Chromatographic characteristics of surfactant-mediated separations: micellar liquid chromatography vs ion pair chromatography. The effects of concentrations of organic solvent and surfactant on elution strength and selectivity in MLC and IPC are studied. It is observed that selectivity between most pairs of solutes used in this study increases in MLC and either decreases or passes through a minimum in IPC, with the volume fraction of organic modifier. In both MLC and IPC, selectivity varies with surfactant concentration; however, the overall variation in selectivity and elution order are more pronounced in MLC. The solvent strength decreases in IPC and increases in MLC as a result of an increase in surfactant concentration. An iterative regression design is used to predict the optimum mobile-phase compositions in terms of solvent strength and selectivity. The correlation between the predicted and measured chromatograms is excellent in MLC and poor in IPC. This is due to a more regular and reproducible retention behavior in MLC which greatly facilitates the development of robust methodologies. For a mixture of amino acids and peptides, a large retention gap between the first and the last eluting solutes is observed in IPC, which makes the use of organic solvent gradient inevitable. However, a better separation for the same mixture of solutes can be achieved in MLC isocratically. Apparently, the general elution problem can be alleviated in MLC by using an optimum eluent composition. It is observed that the efficiencies of MLC and IPC are comparable. The above observations indicate that MLC can be a powerful alternative to IPC in order to achieve optimized separations in shorter analysis time.


Specific base recognition of oligodeoxynucleotides by capillary affinity gel electrophoresis using polyacrylamide-poly(9-vinyladenine) conjugated gel. Poly(9-vinyladenine) was synthesized and utilized as an affinity macroligand entrapped within the gel matrix. Base-specific separation of oligodeoxynucleotides was achieved with high resolution and high speed by electrophoresis, using capillaries filled with conjugated polyacrylamide-poly(9-vinyladenine) gel. Oligothymidylic acids were selectively separated from the mixture of oligothymidylic and oligodeoxyadenylic acids by utilizing a specific hydrogen bonding between poly(9-vinyladenine) and oligothymidylic acids. Migration time and resolution of oligodeoxynucleotides were influenced by several parameters, such as the size of poly(9-vinyladenine), capillary temperature, and concentrations of poly(9-vinyladenine) and urea. Some guidelines are presented, based on the theoretical formulation of the effect of these parameters, in order to find optimum electrophoretic conditions. Analytical capillary affinity gel electrophoresis was developed for the selective and sensitive base recognition of oligodeoxynucleotides with efficiencies as high as several 10(6) plates/m by using a urea-gel capillary with poly(9-vinyladenine) and temperature-programming.


Noninvasive imaging of nanogram quantities of DNA in agarose electrophoresis gels. Image processing techniques which separate the DNA Kerr effect and induced electrokinetic distortion contributions to electric birefringence images of agarose nucleic acid electrophoresis gels are described. Under standard electrophoresis conditions, detection limits of 10 ng of DNA per well are obtained in hydroxyethylated agarose without signal averaging or 7.5 ng with averaging of four measurements. Maintaining constant gel temperature is shown to improve the quality of the images. Monochromatic light (589 nm) is shown to give a small increase in sensitivity compared to polychromatic (650 +/- 20 nm) light.


Kinetic chromatographic sequential addition immunoassays using protein A affinity chromatography. A new type of chromatographic immunoassay based on sequential addition is described. On a protein A column, the antibody, the sample containing the antigen, and then a known amount of antigen are sequentially injected. This assay is designed to shorten analysis times and reduce complexity of dual-column chromatographic immunoassays, circumvent desorption buffer interferences common to affinity chromatography, and eliminate the need for tagged molecules. This new technique is named kinetic immunochromatography sequential addition (KICQA). Because of its kinetic nature, flow rate will have a large effect on KICQA, and the impact of changing flow rate is studied extensively. By use of various amounts of antibody, the dynamic range of KICQA is shown to be selectable over 2.5 orders of magnitude. Finally, KICQA was used to determine transferrin and albumin in human serum. Both analytes show good agreement with their respective reference methods, and an albumin assay was performed in under 1 min.


Time-of-flight mass spectrometry with an electrospray ion beam. An electrospray ionization source has been coupled to a reflection time-of-flight mass spectrometer. By orienting the ion source perpendicular to the field-free drift region, the longitudinal energy spread of the ion packet has been substantially reduced, allowing a mass resolving power of over 1000 to be achieved for both low-mass and high-mass ions of biological interest. In addition, instrument sensitivity allows the routine detection of low-picomole and subpicomole quantities of large multiply charged species such as cytochrome c (MW = 12,360.9). The potential utility of this instrument for conducting rapid screening of chromatographic effluents is discussed in light of its simplicity, rapid scanning speed, and high sensitivity.


Separation and identification of carotenoids and their oxidation products in the extracts of human plasma. Eighteen carotenoids as well as vitamin A and two forms of vitamin E (gamma- and alpha-tocopherol) have been separated from extracts of human plasma by high-performance liquid chromatography (HPLC) on reversed-phase and sillca-based nitrile-bonded columns. In the order of chromatographic elution on a C18 reversed-phase column, the carotenoids were identified as (3R,3'R,6'R)-beta, epsilon-carotene-3,3'-diol [(3R,3'R,6'R)-lutein], (3R,3'R)-beta, beta-carotene-3,3'-diol [(3R,3'R)-zeaxanthin], 5,6-dihydroxy-5,6-dihydro-psi,psi-carotene, 3-hydroxy-2',3'-didehydro-beta,epsilon-caroten-3-ol, 3-hydroxy-beta-carotene,psi,psi-carotene, 7,8-dihydro-psi,psi-carotene, beta,psi-carotene, 7,8,7',8'-tetrahydro-psi,psi-carotene, beta,epsilon-carotene, beta,beta-carotene, 7,8,11,12,7',8'-hexahydro-psi,psi-carotene, and 7,8,11,12,7',8'-11',12'-octahydro-psi,psi-carotene. The polar carotenoids, which eluted in the vicinity of lutein and were unresolved on the C18 column, have been separated on a nitrile-bonded column employing isocratic HPLC conditions. In the order of elution, the carotenoids were epsilon,epsilon-carotene-3,3'-dione, 3'-hydroxy-epsilon,epsilon-caroten-3-one, 5,6-dihydroxy-5,6-dihydro-psi,psi-carotene, 3-hydroxy-beta,epsilon-caroten-3'-one, (all-E,3R,3'R,6'R)-lutein, (all-E,3R,3'R)-zeaxanthin, and (all-E,3R,3'S,6'R)-beta,epsilon-carotene-3,3'-diol (3'-epilutein) followed by several geometrical isomers of lutein and zeaxanthin.


DNA sequencing using capillary array electrophoresis. A DNA sequencing method is presented that utilizes capillary array electrophoresis, two-color fluorescence detection, and a two-dye labeling protocol. Sanger DNA sequencing fragments are separated on an array of capillaries and detected on-column using a two-color, laser-excited, confocal-fluorescence scanner. The four sets of DNA sequencing fragments are separated in a single capillary and then distinguished by using a binary coding scheme where each fragment set is labeled with a characteristic ratio of two dye-labeled primers. Since only two dye-labeled primers are required, it is possible to select dyes that have identical mobility shifts. It is also shown that the ratio of the signal in the two detection channels provides a reliable identification of the sequencing fragment. DNA sequencing results on a 25-capillary array are presented.


Drug of abuse confirmation in human urine using stepwise solid-phase extraction and micellar electrokinetic capillary chromatography. This paper demonstrates that most common drugs of abuse can be absorbed simultaneously onto a mixed-mode bonded-phase matrix and eluted sequentially in two to three steps for subsequent analysis by micellar electrokinetic capillary chromatography (MECC). Having on-column multiwavelength UV absorption detection, this is shown to be an attractive approach for confirmation testing of barbiturates, hypnotics, amphetamines, opioids, benzodiazepines, and metabolites of cocaine in a single aliquot of human urine. For these compounds, no hydroysis of the urine specimen or sample derivatization is required. Under the examined conditions using 5 mL of urine, excellent recoveries (80-90% level) and detection limits (about 100 ng/mL) are obtained. For patient urines which tested positively for different classes of drugs using immunological screening methods, a two-step extraction scheme is shown to provide extracts suitable for rapid MECC confirmation of the drugs of abuse.


Immunohistochemistry for intermediate filaments in the enteric nervous system of the porcine small intestine. Antibodies against the cytoskeletal neurofilament protein 200 and gliafilament acidic protein were used for an immunohistochemical staining of nerve and glia cells in porcine small intestine. In sections as well as in whole mount preparations, the morphological and topographical features of the enteric nerve plexus could be demonstrated. The enteric glia cells are characterized by an abundance of immunoreactive GFAP, which allows a subsequent staining of the plexus. NFP 200 is immunohistochemically recognized only in a part of the neurons. This immunoreactive neuronal population can be identified morphologically as Typ II-neurons, which are defined as adendritic and pseudouniaxonal to multiaxonal. The immunostaining of intermediate filaments is an easy and reproducible means for studying the enteric nervous system and invaluable for the histopathological diagnosis of its morphological abnormalities.


Lectin binding pattern in human retinal pigment epithelium. A comparative lectin histochemical study of human retinal pigment epithelium (RPE) was performed to investigate the lectin binding pattern of normal, reactive and proliferating RPE. Normal RPE with attached sensory retina was found to bind the lectins Con A, WGA, PNA and RCA I. Reactive and proliferating RPE in retinal detachment and in photocoagulation scars revealed the same lectin binding pattern although its cellular topography changed. RPE-macrophages showed an additional reaction with SBA. In periretinal membranes of human PVR the typical lectin binding pattern of Con A, WGA, PNA and RCA I was found in pigmented and in a subpopulation of non-pigmented cells, suggesting that these lectin-positive elements were of RPE-origin. Additionally, single pigmented cells positive for SBA were found indicating macrophage differentiation. Thus lectin histochemistry provides a tool for cytochemical identification of RPE and its morphologic variants by revealing a specific combination of sugar-binding sites.


