Cysteine conjugate beta-lyase in rat liver. A novel enzyme catalyzing formation of thiol-containing metabolites of drugs. A novel enzyme catalyzing cleavage of the thioether linkage in cysteine conjugates of aromatic compounds, such as 2,4-dinitrobenzene and p-bromobenzene, has been purified about 500-fold from rat liver cytosol. Incubation of S-(2,4-dinitrophenyl)cysteine with the enzyme preparation yielded 2,4-dinitrobenzenethiol, pyruvic acid, and NH3 at equimolar ratios, indicating that the thioether cleavage probably proceeds via an alpha,beta elimination reaction. The thiol product was methylated and the methylated derivative 1-methylthio-2,4-dinitrobenzene, was identified by mass spectrometry and proton NMR spectroscopy. The Km value of S-(2,4-dinitrophenyl)cysteine was 0.5 mM at pH 7.4 in phosphate buffer. The enzyme activity was inhibited by hydroxylamine. No cofactor requirement was observed. A combination of the partially purified enzyme and hepatic microsomes that contain thiol methyltransferase (EC 2.1.1.9) converted cysteine conjugates of 2,4-dinitrobenzene and p-bromobenzene to the corresponding methylthio-containing metabolites; S-adenosylmethionine was required. An important role of this novel beta-lyase enzyme in the formation of methylthio-containing metabolites of various drugs is indicated.


Differential effects of estrogen and progesterone on ovalbumin mRNA utilization. Progesterone treatment of estrogen-primed chicks leads to a shift in the oviduct polysome profile and an increase in the proportion of cytoplasmic RNA which is ovalbumin mRNA. To determine whether the progesterone effect is primarily transcriptional or translational and whether it is separable from estrogen action, rapid estrogen withdrawal by the anti-estrogen tamoxifen was compared in the presence and absence of progesterone. After estrogen stimulation, 24 h of tamoxifen treatment causes ovalbumin synthesis and ovalbumin mRNA levels to fall 10-fold. The proportion of ovalbumin mRNA in the monosome and supernatant fractions of the polysome profile increases; however, these sequences can associate with polysomes if elongation is inhibited by cycloheximide. Progesterone prevents the tamoxifen effects, even if administered 9 h after tamoxifen (at which time ov mRNA has decreased by 30%). The progesterone-induced increase of ovalbumin mRNA (from 0.64% of cytoplasmic RNA to 0.79%) and the increased proportion of ribosomes in polysomes (from 65% to 80%) are thus independent of estrogen action. Twenty-four hours of progesterone plus tamoxifen treatment enhances initiation of translation. However, after prolonged treatments of several days, translation becomes inefficient: ovalbumin synthesis falls by 30% without a coordinate decrease in ovalbumin mRNA, and a significant proportion (15%) of the ovalbumin mRNA becomes localized in monosomes. Thus, optimal maintenance of oviduct mRNA utilization requires the presence of estrogen.


Precursor of egg white ovomucoid. Amino acid sequence of an NH2-terminal extension. The translation of ovomucoid mRNA in a reticulocyte lysate protein-synthesizing system yields a precursor form which contains an NH2-terminal extension of 23 amino acid residues. Edman degradation of radioactive translation products (pre-ovomucoid) identified the following sequence: formula : (see text), where the initiator methionine (in parentheses) is the only residue cleaved from the NH2 terminus during cell-free synthesis and the vertical line indicates the site at which pre-ovomucoid is cleaved in vivo to yield ovomucoid. The precursor sequence differs from those of two other proteins (pre-lysozyme and pre-conalbumin) secreted by the same cell, but resembles these and other secretory protein "signal peptides" in both length and hydrophobicity. Pre-ovomucoid does not interact with trypsin in the same manner as mature ovomucoid.


Identification by affinity chromatography of the eukaryotic ribosomal proteins that bind to 5 S ribosomal ribonucleic acid. Rat liver 5 S ribosomal RNA was oxidized with periodate and coupled by its 3' terminus to Sepharose 4B through an adipic acid dihydrazide spacer. The ribosomal proteins that bind to that nucleic acid were isolated by affinity chromatography and identified by electrophoresis in polyacrylamide gels. The eukaryotic 5 S rRNA binding proteins were L6 and L19: small amounts of L7, L23', L27/L27', L35', and L39 also were bound to the affinity column, but whether they associate directly and specifically with 5 S rRNA is not known. The proteins of the 40 S subunit of rat liver ribosomes did not bind to the 5 S rRNA affintiy column, nor did the proteins of either the large or small subparticle of Escherichia coli ribosomes.


Vitamin K dependence of a calcium-binding protein containing gamma-carboxyglutamic acid in chicken bone. Bone contains a small protein, osteocalcin, rich in the amino acid gamma-carboxyglutamate (gammaCGlu), which is dependent on vitamin K for its synthesis. The vitamin K dependence of osteocalcin biosynthesis has been studied both in the developing chicken embryo and in the skeletons of young chicks. Osteocalcin formation, assessed by quantitation of gammaCGlu in whole bone, begins in the 8-day embryonic skeleton and increases dramatically until hatching. Biosynthesis of gammaCGlu is warfarinsensitive; time- and dose-related inhibition of up to 75% is observed. Vitamin K deficiency reduces bone gammaCGlu concentrations by 50% in 3-week-old chicks, and this is totally corrected by including vitamin K1 in the diet. Feeding the vitamin K antagonist dicumarol decreases the bone gammaCGlu level 80% after 6 weeks, but no corresponding alterations are observed in the calcium and phosphorous content of the bone ash. Osteocalcin purified from bones of dicumoral-fed chicks exhibits several electrophoretically variant undercarboxylated protein species containing 2 and 3 gammaCGlu/57 amino acid residues rather than the normal complement of 4 gammaCGlu/57 residues.


Studies with a homogeneous enzyme from rabbit erythrocytes catalyzing the insertion of guanine into tRNA. An enzyme that catalyzes a post-transcriptional modification of tRNA, resulting in replacement of a base from tRNA by guanine, has been purified 2600-fold from rabbit erythrocyte cytosol. The purest preparation migrates as a single protein band on polycrylamide gel electrophoresis and the enzymatic activity co-electrophoreses with this protein. The native enzyme has a molecular weight of 104,000 and is dissociated into two subunits of Mr= 60,000 and 43,000. The Km for guanine is 1.5 x 10(-7) M and for a pure guanine-accepting tRNA is 3.3 x 10(-9) M. The amino acid composition of the pure enzyme has been determined. To our knowledge this is the first study in which the molecular characteristics of a pure enzyme capable of modifying an internal position in tRNA has been reported.


Synergistic regulation of phosphorylase a by glucose and caffeine. Kinetic studies of both liver and muscle phosphorylase a demonstrate that caffeine and glucose inhibit the binding of glucose 1-phosphate to the enzyme in a synergistic competitive and nonexclusive manner. Inhibition studies for numerous other caffeine analogs show that the muscle enzyme has a relaxed specificity for this negative effector. The liver enzyme is more discriminating by preferential binding of methylated oxypurines. Physiological concentrations of AMP and ATP, which affect the enzymic activity at a separate site, prevent glucose from effectively inhibiting the enzyme. The addition of the second synergistic ligand improves the binding of glucose. These data suggest that glucose homeostasis as regulated by phosphorylase may be dependent on a second ligand and that the role of glucose in this physiological process may have been overestimated. A structural rationalization of this synergistic response is discussed with reference to the crystal structure of the muscle enzyme.


Anterior subluxation of the lateral tibial plateau. A diagnostic test and operative repair. Recurrent anterior subluxation of the lateral tibial plateau is a common type of chronic knee instability resulting from trauma. It can be reproduced by the clinical test described and corrected by a surgical procedure called the sling and reef operation, in which a strip of iliotibial tract is used to create a sling and to reef the posterolateral capsule. From 1971 to 1978, eighty-four patients were operated on, of whom fifty had been evaluated at one to six and one-half years after operation. The results were: forty-one good, six fair, and three poor. The lesions found in the thirty-seven knees in which arthrotomy was performed included a tear of the anterior cruciate in every case, a tear of the medial meniscus in fifteen and of the lateral meniscus in eleven, a notch in the articular surface of the lateral femoral condyle in fifteen, and a lateral marginal tibial (Segond) fracture in three. No definite lateral capsular tears were visualized--only stretching comparable to that seen in recurrent dislocation of the shoulder.


Anterolateral rotatory instability of the knee joint. An early analysis of the Ellison procedure. Our experience with the Ellison procedure for the control of anterior subluxation of the lateral tibial plateau includes fifty-two knees in which we used this procedure either alone or in combination with other reconstructive procedures. The results directly attributable to the Ellison procedure itself after follow-ups of six months or more could be evaluated in fifteen patients who had only the Ellison procedure for anterolateral instability and in thirteen who had both this procedure and a pes anserinus transfer for combined anterolateral and anteromedial instability. Of the twenty-eight patients so evaluated, only sixteen (57 per cent) had good or excellent results. The pivot-shift phenomenon, present in all twenty-eight knees preoperatively, had been eliminated in only four at follow-up. A positive anterior drawer sign was present in all patients both before operation and at follow-up. The subjective and functional improvement did not correlate well with the clinical and roentgenographic findings. From this study it is our impression that the results of this operation are disappointing and that it should be used only as an adjunctive procedure in lateral reconstruction in young patients.


Revision with gentamicin-impregnated cement for deep infections in total hip arthroplasties. Seventy-seven total hip arthroplasties complicated by deep infection were treated by reoperation and fixation of a new prosthesis with gentamicin-containing cement. The infection was proved in each case by positive bacterial cultures. Systemic antibiotic treatment was continued for six months postoperatively. Sixty hips were considered healed; the patients had no pain and had normal sedimentation rates and roentgenograms. Nine still had pain or an abnormal sedimentation rate and were classified as doubtful with regard to eradication of the infection. Eight were still infected. The patients were followed for six months to three and one-half years after reoperation.


