Rapid micromethod for flourometric determination of neutral lipids on thin-layer chromatograms. A rapid fluorometric procedure for in situ evaluation of neutral lipids on thin-layer chromatograms is described. The lipids are rendered fluorescent by treatment with perchloric acid, and the intensity of the emitted light is recorded by flying spot scanning. Detection limits (2.8 x SD) are 0.10, 0.11, and 0.17 nmol for triglycerides, cholesterol, and cholesterol esters, respectively. Range of linearity is 0--3.4 nmol for triglycerides, 0--2.0 nmol for cholesterol, and 0--4.0 nmol for cholesterol esters. When standard in triplicate and 2 samples of each unknown are applied to the chromatogram, standard error of the mean is 2.7% for triglycerides, 1.7% for cholesterol, and 1.0% for cholesterol esters. The sensitivity of the method allows simultaneous determination of the three lipid classes in 5 microliter lipid extract representing 0.238 microliter human plasma.


Modified procedures for the determination of hexuronic acids, hexoses and proteins using the Auto Analyzer II system. Previous automated procedures for the determination of hexuronic acids, hexoses and proteins have been modified to suit the Technicon Auto Analyzer II system. The present methods were highly reproducable and the detection limits showed to be in the order of 10 micrograms/l (hexuronic acids and hexoses) and 60 micrograms/l (protein) when 0.13--0.34 ml of the sample was used.


The role of ion shifts in cell injury.

The role of ion shifts in cell injury. The present paper reviews studies which utilize x-ray microanalysis to determine intracellular ion shifts following several types of cell injury. New data from our own laboratory on several cell injury systems are discussed. Concentration estimates are made by comparison of data from tissues with a series of standards prepared in 20% albumin followed by cryosectioning. Hemorrhagic shock in rats is followed by rapid changes of ions in both muscle and liver. These include increased levels of sodium and chlorine and decreased levels of potassium which can be correlated with deficits in the energy charge. Measurements made over hepatocellular carcinomas in the mouse, induced by safrole  show marked changes in comparison with non-transformed cells. These include striking increases in sodium and chloride and decreases in potassium and phosphorus which may be related to growth control. Studies on ischemia produced by arterial clamping in the rat kidney and the dog heart show somewhat similar changes. Moreover, in these models much interest is directed at early increases of cytoplasmic calcium with decreased mitochondrial calcium levels at later intervals. Following reflow, there is a prominent increase of calcium in the cytosol. These changes in calcium may be related to activation of phospholipases producing permeability changes which may contribute to further ion shifts as well as ultimately to cell death. The paper also comments on the use of cryostat sections for some types of routine pathological analysis.


Ultrastructure and x-ray microanalysis of macrophages exposed to cadmium chloride. Macrophages have a direct role in the inflammatory response to cadmium exposure. Cadmium is not only an important air pollutant, but is also one component of cigarette smoke. To study the effects of soluble cadmium on macrophages, two model systems were chosen:rabbit alveolar macrophages (RAMs) obtained by pulmonary lavage and peritoneal macrophages elicited by intraperitoneal injections of 10(7) viable M. bovis, bacillus Calmette Guérin (BCG MACs). Macrophages were maintained in standard tissue culture medium from 4 to 30 hours with concentrations of cadmium chloride (CdCl2) ranging from 0 to 1.0 mM. Attached macrophages and RAMs in suspension were studied by conventional transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy dispersive x-ray microanalysis (EDX). In addition to routine techniques for TEM and SEM, preparatory procedures included snap freezing in liquid propane, and either cryoultramicrotomy or freeze-substitution with 1% osmium tetroxide in acetone. By TEM many macrophages exhibited laminated nuclear inclusions at doses as low as 0.05 mM CdCl2) (at 20 hrs) and as early as 4 hrs (at 1.0 mM CdCl2). Sections of cells fixed by freezing exhibited the same nuclear inclusions as well as mitochondrial densities that were not visible with any other preparative technique. Cadmium was demonstrated in the nuclear inclusions and mitochondrial densities by EDX in snap frozen cells. Lesser amounts of cadmium were also detected diffusely in treated cell cytoplasm and nuclei. Cadmium was only detected by EDX in cells fixed by freezing. These studies document the localization of Cd in nuclear inclusions and mitochondria providing morphological support for the biochemical findings of other laboratories. In addition, the value of fixation by freezing over conventional chemical fixation is illustrated.


Correlated scanning and transmission electron microscopy of developing lens vesicle in the chick. Cells within the lens placode undergo transient surface modifications during the course of invagination. At stage 13, cellular apices bulge outward and display decreased surface diameters when compared to adjacent surface ectoderm. The lens vesicle is well invaginated by stage 14. An area of modified cells can be recognized ventral to the lens vesicle aperture. These cells present a smaller surface area and increased pleomorphism. Normal surface ectoderm extends to the dorsal and lateral edges of the aperture. The apical surfaces of cells which line the lumen of the lens vesicle display a complex topography such that individual cell boundaries are not easily distinguished. At stage 17, cellular processes which partially occlude the closing aperture of the lens vesicle may assist in reestablishing ectoderm over the newly formed lens. Preliminary results of cytochalasin B exposure on the invaginating lens vesicle suggest that the contractile action of microfilaments influences the complex cellular topography.


Laser microprobe mass analysis: achievements and aspects. After a short review of the general state of the art of laser microprobe analysis, recent achievements of the laser microprobe mass analyzer (LAMMA) are presented. The Lamma-instrument, which was primarily developed for biomedical purposes has been recently used on a large variety of specimens: biological, technical, organic and inorganic in nature. Detection limits, mass resolution and reproducibility have been improved so that for many elements measurements can be extended down to the subppm range. For bulk analysis a separate version of the LAMMA-instrument has been developed. The examples of LAMMA-applications given in this paper include muscle tissue (physiological cations), uterine tissue (Fe), retina tissues (Na, Mg, K, Ca and Ba), airborne particles (Pb, Fe, Al, Ti, Mn, Ca, Zn), cerebral tissue (Li), a fluorinated drug, pure crystalline organic materials (including nicotinic acid) and a selection of non biological materials (for instance thin films).


Scanning electron microscopy of hen granulosa cells before and after ovulation. Using scanning electron microscopy, I examined the granulosa cells of the hen (Gallus domesticus) before and after ovulation. Although the granulosa cells were separated during the final 7-8 days of oocyte maturation, they maintained cell-to-cell contact via cytoplasmic processes. The granulosa cells were cuboidal except at the animal pole (the site of the germinal vesicle) where they were columnar and had surface folds. These morphological characteristics were retained until germinal vesicle breakdown, 1-2 hr before ovulation. At that time the surface of granulosa cells at the animal pole was smoother and their cytoplasmic processes were less numerous and appeared atrophic. Similar changes occurred on the granulosa cells overlying the vegetal pole. About 30 min after ovulation, the granulosa, which remained adherent to its basal lamina, consisted of irregularly shaped columnar cells that lacked surface modifications. These smooth surface cells had rounded apical faces and were generally wider at the apical half. Postovulatory granulosa cells do not transform into luteal cells but undergo fatty degeneration and desquamation during the 4 days after ovulation.


The directions of ciliary beat on the wall of the lateral ventricle and the currents of the cerebrospinal fluid in the brain ventricles. The directions of ciliary beat on the wall of the lateral ventricle of the brain have been studied in the mouse by using a scanning electron microscope. As the studies on the directions of ciliary beat on the wall of the third and fourth ventricles were previously completed, a series of studies of the whole ventricular system has now been completed. The pattern of directions of the beat in the brain ventricular system was always the same in every individual examined in the series and this pattern was not contradictory to the anticipated flow of the fluid. Above all, the cilia on the wall of the brain ventricles beat spontaneously. Considering these facts, it was concluded that the ciliary beat has a close relationship with the flow of the cerebrospinal fluid, and conversely the detailed flow of the cerebrospinal fluid within the whole ventricular system was inferred from the ciliary beat.


The vascularization of the pineal gland (epiphysis cerebri) of the rat. The vascularization of the rat pineal gland was investigated with, respectively, latex and resin injection preparations of the cephalic blood vessels in adult Wistar rats. From the dissection of the latex-injected and aldehyde-fixed rat brains under the stereo light microscope it was found that the arterial supply comes exclusively from separate branches of the posterior cerebral artery. The pineal area was dissected out from the resin injected and corroded specimens under the stereo light microscope and was inspected in the SEM. The supplying arteries, identified on the basis of their endothelial cell imprint patterns, were 4 to 6 in number with a diameter of 20 to 40 micron. The venous drainage consisted of 12 to 16 short veins with a diameter of 40 to 60 micron, all draining directly into the great cerebral vein and thereafter via the sinus influens immediately into the systemic venous circulation.


Morphological and metabolic studies on adult cardiac myocytes. We have previously described detailed procedures for isolating rat and canine myocytes from adult ventricular tissue, and have investigated ultrastructural and functional properties of these cells. The effects of isolation on cell morphology were tested by comparative scanning (SEM) and transmission electron microscopy (TEM) of adult canine cardiac myocytes. Cells cleanly separated at the lateral sarcolemmal borders and at the intercalated discs. Since the cells were contracted during fixation, SEM of the lateral cell borders showed longitudinal folds and transverse ridges. There were rows of regular protrusions corresponding to the mitochondria immediately beneath the sarcolemmal surface. Openings in the sarcolemmal membrane occurred at regular intervals at the level of the Z-bands and may represent the sites of externalization of the transverse tubular system. By SEM, the intercalated disc junction appeared as a microvillar border and this corresponded directly to the scalloped appearance seen by TEM. A controlled sonication procedure was developed to allow specific removal of the sarcolemma without disruption of underlying organelles. This allowed visualization of the cellular organization of myofibrils, mitochondria and the transverse tubular system. These cells retain biochemical and enzymatic characteristics of adult dog cardiac tissues such as ouabain and insulin sensitivity, fatty acid and lipoprotein uptake and metabolism, and prostaglandin synthesis. This cytological preparation permits the correlation of these types of metabolic studies with the known functions of isolated subcellular organelles of the myocardium.