Morphological variants of the suprapatellar bursa. Two hundred and ten specimens were examined anatomically to study the different shapes of the suprapatellar bursa. It is possible to classify the morphologic variants of the suprapatellar bursa into three types. A bursa not subdivided was found in 83% and named Type A. A bursa subdivided by a vertical septum at the lateral side of the suprapatellar bursa was seen in 17% of the knee joints. The vertical septum was complete in 9% (Type B), incomplete in 8% (Type C) of the cases. It separated the bursa into a main medial part and a small lateral saccular space. There was no corresponding subdivision at the medial side. A space proximal to the suprapatellar bursa was found in two cases.


Fine structure of the dorsal lingual epithelium of the domestic, newborn kitten, Felis catus. The tip and the body of the tongue of the domestic kitten, Felis catus, were examined by light and transmission electron microscopy. On the tip of the tongue, no filiform papillae were observed, but the connective tissue papillae of the lamina propria were recognized. On the lingual body, there were filiform papillae composed of an anterior, a posterior and interpapillar epithelium. Under the transmission electron microscope, the epithelium on the tip of the kitten tongue was found to be of the stratified squamous type. The epithelium contained no cells filled with keratin fibers. In the lingual body, the interpapillar epithelium contained very few keratohyalin granules, and no cells with keratin fibers. In the epithelium on the anterior side of the filiform papillae, numerous keratohyalin granules appeared in the intermediate layer. In the surface layer, a thin layer of typical keratinized cells was visible. In the epithelium on the posterior side of the filiform papillae, a thick layer of keratinized cells was located on the surface layer.


Prenatal influence of alcohol following a single exposure in two inbred strains of mice. The effect of a single dose of ethanol on embryonic development was investigated in two inbred strains of mice. Ethanol (25% v/v solution, 0.02 ml/g body weight) was administered intraperitoneally on either gestational day 6, 9 or 12 to C57BL/6J and DBA/2J mice. All fetuses recovered near term were alive and appeared normal. The incidence of fetal death was not affected. Fetal weight was significantly reduced in the C57BL/6J strain of mice. Maternal exposure to a single and moderate dose of ethanol did not have any profound effect on the conceptus. For the development of laboratory models to mimic fetal alcohol effects in humans, the selection of appropriate genotype is important.


Embryonic development in the chick following exposure to ethanol, acetaldehyde and cyanamide. The influence of cyanamide, an inhibitor of aldehyde dehydrogenase, on the embryopathic effects of ethanol and acetaldehyde, was investigated in the chick embryo. Both ethanol and cyanamide significantly increased embryonic mortality, but did not affect embryonic growth, compared to treatment with either ethanol or cyanamide. Acetaldehyde combined with cyanamide increased embryonic mortality and retarded embryonic growth. Cyanamide influence on embryonic development was minimal. The extent of acetaldehyde involvement in ethanol teratogenicity remains unclear from the present findings.


Anomalous course of the median nerve medial to the trochlea and anterior to the medial epicondyle of the humerus. It is generally assumed that in the cubital region the median nerve is directly medial to the brachial artery and that it lies anterior to the trochlea of the humerus, from which it is separated by the brachialis muscle. Three instances are noted where the median nerve passed medial to the trochlea and anterior to the medial epicondyle, deep to the pronator teres, unilateral in one subject and bilateral in another. Its possible significance in elbow trauma is commented upon.


Human doubled renal and testicular arteries. Doubled renal and testicular arteries were found in a well-developed 69-year-old caucasian male. The right kidney had two renal arteries, one at its usual midorgan (hilar) position and one inferior polar. One testicular artery arose from the mid-point of the usual renal artery, passed under the renal vein at its junction with the inferior vena cava, and then crossed over the inferior polar renal artery. The second testicular artery arose from the inferior polar renal artery near its origin from the abdominal aorta. The two testicular arteries remained doubled throughout their course and both entered the right testis at separate sites on the organ. The inferior vena cava passed under the inferior polar and over the superior testicular artery and the superior renal artery.


[A rare variation in the zone of the carotid artery]. An extremely rare variation in the angioarchitecture of the carotid arteries consists in the complete missing of the right common carotid artery. The brachiocephalic trunc (innominate artery), forming a bulbus-like extension, branches out into three major stems: internal carotid artery, external carotid artery and the right subclavian artery. The variation is embryologically explainable: The aortic arches III and IV arise together (without ventral aortic root between them) from the persisting initial segment of the right ventral aorta.


Bending strength of upper canine tooth in domestic dogs. The bending strength of the upper canine tooth is analyzed on 51 domestic dogs of various breeds. Data show that canine tooth length is greater among small domestic dogs when compared to small wild dogs, and large wild dogs when compared among large domestic dogs. Tooth strength is also greater among small domestic dogs and large wild dogs. The greater tooth strength of the small domestic dogs may be attributed to the idea that small domestic dogs are closer in ancestry to the grey wolf (Canis lupus) than are small wild canids. Skull foreshortening and tooth strength appear to be correlated in the canidae.


[Fine structure of the raphe nuclei of sheep and goats]. The medullary raphe system of sheep and goat can be divided in the solitary nuclei raphe magnus and pallidus besides the impaired Nucleus raphe paramedianus. The nucleus raphe magnus begins in the caudal end of the medulla oblongata running far to the pons. On the other hand you can follow the nucleus raphe pallidus and paramedianus only in distinct area near the obex. Different types of neurons are isolated and described in light- and electronmicroscopical investigations. All three nuclei are poor of synapses.


Stability of neuron number in the lateral part of the nucleus of the solitary tract in the ageing mouse brain. Neuron number was estimated in the lateral part of the nucleus of the solitary tract of the mouse brain at 6, 15, 22, 25, 28 and 31 months of age. There was no significant variation in neuron number with age with an overall mean of 8500.


Stability of neuron number in the ageing mouse paraventricular nucleus. The number of neurons in the paraventricular nucleus was estimated in brains from male mice aged 6, 25, 28 and 31 months. There was no significant variation with age in neuron number (mean 2715). At all ages most neurons were parvocellular neurons and the neurosecretory magnocellular neurons made up only 10% of the total. There was no significant variation in neuronal nuclear diameter (mean 9.4 microns) with age. The nuclear diameter of both parvocellular and magnocellular neurons was identical. The difference in size between the two types of neurons was due to the difference in volume of their perikarya.


Immunohistochemical localization of relaxin in the reproductive system of the female opossum (Didelphis virginiana). Relaxin-immunoreactivity was demonstrated in the cytoplasm in the luteal cells from pregnant and lactating opossums. Immunoreactivity for relaxin was not demonstrated elsewhere in the ovary, in the reproductive tract or in the placenta. The corpus luteum is thought to be the primary source of relaxin in Didelphis and in this regard is similar to several eutherian mammals including man.


Ultrastructure of the exocrine pancreas in the snake Waglerophis merremii (Wagler). Three types of serozymogenic cells were found in the secretory compartment of the snake exocrine pancreas. Type I cell was the most common and presented a well-developed granular endoplasmic reticulum arranged in cisternal and vesicular forms. The cisternal form was located predominantly in the basal regions of the cell and the vesicular form was found in the supranuclear regions of the cell next to a prominent Golgi complex. Mature secretory granules were seen at the cell apex. The cytoplasmic matrix of the Type II cells was electron dense but had only poorly-developed organelles. Secretory granules were rare. The cytoplasmic matrix of the Type III cells was electron lucent and the granular endoplasmic reticulum in the cisternal form was located predominantly in the supranuclear region, whereas the vesicular form was randomly distributed throughout the cytoplasm. The nucleus appeared pale due to the fine dispersion of the chromatin; the nucleolus was prominent. Centroacinar and intermediate cells were also examined.


An HRP study of the location of the motoneurons supplying the tensor veli palatini muscle of the rabbit. Location of the motoneurons supplying the tensor veli palatini muscle of the rabbit was examined with the retrograde labeling technique following intramuscular injection of HRP. Labeled motoneurons were ipsilaterally located in the ventral or ventromedial portion of the rostral two-thirds of the motor trigeminal nucleus at the level of about 6.0 to 8.5 mm rostral to the obex. The location of the labeled motoneurons was ventromedial to the region supplying the masseter, the temporalis, and the medial pterygoid muscles and ventral to the region supplying the anterior digastric and the mylohyoid muscles, the location which coincided with the lateral pterygoid region. The labeled motoneurons were scattered around or in this region.


Differentiation of oxytalan fibres from elastic fibres with reagents for detection of magnesium. Elastic fibres in various human and animal tissues revealed positive results with reagents for the detection of magnesium. By contrast, no Mg could be detected in oxytalan fibres or in zonular fibres of the ciliary body, which are composed of microfibrils not containing elastin. Mg is therefore concluded to be associated with the elastin core of elastic fibres. Its antagonism to calcium is speculated to play a protective role in maintaining the extensibility of elastin.