In vitro collagen biosynthesis in healing and normal rabbit articular cartilage. To examine the repair collagens produced by cells in injured cartilage, the femoral articular surfaces of three groups of New Zealand white rabbits were injured by making both superficial and deep lacerations and drill holes. Eight weeks after surgery, the rabbits were killed and slices of injured articular cartilage were harvested. The types of collagen being synthesized at the site of these lesions were identified by labeling the recovered specimens in vitro with 3H-proline and by characterizing the collagen using sodium dodecyl sulphate electrophoresis, carboxymethyl cellulose chromatography, and cyanogen bromide peptide analysis. In all cases, tissue-specific type II ([alpha1 (II)]3) cartilage collagen was synthesized. Histological examination using toluidine blue showed that the chondrocytes bordering the cartilage defect produced by deep lacerations and drill holes responded by increased cellular activity, as shown by cell cloning and increased matrix staining. The drilled holes were completely filled by tissue with staining and morphological characteristics similar to those of hyaline cartilage.


Acute compartment syndromes: diagnosis and treatment with the aid of the wick catheter. Intracompartmental pressures were measured by the wick catheter technique in sixty-five compartments of twenty-seven patients who were clinically suspected of having acute compartment syndromes. A pressure of thirty millimeters of mercury or more was used as an indication for decompressive fasciotomy. The range of normal pressure was from zero to eight millimeters of mercury. Eleven of these patients were diagnosed as actually having compartment syndromes and in these patients, twenty-seven compartments were decompressed. Only two patients had significant sequelae. In the sixteen patients (thirty-eight compartments) whose pressures remained less than thirty millimeters of mercury, fasciotomy was withheld and compartment syndrome sequelae did not develop in any patient. Intraoperatively the wick catheter was used continuously in eight patients to document the effectiveness of decompression. Fasciotomy consistently restored pressures to normal except in the buttock and deltoid compartments, where epimysiotomy was required to supplement the fasciotomy. Continuous intraoperative monitoring of pressure by the wick catheter technique allowed us to select the few cases in which primary closure of wounds was appropriate and to decide which patients were best treated with secondary closure.


Complications with use of the Stryker frame. Roentgenograms were made of two patients with acute dislocations of the cervical spine who were undergoing treatment on Stryker frames with cervical traction. Losses of reduction were evident when the patients were turned from the supine to the prone position. The pulmonary vital capacity of patients with complete traumatic quadriplegia consistently decreased when they were turned to the prone position. Prolonged supine positioning of patients on the Stryker frame also caused occipital decubitus ulcers.


Sinding-Larsen-Johansson disease. Its etiology and natural history. In a prospective study of children with pain in the knee, eight patients (ten knees) were seen with the clinical and roentgenographic findings of Sinding-Larsen-Johansson disease. The etiology appears to be a traction tendinitis with de novo calcification in the proximal attachment of the patellar tendon, which had been partially avulsed. The patients were followed through the course of the disease, which was found to be self-limited and benign like Osgood-Schlatter disease.


Retroperitoneal abscess. A potentially fatal complication of closed fracture of the pelvis. Two patients were seen in whom a retroperitoneal abscess developed following closed pelvic fracture. A retroperitoneal abscess is often unsuspected clinically, and presents a difficult diagnostic challenge. Prompt recognition and treatment are essential to patient survival.


Tumoral calcinosis. A clinical and pathological study of eleven unreported cases in Turkey. Clinicopathological observations of eleven cases of tumoral calcinosis are reported. All of the cases showed characteristic clinical and histopathological features of this unusual condition, but pathological findings did not elucidate all of its etiology and pathogenesis. Two of the patients were related. One of the patients showed skeletal changes characterized by diffuse calcium deposits with widespread soft-tissue calcification. The available data of the present study also led to the suggestion that the disease has a genetic background and that the primary defect may well be in the collagen which undergoes calcification.


Leiomyosarcoma of the thorax.

Leiomyosarcoma of the thorax. Leiomyosarcoma within the thorax is a rare lesion. This report describes an endobronchial lesion presenting with atelectasis, a parenchymal lesion presenting as a mass, and a rare extra-pleural lesion arising in the chest wall. The prognosis is generally poor; our 3 patients died within about 1 year of diagnosis.


Education and practice location of Canadian diagnostic radiologist. At the 1977 Annual Meeting of The Canadian Association of Radiologists, a survey of all practicing diagnostic radiologists was made. Information was also obtained from Royal College and Corporation Professionnelle records. The medical school, postgraduate program, and present location of practice were established for over 98% of active radiologists. This information has been collected to assist program directors, medical faculties, provincial governments, and radiologists in recognizing manpower needs.


Composite photoradiography.

Composite photoradiography. A new simpler method of combining photographic and radiographic images to make a composite photoradiograph is described. The matching of the images can now be accomplished in the camera rather than, as formerly, in the darkroom.


The radiologist's opportunity to teach bone dynamics. Activity and muscle strength are important for bone growth and for development and maintenance of bone density. The rediologist should help students to understand the important relationship between muscular activity and bone strength.


Pitfalls in the ultrasonographic diagnosis of the pancreatic pseudocyst. A pseudocyst of the pancreas typically presents ultrasonographically as a cystic structure in the upper abdomen. We have seen numerous cases which presented similar sonographic features suggestive of a pancreatic pseudocyst in which this diagnosis proved false. A knowledge of the ultrasonographic differential diagnosis of the pancreatic pseudocyst therefore becomes important.


Renal arteriography in von Hippel-Lindau Diease. Since 1960, 17 patients with von Hippel-Lindau disease have had cerebellar hemangioblastomas removed at the Victoria General Hospital, Halifax, Nova Scotia. The 4 most recent patients have had renal arteriography as a part of their investigation. Though 3 of these had negative urography, all 4 had positive findings on arteriography. The first patient had bilateral renal cell carcinomas and cysts and failed to respond to 1 year of 'Laetrile' therapy; the second had small cysts in each kidney; the third had bilateral renal cysts, multiple left renal cell carcinomas and a clinically unsuspected right pheochromocytoma; the fourth had a small renal cell carcinoma. It is recommended that all patients with von Hippel-Lindau disease and siblings with any manifestations of the disease have renal arteriography as part of their early investigation.


[Angiomyolipomas (hamartomas) of the kidney. Radiological study apropos of 2 cases. Review of the literature]. Two patients with renal angiomyolipomas studied angiographically are reported:--A case of tuberous sclerosis with bilateral multiple lesions, renal, hepatic and adrenal;--An isolated lesion whose preoperative diagnosis allowed the excision of the tumor. A review of the literature from 1965 to 1977 identified 242 cases of which 177 were isolated angiomyolipomas. There were 54 cases associated with tuberous sclerosis and one with von Hippel-Lindau disease. The radiologic and especially the angiographic signs are illustrated, the hemorrhagic risks are stressed, and the diagnostic features of fetal renal hamartoma are described.


Renal displacement simulating crossed renal ectopia. Displacement of the right kidney, simulating crossed renal ectopia, was associated with massive obesity and a ventral hernia.


Thymic hyperplasia in thyrotoxicosis.

Thymic hyperplasia in thyrotoxicosis. Thymic hyperplasia in a thyrotoxic patient produced an anterior mediastinal mass. Pathogenesis and diagnostic features are discussed.


Pulmonary cystic disease in ankylosing spondylitis: two cases with unusual superinfection. Two ankylosing spondylitis patients with upper lobe fibrocystic disease are reported. The occurrence of pulmonary parenchymal disease as part of the primary pathologic process in ankylosing spondylitis is now accepted as an integral part of the disease. The frequency of superinfection of cavitary lung disease with Aspergillus is noted. Another unusual superinfection, Mycobacterium scrofulaceum, is reported.


Reflux of barium into a maxillary antrum. A patient with achalasia and neurologic deficit permitting intermittent nasopharyngeal reflux developed unilateral maxillary sinusitis. Heavy metal density in the center of the inflamed sinus almost certainly was barium sulfate refluxed during an upper gastrointestinal study.


Chondromyxoid fibroma of lumbar spine.

Chondromyxoid fibroma of lumbar spine. Chondromyxoid fibroma involved the spinous process of L2. To our knowledge, this is the first reported occurrence in the lumbar spine.


Cardiac fibrosarcoma with bone metastases. The diagnosis of a primary cardiac sarcoma with bone metastases was suspected in a previously normal patient because of the development of unremitting pulmonary edema and lytic bone lesions. Echocardiography substantiated the diagnosis, which proved at autopsy to be a primary fibrosarcoma of the heart. This represents only the third such case reported with bone metastases.


Peritoneal calcification in a neonate with imperforate hymen. Contrast medium for vaginography in a premature neonate passed into the peritoneal cavity; calcification in that area apparently represented genital-tract secretions which had accumulated above the imperforate hymen. Ultrasonography of the mother's abdomen had indicated fetal ascites, a finding that may be of value in diagnosing genital-tract obstruction prenatally.


Binding of cytochrome b5 to membranes of isolated subcellular organelles from rat liver. The in vitro incorporation of a well-characterized integral protein cytochrome b5 into membranes of various subcellular organelles was investigated by biochemical and immunochemical methods. Microsomes, peroxisomes, and outer mitochondrial membranes, all containing endogenous cytochrome b5, incorporated large amounts of the hemoprotein in such a way that it was reducible by an inherent NADH cytochrome b5 reductase. Lysosomal membranes did not incorporate cytochrome b5. Inner mitochondrial and Golgi membranes, which do not naturally contain cytochrome b5, bound it in vitro but it was not reduced in the presence of NADH. These results show some discrepancies between the natural localization and the in vitro binding of cytochrome b5. They confirm one aspect of the fluid membrane theory and bring new elements to our understanding of the maintenance of the specific features of the membranes of subcellular organelles with respect to the cell dynamism.