Raman microprobe studies of two mineralizing tissues: enamel of the rat incisor and the embryonic chick tibia. The laser-Raman microprobe developed at the National Bureau of Standards has been applied to the study of the mineralization process in rat incisor enamel and embryonic chick tibia. Cryostat sections were prepared from fresh frozen tissues and allowed to air dry. In these mineralizing tissues two forms of phosphorus compounds have been observed: (1) an inorganic phase identified as apatitic phosphate and (2) an organic phosphate. The distribution of these components from the mineralizing front to regions of higher mineralization has been determined with a spatial resolution of approximately 15 micrometer. The studies suggest the existence of a carbonate, with a Raman band corresponding to that of the mineral huntite, Mg3Ca(CO3)4, and found in regions of low phosphate mineral content.


Comparative investigations on fluorosed enamel. Structural observations upon fluorosed enamel were made on human teeth extracted from Nabeul (Tunisia). Longitudinal sections of enamel were made and microradiographed. Topographical observatoons in the S.E.M. showed alterations of the pattern of mineralization. Fissures and clefts filled with plaque material were observed to reach to the surface of the teeth. They penetrated into enamel along the prisms. Acid etching of the surface by 30% H3PO4 revealed enlarged acid-resistant sheaths while the core of the prisms was irregularly etched. Inner portions of the enamel were observed on fractured fragments. The structure was not significantly perturbated except for areas of porosity. After demineralization of the sections by 10% EDTA at pH 7 and critical point drying, the organic matrix was observed in the S.E.M. : It appeared as a fibrous network. Compared with sound teeth the matrix was dense and irregular. Histochemical stainings showed the presence of organic material. Alcian blue and PAS positive contents were very low except near the enamel-dentine junction. Preliminary observations were made using SMI Cameca ion microanalyser and histochemical studies on undecalcified sections were made in order to compare the observations made with light microscopy with S.E.M. findings.


Colonization of various natural substrates by osteoblasts in vitro. The ability of osteoblasts to transfer from normal rat growing skull bone to small fragments of various biological materials was tested in an organ culture system. The substrates were tested as overlays and included intact or anorganic cut, polished or fractured bone, osteoid, cementum, dentine and enamel. Osteoblasts migrated on to and over all these materials, whether collagenous or not and whether mineralized or not. The culture medium affected the cell shape, elongation, alignment and dorsal ruffling activity of the cells similarly on the original bone surfaces and on the overlays. The position of the cells on the overlay was influenced by constraints due to space available for each cell, sharp edges and corners, and fine textural patterns, in that order. It is concluded that the ability of such substrates to support new bone formation cannot rest with the readiness of the osteoblasts to coat them. The procedures outlined constitute a valuable, simple and rapid means of testing one aspect of the biocompatibility of potential prosthetic bone implant materials.


Application of electron probe X-ray microanalysis to calcification studies of bone and cartilage. The use of electron probe x-ray microanalysis in previous studies of bone and cartilage has been reviewed with emphasis on the results which have contributed to some of the current concepts of the mechanism of mineralization in these tissues. A number of investigations continuing in the author's laboratory utilizing high spatial resolution x-ray microanalysis and anhydrous methods of specimen preparation are described, including aspects concerning the derivation of calibration curves from synthetic calcium phosphate solids, qualitative and quantitative analyses of calcium and phosphorus in bone from embryonic chicks and in growth plate cartilage from rats, and the role of organically-bound phosphorus in mineralizing tissues. The data obtained have helped identify brushite, CaHPO4-2H2O, as the major crystalline solid phase of calcium phosphate in the earliest mineral deposits of bone tissue, brushite and poorly crystalline hydroxyapatite in bone mineral of increasing age, and poorly crystalline hydroxyapatite in the most mature mineral portions of the tissue. Growth plate cartilage examination has revealed calcium and phosphorus in single mitochondrial granules within chondrocytes and in certain extracellular particles distinct from matrix vesicles. These results have provided important information about the possible roles of cells, extracellular components, and the organic matrix in the regulation of mineralization and about the composition, structure, and organization of the mineral phase as a function of progressively increasing age and maturation of the tissues studied.


Labeling of lymphocyte surface immunoglobulins with T4 as marker for scanning electron microscopy. The technique originally described by Kumon et al. for the labeling of cell surface antigens under the SEM has been adapted to the study of surface immunoglobulins on human lymphocytes. Briefly, the peripheral blood lymphocytes are: 1) separated in a Ficoll Hypaque gradient, 2) rinsed with culture medium, 3) allowed to attach to plastic coverslips, 4) prefixed with 1% glutaraldehyde for 10 minutes, 5) incubated with goat anti-human immunoglobulins for 30 minutes at 24 degrees C, 6) rinsed, 7) incubated with a rabbit anti-goat IgG coupled with purified bacteriophage T4, 8) rinsed, 9) postfixed in glutaraldehyde for several hours, and 10) dried from ethanol at -75 degrees C and under 10(-2) Torr. Results from normal human lymphocytes and cells from cases of Chronic Lymphoblastic Leukemia (CLL) indicate that the method makes it possible to recognize cells with surface immunoglobulins and permits some correlation between the surface architecture of the cells and the presence of surface Ig. It also permits the study under the SEM of T-derived lymphocytes and of null cells, the unlabeled cell population, without resorting to techniques for lymphocyte subclass separation which may alter surface morphology.


Studies on carcinogenesis of human prostate. IV. Comparison of normal and neoplastic prostate during long-term explant culture. Morphologic responses of neoplastic human prostate to long-term explant culture were monitored at serial intervals by LM, TEM and SEM, and compared to normal prostate. Explants were cultured at 37 degrees C in CMRL-1066 supplemented with fetal calf serum and antibiotics. At 0-time culture, normal prostate of young adult males obtained at immediate autopsy, consisted of glandular spaces and ducts lined by columnar to cuboidal secretory epithelial cells and basal cells embedded in fibromuscular stroma. Neoplastic tissue was obtained surgically by transurethral resection (TUR), and consisted of stroma widely infiltrated by well-to moderately-differentiated tumor cells arranged in variable sized, gland-like structures. Secretory activity was evident; basal cells were absent in these glands. During early periods of culture up to several weeks, secretory cells of normal prostate became necrotic. Basal cells remained viable, repopulated acinar structures and epithelialized explant surfaces. At these sites, basal cells, or their derivatives, formed a multicellular epithelium. Exaggerated intercellular spaces separated cells, and synthesis of mucus-like material was seen. Epithelial characteristics included microvilli, junctional complexes, and basal lamina. In marked contrast, tumor cells covered explant surfaces forming an irregular, disorganized layer of squamous-like cells with elongated nuclei and prominent nucleoli. Microvilli, junctional complexes, and basal lamina were poorly developed or absent. Intercellular attachments appeared tenous. Some tumor cells accumulated lipid; synthesis of mucus-like material was not seen. At later intervals of culture up to 10 weeks, synthesis of mucus-like material by basal cells, or their derivatives, declined. Surface cells of neoplastic prostate gradually became more anaplastic in appearance; cells contacted neighboring cells with pseudopodia and filopodia.


SEM of human and experimental renal disease. This review concerns the present state of accomplishments in the study of SEM of human and experimental renal disease. Critical techniques of specimen preparation reviewed include perfusion fixation, razor tissue sectioning, alcohol cryofracture, microtome sectioning of paraffin or styrene embedded tissue, ultraplaning with glass knives of hard carbowax embedded tissues and glomerular isolation. Gold-palladium coating and heavy metal impregnation with osmium, uranium, and silver are discussed. A compendium of SEM observations of human glomerular, vascular and tubular disease is presented. Techniques for SEM of experimental renal disease are reviewed. These include latex vascular injection, freeze drying, x-ray microanalysis and use of backscattered electron imaging. Experimental models previously investigated by SEM are puromycin aminonucleoside nephrosis, daunomycin nephrosis, and N,N1-Diacetylbenzedine glomerulopathy, nephrotoxic serum nephritis, and protamine perfusion glomerulopathy. Reviewed are acute tubular necrosis caused either by angiotensin, hypotension, norepinephrine, glycerol, mercury, and unilateral renal artery occlusion, also potassium depletion nephropathy, alloxan diabetes and diphenylamine-induced polycystic disease.


The influence of lectin concanavalin A on the configuration of renal podocytes. Concanavalin A (ConA) was applied directly to renal glomerular podocytes by micropuncture techniques in vivo. Others received a control solution without ConA. To make visible the distribution of ConA, some rats were given fluorescein-isothiocyanate conjugated ConA. The glomeruli undergoing the micropuncture experiments were labeled and then prepared for SEM observation. Comparatively, the effect of application of the same solutions by intra-arterial infusion was studied. The glomeruli of a total of 33 Munich -rats were studied. Our results show that direct contact of ConA with the podocytes is needed to induce retraction of the podocytic cell processes. The pattern of podocytes treated with ConA is comparable to that of the minimal change nephrotic glomeruli. ConA seems to act by blocking anionic surface charges of the podocytes. This reduction in negative charges along the surface coat of the podocytes could be the key for understanding the combined morphological and functional alterations in podocytes of nephrotic glomeruli: The reduction in negative charges along the surface coat of the podocytes can alter the permselectivity for anionic and cationic proteins and can, at the same time, alter the configuration of the podocytes by changing the potential gradient along the cell membrane.


Microanalysis in biology and medicine. A review of results obtained with three microanalytical methods. Three microanalytical methods have been used : X-ray emission, secondary ion emission and electron energy loss analysis. Althouth X-ray microanalysis is the most used method, many biological problems cannot be solved because of its relatively low sensitivity and inability to analyze light elements. New possibilities are offered by Secondary Ion Mass Analysis and Electron Energy Loss Analysis. Analysis by secondary ion emission permits the study of elements at low and even trace element concentration, and even the lightest elements such as hydrogen and beryllium are detected. Electron Energy Loss Analysis makes possible the study of very small volumes, less than 500 A in diameter. These methods have been applied to the study of a variety of biological tissues (mostly lungs, kidneys and biomineralized tissues) and typical applications to pathology and cell biology will be presented.