[History of classification systems of conjoined twins by specific consideration of the classification systems of Louis Bolk (1866-1930)]. Classification systems concerning conjoined twins have been developed in the nineteenth century by many authors, a.o. J. F. Meckel (1816), I. Geoffroy Saint-Hilaire (1832), E. F. Gurlt (1831), Fr. Ahlfeld (1880), C. Taruffi (1881), and in the beginning of the twentieth century, a.o. J. W. Ballantyne (1902) and E. Schwalbe (1907). However, these classification systems were very complex and mainly based on the external morphology of the conjoined twins. In contrast to many other classification systems the professor in Anatomy at the University of Amsterdam Louis Bolk divided conjoined twins in only three main groups: 1 greater than diplopagi simplex caudad; 2 greater than diplopagi simplex craniad; 3 greater than diplopagi simplex mesad. The last group was divided into ventrad and laterad conjuncti. For the study of conjoined twins the Vrolik Collection and many other specimens of the Museum of the Department of Anatomy and Embryology of the University of Amsterdam were very important for Bolk. Three factors were the main reason that the concept of the classification of double monsters of Bolk has not been cited often in the international literature: 1 greater than the publications of the classification in a national journal in Dutch; 2 greater than the coincidence of the publication of E. Schwalbes famous handbook "Die Morphologie der Missbildungen des Menschen und der Tiere II. Die Doppelbildungen" in the same period and 3 greater than the problem that the verification of the classification, mainly based upon morphogenesis, has not been possible because of technical problems in performing these experiments in mammals.(ABSTRACT TRUNCATED AT 250 WORDS)


Distribution of type I collagen, type II collagen and PNA binding glycoconjugates during chondrogenesis of three distinct embryonic cartilages. Previous studies of chondrogenesis have been focused on limb bud cartilage, whereas little is known about chondrogenic processes of other cartilages with different developmental fates. We hypothesize that cartilages with various developmental fates might show identical characteristics of chondrogenesis. The chondrogenic processes in the nasal septum, the mandible, and the limb bud of the mouse were examined by means of PNA-binding glycoconjugate, and types I and II collagen expression. Swiss-Webster mouse embryos of 11 days (E11) to 14 days (E14) gestation were fixed and processed for immuno- and lectin histochemistry. The blastema of mesenchymal cell aggregates stained positively with anti-type I collagen, but very weakly with anti-type II collagen in all three models at E12, whereas PNA bound to the blastema in the limb bud but not in nasal septum or mandible. Types I and II collagens coexisted in cartilages at E13. Type II collagen was predominant in E14; type I collagen was confined to the peripheral region. The synchronized transitional expression of the collagen phenotypes in all three embryonic cartilages may be systemically regulated. The presence or absence of the PNA-binding glycoconjugates may be involved in characterizing the nature of the cartilages.


Cholinergic and non-cholinergic neurons in the rat pedunculopontine tegmental nucleus. Choline acetyltransferase immunhistochemistry was employed at light and electron microscopic levels in order to determine the distribution of cholinergic neurons in two subdivisions of the rat pedunculopontine tegmental nucleus that were previously defined on cytoarchitectonic grounds, and to compare the synaptic inputs to cholinergic and non-cholinergic somata in the subnucleus dissipatus, which receives major input from the substantia nigra. Large cholinergic neurons were found in both the pars compacta and the pars dissipata of the pedunculopontine nucleus. However, they were intermingled with non-cholinergic neurons and did not respect the cytoarchitectural boundaries of the nucleus. Ultrastructural study showed that all cholinergic neurons in the subnucleus dissipatus exhibited similar features. The majority had large somata (largest diameter greater than or equal to 20 microns) containing abundant cytoplasmic organelles and nuclei displaying a few shallow invaginations. Synaptic terminals on the cholinergic cell bodies were scarce and unlabeled boutons containing spherical synaptic vesicles and establishing asymmetric synaptic junctions were the dominant type. In contrast, the non-cholinergic neurons presented prominent differences in the size of their somata as well as in the distribution of axosomatic synapses. Two almost equally represented classes of non-cholinergic neurons which are referred to as large (largest diameter greater than or equal to 20 microns) and small (largest diameter less than 20 microns) were recognized. Large non-cholinergic cell bodies were ultrastructurally similar to the cholinergic ones, but they received rich synaptic input by unlabeled nerve terminals which contained pleomorphic vesicles and were engaged in symmetric synaptic junctions. Small non-cholinergic cell bodies were characterized by deeply invaginated nuclei surrounded by a narrow rim of cytoplasm, and were often found near or in direct apposition to the cholinergic somata. Their major input consisted of axosomatic boutons containing round synaptic vesicles. These results demonstrate that cells in the pedunculopontine tegmental nucleus are differentiated with regard to their axosomatic synaptic inputs which may influence their firing properties. Furthermore, they support previous suggestions that nigral afferents may be preferentially distributed to a subpopulation of the pedunculopontine neurons.


Mast cell populations in the chick embryo lung and their response to compound 48/80 and dexamethasone. Two mast cell populations, connective tissue mast cells (CTMCs) and mucosal mast cells, (MMCs) containing different proteoglycans in their granules, can be distinguished in several animal species by means of histochemical methods. In this study we documented the presence of these two types of mast cell in the chick embryo lung, from the 15th incubation day for the MMCs, and from the 18th incubation day for the CTMCs. Lungs of embryos treated with compound 48/80, which produces degranulation of the CTMCs, showed a decrease in the number of this type of mast cell and an unchanged number of MMCs. In the lungs of embryos treated with dexamethasone, which degranulates MMCs, a reduction in the number of these cells and an unchanged number of the CTMs were found.


Laminar distribution and morphology of NADPH-diaphorase containing neurons in the superior colliculus and underlying periaqueductal gray of the rat. We have studied the laminar distribution of reduced nicotinamide dinucleotide phosphate diaphorase (NADPH-d) activity and the morphology of positive neurons in the superior colliculus (SC) and the underlying periaqueductal gray (PAG) of the rat. The morphology of NADPH-d-positive neurons has been compared to that of Golgi-impregnated cells. The highest activity occurs in the stratum zonale and stratum griseum superficiale, contrasting with the pale neuropil in the stratum opticum, where only a few positive neurons are found. In the stratum griseum intermedium positive neurons are grouped in patches separated by narrow, NADPH-d-negative bands. In the deeper layers, the neuropil is NADPH-d-negative, and few neurons show enzymatic activity. In contrast, numerous neurons in the dorsolateral part of the PAG are intensely positive. They are continuous with the positive neurons in the stratum album profundum, with no clear border between the two centers. In both SC and PAG, only small and medium sized neurons are NADPH-d-positive. In comparison with Golgi material, all types of small neurons in the superficial layers show NADPH-d activity; NADPH-d histochemistry, however, does not visualize the characteristic dendritic appendages of these neurons. The large neurons of the SC and PAG, probably representing the long-projecting neurons of these centers, do not contain the enzyme.


Late postnatal development and differentiation of the ductus epididymidis in a dasyurid marsupial (Antechinus stuartii). The general histology and ultrastructural features of the developing ductus epididymidis were examined in the brown marsupial mouse, Antechinus stuartii, from April, when males were sexually immature, until August, when the adult males were involved in mating activities, just prior to the annual male die-off. Samples were also examined 3 and 6 months after the August die-off period in males kept in isolation from conspecifics during the prebreeding and breeding periods. In April, tubule diameter and epithelial height were largest in the caput and least in caudal segments but the reverse was observed thereafter. Epithelial height increased in caput segments in August and remained high in the post die-off samples. However, caput epithelial height and tubule diameters were low compared with the remainder of the duct from July until February. Luminal shape in caudal segments (10, 11 and 12) changed in June from circular to a narrow slit, and the epithelium became variable in height. The epididymal epithelium was undifferentiated with few cytoplasmic organelles in April. Differentiation occurred mostly from May to June in association with an increased abundance of cytoplasmic organelles, increasing prostatic weight and rising plasma androgen levels. Differentiated principal and basal cells were found in caput and corpus regions in May and in caudal segments in June in association with the de novo development of a brush border of microvilli. Few clear cells were seen in caput and corpus regions of the duct in May but they, and mitochondria-rich cells, were common throughout the duct from June. Development of the unusual structural features of the cauda epididymidis preceded the arrival of spermatozoa in June. The presence of degenerating spermatozoa and cytoplasmic droplets in the cauda at this time suggested that it was not yet capable of supporting sperm viability. There was no evidence to suggest that the presence of spermatozoa has a stimulatory effect on the epididymis. Intact sperm were observed throughout the duct from July. Free cytoplasmic droplets, which showed some evidence of degeneration, collected in large masses in the distal corpus/proximal cauda epididymidis of adult males between aggregates of spermatozoa. Epididymal differentiation appeared complete by mid-July; few ultrastructural changes occurred after this time. Recruitment of spermatozoa into the epididymis ceased by August and was associated with a rapid decline in sperm content in the proximal caput segments. In the November and February samples, spermatozoa were present only in distal corpus and proximal cauda segments.(ABSTRACT TRUNCATED AT 400 WORDS)


Fetal antigen 2 (FA2) in human fetal osteoblasts, cultured osteoblasts and osteogenic osteosarcoma cells. Immunohistochemical staining techniques used on an 11-week-old fetus showed that fetal antigen 2 (FA2) was present intracellularly in endochondral and perichondral osteoblasts, and the immunoreaction was extended into the adjacent bone matrix. Osteoclasts and chondroblasts were found to be FA2 negative. A granular perinuclear intracytoplasmic FA2 immunoreaction was found in cultured osteoblasts and osteogenic osteosarcoma cells, and immunoelectron-microscopical examination revealed a granular immunoreaction product in the rough endoplasmic reticulum. These findings indicate that FA2 is synthesized by osteoblasts and osteogenic osteosarcoma cells. A reaction of immunological identity was found between FA2 purified from second trimester amniotic fluid and serum-free supernatants of cultured osteogenic osteosarcoma cells. This shows that an antigen recognized by the anti FA2 antibody is secreted by these malignant cells. Thus, FA2 may represent a marker for altered bone metabolism, and have a potential in the classification of osteogenic osteosarcoma/chondrosarcoma.


Histochemical distribution of zinc in the brain of the rainbow trout, Oncorhynchos myciss. II. The diencephalon. The distribution of zinc in the diencephalon of the rainbow trout, Oncorhynchos myciss, is described in the present paper, which is the second in a series of three reporting for the first time the distribution of a heavy metal in the fish brain. The Neo-Timm method was used for the histochemical demonstration of zinc. The staining was essentially confined to the neuropil, in all probability representing stained axon terminals, but stained nerve cell bodies were observed in the nucleus lateralis geniculatus and the nucleus cerebellosus hypothalami. Stained fibers were never seen. The staining gave rise to a consistent, specific distribution pattern, which accorded well with the diencephalic nuclei defined on the basis of cytoarchitectural criteria. The diencephalon was in general stained with much higher intensity than the telencephalon, in surprising contrast to the state of affairs in the mammalian, reptilian, and avian brain. In species of these classes, high staining intensities are observed almost exclusively in the telencephalon. The Neo-Timm staining was predominantly distributed in the nuclei of the periventricular zone, but some internal (migrated) nuclei did show a positive staining reaction, namely the nucleus lateralis geniculatus, the anterior thalamic nucleus, the nucleus diffusus tori lateralis, and the nucleus cerebellosus hypothalami. The zinc distribution pattern has been compared with the terminal fields of afferent projections, known from experimental studies, and with the distribution of substance P. The possible function of zinc in synaptic vesicles is considered.