Heterogeneity in sensitivity to cleavage by the restriction endonucleases ECORI and HindIII of circular kinetoplast DNA molecules of Crithidia acanthocephali. Kinetoplast DNA (kDNA) of the protozoan Crithidia acanthocephali consists mainly of an association of approximately 27,000 covalently closed, 0.8-micron (1.58 X 10(6) daltons) circular molecules apparently held together in a particular structural configuration by topological interlocking. The sensitivities of circular kDNA molecules to the restriction endonucleases EcoRI and HindIII have been studied using agarose gel electrophoresis and electron microscopy. Digestion with EcoRI or HindIII of collections of single circular molecules obtained from sonicated kDNA associations resulted in a single cleavage of 9.3 and 12% of the molecules, respectively. Digestion of intact kDNA associations with EcoRI or HindIII resulted in cleavage of 9.2 and 10.4%, respectively, of the component circular molecules, but not in detectable disruption of the characteristic structure of the associations. Analysis of the products of sequential digestion of kDNA with the two enzymes indicated that approximately 8% of the circular molecules each contain a single site sensitive to EcoRI and a single site sensitive to HindIII; 1.5-3% contain only an EcoRI-sensitive site; 3-4% contain only a HindIII-sensitive site; and the remainder (approximately 86%) are insensitive to either enzyme. Further, data obtained from sequential digestion experiments and from studies of the partial denaturation products of the circular molecules digested with EcoRI or HindIII indicated that when they occur the EcoRI site and the HindIII site are each at a unique position in all molecules, 10-13% of the circular contour length apart. Similar digestion products were found for kDNAs from different cloned organisms, suggesting that the four different kinds of circular molecules, in regard to EcoRI and HindIII sensitivity, are found in similar proportions in the kDNA association of different organisms.


Stratification, specialization, and proliferation of primary keratinocyte cultures. Evidence of a functioning in vitro epidermal cell system. A population of neonatal mouse keratinocytes (epidermal basal cells) was obtained by gentle, short-term trypsin separation of the epidermal and dermal skin compartments and discontinuous Ficoll gradient purification of the resulting epidermal cells. Over 4--6 wk of culture growth at 32--33 degrees C, the primary cultures formed a complete monolayer that exhibited entire culture stratification and upper cell layer shedding. Transmission and scanning electron microscopy demonstrated that the keratinocyte cultures progressed from one to two cell layers through a series of stratification and specialization phenomena to a six to eight cell layer culture containing structures characteristic of epidermal cells and resembling in vivo epidermal development. The temporal development of primary epidermal cell culture specialization was confirmed by use of two histological techniques which differentially stain the specializing upper cell layers of neonatal mouse skin. No detectable dermal fibroblast co-cultivation was demonstrated by use of the leucine aminopeptidase histochemical technique and routine electron microscope surveillance of the cultures. Incorporation of [3H]thymidine ([3H]Tdr) was greater than 85% into DNA and was inhibited by both 20 micron cytosine arabinoside (Ara-C) and low temperature. Autoradiography and 90% inhibition of [3H]Tdr incorporation by 2 mM hydroxyurea indicated that keratinocyte culture DNA synthesis was scheduled (not a repair phenomenon). The primary keratinocytes showed an oscillating pattern of [3H]Tdr incorporation into DNA over the initial 23--25 days of growth. Autoradiography demonstrated that the cultures contained 10--30% proliferative stem cells from days 2-25 of culture. The reproducibility of both the proliferation and specialization patterns of the described primary epidermal cell culture system indicates that these cultures are a useful tool for investigations of functioning epidermal cell homeostatic control mechanisms.


Ultrastructural and physiological studies on the longitudinal body wall muscle of Dolabella auricularia. I. Mechanical response and ultrastructure. The physiological properties of mechanical response and the ultrastructure in the longitudinal body wall muscle (LBWM) of the opisthobranch mollusc Dolabella auricularia were studied to obtain information about excitation-contraction coupling in somatic smooth muscles responsible for smooth and slow body movement of molluscans. The contracture tension produced by 400 mM K was not affected by Mn ions (5--10 mM) and low pH (up to 4.0), but was reduced by procaine (2 mM). The K-contracture tension was not readily eliminated in a Ca-free solution containing ethylene glycol-bis(beta-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA). A large contracture tension was also produced by rapid cooling of the surrounding fluid from 20 degrees to 5 degrees--3 degrees C even when the preparation showed no mechanical response to 400 mM K after prolonged (more than 2 h) soaking in the Ca-free solution. These results indicate that the LBWM fibers contain a large amount of intracellularly stored Ca which can be effectively released by membrane depolarization. The fibers were connected with each other, forming the gap junctions, the desmosomes, and the intermediate junctions. The sarcoplasmic reticulum (SR) consisted of vesicular and tubular elements, and was mostly located near the fiber surface. The plasma membrane showed marked tubular invaginations of 600-800 A in diameter, with many branches (surface tubules), extending inwards for approximately 2 micron. These surface tubules were closely apposed to the SR, and the bridgelike structures analogous to those in the triadic junction of vertebrate skeletal muscle were observed in the space between the surface tubules and the SR. It is suggested that the influence of membrane depolarization is transmitted inwards along the surface tubules to cause the release of Ca from the SR.


Ultrastructural and physiological studies on the longitudinal body wall muscle of Dolabella auricularia. II. Localization of intracellular calcium and its translocation during mechanical activity. The localization of Ca-accumulating structures in the longitudinal body wall muscle (LBWM) of the opisthobranch mollusc Dolabella auricularia and their role in the contraction-relaxation cycle were studied by fixing the LBWM fibers at rest and during mechanical response to 400 mM K or to 10(-4)--10(-3) M acetylcholine in a 1% OsO4 solution containing 2% K pyroantimonate. In the resting fibers, electron-opaque pyroantimonate precipitate was mostly localized at the peripheral structures, i.e., along the inner surface of the plasma membrane, at the membrane of the surface tubules, and at the sarcoplasmic reticulum. In the fibers fixed during mechanical activity, the precipitate was diffusely distributed in the myoplasm in the form of numerous particles with corresponding decrease in the amount of the precipitate at the peripheral structures. Electron-probe X-ray microanalysis showed the presence of Ca in the precipitate, indicating that the precipitate may serve as a measure of Ca localization. These results are in accord with the view that, in the LBWM, the Ca stored in the peripheral structures is released into the myoplasm to activate the contractile mechanism.


Distribution of an endogenous lectin in the developing chick optic tectum. We determined the cellular localization of an endogenous lectin at various times during the development of a well-characterized region of chick brain, the optic tectum. This lectin is a carbohydrate-binding protein that interacts with lactose and other saccharides, undergoes striking changes in specific activity with development, and has previously been purified by affinity chromatography from extracts of embryonic chick brain and muscle. Cellular localization in the tectum was done by indirect immunofluoresecent staining, using immunoglobulin G derived from an antiserum raised against pure lectin. No lectin was detectable in the optic tectum examined at 5 days of embryonic development. From approximately 7 days of development, neuronal cell bodies and fibers were labeled by the antibody; and extracts of tectum contained hemagglutination activity that could be inhibited by lactose or by the antiserum. Lectin remained present in many tectal neuronal layers after hatching; but in 2-month-old chicks it was sparse or absent in most of the tectum except for prominent labeling of fibers in the stratum album centrale. The initial appearance of lectin in the optic tectum was not dependent on innervation by optic nerve fibers since bilateral enucleation during embryogenesis did not affect it. Lectin was detectable on the surface of embryonic optic tectal neurons dissociated with a buffer containing EDTA.


Localization of intestinal sucrase-isomaltase complex on the microvillous membrane by electron microscopy using nonlabeled antibodies. Microvillous vesicles isolated from rabbit small intestine showed a trilaminar membrane with a rather smooth surface, which was apparently not affected by papain solubilizing sucrase-isomaltase complex or by trypsin unable to solubilize it. When microvilous vesicles or trysinized ones were incubated with immunoglobulin G against the sucrase-isomaltase complex or monovalent fragments therefrom, an apparently continuous electron-opaque layer approximately 180 A in width appeared around the external surface of vesicles. Such a layer was not formed on papainized vesicles. Microvillous vesicles and trypsinized ones negatively stained with phosphotungate showed a great number of particles protruding approximately 150 A from the membrane surface, but papainized vesicles did not. The particles existed close to one another and appeared to form a particulate layer 150 A in width on the surface. The antibodies, whether they were divalent or monovalent, increased the width of the layer to approximately 200 A and obscured the fine particulate structure of intact and trypsinized vesicles. Papainized vesicles retained their smooth surface upon interaction with antibodies. These results, together with those with the Triton-solubilized sucrase- isomaltase complex (Nishi and Takesue, 1978), J. Ultra-struct. Res., 62:1- 12), indicate not only that sucrase-isomaltase complexes are located close to one another on the membrane, but also that they or at least their protein portions protrude approximately 150 A from the surface of the trilaminar membrane.


Non-reciprocal contact inhibition of locomotion of chick embryonic choroid fibroblasts by pigmented retina epithelial cells. Using light and electron microscopy, we have confirmed an earlier observation that chick embryonic pigmented retina epithelial cells (PRE cells) seeded in vitro on cultured sheets of choroid fibroblasts, are able to spread. Spreading is as rapid (and shows the same dependence on lateral contact between PRE cells) as on a serum-coated culture substrate. After 1 h most cells are spreading on the upper surface of the choroid sheet, but after 4 h, some PRE cells can be found sandwiched between overlapping choroid cells, and thus have invaded the sheet. Choroid fibroblasts underlie PRE in vivo, but the ability of PRE cells to spread on cultured fibroblasts is not specific for choroid, since PRE cells spread also on BKH21 hamster kidney fibroblasts, and on fibroblasts from chick embryonic heart. As reported by others for various fibroblastic cells, choroid cells seeded on to choroid sheets or on to cultured PRE are unable to spread. A possible explanation is that spreading of adherent cells is contact-inhibited by the cells in the sheet, just as their leading edges are paralysed on contact, and thus locomotion is inhibited, when fibroblasts collide on a plane substratum. If spreading of seeded cells and cell locomotion are inhibited by the same mechanism, PRE cells should contact-inhibit choroid fibroblasts with which they collide, but not themselves be so inhibited. Using time-lapse cinemicrography, we have found this to be the case. We first established that in homotypic collisions, choroid fibroblasts do show contact inhibition of locomotion, despite the criss-cross (not well monolayered) appearance of confluent cultures. In heterotypic collisions between choroid and PRE we found the predicted nonreciprocal behaviour: the choroid leading edge is paralysed on collision, and the cell subsequently retracts, whereas the active PRE margin appears to be completely unaffected. Speed measurements from a series of such collisions show that the speed of choroid cells is markedly reduced on collision with PRE, whereas the slight slowing of PRE is not statistically significant. We have observed similar behaviour in heterotypic collisions between various epithelial and fibroblastic cells, and so it seems possible that non-reciprocity may prove general for this interaction. If so, it has important implications for the role of contact inhibition of locomotion in phenomena such as morphogenesis, wound healing and the invasiveness of carcinoma cells. On the one hand, non-reciprocal contact-inhibition of locomotion may permit the spreading of epithelia over mesenchymal cells, thus generating or restoring an epithelial bounding membrane. On the other hand, in the absence of other interactions, it would fail to inhibit the invasion of mesenchymal territory by aberrant epithelial cells, or presumably by epithelial free edges.