Electron probe analysis of muscle and X-ray mapping of biological specimens with a field emission gun. Recent electron probe analytic studies of freeze-dried cryosections of vascular smooth and vertebrate striated muscle are reviewed. The results show that the sarcoplasmic reticulum of striated muscle is not in ionic communication with the extracellular space. Vacuolation by hypertonic solutions and fatigue involves the T-tubule system. The high calcium content of the terminal cisternae of the resting muscle has been quantitated in situ. In smooth muscle, the high Cl content is distributed in the cytoplasm, and mitochondria in rabbit portal vein smooth muscle cells do not contain high concentrations of calcium. Mitochondrial calcium loading in the form of granules is generally due to fiber damage. Nuclear and mitochondrial composition in situ has been quantitated and compared to the composition of the cytoplasm of the same cells. Preliminary phosphorus x-ray maps of smooth muscle show the feasibility of this approach in defining the composition of organelles in thin cryosections. The use of x-ray maps at intermediate resolution is illustrated with tropomyosin paracrystals labelled with Hg-containing dye at the thiol residues. Mercury x-ray maps of such paracrystals show the 40nm periodicity of the thiol groups and their Fourier transforms contain information to a spatial resolution of 10-20nm.


Scanning electron microscopy of human prostatic corpora amylacea and corpora calculi, and prostatic calculi. Ultrastructural studies of human prostatic corpora amylacea and corpora calculi, and prostatic calculi were conducted in order to delineate their etiology and pathogenesis. Scanning electron microscopy was employed in conjunction with histology and transmission electron microscopy in the study of prostatic tissues and concretions obtained from 21 individuals. Results confirmed that desquamated acinar cells contribute to the formation and growth of corpora amylacea. A variation in density of the matrix of the matrix of the corpora produces a laminated structure. The deposition of hydroxyapatite crystallites in corpora amylacea leads to the formation of corpora calculi. Further growth and mineralization of corpora calculi lead to the development of the more clinically significant prostatic calculi. Small spherical aggregates (from 0.5 to 5 micron in diameter) of hydroxyapatite crystallites are a prevalent constituent of the corpora and prostatic calculi. Similar spherical aggregates of hydroxyapatite crystallites have also been identified in urinary calculi and other pathologic tissues suggesting similar mechanisms of mineral precipitation.


Intracellular electrolyte concentrations in epithelial tissue during various functional states. The scanning electron microscope together with an energy dispersive system was used to quantify intracellular elemental concentrations of transporting epithelial cells under a variety of experimental conditions. In the frog skin, the Na transport pool is comprised of the intracellular compartments of all vital cells in the different cell layers, except the mitochondria rich cells. The Na content of this transport pool exchanges easily with the epithelial (outer) bathing solution. Vasopressin increases the Na permeability of the corial cell barrier. Proximal and distal tubular cells of the rat kidney show differences in the pattern of intracellular element concentrations. The distal tubular cell is more resistant to 20 min of ischemia than the proximal cell. The changes in intracellular electrolyte concentrations following 60 min of ischemia are reversible after reperfusing the kidney with blood for 60 min. In the cells of the frog skin and rat kidney, Na and K were equally distributed between the cytoplasm and the nucleus. Differences exist for P, Cl and dry weight.


X-ray analysis of pathological calcifications including urinary stones. Electron-probe microanalysis (EPM) is an ideal technique with which to study biological calcification. It is particularly effective in identification of crystalline or non-crystalline deposits of minute size in tissues, and in detecting artifacts which may occur during tissue processing. Human aorta, aortic valves, tumours, joint fluid, and calculus specimens were analyzed via scanning electron microscopy (SEM), EPM, transmission electron microscopy (TEM), and selected-area electron diffraction (ED). Crystals found in the specimens were definitively identified by combined SEM-EPM. It is apparent that EPM is an invaluable tool that will potentially improve the acuity of 'in house' laboratory diagnosis of many pathological calcifications.


Scanning electron microscopy of collecting lymphatic vessels and their comparison to arteries and veins. The morphology of canine thoracic duct and peripheral collecting lymphatics was determined using light microscopy together with scanning and transmission electron microscopy (SEM and TEM). The thoracic duct was compared to the thoracic aorta and to the vena cava. Luminal surface detail was determined using the secondary imaging mode of the SEM. Subsurface nuclear and connective tissue detail was determined using back-scattered electron imagining combined with Willard's modification of Gomori's Methenamine Silver Stain. Central and peripheral lymphatic vessels have surface morphology distinct from either arteries or veins. The endothelial cell density in lymphatic vessels is less than in arteries or veins. The nuclear chromatin of lymphatic endothelial cells is coarsely granular and evenly distributed. This contrasts with nuclei from arteries or veins in which the chromatin is segmented. The distribution and orientation of lymphatic subsurface connective tissue fibers also differs from that seen in arteries and veins. It is concluded that canine lymphatic vessels have a unique surface and subsurface morphology and can be unequivocally identified by SEM.


Scanning electron microscopy of the parimarginal cavernous sinus plexus of the human spleen. The perifolicular region of the human spleen was studied under the scanning electron microscope. The marginal zone 10 to 50 micron in width was located around the white pulp, and was composed of concentrically arranged reticulum cell networks and intervening lymphocytes, red blood cells and macrophages. The cavernous sinuses 10-150 micron in width were situated between the marginal zone and the red pulp, and were lined by thin, flat endothelia. The perimarginal cavernous sinuses communicated with one another through narrow canals and formed a plexus. Transmural passages of lymphocytes were noted on the side of the white pulp. The perimarginal cavernous sinus plexus was also in communication with the interspaces between reticulum cells in the marginal zone. The human perimarginal cavernous sinus plexus appears to correspond to the marginal sinus in rat and rabbit spleens.


Techniques for quantitative organic analysis in microdroplets. Techniques are described for the chemical analysis of urea by means of X-ray microanalysis. A selective labelling reagent, thioxanthen-9-ol, is used to precipitate urea together with a sulfur label. Reagents are added to and removed from microdroplets of 50 picoliter volume by means of oil-water partition. The sensitivity of the method is less than 1 picomole. The possibility of extension of these techniques to other analyses is discussed.


Metal binding by intestinal mucus.

Metal binding by intestinal mucus. Electron probe microanalysis offers distinct advantages for the study of intestinal mucus. This technique permits analysis of metal binding in situ, requires only a small amount of tissue, allows several experiments to be performed with one animal, and can resolve variations in binding that may occur in different portions of the intestine. We have used electron probe microanalysis to examine the metal binding capacity of intestinal mucus in situ.  We have exposed portions of excised intestine to various concentrations of several metals, rapidly frozen the tissue and freeze dried it. After anhydrous embedding, thick sections were cut and analyzed on silicon discs or carbon coated copper grids. Qualitative analysis shows two distinctive patterns of distribution. The results of this work show clearly that at least three divalent cations are bound by mucus, that mucus exhibits different affinities for different metals, and that binding of metals is not uniform throughout mucus.


Vascular organ culture: prevention of endothelial damage due to removal and reperfusion. Endothelial cell damage is recognized as a complication in blood vessels which are excised and reperfused for grafting or vascular organ culture. We demonstrate that the damage consists of lifting, crenation and detachment of endothelial cells, partially due to contracture and forceful redilation of the vessel wall. We use a mechanical brace and smooth muscle cell relaxation with papaverine to maintain length and circumference of rabbit carotid arteries during removal and reperfusion. With this technique, a considerable degree of morphological preservation of the endothelial cell layer can be achieved in isolated and perfused vessel segments. Furthermore, this principle of endothelial cell preservation by prevention of contracture during vessel preparation may be applicable clinically.


Scanning electron microscope surface morphometry of type II alveolar lung cell response to urethane in strain A mice. Quantitative scanning electron microscope (SEM) examination of the time course of type II cell hyperplasia in mice exposed to a single i.p. injection of urethane is presented. Counts of type II cells from random alveoli suggest that, compared to controls, a significant depression in type II cell numbers occurs during the first week, followed by a doubling in the next 3 to 4 week period. This supports conclusions of earlier studies done with thin sections and light morphometric techniques. The use of a SEM as the morphometric tool is statistically more efficient, however, since initial sample preparation is easier, samples are larger in area, resolution is higher and, most importantly, the amount of surface area available for examination is many times greater than that presented by light microscope examination of cross sections with equivalent magnification.


Human natural killer cell activity is augmented by interferon via recruitment of 'pre-NK' cells. Contact with various cell-line targets increases the natural killer (NK) activity of human lymphocytes. Supernatants of such 20 h co-cultures augment the NK activity of virgin lymphocyte populations, and the augmenting factor penetrates 0.2 micrometer Millipore filters. The supernatants also contain interferon, and partially purified human leucocyte interferon increases NK activity when added to 20 h assays with 51Cr-labelled K-562 target cells. Potent anti-interferon antiserum added to the co-cultures inhibits the target-cell-induced augmentation phenomenon. Both the target-cell contact and interferon-induced augmentation affect a population of human lymphocytes, from which the 'mature' NK cells have been removed by adsorption-elution using fetal fibroblasts as adsorbents. The activity of 'mature' NK cells is not enhanced, and we conclude that the augmentation is mediated by recruitment of 'pre-NK' cells.


Herpes zoster in infancy.

Herpes zoster in infancy. Three cases of herpes zoster occurring in infancy are reported. In each case the mother had chickenpox in pregnancy.


Septicemia I. A prospective study on etiology, underlying factors and sources of infection. In a prospective study, 110 cases of proved and 41 of probable septicemia were analysed. Underlying diseases or factors were common, also in young patients, often narcotic drug or alcohol abusers. In one third, septicemia was probably hospital-acquired. Streptococci, staphylococci and gram-negative enteric rods were most common, followed by pneumococci and haemophilus species. Staphylococci and gram-negative enteric rods dominated in patients hospitalized at or before the onset of disease, and staphylococci in patients with vascular foreign bodies and in drug addicts. Enteric rods were common in elderly patients with urogenital disease, especially after instrumentation. Among streptococci, alpha-streptococci dominated; they did not emanate as often from dental foci as expected. In 2 of 3 asplenic patients with extensive hemorrhages pneumococci were found. Bacteria of low virulence and fungi occurring as opportunists were rare. Mixed infections, present in 4 cases, were severe. Portals of entry could often be proved or assumed. In half of all patients, the same organism was isolated from a primary focus as from blood. The importance of underlying illnesses and factors and of invasive procedures was evident. A tentative etiological diagnosis could often be made, based on case history, underlying factors and the probable portal of entry.