The relationship between the number of nuclei of an osteoclast and its resorptive capability in vitro. This study examined the relationship between the number of nuclei in an osteoclast and its resorptive efficiency, as demonstrated by the size of the pit it can make in a mineralized tissue in 24 h in vitro. Osteoclasts released mechanically from prehatch chick long bones were cultured on dentine slices or on plastic dishes for periods of 6 or 24 h. The frequency distribution of the multinucleate tartrate-resistant acid phosphatase (TRAP)-positive cells with different numbers of nuclei was determined: the mean number of nuclei per cell was 6.92, with a mode of 4.47% had 5 or fewer nuclei and only 11% more than 10 nuclei. The pits associated with 292 osteoclasts with known numbers of nuclei were measured using a confocal laser light microscope (Lasertec) and dedicated image analysis system, and depths, plan areas and volumes determined. There was a positive correlation between the number of nuclei per osteoclast and the volume of the pit made, but a trend for the volume resorbed per nucleus to decrease with increase in the number of nuclei per osteoclast.


Superfluousness of motor innervation for the formation of muscle spindles in neonatal rats. Muscle spindles form de novo in reinnervated muscles of neonatal rats treated with nerve growth factor. Whether the spindles can also form in muscle reinnervated only by afferents was investigated by removing the lumbosacral segment of the spinal cord immediately after crushing the nerve to the medial gastrocnemius muscle at birth, and administering nerve growth factor for 10 days afterwards. As predicted, the medial gastrocnemius muscles were reinnervated by afferents, but not efferents. No motor endplates were visible on any muscle fibers, and extrafusal fibers were atrophied. The reinnervated muscles contained spindle-like encapsulations of one to four fibers at 5, 7, 9 and 30 days after the nerve crush. The number of spindles as well as encapsulated fibers exceeded that of normal medial gastrocnemius muscles. The encapsulated fibers resembled typical intrafusal fibers. They had normal sensory-muscle contacts, but no motor endings. The fibers displayed equatorial clusters of myonuclei and expressed the spindle-specific slow-tonic myosin heavy chain isoform at postnatal day 30. Thus, efferents are not essential for the formation and differentiation of muscle spindles in reinnervated muscles of neonatal rats.


Schwann cells are not required for guidance of motor nerves in the hindlimb in Splotch mutant mouse embryos. The topogenesis of the hindlimb nerves of Splotch homozygous mutant mouse embryos was studied using light and electron microscopy. Homozygous mutants show multiple defects of neural crest-derived tissues. The defects increase along a rostro-caudal gradient. The cervical and upper thoracic segments have small spinal ganglia, and Schwann cells are associated with the spinal nerves. In the lumbo-sacral region neurulation is not complete, and the derivatives of the neural crest are missing. The lumbo-sacral nerve trunks are formed by ventral roots only. They are occasionally associated with presumptive glial cells that have migrated from the spinal cord for a short distance. Beyond the vertebral primordia, the spinal nerves are not accompanied by Schwann cells. No compartmentalization of the axons within the lumbo-sacral nerves was visible, whereas Schwann cells did segment the nerve into the fascicles in brachial nerves. The lumbo-sacral plexus develops, and its branches grow into the hindlimb despite the absence of Schwann cells. On day 13.5 of gestation, the lumbo-sacral nerve trunks extend well into the distal calf. They are topographically correctly positioned. Their branches enter the muscle primordia and form contacts with their mesenchymal cells though the cutaneous branches are missing. Generally, the outgrowth of lumbo-sacral nerves is slower than in phenotypically normal littermates, whose nerves reach the foot plate at corresponding stages of development. These results demonstrate that the lumbo-sacral plexus and the topographically correct position of lumbo-sacral nerve trunks develop despite the absence of Schwann cells. Therefore Schwann cells are not necessary for the outgrowth and guidance of axons within the limb.


A scanning electron microscopic study on the distribution of peritubular dentine in cheek teeth of Cervidae and Suidae (Mammalia, Artiodactyla). Distribution of peritubular dentine was studied in cheek teeth of fallow deer (Dama dama), roe deer (Capreolus capreolus) and wild boar (Sus scrofa). In the two cervid species, especially intense peritubular dentine formation was found in the outer regions of mid and cuspal coronal dentine. In these areas a marked asymmetry occurred, peritubular dentine being predominantly secreted onto the side of the dentinal tubule walls nearest to the dentinoenamel junction. Intensity and asymmetry of peritubular dentine formation decreased cervically. In root dentine, the walls of the dentinal tubules were covered with only a thin peritubular dentine layer of even thickness. Here, in contrast to peripheral coronal dentine, the volume of intertubular dentine far exceeded that of peritubular dentine. In porcine coronal dentine, PTD asymmetry, being of lesser extent than in cervids, was observed only in peripheral areas of cuspal and flank regions of the cheek teeth. Because peritubular is more highly mineralized than intertubular dentine, the relative volume of dentine made up from the two components has an important influence on dentinal wear resistance. The significance of variations in volume and distribution of peritubular dentine between different dentinal regions for achieving and maintaining a functional occlusal surface is shown for cervid cheek teeth. Our results suggest that dentinal structure (in addition to enamel structure) should be taken more into consideration when discussing occlusal surface morphology in herbi- and omnivores from a functional point of view.


Ultrastructural study of postnatal development of the tonsillar crypt epithelium of the musk shrew, Suncus murinus. An ultrastructural study was made of the postnatal development of the tonsillar crypt epithelium in the musk shrew, Suncus murinus. On day 3 after birth, a particular kind of large lymphoid cell was first seen to move through the basement membrane into the epithelium. The next migration was that of lymphocytes, which passed through holes in the basement membrane. On days 5 to 7, the lymphocytes formed clusters, and pale epithelial cells of low electron density appeared. The cell clusters and pale epithelial cells fused on day 10. By day 14, these epithelial cells extended cytoplasmic projections to the surface of the epithelium, which had many heterophagic vacuoles and some microvilli-like structures. These findings suggest that the lymphoepithelial relationship is important for the organization of the immunological microenvironment in tonsillar crypt epithelium of the neonatal musk shrew.


Fiber differentiation of the human laryngeal muscles using the inhibition reactivation myofibrillar ATPase technique. The aim of the present study was to further subdivide the type II fibers of the human thyroarytenoid and posterior cricoarytenoid muscles by means of a modified myosin ATPase reaction. In order to understand the functioning of these highly strained muscles better, it is important to know the respective percentage of fatigue-resistant type IIA fibers and fatigable type IIB fibers. The material comprised the larynges of seven laryngectomized males aged between 45 and 70 years and four laryngectomized females aged between 39 and 72 years. After having been frozen in nitrogen, 10-microns-thick sections were cut from the laryngeal muscles in a cryostat. The pH-lability of the enzyme that can be utilized in a classical myosin ATPase reaction permits a differentiation between fiber types I, IIA and IIB. Evidently, this is not possible with every human muscle. The fiber types IIA and IIB of the thyroarytenoid and the posterior cricoarytenoid muscles could be clearly distinguished by means of the inhibition reactivation myofibrillar ATPase technique. Using this method, the myosin ATPase enzyme was initially inhibited by hydroxymercuribenzoate and subsequently reactivated by cysteine. Regarding the incidence of type I and IIA fibers, there was a statistically significant difference between the thyroarytenoid and the posterior cricoarytenoid muscles. The type IIA fiber content was statistically significantly higher in the arytenoid muscle than in the posterior cricoarytenoid muscle. The percentage of type IIB fibers was low, not only in the thyroarytenoid muscle and the posterior cricoarytenoid muscle but also in the other laryngeal muscles.(ABSTRACT TRUNCATED AT 250 WORDS)


Ultrastructural morphometric characterization of alveolar type II cells of perinatal sheep: a comparison with adults. The quantitative morphologic changes in alveolar type II cells during the perinatal period were characterized morphometrically in the lungs of fetal lambs at 132, 138, and 147 days gestational age (DGA) and in newborns at 2 days postnatal age (2 DPN). Ultrastructural features were compared with those of type II cells of ewes 365 days old. Lamellar body profile number per type II cell profile was highest at term (147 DGA) and 2 DPN. In adults, the number of lamellar body profiles and volume density of lamellar bodies were equal to those of the 132 DGA fetus. Multivesicular bodies were most common at 138 DGA and in adults. The volume density of cytoplasmic glycogen fell dramatically during the latter part of gestation. The volume density of many cellular organelles increased to the level observed in adults by term (147 DGA). Subcellular composition of type II cells of adult sheep differs from that reported for adult rats chiefly by the volume density of lamellar material within the cytoplasm. Plate-like or globe-like inclusions were present only in the type II cells of adults. Cytoplasmic extensions of the type II cell crossing the basal lamina were most abundant in the 132 and 138 DGA fetal sheep. Cytoplasmic extensions were rare in adults. We conclude that morphologic changes of the alveolar type II cell associated with gestational age follow a species-specific time course. In the sheep, this occurs during the later part of gestation and extends into the neonatal period. Morphologic and morphometric changes appear to correspond with cellular interactions between alveolar type II cells and mesenchymal cells of the interstitium.


Retinal photoreceptor fine structure in the velvet cichlid (Astronotus ocellatus). The structure and arrangement of the retinal photoreceptors of the velvet cichlid fish (Astronotus ocellatus) have been studied by light and electron microscopy. Rods, single cones and double (twin) cones are present. In the light-adapted state, rods are very tall cells that reach deep into the retinal epithelial (RPE) layer. The long outer segment is composed of discs of uniform diameter displaying one or two incisures. The rod inner segment shows a distal ellipsoid of mitochondria, and then narrows dramatically in the myoid region. Rod nuclei are electron dense and located deep in the outer nuclear layer. Rod synaptic spherules are small and show two to three invaginated synaptic sites as well as superficial synapses. Single cones are similar to the individual members of a double cone and all display a short tapering outer segment, a large ellipsoid of mitochondria and a myoid rich in rough endoplasmic reticulum, polysomes, Golgi zones and autophagic vacuoles. Double cones have extensive subsurface cisternae along their entire contiguous surfaces. Cone nuclei are large and vesicular and located close to or through the external limiting membrane. The synaptic pedicles of cones are larger, more electron lucent, and display more invaginated (ribbon) synapses as well as conventional (superficial) synaptic sites than do the rod spherules. Rod photoreceptors certainly undergo retinomotor movements and it is probable that cones do as well. In the light-adapted state the cone photoreceptors are arranged in a repeating square mosaic pattern with one single cone surrounded by four double (twin) cones.