Selective adhesion and impaired adhesive properties of transformed cells. Quantitative studies on the adhesive properties of transformed cells have yielded inconclusive and sometimes contradictory results. The present investigation has examined adhesive interactions between normal human fibroblasts, established as well as virus-transformed animal cell lines, and human tumour-derived cell lines by the cell-cell layer binding assay. The results of these investigations indicate that adhesive selectivity can be observed between normal human fibroblasts and 2 human tumour-derived cell lines, providing an  in vitro system to study cell surface components involved in cellular interactions between normal and malignant cells. In addition it is demonstrated that cell layers of transformed cells form a poorly adhesive substratum for both trypsinized normal and transformed cells. Furthermore, it is confirmed that the adhesive properties of transformed cells, including adhesive selectivity, are affected by the dissociation procedure (trypsin or EDTA). In view of the observations made by other investigators, the present results suggest that transformed cells display adhesive properties which can be quantitatively and reproducibly measured but which are modulated by the dissociation procedure as well as by the configuration in which the cells are at the time of the assay.


Mechanisms of invasiveness of epithelial tumours: ultrastructure of the interactions of carcinoma cells with embryonic mesenchyme and epithelium. Three different kinds of carcinoma cell, human breast tumours, mouse lung tumour and rat bladder tumours have been implanted into the developing chick wing to assay their invasive behaviour. We found that the majority of the carcinoma cells did not invade the mesenchyme but were positioned in the ectoderm. We examined the ultrasturcture of the interaction of the carcinoma cells with both the ectoderm and the mesenchyme to see if there are any differences. We found no specialized adhesive junctions between carcinoma cells and mesenchyme cells. In contrast, the carcinoma cells seem to be adhesive to the ectoderm cells; desmosomes between the two have been found and also frequent parallel alignment of membranes over long distances. This difference between the interaction of the carcinoma cells with the mesenchyme and the ectoderm may explain why the carcinoma cells can penetrate into the ectoderm but rarely do so into the mesenchyme. The carcinoma cells formed a smooth border with the mesenchyme even in the absence of a basal lamina. These results are discussed in relation to the mechanisms of invasion of epithelial tumours in vivo. In particular, the relative importance of the basal lamina, and cell adhesion within the tumour, in limiting invasiveness are assessed.


Suppression of the transformed phenotype in somatic cell hybrids. Somatic cell hybrids between mouse mammary tumour cells (TA3B) and diploid rat embryo fibroblasts (REF) or between TA3B and Syrian hamster sarcoma cells (BI) were examined for the in vitro characteristics of transformed cells as soon as possible after cell fusion. Unlike the parental tumour cells as three of four TA3B X REF and five BI X TA3B independent hybrid lines had low colony-forming efficiencies in agar, exhibited density-dependent inhibition of growth and did not form colonies on confluent monolayers of 3T3 cells, demonstrating that the transformed phenotype was suppressed in these hybrids. In addition tests of some of the hybrid lines for tumour production in nude mice showed that this was also suppressed. Suppression was more stable in the TA3B X REF than in the BI X TA3B hybrids, variants of the BI X TA3B hybrids with the properties of transformed cells could be readily isolated by subculturing cells that had grown in agar. Tumour growth selected for hybrids with the characteristics of transformed cells, and derivatives of the hybrids selected to show the transformed phenotype readily produced tumours. These correlations suggest that the transformed phenotype and malignancy may be under the same control in these cells. The phenomenon of suppression may be explained by the hypothesis that neoplastic transformation results from recessive mutations in genes which control the normal phenotype. On this model the finding of suppression in hybrids between two different tumour lines is interpreted as complementation and indicates that the mutations are not the same in all cell lines.


Directionality of microtubule assembly: an in vivo study with the ciliate Tetrahymena. A temperature-sensitive mutant homozygous for the recessive gene molb in Tetrahymena thermophila offers opportunity for studying the direction of microtubule assembly in vivo. At 39 degrees C the mutant fails to divide properly; the 2 daughter animals remain attached and bend over each other. As revealed by protargol staining, the bending results in acute turning and breaking of some of the longitudinal microtubular bands close and parallel to the surface. Hence, 2 broken microtubular ends are available for study of the problem of directionality of microtubule assembly, by assessing which of the 2 ends regenerates. In most cases the posterior portion of the longitudinal microtubular band regenerates. The present study hence supports the conclusion based on in vitro observation in other systems that microtubule assembly is predominantly unidirectional.


The packaging unit: a basic structural feature for the condensation of late cricket spermatid nuclei. The alignment, folding and packaging of cricket chromatin was examined during late spermiogenesis by an electron-microscope study of nuclei dispersed by air--liquid surface tension forces after detergent treatment. Late developing spermatid genomes arrange themselves in multiple packaging units in a stepwise process which includes: (1) a loss of the beaded repeating structure of chromatin as nucleoprotein fibres become smooth and gradually assume a uniform diameter; (2) a side-by--side alignment of structurally modified chromatin fibres; and (3) a regular folding into packaging units. Alignment and folding of chromatin fibres are presumably mediated by intermolecular bonds easily disrupted by spreading forces. In very late spermatids, interfibre binding forces are difficult to overrride by spreading alone, indicating a stronger cross-linking of increasingly coalescent packaging units. 'Unit to unit' coalescence stabilizes the nuclear structure, first limiting and afterwards denying penetration of phosphotungstic acid, as displayed in thin sections of extremely cricket spermatid nuclei. Binding of phosphotungstate by nuclear basic proteins can be facilitated by limited protein solubilization after disulphide reduction of unfixed cricket tests with sodium dodecyl sulphate and dithiothreitol. Results of this study permit the proposal of model experiments useful for clarifying the organization of highly condensed spermatid genomes and for evaluating the structure of genome segments in systems wherein changes of chromatin-associated protein occur.


Membrane changes associated with mucus production by intestinal goblet cells. Changes in the structural organization of membranes of mucous bodies and the plasma membrane that occur during mucus production in goblet cells of rat rectum have been studied by thin-section and freeze-fracture techniques. Immature mucous bodies are bounded by a trilaminar membrane and fracture faces of the membrane have randomly distributed intramembrane particles. During maturation, mucous bodies become packed tightly together and changes in the structure of their membranes include (1) fusion of apposing membranes of adjacent bodies to form a pentalaminar structure, (2) a reduction in the density of particles on membrane fracture faces, and (3) exclusion of particles from regions of membrane apposition. Some trilaminar membranes of mucous bodies fuse with the lumenal plasma membrane to form a pentalaminar structure. Sites of apposition between mucous body membranes and the lumenal plasma membrane are seen as particle-cleared bulges on fracture faces of the plasma membrane. Our results indicate that membrane reorganization associated with mucous production in goblet cells includes a reduction and redistribution of some membrane proteins and that membrane fusion occurs between portions of membranes from which proteins have been displaced.


Intercellular bridges between follicle cells and oocyte during the differentiation of follicular epithelium in Lacerta sicula Raf. Intercellular bridges first appear during lizard oogenesis when follicles are rather small (150 microgram in diameter); at this stage they form connecting links between the oocyte and follicle cells, which have not yet differentiated into pyriform cells. Later on, when the follicles have become larger (1 mm) and the follicular epithelium appears constituted by 3 types of cells (small, intermediate and pyriform cells) they form connecting links between the oocyte and both intermediate and pyriform cells. The establishment of intercellular bridges between pyriform cells and the oocyte precedes the complete differentiation of the former, which excludes the possibility that the fusion between pyriform cells and oocyte occurs only after these cells are completely differentiated. In still larger follicles (up to 2 mm in diameter), during the degeneration of the pyriform cells, the occurrence, inside the bridges, of mitochondria and other cytoplasmic material suggests that these cells at the end of their function transfer their contents into the oocyte.


Labelling of human resting lymphocytes by continuous infusion of [3H]thymidine. I. Characterization of cytoplasmic label. After continuous 3H-TdR infusion in vivo or incubation with 3H-TdR in vitro human blood lymphocytes were examined by light-microscopic and electron-microscopic autoradiography. Using relatively long autoradiographic exposure times (50--300 days) not only nuclear but also cytoplasmic labelling was visualized, the cytoplasmic label being present in up to 96% of the cells. The cytoplasmic label was predominantly associated with the mitochondria and was removed from the cells nearly completely by treatment with DNase but not with RNase or cold perchloric acid. It is concluded that this cytoplasmic label mainly represents 3H-TdR incorporated into mitochondrial DNA which is continuously renewed in an average turnover time of 14 days or less. This value is compatible with a turnover time of 11 days for mitochondrial DNA in mammalian cells reported in the literature.


In vivo phagocytosis by developing myocardial cells: an ultrastructural study. Phagocytosis of naturally degenerating cells, by healthy myocardial cells of the bulbus cordis of 5--7-day chick embryos, was studied by electron microscopy. Myocardial cells showed cell processes surrounding dead cell fragments. The cell fragments appeared to be internalized later and digested within phagosomes. Ruthenium red was employed to ascertain whether the cell fragments were in fact internalized or located in pockets of the cell membrane. The results are discussed in the light of present knowledge of phagocytosis.


Cell motility in fibroblast aggregates.