Anthrax in children: a long forgotten, potentially fatal infection. After a brief account of anthrax morbidity in northern Greece in recent years, 4 cases in 1977 of cutaneous anthrax in 10 to 13-year-old children are presented. Two had an atypical cutaneous lesion. In 1 of these, a 13-year-old girl, the disease was complicated by severe, eventually fatal meningitis. Death ensued despite intensive treatment with high doses of penicillin and hydrocortisone. This case is the first report of anthrax meningitis in a child in Greece and the third reported in the last 15 years. We stress the diagnostic difficulties in atypical cases of cutaneous anthrax and the need for early diagnosis and treatment to avoid spread of infection and appearance of complications such as the usually fatal meningitis.


Evaluation of spiramycin in meningococcal carriage. Spiramycin was administered to healthy carriers of meningococci in doses of 1.5 g twice daily or 2.5 g once daily for 3 days, or 1.5 g once daily for 10 days. Meningococci were eliminated from the nasopharynx and throat in approximately 50% of the carriers with all dose schedules. In the remaining carriers only a temporary suppression of the strain was achieved. Elimination occurred only in carriers where spiramycin saliva concentrations reached or surpassed the MIC value of respective strain for 42 h or more. Evidence of reinfection from close contacts was not obtained. Due to side effects the dose of spiramycin could not be increased. Further trials with spiramycin as the sole agent for eliminating meningococci in carriers do not seem warranted.


The influence of different footwear on floor contamination. The importance of walking as a source of floor contamination and the influence of different footwear routines on floor contamination was evaluated. The contamination of a floor due to walking was about 16 times higher than contamination of the floor due to sedimentation only, measured with impression plates. No significant difference in floor contamination could be seen with different footwear (ordinary shoes, clean shoes, shoe covers). In all experiments the amount of bacteria, was low in the morning and increased in the middle of the day when a steady state seemed to be reached. The number of Staphylococcus aureus continued to increase. A footwear regimen is only recommended in areas where floors are cleaned frequently during the day as in operating wards.


Doxycycline prophylaxis in fracture surgery. Doxycycline was given as preventive preoperative treatment to 64 patients with fractured collum femoris. 46 patients, not administered doxycycline, served as controls. The overall incidence of postoperative infection was 6.3 and 6.5% in treated and control groups respectively. It is concluded that doxycycline preventive treatment offers no advantage in this type of operative procedure.


Yersinia enterocolitica infection and sarcoidosis. A report of seven cases. Sarcoidosis of recent onset was diagnosed during a Yersinia enterocolitica infection in seven patients at the Copenhagen County Hospitals during a 3 1/2 year period. In all cases the diagnosis was made from clinical symptoms and confirmed by chest X-ray and biopsy. In five of the patients a fourfold rise or decrease was measured in the antibody titre against Yersinia enterocolitica, serotype 3, confirming an acute infection. In another two patients this titre remained significantly elevated greater than or equal to 80, indicating chronic infection. Five patients experienced joint symptoms and four erythema nodosum, symptoms common to both yersiniosis and sarcoidosis. A follow-up examination 14 to 41 months after the debut of sarcoidosis showed total remission (including chest X-ray and respiratory function) in five of the seven patients. Recent research has shown that Yersinia enterocolitica following the early stage of the infection causes granulomatous lesions in lymph follicles, and this aetiology should thus be considered as a differential diagnosis to sarcoidosis.


Depression of nasal mucociliary transport by an aerosol hair-spray. The nasal mucociliary transport was measured on 15 healthy test subjects, using the saccharine method, before and after exposure to hair-spray and Freon. The hair-spray reduced the mucociliary transport in the nose for more than 1 h, whereas Freon had no effect. Hair-spray must be considered potentially dangerous, and investigations of the mucociliary transport in persons subject to prolonged exposure to hair-spray (ladies' hairdressers) must be relevant.


Prevalence and manifestations of obstructive lung disease in the city of Oslo. A stratified random sample of 1209 subjects aged 16--69 was examined in Oslo, Norway in 1973--74 by a team of 11 chest physicians. They used information from a standardized interview and clinical examination, radiographs and ventilatory function tests to make a diagnostic decision. The within-observer agreement for the diagnosis bronchial asthma and the composite group obstructive lung disease (OLD) was 87% and 80%, respectively. More than 80% of maximal between-observer agreement was observed for these two entities. The point prevalence estimate of chest physicians' diagnosis of obstructive lung disease (OLD) was 5.5%, comprising bronchial asthma 1.4% and chronic obstructive lung disease (COLD) 4.1%. The prevalence of COLD increased with age, and it was four times more frequent in smokers/ex-smokers than in non-smokers. The prevalence of bronchial asthma showed no relation to age or smoking. Individuals aged 20--69 years with bronchial asthma, COLD and OLD, had 1 sec forced expiratory volumes of 83%, 71% and 74% of predicted values, respectively. A previous history of hay fever was reported five times more frequently by those with bronchial asthma than in the survey population. Disability pension was received by 15% of those with OLD.


Survival in lung cancer after surgery.

Survival in lung cancer after surgery. In 1053 patients hospitalized for primary lung cancer the overall crude survival rate (5 years) was 15.5%. Prognostically favourable indices were: absence of clinical symptoms (27% survival) and a peripheral site of tumour in the lung (28% survival). Duration of symptoms, when present, had no consistent influence on prognosis. Resection could be done in 419 cases. In this group the 5 years survival rate was strongly related to the extent of surgery and the completeness of resection. Cases with a radical lobectomy had a survival rate of 52% against 9% only in patients with a pneumonectomy that might not have been radical. Of 33 cases of resected small cell anaplastic cancer, 10 survived more than 5 years. This unexpectedly high survival rate persisted after revision of the histological typing and may justify an active surgical attitude even in this group of patients. Although hardly decisive in this context, confusion between the comprehensive term small cell carcinoma and its subtype, oat cell cancer, should be avoided.


Adjustment and behaviour of Finnish and Southern European immigrant children in Stockholm. I. The teachers' assessment. The adjustment and behaviour of immigrant schoolchildren were studied by means of teacher interviews. The material consisted of 50 Finnish and 37 southern European children. For comparison, 44 Swedish migrant children were also sampled. All had settled in the County of Stockholm three years prior to the study. Swedish children who had lived in the county for more than four years served as controls. In the teachers' opinions the immigrants as a whole had adjustment difficulties more often than the controls, but the proportion of children with such difficulties was no higher among the immigrant children. Compared with the controls the immigrant children showed a higher frequency of symptoms relating to a disordered self-esteem. The immigrant children were also considered to have a lower status and to be less trustworthy than the controls. The schooling of these children demands serious attention in order to prevent discrimination and to promote a feeling of personal worth among the children.


The mortality of male old-age pensioners following spouse's death. The effect of the death of the spouse on mortality of retired men was investigated. The material comprised 939 married men retired on old age pension, 174 of whom had lost their spouse after retirement. During the first six months following the wife's death, more than the expected number died, though no increase in mortality could be shown over a longer term. Circulatory diseases and tumours were over-represented as causes of death among those who died within six months of the wife's death. Loss of an important object relationship may lead to sudden cardiac death, or the development of a disease may be accelerated.


Work and disability at the age of 30 years. A sociomedical study of a birth-cohort from Bergen. VI. Education, intellectual ability and occupation. The relationships between occupation, education and intellectual ability at 30 years of age are analysed with particular reference to type of school attended at the age of 14 years. Data utilized are derived from interviews, psychological tests, local files of various schools, and journals of the National Services for Mentally Retarded. The study comprises all live births in the year 1940 of mothers then residing in Bergen, a total of 1570 persons. A sample was taken from this cohort after stratification according to type of school attended at age 14 years. The sample was supplemented with persons who had either attended Special Schools for the educable Mentally Retarded (EMR) or received care from the Services for the Mentally Retarded (SMR). The final sample consisted of 262 persons. A relationship was found for both sexes between type of school attended at the age of 14 years and level of general education at the age of 30. For men, both occupational training acquired and intellecutal ability at 30 years were also clearly related to type of school attended at age 14 years. The test performance of the male group was superior to that of the female group. Differing careers in the two sexes may provide a clue as to the reason underlying this finding.


Seasonal variations in the incidence of acute myocardial infarction in Stockholm. The aim of the present paper was to ascertain whether or not there is a seasonal trend in the incidence of acute myocardial infarction (AMI) in Stockholm. The material consisted of hospitalized patients with a diagnosis of AMI, recorded in a medical information system, and of deaths recorded in the cause-of-death register. The hospital stays and the deaths together totalled 4020 cases of AMI occurring in 1973. In contrast to the results of other studies, no seasonal trend was found.


Health complaints and work conditions among lacquerers in the Danish furniture industry. In the present work, morbidity rates were based upon 418 replies to postal inquiries sent to 478 members of the carpenters/cabinet makers trade union in the Arhus area of Denmark. Lacquerers, when compared with former lacquerers and non-lacquerers, have an excess of neurophysiological symptoms and symptoms from the respiratory tract. The statistical association between exposure to lacquers and lung symptoms persists after checking for differences in age, smoking habits, and exposure to dust. Organic solvents are an important component of lacquers. Far too little is yet known of solvents' long-term effects on the human body. Most of our present knowledge concerns the short-term toxic effects on the nervous system. This study further points out a relation between lung symptoms and exposure to organic solvents, and it is suggested that further research into this association must be carried out.