Retinal pigment epithelial fine structure in the velvet cichlid (Astronotus ocellatus). The morphology of the retinal pigment epithelium (RPE), choriocapillaris and Bruch's membrane (complexus basalis) have been studied by light and electron microscopy in the velvet cichlid (Astronotus ocellatus). The RPE is composed of a single layer of large columnar cells. The basal (scleral) border of these cells is minimally infolded, whereas the apical (vitreal) surface displays numerous pigment-laden processes which in light-adaptation surround both rod and cone outer segments. Laterally the RPE cells are joined by a series of basally located tight junctions. Wandering phagocytes are a constant feature within this epithelial membrane. The RPE cells display a large, vesicular nucleus, numerous mitochondria, much smooth endoplasmic reticulum, polysomes, myeloid bodies, phagosomes and melanosomes. Rough endoplasmic reticulum is relatively scarce within these cells. Although only light-adapted specimens were examined, it is thought that the melanosomes are capable of extensive retinomotor movement. The endothelium of the choriocapillaris facing Bruch's membrane is typically very thin but shows few fenestrations. Bruch's membrane is typical of other teleost species in that it is composed of only three layers.


The development of pericardial villi in the chick embryo. The development of pericardial villi and their relation to the development of the cardiac surface was studied in chick embryos from the 3rd to 10th day of incubation by scanning electron microscopy. During the 3rd day of incubation (stage 14-17 HH) the coelomic epithelium covering the ventral wall of the sinus venosus forms villous protrusions. By the end of the 3rd day (stage 17 HH) these protrusions contact the dorsal wall of the heart, so that a secondary dorsal mesocardium is formed. This bridges the pericardial cavity between the ventral wall of the sinus venosus and the dorsal base of the ventricles. This sinu-ventricular mesocardium exists only temporarily, as on the 8th day of incubation it becomes a part of the cardiac wall due to fusion with the epicardium of the coronary sulcus. During the 4th and 5th day of incubation (stage 17-25 HH), the formation of the epicardium proceeds from the point of attachment of the sinu-ventricular mesocardium. Although these findings suggest that the epithelium of the villous protrusions spreads over the surface of the embryonic heart, one cannot exclude other hypotheses on epicardial origin. The impression of a spreading epicardium could also be created if epicardial cells were to delaminate from a local epithelium in a temporally and spatially organized pattern.


Cellular distribution of the new growth factor pleiotrophin (HB-GAM) mRNA in developing and adult rat tissues. Pleiotrophin (PTN), also known as HB-GAM, belongs to an emerging cytokine family unrelated to other growth factors. We report here the first comprehensive study using in situ hybridization on the cellular distribution of this new heparin-binding growth factor mRNA in rat tissues. PTN mRNA was developmentally expressed in many--but not all--neuroectodermal and mesodermal lineages, whilst no PTN mRNA was detected in endoderm, ectoderm and trophoblast. PTN mRNA was found in the nervous system throughout development, with a post-natal peak of expression. In the adult nervous system, significant expression persisted in hippocampal CA1 pyramidal neurons and in cortical neurons, but also in different non-neuronal cells types in various locations (olfactory nerve, cerebellar astrocytes, pituicytes, Schwann cells surrounding the neurons in sensory ganglia). PTN mRNA was also found during development in the mesenchyme of lung, gut, kidney and reproductive tract, in bone and cartilage progenitors, in dental pulp, in myoblasts, and in several other sites. Expression was differently regulated in each location, but usually faded around birth. In the adult, PTN mRNA was still present in the meninges, the iris, the Leydig cells of the testis and in the uterus. PTN mRNA was also strongly expressed in the basal layers of the tongue epithelium, which is the only epithelium and ectodermal derivative to express PTN mRNA, and this only after birth. PTN is known to be a growth factor for perinatal brain neurons and a mitogen for fibroblasts in vitro. Recently, trophic effects on epithelial cells and a role as a tumour growth factor have been reported. The mechanisms of regulation and the functions of PTN are however still uncertain. Its expression pattern during development suggests important roles in growth and differentiation. Moreover, the presence of PTN mRNA in several adult tissues and the up-regulation of PTN mRNA expression in the gravid uterus indicate that PTN also has physiological functions during adulthood.


Bone and cartilage resorption in relation to tooth development in the anterior part of the mandible in cichlid fish: a light and TEM study. This paper presents ultrastructural features of the contact region between particular tooth germs and Meckel's cartilage prior to, during, and after initial resorption of the perichondral bone and of the cartilage in the cichlids Hemichromis bimaculatus and Astatotilapia burtoni. Imminent resorption opposite such teeth is announced by the presence, in this region, of a particular cell type, considered to be a stage in the cytodifferentiation of osteoclasts. Slightly later, an osteoclast with typical ruffled border is seen to open a fenestra in the perichondral bone which surrounds Meckel's cartilage. Although the action of the osteoclast is directed primarily towards the bone, it may also affect, to a much lesser extent, the underlying uncalcified cartilage. Typically, fibroblast-like cells invade the resorption cavity along with the osteoclast; the tooth germ soon follows. Capillaries are seen to invade the cartilage only at a later stage when a large cavity has been established. It is proposed that the fibroblast-like cells may have a dual function: degradation of cartilage and deposition of new bone. Although these processes are normally limited to the area surrounding tooth germs at specific loci, tooth germs in other positions may sometimes be seen invade the cartilage. They do so either passively, because of the existence of such a cavity, or as a result of their own resorption-inducing activity. Whatever the mechanism, attachment bone is being deposited within the erosion cavity and on the surface of the exposed perichondral bone. The stimuli possibly eliciting resorption of Meckel's cartilage are discussed. It is hypothesized that pressure exerted by the growing tooth germ may stimulate the osteoblasts covering the bone surface and, in this way, provoke osteoclastic bone resorption.


Fine structural characteristics of the zone of contact between the lower infundibular stem and the pituitary pars distalis in the little brown bat, Myotis lucifugus. The purpose of this study was to examine the morphological characteristics of the pituitary gland in the little brown bat that might influence mechanisms of hypothalamic releasing hormone transport. Paraffin sections were prepared from whole crania to examine in situ the orientations of the three parts of the adenohypophysis (pars distalis, pars intermedia, and pars tuberalis) relative to the components of the neurohypophysis (pars nervosa and infundibular stem) and the basal hypothalamus. Of particular interest was the observation that the axis of the infundibular stem is directed posteriorly from the median eminence and occupies a depression in the dorsal surface of the pars distalis as it approaches the pars nervosa. Previous studies have revealed that neuronal projections containing releasing hormones extend into the infundibular stem in this species. Therefore, we conducted a fine structural study to determine whether the zone of contact between the infundibular stem and the pars distalis could represent a site of specialized interaction between hypophysiotropic hormones and their target cells. The results show that the sparse connective tissue along this boundary contains abundant fenestrated capillaries that are exposed on one side to neurosecretory axons and on the other to cells of the pars distalis. Furthermore, secretory cells nearest these capillaries exhibit ultrastructural evidence of heightened secretory activity. We conclude that the fine structural characteristics of this zone are consistent with localized mechanisms of releasing hormone transport.


Quantitative morphology of the embryonic heart: an approach to development of the atrioventricular valves. In 32 human embryos ranging from 4.0 to 42 mm CR-length, the volumes of the atrioventricular endocardial cushions and of the ventricular myocardium were estimated by the point counting method. The surface densities of the left and right ventricular apical trabeculations were estimated by the point and intersection counting method. It is concluded that the cushions do not grow after the 25 mm stage, by that time having reached the maximal value of 0.074 mm3. This supports the concept that the cushions do not materially contribute to the definitive atrioventricular valves. In young embryos, the left ventricular trabeculations are thicker (as concluded from their higher surface density) than the right ventricular trabeculations. Only around the 25 mm stage, the ratio becomes 1. After this stage, right ventricles have thicker trabeculations than left ventricles have. This supports the concept that the trabecular pattern is changed during the period of valve formation, which process is characterized by delamination of the inner myocardial layers.


Development of the vestibular and auditory system of the northern native cat, Dasyurus hallucatus. The developing vestibular and auditory system of the native cat Dasyurus hallucatus was examined from birth to day 55 postpartum to determine when the six sensory regions had an adult structure. The utricle, a sensory epithelium with an overlying discrete population of otoliths, was present in the newborn native cat. The saccule, which has a similar structure to the utricle, and the three crista ampullaris of the semicircular canals, were present by day 21 postpartum. The organ of Corti was formed by day 50 and the external ear duct was patent by day 55 postpartum. Hair is first seen on the native cat by day 45 when the young first leave the pouch. They are left in the nest and travel on the back of the lactating mother until about day 80. The eyes open on approximately day 75. Thus, the native cat has fully functional visual, auditory, and vestibular systems when the juvenile leaves the nest.


Anatomy and developmental chronology of the rat inferior alveolar nerve. This report describes the anatomy of the inferior alveolar neurovascular bundle in the adult rat and provides a quantitative analysis of the developing inferior alveolar nerve (IAN). Soon after its entrance in the mandibular canal, the IAN splits into a mental nerve (MN) and an inferior dental nerve (IDN), which course in separate bony compartments. The MN passes unbranched through the mandibular canal. The IDN sends branches to the incisor, the first molar, and the second molar. The third molar (M3) is supplied by a separate IAN branch. The adult rat IAN contains 8,000-10,000 axons, 70% of which are myelinated. The MN accounts for 70% of all IAN axons, the IDN 26%, and 4% form the M3 branch. The proportion of large myelinated axons is lower in the MN than in the IDN. Following chemical sympathectomy, the IAN axon number does not change in a statistically significant way. The total number of IAN axons, which is high prenatally and neonatally, has decreased to the adult level about 1 week after birth. De novo myelination commences at birth and is complete 3-4 weeks later. The size spectrum of the myelinated fibres is narrow and unimodal during the first postnatal weeks. By 1 month, the largest fibres reach diameters of approximately 6 microns, and a bimodal pattern is emerging. From 3 months and on, the size range reaches up to 10-12 microns, and the distribution is bimodal. These data provide a basis for further studies on developmental tooth-nerve interactions.