Cell motility in fibroblast aggregates. The stable arrangements of cells in tissues and organs require that most tissue cells, whatever their inherent abilities for active pseudopod-directed locomotion, remain confined to the boundaries of their parent tissues and organs for the lifetime of the organism. Contact inhibition of motility represents a potential mechanism for immobilizing cells in tissues and thus in promoting positional stability. In order to assess the effectiveness of contact inhibition in immobilizing cells in 3-dimensional tissue-like masses, spherical aggregates of chick heart, sclera and skin fibroblasts were fused with tritiated thymidine-labelled aggregates of the identical cell type. After being placed in contact, the 2 aggregates cohere and broaden the area of mutual contact to form a single aggregate with a planar interface between the two. Mobility of cells was detected by the migration of labelled cells across the interface and into the unlabelled aggregate. Based on measurements of distances moved in day-old aggregate pairs, it was found that fibroblasts moved up to 17 cell diameters and up to 36% as far in solid tissue masses as in monolayer culture under optimal conditions for directed migration.


Interaction of leukocytes with vascular cells in culture. The nature, kinetics and cofactor requirements of leukocyte adhesion to cultured vascular cells have been investigated in vitro, using a model system in which leukocyte suspensions are in continuous motion over the cultured cells. Adhesion is assessed by histological examination or by using 51Cr-labelled leukocytes. Leukocytes adhere preferentially to endothelial cell monolayers rather than to smooth muscle cells, adventitial fibroblasts or serum-coated glass. Arterial and venous endothelium are equally good substrates for leukocyte adhesion, and lymphocytes and granulocytes at the same suspension density adhere to endothelial cells in similar numbers. Adherence is greatest during the first 10 min, and is inversely related to flow rate. Numbers of leukocytes adhering to the cell monolayer are proportional to the initial concentration of leukocytes in suspension, although only 2--10% of the leukocytes adhere. Leukocytes that have adhered to the cell monolayer spread over the cells and then migrate, apparently through intercellular junctions. Adhesion requires Ca2+ or Mg2+ ions, but not plasma protein cofactors. Fewer leukocytes adhere to endothelial cells grown for 48 h in rapidly stirred medium than to cells grown under conventional, static conditions.


[Jejuno-gastric retrograde invagination after vagotomy: a case of spontaneous desinvagination (author's transl)]. The authors report a case of retrograde invagination of the duodenum 8 years after vagotomy and gastroenterostomy: gastric aspiration then reoperation permitted verification of the disinvagination. Cure by gastro-enterostomy and excision of the remaining duodenal ulcer.


Quantitative analysis of steroid profiles from urine by capillary gas chromatography. I. Accuracy and reproducibility of the sample preparation. A method is described for the determination of steroid profiles from urine by means of gas chromatography using high-efficiency glass capillary columns. The accuracy and reproducibility of essential steps in the sample preparation (extraction of steroids and steroid conjugates by means of XAD-2, enzymatic hydrolysis with Helix pomatia juice, solvolysis in acidified ethyl acetate and alkali wash) are established using different endogenously labelled urine samples, obtained from normal subjects to whom labelled steroids had been administered. Preliminary results are given on the reproducibility of the derivatization procedure (formation of methoxime-trimethylsilyl (MO-TMS) ethers), the gas chromatographic analysis and the whole method. Two procedures for the purification of MO-TMS steroid derivatives are compared. Application of the method to urine samples of patients with various endocrine disorders is included.


[What can be expected from suspension of the bladder neck in effort incontinence of urine? 85 operated cases (author's transl)]. Among the large number of technics proposed to cure effort urinary incontinence in women, two types of operation are usually considered: --indirect cervico-cystopexy or colopexy (Marshall-Marchietti-Krantz operation or its derivatives); --operations of catapult type or suspension of the bladder neck, e.g. Goebell-Stoeckel's operation. The latter was chosen exclusively by us over the last ten years. We have thus grouped together 85 cases of effort urinary incontinence operated and followed up for a period of 1 to 11 years. At five years, this operation gave 82.85% good results and a delay of two years seemed to us the minimum necessary to assess these results. We noted 6 failures (7.05%) and 4 relapses (4.7%). Suspension of the bladder neck is often disputed and even denigrated. Some authors consider it is the best operation available especially in cases of relapse of effort incontinence [21]. Its technical difficulty has been reproached. Our objective thus double: --describe our technic of musculo-aponeurotic suspension of the bladder neck; --give our results emphasising the long term results. We thus hope to prove that this operation maintains its place in the treatment of effort incontinence.


[Report of 39 traumatic and iatrogenic lesions of the extra-cranial carotid artery (author's transl)]. The authors present 39 carotid lesions from various and sometimes unusual origins: ski accident, cervical irradiation. The lesions have a preferential topography according to etiology: internal carotid for blunt trauma and arteriographic lesions, common carotid for penetrating injuries, common and bifurcation for X-ray lesions. Arteriography and surgical exploration confirm this notion. Macroscopic lesions don't always accord to the classical aspect: thus, only one intimal lesion has been found in blunt trauma. Symptomatology is orientated by etiology: neurological symptoms in blunt trauma and X-ray lesions, vascular symptoms in penetrating injuries. The diagnosis has sometimes been made at a distance from the initial accident and in unusual circumstances, mainly in blunt trauma. The treatment has been a surgical one except for usual countra-indications. The results are favourable on young subjects and above all for particular lesions such as penetrating injuries and X-ray lesions. Lastly, the authors insist on the post-operative evolutivity which is often misunderstood, and on the interest of an angiographic control.


Comparative determination of plasma cholesterol and triacylglycerol levels by automated gas--liquid chromatographic and autoanalyzer methods. Plasma samples obtained during a prevalence study of hyperlipemia in a free living urban population were analyzed for total cholesterol and triacylglycerol content by automated high-temperature gas--liquid chromatographic (GLC) and automated colorimetric (Auto-Analyzer, AA11) methods. The analyses were done over a three-year period. The methods gave excellent overall correlation for both total cholesterol (r = 0.9811) and total triacylglycerols (r = 0.9739). Detailed comparisons of the results obtained by the two methods with natural samples over the entire concentration range, indicated that the GLC method gave cholesterol values 5--10 mg% lower and triacylglycerol values 10--20 mg% lower than the corresponding AA11 determinations. The differences between the two methods are attributed to an overestimation of the cholesterol and triacylglycerol levels by the AA11 method due to presence of variable amounts of interfering chromogens in the plasma extracts. The between-method relative error ranged from 3 to 5% for cholesterol and from 5 to 10% for triacylglycerols. The within-day standard deviation of GLC averaged 2.3 mg% for cholesterol and 3.5 mg% for triacylglycerols. The between-day standard deviation of the GLC method averaged about 6 mg% for both cholesterol and triacylglycerols. The within-day, within GLC, relative error averaged 1.12% for cholesterol and 2.66% for triacylglycerols. The apparent high precision and high accuracy of the GLC method recommend it as an alternative to the indirect methods of plasma cholesterol and triacylglycerol analysis, especially where a smaller throughput of samples is not a limitation and where both total amount and composition of the lipids is of interest.


[Radiological anatomy of the spermatic veins. Report of 152 retrograde spermatic phlebographies (author's transl)]. The authors analyse the results of 152 retrograde phlebographic explorations in the investigation of varicocoele: anatomical variations of the right and left spermatic veins (site of arrival, number of venous trunks, existence or not of valvules). They emphasise the importance of anastomoses with other venous systems, not only the inferior vena cava, but also the portal veins. These anastomoses which are essential, explain certain relapses of varicocoele after surgery, and explain the spontaneous portacaval anastomoses which form through the genital veins in certain cases of portal hypertension.


Simplified method for monitoring tricyclic antidepressant therapy using gas--liquid chromatography with nitrogen detection. A simplified gas chromatographic method for the rapid measurement of tricyclic antidepressant drugs in plasma using a nitrogen-sensitive detector is described. All drugs are extracted and chromatographed under identical conditions. Tertiary amines are separated from their secondary amine metabolites, which are determined simultaneously without the need for derivatisation. The lower limit of accurate determination for most drugs is 10 microgram/1. The method has been applied to the routine measurement of amitriptyline and nortriptyline in plasma from patients receiving antidepressant treatment. Large and important interindividual differences in plasma concentrations in the patients investigated have been found, and the significance of these results is discussed.


[Extra-corporeal circulation. Experimental study. Comparative study of various oxygenators (author's transl)]. An experimental study of three different types of oxygenator was carried out in the dog during two hours extra corporeal circulation. The hemodynamic aspects and the characteristics of pulmonary elasticity that a previous study permitted us to choose as controls of the quality of extracorporeal perfusion, were studied. The results show that the pulmonary arterial resistances and the indices of pulmonary compliance do not recover during E.C.C. to their initial values, in the case of an E.C.C. using a bubble oxygenator. These results should be taken into consideration in the choice of the most suitable oxygenator.


[Volvulus of the gall bladder. Report of one case (author's transl)]. A case of torsion of the gallbladder is presented. Torsion of the gallbladder is uncommon and should be remembered in right upper quadrant acute abdomens, especially in elderly, asthenic individuals.


Improved method for the rapid determination of isosorbide dinitrate in human plasma and its application in pharmacokinetic studies. A rapid, accurate and highly sensitive method was developed for the determination of isosorbide dinitrate in human plasma. Concentrations in the lower nanogram and subnanogram range are determined by a one-step extraction of 2 ml plasma, containing 4 ng/ml nitroglycerine as internal standard, with 5.5 ml n-pentane. The extract is subjected to gas--liquid chromatography--electron capture detection analysis. The lower limit of quantitation is 200 pg/ml, but concentrations as low as 50 pg/ml are still detectable. The method allows the quantitative determination of isosorbide dinitrate plasma levels in man following a 5 mg sublingual administration up to four hours after application.


[Coagulation disturbances after subtotal or total hepatectomy in the dog (author's transl)]. The authors relate 15 cases of blood coagulation disturbances among three groups of animals after 80%, 90% or total hepatectomy. Animals in group I, after 80% hepatectomy, were sacrified between the 10th and 30th postoperative days. Group II animals, with 90% hepatectomy, survived for periods ranging from 18 to 24 hours. In group III, after total hepatectomy, survival varied between 30 and 40 hours. The coagulation disturbances which were observed following 80% hepatectomy were spontaneously compatible with the survival of the animals. Following 90% and total hepatectomy the resulting disturbances were constantly severe enough to limit survival time. Supportive procoagulant therapy associated with sequential heparin administration seems justified by the insufficient biosynthesis of clotting factors and their unavoidable consumption as a result of localized then disseminated intravascular coagulation and secondary fibrinolysis.