Municipal occupational health services for small workplaces. Background and general methodology of the study. This paper deals with the new Occupational Health Service Act of Finland, as well as the background, scope and general methodology of a walk-through survey. The study comprised 163 small places of work within two municipal health center areas, and the primary objective was to determine their need for actual occupational health services. The workplaces were surveyed for a comprehensive picture of their physical and chemical hazards, as well as for knowledge of first-aid preparedness, need for job-related health counseling, personnel facilities, personal protection, and the required ergonomic and safety activities at the workplaces within the community. This ad-hoc information was considered essential for the planning of a nationwide occupational health program, and, more specifically, an assessment could be made of the utility of health personnel in reducing and preventing occupational health and safety risks at small places of work. More specific details of the sample and methodology are reported in other communications.


First-aid preparedness in small workplaces with special reference to occupational health services. First-aid preparedness was surveyed in 163 small firms employing 2,400 persons. Although 11% of the employees had received first-aid training, half of the firms lacked first-aiders. Only one-fourth of the first-aid kits were equipped according to instructions. Checking and maintenance of the equipment, as well as information about first aid at the place of work, were generally inadequate. Instructions were commonly lacking for the safe use of chemical substances. Only 9% of the firms fully complied with the requirements for optimum first-aid preparedness. Employers at small firms seem not to be sufficiently aware of their liabilities with regard to first aid. Yet, most of the faults and defects could be corrected by means of arrangements within the firms once the instructions were better known. During their visits to places of work occupational health personnel should inspect the first-aid preparedness and advise the employer on first-aid requirements. In addition the expertise of the insurance company, the local Red Cross or the regional institute of occupational health may be utilized.


Personnel facilities in small workplaces with special reference to occupational health services. The personnel facilities of small workplaces generally complied with the requirements. Yet, defects were detected in 20% of the 163 firms surveyed. Most of the faults were insignificant from the point of view of health. The failures were due to the fact that the facilities had not originally been planned for their present purpose. The need for the inspection of sanitary installations such as toilets and washrooms and facilities for storing and drying clothes was noted in dirty jobs and in places of work with toxicologic hazards. The management of small firms rarely needs medical advice on sanitary, catering and welfare amenities.


Need for job-related health counseling in small workplaces. Health counseling aimed at maintaining health and safety on the job is required in most small workplaces. From a labor protection point of view, the most important advice deals with environmental hazards such as protection against physical and chemical factors, ergonomic problems, and safety risks. Information provided by the employers was considered insufficient in three-fourths of the 163 firms surveyed in this study. The staff providing local occupational health services could perform two broad types of counseling. First, they could educate management as to its responsibilities for the health and safety of employees; health personnel can instruct management during their workplace visits. Second, during health examinations and other personal contacts, the worker can be advised about ways to improve or maintain his health. Moreover, such occasions also provide an opportunity for more extensive health counseling about personal, social emotional, and other problems although, in municipal health centers, these subjects belong within the scope of public health rather than occupational health.


Need for protective clothing and equipment in small workplaces. Some kind of protective clothing and personal safety items was needed in 90% of the 163 workplaces surveyed. Protective clothing was used in 31%, ear protectors in 28%, eye protectors in 22%, respiratory tract protectors in 18% and other safety items in 1% of the firms. Gloves were the most commonly used type of protective clothing. Protection against risks was proper in one-fourth of the firms, satisfactory in half, and poor or lacking in the rest. Ninety-five percent of the protectors available were appropriate, but only in a few companies were they properly cared for. Seven (12%) industrial firms, 15 (20%) service establishments and 1 (7%) other firm lacked proper protective clothing or safety items. Faults were generally attributed to insufficient knowledge or negligence of responsibilities on the part of the employers. Management should provide more appropriate instruction for workers in the use and, particularly, the maintenance of personal protectors. To be effective, the inspection of the appropriateness of the protectors requires suitable training. This training should also be received by the staff providing occupational health services. Visits to workplaces, as well as health examinations, provide an opportunity to discuss protection and to advise employers and employees on protection. The problems of protection against harmful exposures is closely connected with health counseling. Yet, the provision of proper protectors and the supervision of their use is still the responsibility of the employer.


[The course of diabetes and clinical findings in glucagonoma]. A 66-year-old male patient with non-insulin-dependent diabetes of probably 20 years' duration presented with necrolytic migratory erythema, stomatitis, anemia and weight loss. Plasma-glucagon concentration measured with Unger's antibody 30-K was 8500 pg/ml, representing a hundredfold elevation. Two thirds consisted of high molecular glucagon fractions (10 000--40 000 Dalton). This may be an important index for detection of glucagonoma with endocrine activity. After excision of the glucagonoma the clinical syndrome was reversed and the patient recovered completely. Histological and histochemical investigation confirmed that the tumor was a glucagonoma. Despite complete removal of the tumor and a normal plasma glucagon concentration, the diabetes remained unchanged. Excessive hyperglucagonemia does not appear to play a primary role in the pathogenesis of this patient's diabetes.


[Bile acid concentration in serum after a test meal in hepatobiliary diseases. A comparison with quantitative liver function tests]. The concentration of bile acids in serum was measured by an enzymatic-fluorometric method under fasting conditions and 2 hours after a standardized meal in 26 patients with chronic liver disease (chronic hepatitis, liver cirrhosis, primary biliary cirrhosis) and compared with other tests of liver function. Postprandial bile acids and transaminases were false negative in only 12% and are thus the most sensitive tests after the BSP-retention test (3% false negative results). In comparison, fasting bile acids proved to be a relatively insensitive screening test for liver disease (38% false negative results). Postprandial bile acids were more closely correlated with BSP retention and BSP disappearance rate constant (Ki) than fasting bile acids. In view of these findings postprandial serum bile acid concentrations should be preferred to fasting bile acid concentrations in screening for liver disease and monitoring liver function.


[Basis of respiratory insufficiency].

[Basis of respiratory insufficiency]. Setting out from the components of respiratory function, i.e., ventilation, distribution, diffusion, circulation, respiratory mechanics, and regulation of breathing, the pathogenic mechanisms leading to respiratory failure are discussed. In every case, the vital capacity is decreased by 4 factors, namely loss of ventilated lung parenchyma, diminished compliance of lungs, thorax or both, airway obstruction, and insufficient respiratory airflow. With few exceptions, these alterations can be attributed to the two general groups of obstructive and restrictive disturbances of ventilation. Essential for the understanding of airway obstruction from the viewpoint of mechanical ventilation is the dependence of the airway caliber on lung volume, thoracic pressure, and bronchial gas flow. The functional differentiation of restrictive disorders between forms with lung retraction(fibrosis, scarring) and with lung fettering (pleural thickening) is important for adequate correction of complications during the intensive care phase. Respiratory failure is the consequence of these alterations which usually impede pulmonary gas exchange. Hypoxemia results in most situations through disturbance of ventilation/perfusion ratio, especially increase of anatomical or functional pulmonary right-to-left shunting. Disturbance of diffusion or alveolar hypoventilation are far less frequently leading mechanisms for hypoxemia. The differential diagnosis of these hypoxemic mechanisms is generally by arterial blood gas analysis under resting conditions breathing air and 100% oxygen, and during exercise. Respiratory failure often leads to hypertension in the lesser circulation. Pulmonary arterial hypertension must be subdivided into the active, the passive and the hyperdynamic forms, of which only the active component is important for the evaluation of pulmonary insufficiency since only this kind of elevated pressure in the pulmonary circulation is connected with increased vascular resistance due to thoracopulmonary disease. By restoration of normoxic conditions, the functional variant of active pulmonary hypertension can be efficiently improved by correction of respiratory disease or directly by treatment with oxygen and by mechanical ventilation. Finally, disturbances of gas transport in the blood may have an essential bearing on respiratory failure, but are often overlooked in diagnostic and therapeutic considerations. Shifting of the oxygen-dissociation curve to the left may, by increased oxygen affinity of hemoglobin, create a lack of oxygen in the peripheral tissue, while right wardshifting impedes oxygenation of hemoglobin in the lung. Thus, the correction of acidosis and elevated body temperature may become an important factor in the treatment of respiratory failure.


[The respiratory intensive unit. Requirements for the treatment of patients with acute respiratory insufficiency]. Patients with acute severe respiratory insufficiency can only be correctly monitored, treated and nursed in a well-equipped intensive care unit staffed with well-trained personnel. Qualified doctors and nursing staff are more important for the monitoring and assessment of the patient's symptoms than highly sophisticated electronic monitoring systems. Continuous controls of vital and mechanical functions are however essential , as are the services of well-run laboratories. Treatment calls, above all, for the use of efficient and reliable respirators capable of adjustment to suit the impaired breathing. The quality of nursing and the success of intensive medical care measures are best ensured by well-designed layouts, strict adherence to sensible and appropriate hygienic techniques, continuous and practice-related further education for personnel, and unrestricted communication inside and outside the intensive care unit.


[The role of respiratory physiotherapy in an intensive care unit]. In an intensive care unit an important role is assigned to respiratory physiotherapy. Its principal task is efficacious toilet of the bronchi by fluidifying the secretions, promoting their ungluing from the respiratory tree and facilitating their evacuation by cough or by aspiration with a catheter or bronchoscope. The technique comprises the inhalation of a secretolytic (e.g. Bisolvon, NaCl 9%) and, in the case of asthma, bronchospasmolytic (e.g. Ventoline) aerosol followed by breathing exercises. The other objectives of physiotherapy are to ensure a better distribution of inspired air, increase failing ventilation, ameliorate disturbed gas exchange, relax the contracted respiratory muscles and prevent bronchiolar collapse in emphysema during expiration. The field of application of respiratory physiotherapy is large; its purpose is prophylactic and therapeutic. The method is prophylactic in all patients confined to bed, where there is a risk of bronchial obstruction or ventilatory failure, especially in those with severe operation, traumatism or consciousness disorder. Physiotherapy has a therapeutic role in several, principally broncho-pulmonary diseases, such as asthma, obstructive emphysema, pneumonia, bronchiectasis, pulmonary abscess, atelectasis, and pulmonary and pleural fibrosis. Myocardial infarction and pulmonary embolism in the acute state, acute pulmonary edema, pneumothorax and pulmonary hemorrhage are contraindications for physiotherapy. If the method is to be effective the intensive care unit should have a specialized physiotherapist attached to it working there on a daily basis.