Nuclear membrane modifications in polytene nuclei of Drosophila melanogaster: serial reconstruction and cytochemistry. The nuclear envelope of polytene nuclei of salivary glands of Drosophila melanogaster displays modifications consisting of nuclear envelope invaginations (NEI) and evaginations (NEE). Ultrastructural analyses combined with three-dimensional reconstruction and cytochemistry show that NEI are bounded by a single membrane and that they may arise as invaginations of the inner nuclear membrane. NEI extend deeply into the nucleus. The lumens of NEI may collapse resulting in membranous sheets which may combine with those arising from adjacent NEI to form intranuclear structures resembling annulate lamellae. All NEI are associated with NEE. In contrast to NEI, NEE are enclosed in a double membrane morphologically identical to the nuclear envelope. While NEI and NEE share wheat germ agglutinin binding properties with the nuclear envelope, they differ in their ability to localize lanthanum. Pore annuli of NEI display complete lack of lanthanum binding, while those of NEE exhibit minor deposition of this cation. In contrast, pore annuli of the nuclear envelope are specifically and significantly decorated by lanthanum. A conceptual model based on the results obtained suggests that NEI are formed by invaginations of the inner nuclear membrane, together with accompanying modifications of pore complexes.


Lymphatic vessels of the human dental pulp in different conditions. The characteristics of the lymphatic vessel endothelial wall have been investigated in human normal and inflamed dental pulps. In normal pulps the endothelial wall is characterized by the presence of micropinocytotic vesicles and intraparietal channels. In the inflamed pulpal tissue, where an increase in interstitial fluid pressure occurs, the distended endothelial wall presents open junctions between endothelial cells and the openings of the intraparietal channels. Moreover the micropinocytotic vesicles disappear. The cytoplasm of the endothelial cells is characterized by the presence of numerous Weibel-Palade bodies, which increase in number in the dilated vessels. In the fibrillar apparatus surrounding the lymphatic vessel wall collagen fibrils are the prevalent component, while elastic fibers are not present. The different morphological properties of the lymphatic vessels are compared and discussed with regard to the variation of the functional conditions of the tissue.


Capillaries measured in canine diaphragm by two methods. We have measured capillary distribution in costal and crural canine diaphragm using two methods: histochemical processing and perfusion fixation. Each of 18 dogs was deeply anesthetized, the abdomen opened, and the left inferior phrenic artery cannulated. The animal was heparinized and overdosed with pentobarbital. The right hemidiaphragm was frozen, either postexcision (Protocol 1) or intact with no preload (Protocol 2), for histochemical processing. The left hemidiaphragm was fixed by perfusion in situ using 2% glutaraldehyde, either with preload (Protocol 1) or without (Protocol 2). Costal and crural regions of each hemidiaphragm were sampled for analysis. Frozen samples were sectioned and processed for acid-stable (pH 4.0) ATPase activity; perfusion-fixed samples were postfixed, stained, embedded in Epon, and sectioned. Measurements were made using a digital imaging system. We found that muscle fibers had smaller cross-sectional areas in costal than in crural diaphragm; capillary-to-fiber ratio (C:F) did not differ by region and regional differences in capillary density could be attributed to differences in fiber size. Results depended critically on methodology. In perfusion-fixed muscle, fiber area was less, C:F was greater, and capillary density was greater than in histochemically-processed tissue. We conclude that capillary distribution is similar in costal vs. crural diaphragm and that perfusion fixation identifies capillaries more effectively than histochemistry.


Immobilization-induced muscle atrophy is not reversed by lengthening the muscle. In clinical practice, repaired tendocalcaneus (Achilles tendon) ruptures are often protected in immobilization casts for 4 weeks in the fully plantar flexed position and for up to another 4 weeks after returning the ankle to joint neutral. Moderate to severe muscle atrophy occurs within 4 weeks of immobilization in plantar flexion, but it is not known if this atrophy is minimized or reversed following restoration of joint neutral position. We tested the hypothesis that the extent of atrophy could be reduced by returning the ankle to joint neutral after 4 weeks of immobilization. Eighteen rabbits were anesthetized, and their right hind-limbs were casted with the knee flexed 90 degrees and the ankle fully plantar flexed. Three animals each were studied after 3, 4, 6, or 8 weeks of immobilization. After 4 weeks of immobilization, the immobilization casts of the remaining six rabbits were modified to return the ankle to joint neutral for another 2 or 4 weeks. For muscle studies, the animals were anesthetized, and the soleus (SOL), plantaris (PLN), and gastrocnemius (GST) muscles were removed and weighed; the SOL and PLN were quick frozen and processed for histochemical fiber typing and fiber cross-sectional area measurement. All three muscles showed significantly reduced muscle weight to body weight ratios after 3 weeks of immobilization. SOL was the most affected, and GST was least affected. There was no significant further atrophy through 8 weeks of immobilization. The atrophy correlated with a significant reduction of mean fiber area (MFA) for Types I, IIo, and IIc fibers in SOL and PLN. In PLN, Type IIg fiber area was not significantly reduced.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of cyproterone acetate on structure and function of rhesus monkey reproductive organs. A low dose of Cyproterone acetate (CPA; 1 mg/kg body weight/day for 70 days) was administered to adult male rhesus monkeys to assess its effects on testicular and epididymal structure and function in a nonhuman primate species. CPA caused extensive degenerative changes in morphology of seminiferous, efferent duct, and epididymal epithelia, including decrease in diameter of seminiferous and epididymal tubules and their lumen, height of epididymal epithelium, and an increase in intertubular connective tissue. The protein profile of spermatozoa showed alterations during their epididymal transit in control and CPA-treated monkeys. In CPA-treated animals, 19 polypeptides were acquired and nine were eliminated during epididymal transit in contrast to acquisition of 12 and loss of 14 polypeptides in control animals. Treatment with CPA also resulted in the appearance of 14 new polypeptides in epididymal cytosol and luminal fluid, probably of lysosomal origin. The protein pattern of caput and cauda epididymal tubule cytosol, maintained in organ culture and exposed to 100 microM CPA for 3 days, showed absence of eight polypeptides. These results indicate that even at the low dose used in this study, CPA has caused spermatogenic arrest, degenerative changes in the epididymal structure, and alterations in epididymal and sperm protein profile. Suppression of serum testosterone levels indicates the need for androgen supplementation if CPA is to be used for male contraception.


Reproduction in a South American mouse, Abrothrix longipilis. Approximately 700 Abrothrix longipilis were trapped in Argentina, and their reproductive organs were studied. Some were kept briefly in captivity. Males have the same body length as females but are 8% heavier. Sexually immature males are shorter and lighter than breeding males of the same age. Few individuals in the wild live as long as two years. All overwintering males come into breeding condition in the spring and become infertile in the autumn. All overwintering females have open vaginas in the spring, and most undergo several infertile ovulatory cycles, probably without copulating, before they become pregnant. Corpora lutea of the sterile cycles accumulate; there may be as many as 28 in a pair of ovaries. The average number of ova shed per cycle is 4.66; average litter size is 3.85. Ovulation is spontaneous. There is postpartum ovulation and fertilization without intervening sterile cycles but with maintenance of old corpora lutea. Unusual architecture of the penis and vagina may result from unusual timing of hormone production during development, and the disjunction of ovulation and fertilization may similarly result from hormonal heterochrony at the beginning of the breeding season. The apparently nonadaptive wastage of ova in sterile cycles may be a novel example of kin selection.


Macrostructure differences of polar fox and dog lungs. The lungs of the polar fox and dog have the typical form and lobular structure characteristic of beasts of prey. Both display secondary fusion of the cranial and middle lobes in the left lung, but this is more extensive in the fox. A consistent relationship between the beast body mass and the mass and volume of the lungs is present. The relative weight of dog lungs independent of body size (weight index) is 1.7 times that of the polar fox. In the latter the parenchyma is much more subdivided than in the dog. There are 23 segments per lung pair in the polar fox, compared to 19 in the dog. Although these are of unequal size throughout the lungs of both species, corresponding segments in the fox are about half as large as those in the dog. The greater segmentation of polar fox lungs may be of assistance in restricting the spread of inflammatory processes.


Morphogenetic and functional activity of type II cells in early fetal rhesus monkey lungs. A comparison between primates and rodents. To evaluate further the role of type II alveolar epithelial cells in primate lung development, lungs of fetal (46 to 155 days gestational age [DGA]), postnatal, and adult rhesus monkeys were investigated with antibodies against surfactant protein A (SP-A), Alcian blue (AB) staining, and periodic acid-Schiff (PAS) staining with/without alpha-amylase pre-treatment. In adult and postnatal lungs, type II cells (cuboid shape; large, roundish nucleus) displayed a unique cytoplasmic staining for SP-A. In prenatal lungs, a low-columnar to cuboid type of cell with a large, roundish nucleus was first detectable by 62 DGA. It was the only cell type to line the distalmost tubules or buds of the prospective respiratory tract. It exhibited (initially partial) cytoplasmic staining for SP-A. AB and PAS stainings showed the presence of acid glycoconjugates and large apical and/or basal glycogen fields. After 95 DGA, the lining of the distal respiratory tract additionally displayed flatter cells with immunoreactivity for SP-A and non-reactive zones. Columnar epithelium (pseudostratified or simple) never stained for SP-A. We conclude that morphologically identifiable type II cells first appear in fetal rhesus monkey lungs by 62 DGA (pseudoglandular period). The cells may already synthesize surfactant and extracellular matrix components. They generate type I cells, and thus the entire pulmonary acinus lining. These conclusions for the rhesus monkey fully agree with our earlier conclusions for another primate, the human, and for rodents. However, as presently shown, primates differ greatly from rodents with respect to the timing of type II cell differentiation (at 29-38% versus 73-75% of gestation or at 22-25% versus 48-49% of prenatal lung development).