[Ileal and appendicular endometriosis (author's transl)]. A bibliographical review it's been made on the base of a double rara localization of endometriosis: ileal and appendicular, marking the diagnostic difficulties found appendicular, marking the diagnostic-difficulties found before and after surgery, emphasizing the interest of the histological integrity and motivity of intestinal mucosa.


Selective ion monitoring of tryptophan, N-acetyltryptophan and kynurenine in human serum. Application to the in vivo measurement of tryptophan pyrrolase activity. A specific method is described for the determination of deuterated and non-deuterated N-acetyltryptophan, tryptophan and kynurenine in human serum and urine using gas chromatography-mass fragmentography. N-Acetyltryptophan was analysed as the N-trimethylsilyl methyl ester derivative; tryptophan and kynurenine were converted into their N-pentafluoropropionyl methyl esters. N-Acetyl-DL-tryptophan-d11, tryptophan-d8 and kynurenine-d2 were used as internal standards. The coefficients of variation were found to be about 8% (n = 9) for tryptophan and N-acetyltryptophan and about 2.4% (n = 9) for kynurenine. Using this method, an in vivo determination of the tryptophan pyrrolase activity [L-tryptophan oxygen 2,3-oxidoreductase (decyclizing), E.C. 1.13.11.11] is possible by loading the subjects with deuterated L-tryptophan-d5 and subsequently measuring the deuterated L-kynurenine-d4 formed and the residual L-tryptophan-d5.


Fluorimetric determination of erythromycin and erythromycin ethylsuccinate in serum by a high-performance liquid chromatographic post-column, on-stream derivatization and extraction method. The method described is capable of detecting less than 0.01 microgram/ml of erythromycin and/or erythromycin ethylsuccinate in serum. Recoveries of erythromycin and erythromycin ethylsuccinate, when added to the level of 0.6 microgram/ml, were 102 and 97% with relative standard deviations of 6.0 and 4.5%, respectively. Erythromycin ethylsuccinate in serum at 37 degrees was shown to degrade rapidly to erythromycin in a first-order rate. When stored at -20 degrees, however, only 10% of the erythromycin ethylsuccinate in serum was hydrolyzed even after storage for 36 days. The method was used to analyze erythromycin and erythromycin ethylsuccinate in sera from ten subjects administered with an oral dose of erythromycin ethylsuccinate. In addition to a small amount of anhydroerythromycin ethylsuccinate and 8,9-anhydro-6,9-hemiketal erythromycin ethylsuccinate, at least two other metabolites were detected in sera and one was tentatively identified as erythralosamine.


Techniques for detecting enzymes in high-performance liquid chromatography. Techniques are described for the automated detection of a series of enzymes in a high-performance liquid chromatographic system. Detection was achieved by either a direct or a coupled enzyme assay using photometric detectors. In direct detection the immediate enzymatic product was monitored. Coupled enzyme assays required additional enzyme(s) to convert the product of the primary enzyme reaction into a more easily detectable form. The efficiency of both free and immobilized coupling enzyme(s) was evaluated. The detector sensitivity could be increased three-fold by increasing the reaction temperature. This system is particularly suitable for isoenzyme profiling in biological materials.


Simultaneous determination of antiarrhythmia drugs by high-performance thin-layer chromatography. A method is described for the determination of antiarrhythmia drugs in serum by high-performance thin-layer chromatography. Baseline separations are achieved for all the drugs and clozapine, an internal standard, in two developments with solvents of different polarity. Lidocaine and diphenylhydantoin are scanned at 220 nm after the first development. Procainamide, propranolol and quinidine are scanned at 290 nm after the second development. The relative standard deviation of the determination varies between 3 and 14% depending on the nature of the drug and its concentration.


The determination of sucrose in molasses by high-performance thin-layer chromatography. A method is described for the determination of sucrose in molasses by high-performance thin-layer chromatography. A baseline separation of sucrose is achieved in about 40 min using acetone--water (90:10) as developing solvent. The relative standard deviation of determinations by this method is about 2.2%. Results agree with those obtained by isotope dilution analysis of the same molasses samples.


Study of water treatment effects on organic volatiles in drinking water. Volatile organics were determined during all stages of the water treatment process in a municipal water works. Analytical parameters for a quantitative procedure were investigated. The method used allows simultaneous determination of both halogenated and non-halogenated purgeable organics. Approximately 70 substances were identified by gas chromatography--mass spectrometry with capillary columns. Well water contained considerable quantities of aliphatic and aromatic substances. Halocarbons were formed during both steps of a double chlorination procedure, but the largest quantity was generated during the first step. Only tetrachloroethylene was present in untreated surface water in significant quantities. Most hydrocarbons were carried through the treatment process from the raw water to the finished water with little change in concentration.


Assessment of availability and efficacy of commercial Salmonella grouping antisera. Salmonella somatic antisera for groups A to E were purchased from four commercial producers directly by the Center for Disease Control (CDC) and indirectly through two hospitals. CDC specifications and methods were used to evaluate antisera shipped directly to CDC. To assess the performance of the products under simulated user conditions, we used the commercial antisera purchased indirectly through the hospitals to group coded cultures. Of the 23 antisera ordered by CDC and the hospitals, the CDC received all 23, a large medical complex received 20, and a private hospital received 9. Similar results were obtained with producer and CDC test methods. Forty-five different lots of antisera were evaluated, of which 20% did not meet CDC specifications. The CDC specifications and latest revisions are discussed.


Argon detector: alternative detection system for gas-liquid chromatographic analysis of short-chain organic acids. Two detection systems for gas-liquid chromatography were compared for the identification of metabolic end products (short-chain organic acids) from anaerobic bacteria. Argon and flame ionization detectors were connected in series with inert argon as the carrier gas for analysis of 35 stock strains and 148 clinical isolates. There was an excellent correlation between the argon and flame ionization chromatograms, and both were readily comparable to the thermal conductivity detector tracings published in the Virginia Polytechnic Institute manual. The linear response of the argon detector was examined by analyzing twofold serial dilutions of the short-chain organic acids and comparing the results to the very linear flame ionization detector. The argon detector was found to react with sufficient linearity within the organic acid concentration range normally obtained from anaerobic bacterial broth cultures. The argon detector, therefore, appears to be a viable alternative to the flame ionization and thermal conductivity detector systems in the identification of anaerobic organisms.


Identification of atypical mycobacteria by thin-layer chromatography of their surface antigens. The knowledge that the surface (Schaefer) antigens of certain smooth-colony atypical mycobacteria are multiglycosylated C-mycosidic peptidoglycolipids was used to devise a sensitive thin-layer chromatographic (TLC) procedure for the identification of Mycobacterium avium/M. intracellulare/M. scrofulaceum serotypes. TLC maps of the type-specific peptidoglycolipids from 17 of the 31 serotypes are presented. The primary use of the technique is to corroborate results obtained by seroagglutination. Without the aid of seroagglutination, the TLC procedure almost invariably requires the availability of reference strains or the specific peptidoglycolipids derived therefrom.


Electron capture gas-liquid chromatographic study of metabolites produced by some arthritic transudate-associated organisms in vitro and in vivo in rabbit models. Computerized, frequency-pulsed, modulated electron capture gas-liquid chromatography was used to study the acid metabolites produced in vitro in fetal calf serum and in vivo in an animal chamber model. Several strains of Diplostreptococcus agalactiae, Propionibacterium acnes, Staphylococcus aureus, and Streptococcus serogroups A, B, and G were studied. All of these organisms have been reported to be associated with arthritic transudates in humans. Metabolites were detected by this method from derivatized extracts of both spent fetal calf serum and chamber fluids. Since there was little host response to the organisms cultured in the chambers, it is highly probable that the products detected represent metabolites produced in an in vivo type of environment. The metabolic patterns were reproducible and exhibited many similarities in vitro and in vivo. Production of the acids detected was reproducible, and these acids were useful identification markers. The data support published reports (J. B. Brooks, C. C. Alley, and J. A. Liddle, Anal. Chem. 46: 1930-1934, 1974; J. B. Brooks, G. Choudhary, R. B. Craven, D. Edman, C. C. Alley, and J. A. Liddle, J. Clin. Microbiol. 5:625-628, 1977; J. B. Brooks, R. B. Craven, A. R. Melton, and C. C. Alley, in H. H. Johnson and W. B. Newson, ed., Second International Symposium on Rapid Methods and Automation on Microbiology, 1976; J. B. Brooks, R. B. Craven, D. Schlossberg, C. C. Alley, and F. M. Pitts, J. Clin. Microbiol. 8:203-208, 1978; J. B. Brooks, D. S. Kellogg, C. C. Alley, H. B. Short, and H. H. Handsfield, J. Infect. Dis. 129:660-668, 1974) that bacterial metabolites might be detectable in diseased body fluids. The growth characteristics of the organisms in the animal model and fetal calf serum are discussed, and a moderately priced computer for performing data manipulations is evaluated.


Infections caused by Klebsiella ozaenae: a changing disease spectrum. A total of 64 isolates of Klebsiella ozaenae were recovered from 36 patients during a 40-month period. Over 7,500 isolates of K. pneumoniae were isolated during the same time period. Before this decade, K. ozaenae was considered to be only a colonizer of the nasopharynx or a putative cause of ozena (atrophic rhinitis). K. ozaenae was recovered most frequently from sputum in mixed culture but was associated with infection in 12 patients (2 with bacteremia, 3 with urinary tract infection, 1 with soft tissue infection, and 6 with mucopurulent nasal discharge). The spectrum of disease caused by this organism is more extensive than has been appreciated previously.


Thermotolerance of Wangiella dermatitidis. A variety of diagnostic tests used by many laboratories to identify isolates of Wangiella dermatitidis (= Fonsecaea dermatitidis) were evaluated. Thirteeen isolates of W. dermatitidis were studied with respect to their ability to grow at 25, 37, 40, 45, and 50 degrees C, colonial and micromorphology, gelatin liquefaction, and hydrolysis of casein, xanthine, hypoxanthine, and tyrosine. All 13 isolates showed growth at 25, 37, and 40 degrees C but failed to grow at higher temperatures. The ability of W. dermatitidis to grow at 40 degrees C can be useful in its identification.