[Continuous monitoring of intracranial pressure and new aspects of neurologic intensive care for children]. A new concept of neurointensive care is presented which is based on earily measured parameters such as intracranial pressure (ICP), mean arterial pressure (MAP) and cerebral perfusion pressure (CPP) (CPP = MAP - ICP). ICP should preferably be measured by a subarachnoid hollow screw (Richmond screw). Of chief importance in neurointensive care (after adequate neurodiagnosis) is the avoidance or treatment of cerebral edema and maintenance of sufficient CPP, which should be above 50 mm Hg in older children. This is achieved by controlled hyperventilation under curare, generous oxygenation, control of body temperature, dexamethasone and possibly barbiturates in very high doses (phenobarbital and thiopentone). This kind of neurointensive care should be administered in all types of severe CNS accidents which are followed by substantial cerebral edema (head injuries, near drowning, Reye's syndrome, hypoxic encephalopathy, encephalitis, meningitis and intracranial bleeding). To obtain the indication for ICP monitoring, the depth of the disturbance of consciousness is measured by the Glasgow Coma Scale. Children with the aforementioned affections and a Glasgow Coma Scale below 6 to 8 should be treated as outlined above. The data published in the literature and our own experience point very much in this direction, especially for severe head injury, Reye's syndrome and near drowning.


[Edema, heavy legs and popliteal compression]. Fifty patients with edema and heaviness of the lower limbs lacking any general or local explanation were investigated by a plethysmographic method to detect venous stasis of the calf. In 27 of them, hyperextension of the knee produced venous stasis, which was only elicited in one of 39 normal subjects. In 9 patients with edema of systemic origin there was no stasis when the knee was extended. Intermittent venous stasis due to hyperextension of the knee therefore appears to be a causal phenomenon in many cases of edema of unknown origin. A popliteal cyst was found in 9 of 16 patients who underwent knee arthropathy. Further studies are necessary to determine what mechanisms may produce venous stasis when the knee is placed in hyperextension.


[Iatrogenic chromodermatoses].

[Iatrogenic chromodermatoses]. Generalized hyperpigmentation may be caused by internal diseases, by occasional or occupational intoxications, and by a group of drugs. Seven cases occurring after treatment with amiodarone (Cordarone) are reported, together with one case of argyria and one of mercury poisoning. Several of these cases were initially mistaken for cyanotic heart disease. Silver intoxication may occur after indiscriminate use of currently available silver-containing drugs. It may become even more frequent as a result of antismoking preparations.


[Clinical and immunological studies on a patient with myasthenia gravis before and after repeated plasma exchange]. A 56-year-old patient with severe acquired myasthenia gravis refractory to conventional therapy underwent a series of plasma exchanges. Three single exchanges did not improve her status. A series of intensive plasma exchanges of 6 liters each on 4 consecutive days reduced her anti-acetylcholine receptor antibodies to one-tenth of the initial value. The reduction was accompanied by an impressive clinical improvement. This study confirms that antibodies can be removed by plasma exchange. It further shows that the clinical improvement appears to be closely related to the quantity of antibodies removed.


[2 cases of multiple myeloma with osteosclerosis]. A report on two patients with osteoclerotic myeloma is presented (myeloma with osteoblastic lesions). In case one, metastatic bone tumor of unknown origin was wrongly diagnosed initially. Because of increasing neurological symptoms laminectomy was performed. Biopsy led to the correct diagnosis of multiple myeloma of IgA-type. Patient also had a very severe peripheral neuropathy. The second patient had pancytopenia and osteosclerotic lesions of the pelvis. Bone marrow aspiration revealed so-called "empty marrow". Based on these findings, myelofibrosis was wrongly diagnosed at another hospital. Bone marrow aspiration, paper- and immunoelectrophoresis subsequently produced the correct diagnosis of multiple myeloma of IgG-type. Multiple myeloma usually is characterized by osteolytic lesions of the bones. However, the literature contains some 50 cases with osteosclerotic multiple myeloma, three different forms of which are described. In a fairly large percentage osteosclerotic multiple myeloma is combined with periphereal polyneuropathy. It would appear that in IgE-myeloma the incidence of sclerotic lesions is higher. Osteosclerotic multiple myeloma is very rare. It should however be considered if the differential diagnosis of osteosclerotic bone lesions is established.


Breast-feeding and infant health.

Breast-feeding and infant health. A mother who breast-feeds makes a significant contribution to her infant's health. Currently researchers are delineating many of the unique nutritional and antiinfective factors in human milk. The psychological advantages that accrue to both the breast-feeding mother and her infant are harder to quantify. Contraindications to breast-feeding are few and relate primarily to maternal illness and need for medications, or to those rare infants who have inborn errors of metabolism. Physicians and other health personnel should advise the mother knowledgeably on breast-feeding and seek imaginative solutions to any problems so that breast-feeding can be continued well into the first year of the infant's life and into following years if both mother and child desire it.


Unexpected red cell antibodies in donors and patients at Siriraj Hospital. The incidence of unexpected red cell antibodies in patients and donors at Siriraj Hospital were found to be 4.91% and 4.33% respectively. Anti Lewis, anti P1 and anti Mia were the most common antibodies detected. Antibodies of Rh system were the next, in which anti E was more common than anti D. Anti Jka, Jkb, Fyb, Dia, S and rare antibodies such as anti K, Ge1, Mta and H were also detected in this study.


The HLA antibodies in Thai pregnant women at Siriraj Hospital. The HLA antibodies in Thai pregnant women was determined. From 4,000 sera, only 577 cases (14.4%) showed positivity, and only 21% were monospecific antisera.


Recent advances in hemophilia.

Recent advances in hemophilia. Classical sex-linked hemophilia (Hemophilia A) has been described as due to deficiency in the synthesis of Factor VIII procoagulant activity (VIII:C). The availability of immunological techniques provided the means of identifying Factor VIII-Related Antigen(VI-IIR:Ag) detectable by rabbit antibodies to F VIII, which is distinct from VIII:C detected by human anti-F VIII available from multitransfused patients. Hemophilia A is lacking in VIII:C but not VIIIR:Ag. Recently, a third function of the F VIII "complex" was discovered with the help of ristocetin (von Willebrand's Factor, VIIIR: RCo). This activity is reduced in von Willebrand's syndrome. Estimation of the titers of VIII:C and VIIIR:Ag provides a method for more accurate detection of hemophilic carriers. Newly available chromogenic substrates perhaps will give rise to more simplified assays of VIII:C. The development of cryoprecipitates and stable lyophilized concentrates of F VIII has greatly simplified and intensified maintenance therapy, and has opened a new era in treatment. Prophylactic therapy has been shown to be very helpful in certain "high risk" cases. The impact and benefits of home care and self-administration has been tremendous. However, the varying quality of cryoprecipitates and the high cost of more purified concentrates are still stumbling blocks in treatment regimes. Other problems exist. Spontaneous bleeding, especially central nervous system bleeding, account for the majority deaths by haemorrhage. Inhibitor kinetics have been well characterized. It is clear that there exists "low" and "high" responders. For the "high" responders, plasmapheresis, immunosuppressives and the infusion of Factor IX concentrates have been utilized with varying success. The prevention of hemophilic arthropathy and its progression by maintenance therapy seems to be still inadequate. The results of trials with more vigorous regimes are awaited. The complications of therapy still remain to be solved. Apart from the well-known complications wuch as hepatitis, haemolytic disease and F VIII inhibitors, the existence of previously unnoticed complications as splenomegaly, hypertension, renal disease and paradoxal bleeding have been recently realized. The role of altered fibrinogen, fibrin degradation products (FDP) and unclassified fibrinogen derivatives (UFD) present in cryoprecipitates and F VIII concentrates in the above complications needs to be further clarified. In conclusion, tremendous progress in various aspects of hemophilia has been achieved in developed countries. Comprehensive care can now be carried out in various centers. On the other hand, developing countries still face a number of basic problems. The concept that hemophilia is a "manageable" disease and that chronic crippling and death from exsanguination can be prevented, should be disseminated widely by various means...


Functional platelet defects and their laboratory evaluation. Primary diseases of platelet function include Glanzmann's thrombasthenia, hereditary platelet release abnormalities (storage pool disease and release defect), Bernard-Soulier giant platelet syndrome, and platelet factor 3 defects. Qualitative defects of platelets are associated with many diseases, notably of the liver and kidney, and with the use of many drugs, particularly aspirin.


Incidence, clinical manifestation and general management of snake bites. There are regional patterns in snake-bites. Bites by cobras have a high incidence in Thailand and in the Philippines with a high case fatality rate. Among the venomous snakes of haemorrhagic nature, bites by Trimeresurus species such as green pit viper, Taiwan habu and Taiwan bamboo viper are important in Thailand and Taiwan for their high incidence of bite, although the case fatality rates are low. Bites by Malayan pit vipers are also important in Thailand and Malaysia because of their high incidence. Bites by sea snakes are more common in Malaysia than in the Philippines and Thailand.


Bleeding manifestations in snake bite.

Bleeding manifestations in snake bite. Bleeding following bites by the Malayan Pit Viper can either be local or systemic. Bleeding at the site of the bite is due to the local action of the venom as a vasculotoxin. Systemic bleeding occurs with severe poisoning and appears to be mainly dependent on platelet deficiency and the co-existing defibrination syndrome appears to play a minor role in the initiation of bleeding. Thus in the clinical situation non-clotting blood with no overt bleeding can continue up to weeks when specific antivenene is not given. Assessment of the severity of poisoning can easily be made at the bedside. Specific viper antivenene rapidly corrects the spontaneous bleeding and clotting defect of severe systemic poisoning but has no effect on local poisoning.


Effect of five Thai snake venoms on coagulation, fibrinolysis and platelet aggregation. The effects of five Thai snake venoms--Malayan pit viper--MPV (Ancistrodon rhodostoma); Russell's viper--RVV (Vipera russelli); cobra--CV (Naja naja); king cobra--KCV (Naja hannah) and banded krait--BKV (Bungarus fasciatus)--on blood coagulation fibrinolysis and platelet aggregation were studied. MPV has strong thrombin-like coagulant action. RVV has activating effect most probably on factor X. Other three venoms--CV, KCV and BKV have been shown to be anti coagulant, on both intrinsic and extrinsic clotting pathways. MPV has strong lytic activity on normal euglobulin clot while CV has mild accelerating action. RVV and BKV inhibit the lysis of euglobulin clot. KCV has no effect on fibrinolysis. None of the five venoms has direct aggregating activity on platelets. All except MPV inhibit ADP-induced platelet aggregation at their higher concentrations.


