol 0.92; S Vol 0.86. Mean angio and autopsy values were not significantly different for AV, S inflow, and S Vol. Autopsy values were lower than all D angio values and MV (P less than 0.001 for all), and higher than angio W and S outflow (P less than 0.001 for both). The data show that changes of LV morphology with this fixation method are predictable, making estimation of in vivo values from autopsy measurements possible using the derived linear regression equations.


The genesis of the diastolic murmur of complete heart block. Phono-echocardiographic observations. A 72-year-old man with calcific aortic stenosis and complete heart block was noted to have an intermittent diastolic murmur. Using combined echocardiography and phonocardiography, it was demonstrated that the murmur occurred while the mitral valve was closing. This suggested that antegrade flow across a closing mitral orifice is of etiologic importance in the production of the murmur. Atrial sounds were also recorded and probably have a similar etiology. It is postulated that reduced ventricular compliance resulting in a more rapid rate of mitral valve closure was of etiologic importance in the genesis of the murmur.


Isolated drainage of the superior vena cava into the left atrium in a 52-year-old man: a rare congenital malformation in the adult presenting with cyanosis, polycythemia, and an unsuccessful lung scan. This report describes a 52-year-old black male with the isolated finding of an anomalous superior vena cava draining into the left atrium. The patient presented with dizziness, mild cyanosis, polycythemia and normal cardiac and pulmonary findings. The first major diagnostic clue in this confusing clinical presentation was an unsuccessful lung perfusion scan in which intravenous tracer consistently bypassed the lungs. This appears to be the first adult presenting with this rare anomaly. This condition should be suspected if cyanosis, clubbing, and a "normal" cardiac examination coexist and if the more common pulmonary and hematological causes of this triad have been excluded.


Determination of drugs of abuse in body fluids by radioimmunoassay. Only recently has radioimmunoassay been used for the detection of drugs of abuse in body fluids. The radioimmunoassay method is rapid, sensitive, specific, and can be performed with a minimum of sample, while conventional assay methods are time-consuming, relatively insensitive, and require a larger sample volume. Performance of individual radioimmunoassays is identical, and requires about one hour to complete. The radioimmunoassays which have been developed for measurement of drugs of abuse in humans are reviewed in this paper. These new techniques are of importance in individual and/or emergency testing, in screening, and in further research into the effect of these drugs on the functions of the human body.