Comparison of immunogenicity of a whole virion and a subunit influenza vaccine in adults. The immunogenicity and reactogenicity of a whole virus (Merck Sharp & Dohme) and a subunit (Wyeth) influenza A/New Jersey/76 vaccine were compared in a group of 214 normal adult subjects. Both the seroconversion rate and the magnitutde of hemagglutination inhibition antibody response were significantly (P less than 0.01) lower in the recipients of the subunit vaccine, whereas there were no significant differences in local or systemic reactions between the two preparations. On the basis of these data, we question the previous Public Health Service recommendation that one dose of either preparation of the influenza A/New Jersey/76 vaccine is equally efficacious in individuals over 24 years of age.


Clinical pharmacologic observations on timolol. I. Disposition and effect in relation to plasma level in normal individuals. The disposition profiles of a new beta-adrenergic blocking drug, timolol, were investigated at 11 different times in normal individuals after a single oral dose. The disposition of timolol follows first-order kinetics and may be adequately described by a one-compartment model. The pharmacokinetic data were not significantly different among the three dose levels examined. After the single oral doses, overall elimination half-life was 3.2 +/- 0.2 hours (mean +/- S.E.M.), with an observed peak time of 2.0 +/- 0.2 hours; extrapolated volume of distribution was 1.81 +/- 0.15 liter/kg; and the total plasma clearance was 557 +/- 61 ml/min. Approximately 20 per cent of elimination from the human body was dependent on the kidney. The area under the curve from zero to infinity and the peak concentration observed were dose dependent. A linear relationship was found between timolol plasma concentrations and beta-adrenergic blocking effects (per cent inhibition), as estimated from exercise-induced tachycardia. Timolol is a beta blocker which must await further clinical trials for the assessment of therapeutic implications in relation to plasma levels.


Clinical pharmacologic observations on timolol. II. Antihypertensive effect and kinetic disposition on twice-daily dosing in patients with mild or moderate hypertension. Timolol given on a twice-daily schedule has shown both antihypertensive effectiveness and plasma renin-suppressing action in eight patients with mild or moderate hypertension. However, the causal relationship between the drug plasma level, blood pressure fall, and change in plasma renin activity was not so clearly demonstrated in the present study. The disposition profiles of timolol at the steady state show an elimination half-life of 4.7 +/- 0.7 hours and a total plasma clearance of 225 +/- 21 ml/min. These values are found to be different from those of our previous observations obtained in normal individuals. Although the range of mean timolol concentrations at steady state varies to a certain extent among different patients, the dosage regimens for patients who will receive treatment for certain chronic disease states (e.g., arrythmias, obstructive cardiomyopathy, and angina pectoris, but not hypertension) in relation to plasma levels should be based on the disposition data obtained under steady state conditions. Bronchospasm developed in one of eight patients whose timolol level was found to be higher than the average of other patients given the same dosage.


Interrelationships among thiazide diuretics and calcium, magnesium, sodium, and potassium balance in normal and hypertensive man. The benzothiadiazide diuretics are known to cause magnesium and potassium wasting and retention of calcium and uric acid. The effect of magnesium replacement on diuretic-induced changes in sodium, potassium, calcium, and magnesium balance, weight, and blood pressure was studied in 12 normal subjects and 13 hypertensive patients to determine whether oral magnesium would reduce thiazide-induced kaliuresis. The groups differed in their response to diuresis in that the normotensive subjects did not show a fall in blood pressure (despite an equivalent weight loss), returned to a state of sodium balance earlier, and developed a greater net negative potassium balance over 12 days of diuretic therapy. Both groups displayed calcium and uric acid retention and magnesium wasting during diuretic therapy. The addition of oral magnesium replacement during diuretic therapy had no effect on any measured values beyond that seen in subjects who took diuretics without magnesium replacement. Thus, magnesium replacement did not reduce urinary potassium loss. However, urinary calcium losses increased when magnesium was given to subjects who were not receiving diuretics. The data provide the first report that small doses of oral magnesium enhance calcium excretion in hypertensive subjects.


Direct and indirect measures of patient satisfaction with physicians' services. This research examines alternative measures of patient satisfaction. Three measures were compared: (1) a direct measure to evaluate how the patient felt about his own personal physician, (2) an indirect measure that assessed attitudes about physicians in general, and (3) a measure designed to be intermediate between these two. Responses to the three measures were found to differ: the levels of satisfaction increased with the directness of the measure used; indirect evidence that this relationship could not be attributed solely to a patient's reluctance to criticize his own physician is also provided. The three measures were compared in terms of their association with other assessments of outcome and with indicators of the process of care. Although none of the associations was statistically significant, high scores on the intermediate measure tended to correspond with better outcomes and higher scores on the process of care. These findings are at least compatible with the contention than an intermediate measure provides the most valid assessment of patient satisfaction.


Plasma concentrations of isosorbide dinitrate after cutaneous and sublingual doses to human subjects. After application of 100 mg (nominal dose) isosorbide dinitrate as an ointment to the skin of human subjects, mean drug concentrations were 1 to 2 ng/ml for 1.5 hour and reached a peak of 6.2 ng/ml at 6 hours. Thereafter, mean concentrations declined slowly to 2.9 ng/ml at 12 hours and 1.2 ng/ml at 24 hours. After a sublingual dose of 5 mg isosorbide dinitrate, mean drug concentrations reached a peak of 15.9 ng/ml at 0.5 hour and declined with a half-life of about 50 minutes. The mean bioavailability of isosorbide dinitrate from the ointment was estimated as 30 per cent of that from the sublingual tablet when corrected for differences in dose/body weight ratio. The results demonstrate that concentrations of isosorbide dinitrate in plasma can be maintained for relatively long periods when the drug is applied to the skin.


An assessment of emergency medical technicians' performance as related to seasonal population  influx. The periodic influx of large numbers of people into resort areas substantially increases the use of emergency medical services. This investigation assesses the effects of such a threefold increase in the summer population of the Cape Cod area upon the accuracy of emergency medical technicians' diagnoses and treatments. The technicians' diagnoses for ambulance patients were evaluated against those given by the emergency room physicians during the months of August 1975 and February 1976. The distribution of conditions was similar for both months and the observed frequency of correct diagnoses for common conditions was more than 90% in both months. The overdiagnosis rate of 25% to 50% for common conditions and the correct treatment rate for suspected myocardial infarction of 65% did not vary significantly between summer and winter. Thus, a large influx in population does not seem to affect adversely EMT diagnosis rates. Although misdiagnoses were uncommon, a failure to follow through with a correct treatment for patients with suspected myocardial infarctions, thus indicating the need for better quality control on EMT performance.


Assessing the social impacts of medical technologies. The introduction of a new medical technology can have indirect, unintended, or unanticipated effects on individuals or on social systems. Although these impacts result from the widespread use of the technology, many of them can be predicted while the technology is being developed. A method for systematically identifying and evaluating these impacts is technology assessment. Such an assessment, made while a technology is being developed, could provide useful information for decision making about research and development and in planning for the technology's eventual introduction. Because only a few medical technologies have been formally assessed and because the process of medical technology development is porrly understood, one must be cautious in recommending widespread use of technology assessment. Nevertheless, enough is now known to permit the formulation of approaches that could be used in preliminary attempts at medical technology assessment.


Consumer health education in New Jersey community hospitals: a medical school initiative. The Office of Consumer Health Education (OCHE) of the College of Medicine and Dentistry of New Jersey (CMDNJ) has developed a statewide network of hospital-based consumer health education programs. The rationale for a hospital-based program and the criteria for hospital selection emphasize the responsibility of health care institutions in providing patient and community health education. OCHE offers technical and consulting services to these and other hospitals in the areas of adult education, community programming, evaluation, and resource materials: it also participates in the teaching of medical students at CMDNJ. The need for each hospital to individualize its programs has been demonstrated, and eight problem areas have been revealed by OCHE'S experiences. Future financing will depend largely on third-party reimbursement for in-hospital patient education, while some community projects will be able to receive support from tuitions and grants.


The preceptorship, an integral unit of the curriculum in community and family medicine. A curriculum in community and family medicine at the University of Massachusetts Medical School, planned to include a primary care preceptorship as an integral unit, is described. The curriculum has been planned to allow for repeated and increasing exposure of medical students during their undergraduate years to a variety of health care settings within the state. An extensive program of courses is required for all students and consists of a three-week field clerkship in the first year, a field-oriented epidemiology course in the second year, and a six-week field clerkship in the third or fourth year. A preceptorship elective is available to all medical students after they have completed their first two years. Field visits by community and family medicine faculty to the preceptorship site provide overall guidance, facilitate the implementation of objectives, and provide opportunities to strengthen bonds between the practicing physicians of the Commonwealth of Massachusetts and the state medical school. The results to date are discussed.


Assessing technical performance at diverse ambulatory care sites. The purpose of the large study reported here was to develop and test methods for assessing the quality of health care that would be broadly applicable to diverse ambulatory care organizations for periodic comparative review. Methodological features included the use of an age-sex stratified random sampling scheme, dependence on medical records as the source of data, a fixed study period year, use of Kessner's tracer methodology (including not only acute and chronic diseases but also screening and immunization rates as indicators), and a fixed tracer matrix at all test sites. This combination of methods proved more efficacious in estimating certain parameters for the total patient populations at each site (including utilization patterns, screening, and immunization rates) and the process of care for acute conditions than it did in examining the process of care for the selected chronic condition. It was found that the actual process of care at all three sites for the three acute conditions (streptococcal pharyngitis, urinary tract infection, and iron deficiency anemia) often differed from the expected process in terms of both diagnostic procedures and treatment. For hypertension, the chronic disease tracer, medical records were frequently a deficient data source from which to draw conclusions about the adequacy of treatment. Several aspects of the study methodology were found to be detrimental to between-site comparisons of the process of care for chronic disease management. The use of an age-sex stratified random sampling scheme resulted in the identification of too few cases of hypertension at some sites for analytic purposes, thereby necessitating supplementary sampling by diagnosis. The use of a fixed study period year resulted in an arbitrary starting point in the course of the disease. Furthermore, in light of the diverse sociodemographic characteristics of the patient populations, the use of a fixed matrix of tracer conditions for all test sites is questionable. The discussion centers on these and other problems encountered in attempting to compare technical performance within diverse ambulatory care organizations and provides some guidelines as to the utility of alternative methods for assessing the quality of health care.