Fibroblast-mediated collagen gel contraction does not require fibronectin-alpha 5 beta 1 integrin interaction. Fibroblasts cultured within free-floating collagen gels can bind to and reorganize the surrounding collagen fibrils into a more dense and compact arrangement. Collagen gel contraction provides an in vitro model for studying fibroblast-collagen interactions important in wound healing, fibrosis, scar contraction, and connective tissue morphogenesis. We have assessed the role of fibronectin and its interaction with the alpha 5 beta 1 "high affinity" fibronectin-specific integrin receptor in collagen gel contraction. A variety of agents, which specifically inhibit fibronectin-alpha 5 beta 1 interactions, were tested for their abilities to inhibit fibroblast-mediated collagen gel contraction. These included anti-alpha 5 beta 1 monoclonal antibodies, the synthetic peptide GRGDSP, the cell adhesive fragment of fibronectin, and an antibody against the cell adhesive region of fibronectin. None of these agents inhibited collagen gel contraction. Therefore, it is concluded that fibronectin-alpha 5 beta 1 interactions are not necessary for collagen gel contraction. However, collagen gel contraction is dependent on a member or members of the beta 1 subfamily of integrin matrix receptors. A polyclonal antiserum and a monoclonal antibody, both directed against the beta 1 subunit of integrin matrix receptors, inhibited the spreading of fibroblasts in the collagen gel and inhibited collagen gel contraction. This study demonstrates that fibroblast-mediated collagen gel contraction is independent of fibronectin-alpha 5 beta 1 interactions but dependent on an interaction of beta 1 integrin matrix receptors with collagen fibers.


Blood cells of the gilthead seabream (Sparus aurata L.): light and electron microscopic studies. The peripheral blood cells of the gilthead seabream (Sparus aurata L.) were studied by light and transmission and scanning electron microscopic methods. Acidophilic erythroblasts and mature erythrocytes, round, oval, and fusiform thrombocytes, neutrophils, acidophils and basophils, lymphocytes, plasma cells, and monocyte-macrophages were characterized. A comparison of our light and electron microscopic results was carried out. The results were discussed with those for other fish species and the main modifications from the common vertebrate haematological pattern observed being considered.


Effects of daily administration of prostaglandin E2 and its withdrawal on the lumbar vertebral bodies in male rats. The effects of daily prostaglandin E2 (PGE2) treatment (on) and PGE2 treatment followed by withdrawal (on-off) on cancellous bone in lumbar vertebral bodies were studied in 7-month-old male Sprague-Dawley rats. The first groups of rats were given daily subcutaneous injections of 0, 1, 3, and 6 mg PGE2/kg/d for 60, 120, and 180 days, and the second group of rats were given PGE2 for 60 days followed by withdrawal for 60 and 120 days. Histomorphometric analyses were performed on double-fluorescent labeled undecalcified sections of fourth lumbar vertebral bodies. Systemic PGE2 treatment elevated cancellous bone mass of lumbar vertebral bodies 26-60% above control levels within 60 days and continued treatment maintained it for another 120 days, but the excess bone was lost after the treatment was withdrawn. PGE2 treatment for 60 days increased trabecular bone area, trabecular width, and bone formation parameters, and shortened remodeling periods in a dose-response manner. These changes were sustained at the levels achieved by 60-day treatment in the rats treated for 120 and 180 days. The eroded perimeter increased at day 60 and further at day 120 and then plateaued. In the on-off treated rats, the cancellous bone area, bone formation, and resorption parameters returned to near age-related controls by 60 days after withdrawal and were maintained there after 120 days of withdrawal. Therefore we conclude that the continuous treatment is needed in order to maintain the PGE2-induced bone gain. When these findings were compared to those previously reported for the proximal tibial metaphyses, we found that the proximal tibial spongiosa was much more responsive to PGE2 treatment than the fourth lumbar vertebral body.


Differences in fiber number and fiber type proportion within fascicles. A quantitative morphological study of whole vastus lateralis muscle from childhood to old age. Data are presented on the number of fibers and the proportion of different fiber types within fascicles of whole vastus lateralis muscles from 5 male children, 5 to 15 years of age, and compared with results from 25 male adults, middle aged and old individuals. The results verify a difference in the proportion of fibers with different properties within a fascicle. The proportion of type 2 (fast twitch) fibers on the border of fascicles is larger than the proportion internally: the children have (P less than 0.01) greater difference than the adults and the middle aged, whereas the old have (P less than 0.001) less difference than the other age groups. The mean number of fibers per fascicle increases (P less than 0.05) from childhood to adult age, and thereafter reduces (P less than 0.01). The results imply that fascicles continuously rearrange during the normal growth and development and the normal aging process. It is argued that the causes of the difference in fiber type proportions within fascicles are local factors in the muscle, secondary to the overall age related functional demands put on the fiber population.


Morphology of the bile ducts of the brook lamprey, Lampetra lamottenii (Le Sueur) before and during infection with the nematode, Truttaedacnitis stelmioides (Vessichelli, 1910) (Nematoda: Cucullanidae). Routine light microscopy and transmission and scanning electron microscopy were used to describe and compare the biliary tree of larval Lampetra lamottenii before and during infestation of the bile ducts with the nematode, Truttaedacnitis stelmioides. The most prominent changes to the biliary tree following infection by the parasite are the dilation of the bile ducts, alterations to their epithelial cells, and an increase in periductal fibrous tissue. In recently infected animals, the simple epithelium of dilated bile ducts often contains many mitotic figures. In long-term infestations, the epithelium is stratified or pseudostratified. Changes to the fine structure of the biliary epithelial cells include increase and/or dilation of the RER and SER, and increases in microfilaments, intermediate filaments, and microtubules. The abundance of dense bodies may reflect enhance reabsorption of biliary constituents, and their accumulation may ultimately result in cytolysis. There are increased mucous granules in the apical cytoplasm of biliary epithelial cells and an abundance of mucinous material within the bile duct lumen, and the basal lamina appears thickened. The changes to the liver of L. lamottenii following infection are discussed and compared to those reported in small mammals following bile duct ligation, in patients with extrahepatic biliary obstruction, and in parasitic infection of the biliary tree.


Immunocytochemical localization of rat intestinal 15 kDa protein, a member of cytoplasmic fatty acid-binding proteins. Rat intestinal 15 kDa protein (I-15P) is a member of the family of cytoplasmic fatty acid-binding proteins. Using a specific antiserum against I-15P, we studied the tissue distribution and subcellular localization of this protein in the entire rat body. By immunoblot analysis of cytosolic proteins, I-15P was detected not only in the distal portion of small intestine but also in the ovary and adrenal gland. Immunohistochemically, I-15P was localized to the absorptive epithelial cells as well as a subpopulation of enterochromaffin cells in the intestine, the lutein cells in the ovary, and subpopulations of cortical cells in the adrenal gland. Furthermore, I-15P-like immunoreactivity was also demonstrated in the surface mucous cells of stomach and the granular convoluted tubule cells of submandibular gland. Immuno-electron microscopy showed that the immunoreactivity was confined to the cytoplasmic matrix region, except in the enterochromaffin cells and granular convoluted tubule cells, where it was localized in the secretory granules. The present findings suggest that I-15P plays a role in the cellular metabolism of steroids.


In vivo monastral blue-induced lamellar-bodies in lysosomes of pulmonary intravascular macrophages (PIMs) of bovine lung: implications of the surface coat. We previously reported that the pulmonary intravascular macrophages (PIMs) of sheep, goat, and calf lung contained a heparin and a lipolytic lipase sensitive surface coat by using tannic acid as a component of paraformaldehyde-glutaraldehyde-based fixative. The implication of this sensitivity was that the surface coat was predominantly comprised of lipoprotein-like substance. In this study we report that monastral blue (MB) used as a vascular tracer interacted with the coat globules and lost its original particulate appearance. Its precise localization in the PIMs was in combination with altered macromolecules of the surface coat in the form of lipid droplets, which conformed to the conventional view of neutral lipids. In contrast, pigment particles examined in their native state resembled metallic particles as electron-dense elliptical rods. The lipid droplets were subsequently internalized through endocytic route and found their access into the lysosomal compartments of PIMs at the electron microscopic level. Lamellar bodies (LLBs) arose from the lysosomal matrix after the entry of lipid droplets in the secondary lysosomes. Acid phosphatase activity was located in secondary lysosomes as well as in endosomes. These observations suggest that coat granules of the PIMs acted as a carrier of exogenous MB particles to deliver the complex to the lysosomal compartment where partial digestion lead to the formation of lamellar bodies. The implications of MB (cationic dye) as a vascular tracer for studying phagocytic index of PIMs in the light of their coat and the rapid development of LLBs are discussed. It is proposed that MB by initially combining with the surface coat provokes mobilization of intracellular lipid pools. In this way metabolism of vasoactive lipid in the PIMs is stimulated to influence the dynamics of pulmonary circulation in the calves.


Anatomical study of the bronchial system and major blood vessels of the chicken lung (Gallus gallus) by means of a three-dimensional scale model. The bronchial and vascular patterns of the chicken lung, from specimens age 8-10 days, have been studied by serial, paraffin sections of the whole organ. According to the histological structure, the bronchial system consists of three airway types: primary bronchus or mesobronchus, secondary bronchi, and tertiary bronchi or parabronchi. The mesobronchus gives rise to three sets of secondary bronchi: four dorsomedial, four dorsal, and three lateral ones. The total number of secondary bronchi is 11, which is less than the number reported in adult birds by other authors until now. Nevertheless, the number and distribution of the major vessels, arteries and veins are in basic agreement with previous descriptions.