The impact of severe illness on the individual and family: an overview. The course and outcome of severe chronic illness are decisively affected by social and emotional variables. The conceptual approach of this article emphasizes an understanding of the individual, the family, and health care personnel as all facing a series of adaptive tasks in relation to the illness. Each of the three phases of the illness--the diagnostic, chronic, and end stage--poses special tasks and requires different defenses and coping capacities from the patient, family, and health care personnel. These are discussed as interdependent factors in a holistic framework.


Adjustment and coping strategies among the caretakers of cancer patients. The problem of emotional "burn-out" among health care professionals who work with cancer patients is a significant clinical issue. An interdisciplinary group of these health care service providers participated in a workshop aimed at identifying the major stresses which contribute to emotional "burn-out." While some of these stresses are unavoidable aspects of cancer treatment programs, participants also identified a set of coping strategies which seem helpful in reducing caretaker vulnerability to severe emotional exhaustion in oncology treatment settings.


Counseling for women requesting sterilization: a comprehensive program designed to insure informed consent. This paper describes the counseling program implemented by a social worker and a family planning counselor for female clinic patients requesting sterilization. Rather than limiting counseling to the guidelines for informed consent established by New York City law, a comprehensive approach is utilized. The program combines the provision of information regarding the procedure and contraceptive alternatives, with an opportunity for individuals to examine the psychosocial components of their request for sterilization. It is the authors' assessment that such comprehensive counseling is essential to provide individuals with the full assistance they need in making the decision to eliminate reproductive capacity, and that legal guidelines should be expanded accordingly.


Social work and dentistry in clinical, training and research collaboration. The knowledge and knowledge building skills of social workers can improve in three ways the quality and type of services provided to dental patients. First, social workers can be involved in the training of dental professionals so as to improve their interactions with patients. Second, social workers may provide social work clinical services to dental patients. Finally, social workers can conduct research that can add to the body of knowledge on patient behaviors relevant to dental treatment. This paper outlines a program conducted by social workers that incorporates all three of these activities.


Excision of intramedullary arteriovenous malformation using intraoperative spinal cord monitoring. We are presenting a patient whose intramedullary cervical arteriovenous malformation was successfully excised. The importance of monitoring somatosensory cortical evoked responses secondary to peripheral nerve stimulation is stressed.


Superficial temporal artery to superior cerebellar artery anastomosis for distal basilar artery stenosis. The conceptualization, rationale and surgical technique involved with a superficial temporal artery to superior cerebellar artery anastomosis for midbrain ischemia in a patient with symptomatic mid-basilar artery stenosis is described and discussed.


Surgical treatment of giant aneurysm of vertebral-basilar junction. A giant aneurysm of the vertebro-basilar junction is reported. It was successfully treated by ligation of the left vertebral artery, temporary occlusion of the right vertebral and basilar arteries, and evacuation of the aneurysm, following which the neck of the aneurysm was clipped.


Intraventricular pressure and CSF dynamics in chronic adult hydrocephalus. The authors present their experience with continuous intraventricular pressure measurements and the constant infusion test in 50 adult patients suffering from chronic communicating hydrocephalus. A good correlation has been found in 27 cases between the response to CSF shunting and the actual state of the CSF hydrodynamics. The importance of such investigations in selecting adult hydrocephalic patients for shunt therapy is emphasized.


Mechanical properties of human cerebral arteries. Part 1: Effects of age and vascular smooth muscle activation. Mechanical properties of the smooth muscle component of human arteries were studied under passive and active conditions. There exists a considerable difference in the development of sclorosis in intracranial and extracranial arteries. Intracranial arteries were already stiff at birth and become stiffer with age. Their increase in stiffness is attributable to the marked thickening of their walls, which compensates for the decrease of their elasticity with age. Intracranial arteries constrict more than extracranial arteries, although the maximum active stress developed is essentially the same. This competency of intracranial arteries for vaso-constriction may be related to cerebral vasospasm.


Pseudoaneurysm complicating superficial temporal artery--middle cerebral artery bypass. A patient with transient ischemic attacks from occlusion of a left common carotid artery was treated with common carotid endarterectomy immediately followed by superficial temporal artery-middle cerebral artery bypass. Subsequent angiography demonstrated a pseudoaneurysm formation at the anastomotic site. To our knowledge, this is the first report of this complication occurring in a human being.


Cranial CT in multiple sclerosis showing a mass effect. In a patient with histologically confirmed multiple sclerosis, cranial computerized tomography revealed contrast-enhanced lesions showing a mass effect.


Intracranial rhabdomyosarcoma producing bulbar palsy. Two cases of intracranial rhabdomyosarcoma invading the meninges at the base of the skull, causing progressive bulbar palsy and rapid demise, are presented. Discussed are the clinical aspects of this uncommon tumour of interest to the neurosurgeon.


Intracranial tumours among Chinese in Hong Kong. Fifty-six surgical biopsies of intracranial tumours were seen within two years in a newly established neurosurgical unit in Hong Kong. Among the 52 cases of primary intracranial tumours, glioma is the most common, followed by meningioma and pituitary tumour. This is similar to findings in Caucasians. When the distribution of glioma is analyzed, Chinese, or possibly Orientals, have a much higher incidence of ependymoma and pinealoma than do Caucasians. Similar findings are found in other Chinese series reported in the literature.


Intracranial aneurysms causing subarachnoid haemorrhage in the Chinese. Thirty-seven cases of ruptured intracranial aneurysms in the Chinese were retrospectively studied to define their clinical characteristics. The incidence is shown to be low, and a high proportion of anterior communicating artery aneurysms was encountered. Factors contributing to the low incidence are briefly discussed.


Osmotic opening of the blood-brain barrier to methotrexate in the rat. Osmotic opening of the blood-brain barrier in the rat, by intracarotid infusion of 1.6 molal (m) arabinose solution, increases cerebrovascular permeability to 3H-methotrexate by a factor of about seven. PA (product of permeability and capillary surface area) increases from a mean of 3.3 X 10(-5) sec-1 in brains of control rats to as much as 28 X 10(-5) sec-1 at the cerebral hemisphere ipsilateral to carotid infusion. If tracer is introduced into the carotid artery after osmotic treatment, brain uptake is increased by a net factor of 50 (a factor of 70 due to elevation of PA, multiplied by 7 due to infusion by the carotid route) as compared to uptake by normal, untreated brain with infusion into a peripheral vein. Osmotic barrier opening followed by carotid drug administration may be of use in experimental chemotherapy of nervous system disease.


Erythrocyte sedimentation rate following uncomplicated lumbar disc operations. To determine the normal response of the erythrocyte sedimentation rate following uncomplicated lumbar disc operation, 16 patients were studied with serial erythrocyte sedimentation rates during the first six postoperative weeks. The erythrocyte sedimentation rate should be no higher than 25 mm/hr after the first week following an uncomplicated lumbar disc operation in a patient with a normal preoperative erythrocyte sedimentation rate. In patients with elevated preoperative erythrocyte sedimentation rates, further postoperative elevation may be noted. This value may be incorrectly interpreted unless a preoperative erythrocyte sedimentation rate has been determined for baseline purposes.


Occipital encephalocele with holoprosencephaly and aqueduct stenosis. A case is described in which an occipital encephalocele was removed in a newborn and in which a midline monoventricle did not appear until some months later, suggesting the diagnosis of holoprosencephaly. The relationship of encephaloceles and holoprosencephaly is discussed.


Spontaneous extracranial metastasis of cerebral neuroblastoma. A case of extracranial metastasis of cerebral neuroblastoma without previous craniotomy is presented. The tumor in the left posterior frontal region penetrated the dura mater and the skull, formed a lump under the galea and metastasized to the lymph nodes in the infraparotid and superior cervical regions.


Treatment in tumours of the pineal region: value of sterotaxic biopsy. The treatment of tumours of the pineal region is the subject of keen controversy between advocates of surgical excision and those of radiotherapy with ventricular shunting. The proponents of these extremes often seem to ignore the wide pathological variety of tumours of this site. Attempted excision of a germinoma invading the walls of the 3rd ventricle is as futile and dangerous as irradiation of a teratoma. Stereotaxic biopsy provides precise pathological diagnosis and thus allows more specific management; the procedure is safe and no serious complications were experienced in 25 cases.


Endothelial damage in microvascular anastomosis: to splint or not to splint? When performing a microvascular anastomosis, some surgeons use a splint and others do not. We compared the difference in intravascular reactions observed in the carotid artery of the rat anastomosed with these two techniques by observation with a scanning electron microscope. Thrombus formation was more prominent in the artery anastomosed using a splint. The difference was most remarkable 20 minutes after anastomosis. With time, the difference became less remarkable and was no longer present by seven days. By fourteen days, the anastomotic site was covered with neoendothelium. It is concluded that there is little difference in the final outcome achieved by the two methods.


Intramedullary epithelial cyst of the spinal cord. A female patient with an intramedullary cyst presented with a paraplegia. At operation an intramedullary cyst at the fourth dorsal level was found. The microscopic features of the cyst wall were suggestive of ependymal origin.


Supranuclear ophthalmoplegia with basilar artery aneurysms. In two patients, large aneurysms of the basilar artery produced a supranuclear gaze paresis, involving horizontal gaze in one case and vertical gaze in the other. In both cases the diagnosis was suggested by computed tomography and confirmed by vertebral angiography. Autopsy findings in one case confirmed the diagnosis. Although supranuclear gaze palsies must reflect damage to structures deep within the brain stem, these cases illustrate that an extrinsic lesion may produce such damage indirectly. Basilar artery aneurysm should be considered in patients presenting with supranuclear gaze paresis and long tract signs.