Nonmetropolitan consumers attitudes toward physician advertising. The debate over whether physicians should be allowed to advertise their services has not yet included a consideration of consumer attitudes based upon empirical data. The results of a telephone survey of residents of a nonmetropolitan area reveal that the respondents are almost evenly divided between opposition to and support of physician advertising. Most respondents do not specify a reason for their position. Attitudes do not differ when several background variables, including residence, age, education, sex, usual source of care, and preferred source of care, are controlled. However, a significant negative association is observed between attitude toward physician advertising and length of residence. This finding suggests that if physician advertising is permitted it may have its greatest impact in areas with high rates of residential turnover.


Affiliations with medical care providers. Two analytical techniques are applied to a household survey of Rhode Island residents to develop the concept of an affiliation with medical care provider. A modification of a deviant case analysis is used to examine those people who are extreme in terms of the numbers of affiliations that they have with medical care providers. Overall, 4.1% of the sample have no affiliations and 7.3% have four or more affiliations. One of the most important variables in distinguishing those people with an unusual number of affiliations is a subjective variable of health status--whether they worry about their health. Nonparticipants rarely worry about their health and are generally in good health, while those with a high number of affiliations worry a great deal, regardless of their actual health status. A multiple regression analysis reveals that the variables of number of health conditions, sex, insurance, worry about health, income, disability days, and family structure are significantly related to mean number of affiliations with an R2 of 19.5%. Discussion includes the study's implications both for greater understanding of how and why people seek care and for application in the further development of a social psychological model of health behavior.


Epidemiological applications to health services research. A model is presented to illustrate some areas in which epidemiological thinking may be applied to health services research. The model is neither exhaustive nor exclusive, for other areas not specifically described as within the model are also a part of the domain of epidemiology, and several aspects of the model are shared with researchers in other disciplines. Selected examples of epidemiological applications are reviewed; these include: (1) the need for health care services, (2) the use of health services, (3) the impact of health programs on utilization patterns and on health status, and (4) the impact of therapeutic services on outcomes for specific diseases. Additional health services issues that are relevant to epidemiology are suggested.


The formation and evaluation of a graduate program in health administration within a medical school. Several aspects of the program in health administration that was established in 1968 by the faculty of the University of Colorado School of Medicine are described. Based on the premise that knowledge about the organization and the delivery of health services had not kept pace with the advances made in the scientific foundations of medicine, the program was designed to provide training in management, accounting, economics, medical care organization, quantitative methods, and in the other skills required for health administration. The results of a survey to identify the extent to which the program's graduates have fulfilled the original objectives are presented, including a description of the graduate's backgrounds and current job situations. The lessons learned from five year's experience teaching health administration within a medical school are also reviewed.


Termination of electroreceptor and mechanical lateral line afferents in the mormyrid acousticolateral area. The projection regions of electroreceptor and mechanical lateral line afferents in electric fish of the mormyridae family are described. Electroreceptor afferents from the posterior dorsal skin run in the dorsal branch of the posterior lateral line nerve. Electroreceptor afferents from ventral skin and mechanical lateral line afferents and efferents run in the ventral branch of the nerve. Horseradish peroxidase (HRP) injections into each branch resulted in filling of its central terminals with the marker enzyme. The method yields a Golgi-like staining of afferent terminals, allowing some aspects of their morphology to be described. Comparison of results from dorsal and ventral branch injections shows the separate medullary regions to which electroreceptor and mechanical afferents project, and also demonstrates four separate somatotopic maps within the electroreceptor region. Mechanical afferents end predominantly ipsilaterally in nucleus anterior and eminentia granularis as has been suggested by others. Ipsilateral endings in nucleus octavius are also seen. Electroreceptor afferents end exclusively in the cortex and nucleus of posterior lateral line lobe (PLLL). Within the cortex there are three distinct maps of the skin surface which are separated from each other by discontinuities in the cellular layers. Somatotopic mapping is also present in the nucleus of PLLL though it is less precise than in the cortical zones. Large club endings of the cells of this nucleus are filled with HRP. Labeled cells are seen within a small midline nucleus located at the level of the eighth nerve just above the medial longitudinal fasciculus. These are probably the cell bodies of lateral line efferents.


A Golgi study on the subthalamic nucleus of the cat. The subthalamic nucleus (ST) of kittens was studied by means of the rapid Golgi silver impregnation method. The neurons of the ST were classified into three types. Type I neurons, the main constituents of the ST, had oval or polygonal cell bodies with a mean diameter of 26 micrometer by 36 micrometer and four to six primary dendrites. Type II neurons had multipolar or polygonal cell bodies, which measured an average 31 micrometer by 43 micrometer and emitted four to seven primary dendrites. The cell bodies of the type III neurons were polygonal in shape, measured 23 micrometer by 26 micrometer in average and emitted four to six primary dendrites. The dendrite bundle and the dendrite pallisade were observed. Frequently dendrites of the ST extended into the cerebral peduncle (CP), and even cell bodies of some ST neurons were located within the CP. All of the parent axons of the ST neurons coursed rostrally, although intra- and extra-nuclear axon-collaterals arising from the ST neurons travelled rostrally, caudally or caudomedially. The afferent fibers to the ST were divided into three groups; afferents via the Meynert's commissure (MC), decending and ascending afferents. The MC fibers, which ran across the CP, gave terminals to the ST. The descending afferents were axon-collaterals of fibers descending in the CP and those of fibers running through the ST. The ascending afferents were also axon-collaterals arising from ascending fibers in the CP. In addition to these afferents, many descending and ascending fibers of passage ran through the ST without emitting axon-collaterals.


The organization of monoamine-containing neurons in the brain of the sunfish (Lepomis gibbosus) as revealed by fluorescence microscopy. The morphological organization of the monoamine-containing neurons in the brain of the sunfish (Lepomis gibbosus) was studied by means of the Falck-Hillarp histofluorescence method. No attempt was made to distinguish between norepinephrine and dopamine, both primary catecholamines (CA) yielding a similar yellow-green fluorescence after paraformaldehyde treatment. In the brain stem of this teleost fish, three groups of CA-containing neuronal somata have been found. First, there is a small collection of CA perikarya located just caudal to the obex of the fourth ventricle. The neurons of this medullo-sinal group give rise to numerous CA fibers many of which ascend within the central portion of the medulla. Intermingled with these CA fibers are some CA cells that constitute the central medullary group. The CA perikarya of this group are scattered between the levels of cranial nerves X and VIII. The tegmentum of the isthmus also contains a small group of very closely packed CA neurons. The large-sized CA cells of the isthmal group are located dorsolateral to the medial longitudinal fasciculus, partly within the periventricular gray. High densities of CA varicosities were also disclosed in various brain stem structures such as the optic tectum, the torus semicircularis and the cerebeller valvula. In addition, numerous serotonin (5-HT)-type neuronal somata were found in the raphe region of the brain stem, particularly at caudal mesencephalic, isthmal and rostral medullary levels. A large number of CA cell bodies were visualized in the sunfish hypothalamus. Most of them form two populations of small, round cells that are located along and partly within the ependymal walls of the posterior and lateral recesses of the third ventricle. These bipolar cells possess one short club-like process protruding into the ventricle and their thin ependymofugal processes contribute to the CA innervation of numerous hypothalamic regions. Large CA neurons apparently without direct CSF contact also occur in the area of nucleus posterior tuberis, at the level of the mesodiencephalic junction. Although the hypothalamic inferior lobes are devoid of CA cell bodies they are heavily innervated by CA axons. The sunfish telencephalon also receives a strikingly massive and complex monoaminergic innervation. Numerous CA fibers which are first observed at the level of the preoptic area, ascend through the central zone of the telencephalon and arborize profusely particularly within the medial zone of area dorsalis telencephali. Other CA fibers, as well as abundant fine 5-HT varicosities were found in the lateral zone of area dorsalis. Although the exact origin of the telencephalic CA afferents in Lepomis is not known, part of it may arise from the isthmal CA cell group which appears similar to the locus coeruleus of reptiles, birds and mammals.


Ascending connections to the forebrain in the Tegu lizard. The ascending connections to the striatum and the cortex of the Tegu lizard, Tupinambis nigropunctatus, were studied by means of anterograde fiber degeneration and retrograde axonal transport. The striatum receives projections by way of the dorsal peduncle of the lateral forebrain bundle from four dorsal thalamic nuclei: nucleus rotundus, nucleus reuniens, the posterior part of the dorsal lateral geniculate nucleus and nucleus dorsomedialis. The former three nuclei project to circumscribed areas of the dorsal striatum, whereas nucleus dorsomedialis has a distribution to the whole dorsal striatum. Other sources of origin to the striatum are the mesencephalic reticular formation, substantia nigra and nucleus cerebelli lateralis. With the exception of the latter afferentation all these projections are ipsilateral. The ascending connections to the pallium originate for the major part from nucleus dorsolateralis anterior of the dorsal thalamus. The fibers course in both the medial forebrain bundle and the dorsal peduncle of the lateral forebrain bundle and terminate ipsilaterally in the middle of the molecular layer of the small-celled part of the mediodorsal cortex and bilaterally above the intermediate region of the dorsal cortex. The latter area is reached also by fibers from the septal area. The large-celled part of the mediodorsal cortex receives projections from nucleus raphes superior and the corpus mammillare.


Relative numbers of several types of synaptic connections in the substantia gelatinosa of the cat spinal cord. The relative numbers of axo-dendritic, axo-axonic, dendro-axonic and dendro-dendritic synapses were determined by classifying and recording all such specialized contacts in sample areas of the substantia gelatinosa. The samples were taken from segments L1-L5 of the cat spinal cord. In the glomerular complexes 97% of all synapses were recorded as axo-dentritic. In substantia gelatinosa deprived of glomerular complexes by dorsal root section, 96.5% were axo-dendritic. The remainders were about equally divided between axo-axonic, dendro-dendritic and dendro-axonic synapses.