Extravasation from aneurysms during angiography. Six cases of extravasation from ruptured intracranial aneurysm during angiography were encountered during the past four years. Cerebral angiography was performed within several hours after the attack in each case. All had emergency radical operations and three of them survived.


Vein graft bypass in treatment of giant aneurysm. A case of giant internal carotid artery aneurysm which was successfully treated by trapping and internal decompression of the aneurysm is presented. Proximal vascular occlusion of the involved internal carotid artery and long vein bypass graft were performed under hypothermia. This is the first long vein bypass graft reported for the treatment of a giant aneurysm.


Metastatic chordoma.

Metastatic chordoma. The authors present two cases of vertebral chordomas with widespread metastases. The literature is reviewed and the metastatic potential of chordomas is analysed.


Extracranial bone metastases from intracranial meningiomas. Distant bone metastases in cases of intracranial meningiomas are uncommon. Two patients with meningiomas showing malignant histological features are described, in both of whom bone metastases were observed many years after cranial surgery.


Prognostic value of carotid angiography in severe head injury. Seventy-seven carotid angiograms in patients with severe head injuries were reviewed. The prognostic value of angiographic findings was evaluated. We were particularly concerned with midline shift, extracerebral avascular area, cerebral circulation time, phlebographic index, and distance between insula and inner table of the skull. Poor visualization of the internal cerebral and the thalamostriate veins, and prolongation of the arterial circulation time were significantly related to a poor prognosis.


Revascularization of the spinal cord by micro-anastomoses in dogs. The severity of ischemic lesions in the spinal cord justifies attempts at its surgical revascularization. The experiments consisted of: (1) creating devascularization of the conus medullaris by ligation of all the lumbo-sacral collaterals of the aorta, and (2) revascularizing the lumbo-sacral rachidian circulation by performing an end-to-side anastomosis between the caudal mesenteric artery and the 5th left lumbar artery (from which in the dog the Adamkiewicz artery generally arises). After two-months, the patency rate of the 20 cases was 85%. Such a procedure of revascularization could be useful in man in cases of interruption of the arterial supply of the spinal cord.


Intracranial hypoglossal neurilemmoma.

Intracranial hypoglossal neurilemmoma. In a patient presenting with long tract motor and sensory signs accompanied by unilateral 12th nerve cranial palsy, we report the successful complete resection of an intra-cranial hypoglossal neurilemmoma. Our case marks the 15th reported observation of such a lesion. The value of computerized tomography in the diagnosis and localization of this type of tumor is emphasized.


Microsurgical excision of a strio-insular arteriovenous malformation. The complete microsurgical removal of a right strio-insular arteriovenous malformation is described. The surgery was performed in two successive operations. The rationale for these operations is discussed.


Association of cerebral aneurysm with pituitary adenoma. Seven cases of coincidental aneurysm with pituitary adenoma found through the review of our personal series of 95 pituitary adenomas over a period of five years are reported. The incidence of coexisting aneurysms in our series of pituitary adenomas (7.4%) was significantly higher than that in other brain tumors (1.1%) (p less than 0.001). Its clinical significance is discussed including the indications for four-vessel angiography, the surgical approaches to these pituitary adenomas, and the management of the associated aneurysms.


Translocation of proteins across membranes: the signal hypothesis and beyond. Proteins are translocated across membranes either coupled to translation (co-translationally) or after translation (post-translationally). The information for both modes of translocation is encoded in the protein in the form of a short-lived sequence extension (signal sequence). Additional information resides in the ribosome in the case of co-translational translocation, which proceeds via a ribosome--membrane junction. Translocation is mediated by specific receptors (ribosome and/or signal receptors) which are restricted in their location to distinct cellular membranes. In most cases the signal sequence is removed by a signal peptidase operating in an endoproteolytic mode. Membranes endowed with receptors for co-translational translocation are: the rough endoplasmic reticulum (RER) including the outer nuclear envelope membrane, the inner mitochondrial membrane and the thylakoid membrane of chloroplasts, in eukaryotic cells; and the plasma membrane in prokaryotic cells. Each of these membranes presumably contains a single distinctive signal receptor, ribosome receptor and signal peptidase. Membranes endowed with one distinct receptor each for post-translational translocation are both mitochondrial membranes, the chloroplast envelope membrane and the peroxisomal membrane. A signal sequence for co-translational translocation across the RER membrane that is identical in its secondary structure is shared by secretory, lysosomal and certain bitopic integral membrane proteins. Some integral membrane proteins presumably share another common sequence--referred to as stop-transfer sequence--which serves to interrupt translocation and thereby to orient the polypeptide chain in the lipid bilayer. Furthermore, the existence of a few specific 'sorting' sequences is postulated. These would be common to many proteins and would serve to route them to their final destination following translocation across or orientation within the membrane. Thus, the topological information which determines the intracellular pathway and the final location of a great number of proteins appears to reside in a small repertoire of specific sequences which are either a transient or a permanent part of the protein.


The medical profession and congenital malformations (1900-1979). Physicians' interest in congenital malformations has varied greatly during the last two centuries. After an acme in the 19th century, teratology was of little inteerest to the medical profession during the first four decades of the 20th century. Since then a variety of events have again made birth defects important. An attempt is made to explain the waxing and waning of the physicians' attention to this age-old problem.


The evolution of teratological testing.

The evolution of teratological testing. The beginnings of mammalian experimental teratology in this century are briefly reviewed and it is noted that prior to 1960 a degree of sophistication in concept and technology had already been achieved. Thus, contrary to claims that teratology had its beginning with the thalidomide catastrophe, a modest but expanding activity and body of knowledge already existed before this unfortunate event. This activity and this knowledge, however, were largely confined to academic and research institute laboratories and made little impact on the agencies in medicine, government and industry which oversaw public health and safety and set policies intended to preserve them. No individual, group, or agency can rightly be blamed for not having sooner brought together the concepts and methodology needed for meaningful animal testing and the regulatory insignt and experience needed intelligently to apply test data to human safety evaluation and experience needed intelligently to apply test data to human safety evaluation. To accomplish this liaison seems to have required the largest toxicological catastrophe yet recorded in human history. The major events leading to formulation of the first standardized guidelines are reviewed, but it is emphasized that even today the best animal testing can only provide a limited statement of probability regarding human risk vis-à-vis safety.


Prune perineum.

Prune perineum. An infant, born to unrelated parents, who had a rugated perineal mass which measured 17 cm in diameter is reported. No external genitalia or anal orifice was identified although the infant voided from a 5 mm crevice on the caudal surface of the mass. The patient died at four weeks of age. The perineal mass was made up of two separate sacs. The anterior sac resembled a urinary bladder in which two ureteral and a single vaginal orifice were identified. The posterior sac was continuous with the peritoneal cavity and contained bowel, left ovary, uterus and right kidney. The left kidney was small, polycystic and the right gonad a streak.


The extent of fetal ossification as an index of delayed development in teratogenic studies on the rat. In teratogenic studies toxic effects may manifest themselves in retarded fetal development, such as a reduction in fetal weight. In searching for an additional index, the number of centers of ossification in seven skeletal districts (sternum, metacarpus, metatarsus, cervical and caudal vertebrae, anterior and posterior proximal phalanges) of rat fetuses delivered on days 19, 20 and 21 of gestation were counted and compared. Results showed uneven ossification in day-19 and -20 fetuses, but sufficiently advanced, homogeneous and uniform ossification in day-21 fetuses to provide a reliable quantitative index for evaluating retarded fetal development. It is therefore proposed that the stage of skeletal ossification in day-21 fetuses be used in teratogenic studies in the rat to evaluate retarded fetal development.


Palate morphogenesis. III. Changes in cell shape and orientation during shelf elevation. The process of palate shelf elevation has been analyzed by light microscopy in mouse embryos cultured in vitro. The observations presented correlate changes in cell shape and orientation in the palate with the morphogenetic movement of the shelf. These studies suggest that in addition to any physical-chemical force elevating the shelf an active contraction of specific palate cells could also aid the process. Contribution to elevation could be derived from masses of contracting cells from the previously described non-muscle contractile systems in posterior (region 2) and mid-anterior (region 3) palate as well as other peripheral mesenchymal cells. Finally, elongation and contraction of the tongue side epithelial cells may also play a role in palate elevation.


Morphological alterations in the developing fetal rat limb due to maternal injection of chlorambucil. On day 14 of pregnancy, rats were injected i.p. with 9.5 mg/kg of chlorambucil. At term, all the fetuses were stunted and had obvious skeletal malformations. From days 15-20 of gestation there was a steady progression of cell death and histologic derangement in long bone formation. This began on day 15, the day following exposure to chlorambucil, with extensive cell death of chondroblasts in the long bone primordia. By day 16, dead or dying cells were located in the cartilage model and became associated with a higher than normal matrix to cell ratio. In day 17 limbs, many cells of the cartilage model were irregular in size and shape so that the normally precise cellular arrangement was lacking. On day 18, periosteal and enchondrial calcification was delayed and scanty. The fibular hemimelia observed in all drug-treated term fetuses stained with alizarin appeared to result from lack of development of the proximal half of the cartilage model. The observed alterations, particularly bowing of the ulna, in the teratogen-treated fetuses appeared to result primarily from specific cell death and formation of aberrant cell types but abnormal cell products may also play a role.


The early limb deformity caused by acetazolamide. Histologic sections were made of the forelimbs of day 10 to 12.5 embryos removed 3, 16, 30, 36, 42, 45, 63 and 75 hours after the intraperitoneal injection of acetazolamide into C57Bl/6J pregnant females. A postaxial deformity was first visible in the right forelimb of day 10.9 embryos 36 hours after the second dose of acetazolamide 1,000 mg/kg on day 9 of gestation. The deformity was a postaxial deficiency of mesenchyme tissue without any evidence of cell necrosis. The deformity was present before aggregation of mesenchyme cells occurs as the first step in chrondrogenesis. The pattern of cell orientation in the adjacent mesenchyme aggregate of a deformed day 12.5 forelimb was normal. This means that the primary effect of acetazolamide is not on the formation of precartilage aggregates in the postaxial region, but involves other aspects of early limb development.