SEM of capillary pericytes prepared by ultrasonic microdissection: evidence for the existence of a pericapillary syncytium. Retia mirabile of the eel swimbladder were exsanguinated, perfusion-fixed and subjected to prolonged osmication. They were then microdissected by ultrasonication which delaminated the capillary bed along planes which revealed the surfaces of arterial and venous capillaries. This procedure resulted in cleaned capillary surfaces largely free of connective tissue elements and basement membrane material. The arterial capillary segments were heavily invested with pericytes characterized by plump cell bodies containing nuclei and an extensive system of processes encircling the capillary wall. These processes exhibited a hierarchical organization consisting of primary, secondary, and tertiary elements arising roughly at right angles to each other. Primary and secondary processes exhibited frequent anastomoses and resulted in cytoplasmic continuity between adjacent cell bodies. Processes were also observed to form connections between pericytes on adjacent capillaries. These observations are evidence for the existence of a pericapillary syncytium in which cell bodies may be connected in series and in parallel throughout the arterial capillary bed. This syncytial organization would provide for a coordinated and global contractile response of pericytes to vasoactive hormones and other effectors. It may also provide for synchrony of nuclear division during developmental spread of pericytes along capillary surfaces.


Structure and dynamics of adrenal mitochondria following stimulation with corticotropin releasing hormone. The mitochondria of rat adrenals were investigated qualitatively and quantitatively in different functional states of the adrenal cortex. Following stimulation of the animals with corticotropin releasing hormone (CRH), the corticosterone serum levels reached a maximum 1 hour after stimulation with CRH. The amount of inner mitochondrial membrane within the zona fasciculata increased showing a biphasic time course, with a first maximum 2 hours and a second maximum 8 hours after stimulation. In contrast, a significant rise of mitochondrial volume occurred only 24 hours after CRH stimulation. Therefore, the dense vesicularization of mitochondrial cristae may constitute an early process to enhance the steroidogenic capacity of these cells. Within cells of the transition zone between zona glomerulosa and zona fasciculata, we could depict a special type of mitochondria with characteristic crescent-like cristae only seen after stimulation with CRH. This type of mitochondria may represent an intermediate form between mitochondria of zona glomerulosa and zona fasciculata underlining the impressive transformational capacity of adrenocortical mitochondria. After hypophysectomy, zona fasciculata cells contained mitochondria with tubular inner membranes, representing a hypofunctional state. In contrast, the hypofunctional state after hypophysectomy and the hyperfunctional state after stimulation of the adrenal cortex via CRH injection did not appear to correlate with the morphology of mitochondria from the zona reticularis and adrenal medulla.


Transplantation of fetal mouse colon under the kidney capsule of an adult mouse: a model for the study of colonic development. Fourteen-day fetal mouse colon was transplanted under the kidney capsule of an adult mouse to determine whether this system could be used as a model of embryonic colonic development. The 14-day fetal colon was transplanted and left for a period of 7 days. Comparisons of the normal one day postnatal colon and the transplanted colon were made morphologically and morphometrically. It was found that the transplanted fetal colon resembled its postnatal counterpart with respect to morphology; the cell types seen in the transplanted colon were similar to those observed in the in situ colon of the same age. However, morphometric analysis showed that the transplanted colon was significantly smaller than its postnatal counterpart, suggesting that conditions in the host were not optimal to support the full growth of the colon. In spite of this, it appears that the fetal colon can differentiate normally under the kidney capsule and this model can be used to study both epithelial-mesenchymal interactions and the role of hormones in fetal colonic development.


Coronary artery development in the chick: origin and deployment of smooth muscle cells, and the effects of neural crest ablation. Previous studies of coronary artery ontogeny have stressed early development and therefore have dwelt mainly upon the origin of the endothelium of the nascent coronary artery stem. This study has analyzed the ontogeny of the vascular smooth muscle cells (VSMC) in the coronary arteries of the domestic chicken, by establishing the timing and deployment of smooth muscle alpha-actin (SMAA). Anti-SMAA was applied to sections of normal embryos, and to sections of experimental embryos that had undergone surgical ablation of the neural crest over somites 1-3. The results show an orderly symmetrical deployment of SMAA in control coronary arteries. SMAA was expressed significantly earlier in the coronary artery VSMC compared with those of the cardiac outflow vessels; this early expression may indicate a unique responsiveness to induction of the smooth muscle phenotype. The normal orderly development of coronary artery VSMC was dependent upon the presence of the neural crest, and therefore was disrupted in the experimental embryos whose neural crest was ablated.


Effect of preoperative normovolemic hemodilution on left ventricular segmental wall motion during abdominal aortic surgery. Preoperative normovolemic hemodilution (PNH) has been proposed for patients scheduled to undergo aortic surgery. Coronary artery disease is frequent in these patients. The aim of the study was to assess the effect of PNH on hemodynamics and segmental wall motion (SWM) evaluated by transesophageal echocardiography in such patients. Twenty patients with coronary artery disease were allocated randomly to either PNH or no PNH; PNH was performed after anesthetic induction using dextran 60,000. Patients were operated on under general anesthesia and monitored intraoperatively with electrocardiographic lead CM5, radial and pulmonary artery catheters, and transesophageal echocardiography positioned to obtain a short-axis view. Hemodynamic and transesophageal echocardiographic data were collected after anesthetic induction and after PNH, before and 5 min after aortic clamping, after unclamping, and at the end of surgery. Aortic clamping induced a significant increase in systemic vascular resistance and arteriovenous difference in oxygen and a decrease in cardiac index (P < 0.05), but the effect of aortic clamping was inversely related to hemodilution. The SWM score (graded from 1 = normal to 5 = dyskinesia) was significantly increased after aortic clamping, mainly in the anterior segment (P < 0.05). Four patients in the control (no PNH) group and one in the PNH group developed new SWM abnormalities indicative of myocardial ischemia during surgery (P = NS). This study suggests that PNH may improve hemodynamic tolerance to aortic clamping in patients with coronary artery disease. The observed changes in SWM indicate that PNH may not worsen myocardial ischemia in patients scheduled to undergo aortic surgery.


Splanchnic hemodynamics and oxygen supply during acute normovolemic hemodilution alone and with isoflurane-induced hypotension in the anesthetized pig. Safety of combined hemodilution and isoflurane-induced hypotension was assessed by studying their effects on splanchnic hemodynamics and oxygenation in nine anesthetized (ketamine/flunitrazepam) pigs. Acute normovolemic hemodilution (decrease in hematocrit from 29% to 15%) with 6% hydroxyethyl starch decreased (P < 0.05) O2 delivery despite increases in cardiac output and all splanchnic flows, as measured by electromagnetic flow probes. Superimposed isoflurane (1.45% end-tidal concentrations) caused marked decreases in arterial blood pressure, cardiac output, and splanchnic flows. Oxygen uptake of the liver became O2 supply dependent. A liver surface PO2 histogram (Clark-type electrode) showed 38% of PO2 values between 0 and 5 mm Hg. Hepatic lactate uptake decreased by 75%. These data suggest that depression of cardiovascular function during reduced O2-carrying capacity adversely affects hepatic perfusion, oxygenation, and function. Combined hemodilution and isoflurane-induced hypotension may not be appropriate in patients.


Greater neuromuscular blocking potency of atracurium during hypothermic than during normothermic cardiopulmonary bypass. Previous studies drew attention to the greater neuromuscular blocking potency of atracurium during, than before or after, hypothermic cardiopulmonary bypass (CPB) while disregarding the periods of normothermic perfusion. We recorded the evoked twitch tension in 15 patients during nitrous oxide/narcotic anesthesia who were undergoing open heart surgery. Atracurium was injected as an initial bolus dose of 460 micrograms/kg. Waning neuromuscular blockade was enhanced by repeat injections of 138 micrograms/kg whenever the twitch tension attained 25% of control. During hypothermic (< 32 degrees C) and normothermic (> 34 degrees C) CPB, the times of onset of the maintenance doses were 57% and 18% longer, respectively, than before CPB (P < 0.05). Maintenance doses of atracurium were required every 24 +/- 4 min (mean +/- SD) before CPB, every 45 +/- 8 min (P < 0.05) during hypothermia, every 22 +/- 3 min during normothermic perfusion, and every 23 +/- 3 min after CPB. In conclusion, the patients' changing demand of atracurium paralleled the changes of temperature rather than the institution and cessation of CPB.


Effects of surgical stress and volatile anesthetics on left ventricular global and regional function in patients with coronary artery disease. Evaluation by computer-assisted two-dimensional quantitative transesophageal echocardiography. We investigated the effects of halothane, enflurane, and isoflurane on central hemodynamics and left ventricular global and regional function when used to control intraoperative hypertension in 39 patients with coronary artery disease. Left ventricular short-axis, midpapillary images were obtained by transesophageal echocardiography. Using a centerline algorithm, we analyzed left ventricular images for global area ejection fraction (GAEF) and segmental area ejection fraction (SAEF). The SAEF/GAEF ratio was calculated for each of eight segments. Measurements were performed after induction of anesthesia but before skin incision; 1 min after sternotomy; and during administration of the inhaled anesthetic. The increase in arterial blood pressure during sternotomy was due to an increase in vascular resistance accompanied by increases in heart rate and filling pressures while GAEF decreased. No changes in the SAEF/GAEF ratio appeared during sternotomy. The inhaled anesthetics restored arterial blood pressure by a similar decrease in vascular resistance. Isoflurane caused an increase in cardiac index that was not seen with halothane or enflurane (halothane vs isoflurane, P < 0.05). The GAEF was decreased by halothane but unaffected by isoflurane and enflurane (halothane vs enflurane; P < 0.05). Isoflurane induced a decrease in the SAEF/GAEF ratios of two segments corresponding to the inferolateral wall of the left ventricle that was, in one of these segments, significantly more pronounced compared with both halothane and enflurane. Halothane or enflurane did not cause any change in regional wall motion. We conclude that isoflurane is more likely to cause regional wall motion changes than halothane or enflurane in patients with coronary artery disease.


Effects of halothane and quinidine on intracardiac conduction and QTc interval in pentobarbital-anesthetized dogs. To confirm in vitro data that halothane and quinidine depressed cardiac conduction and prolonged action potential (AP) duration, the electrocardiogram and His bundle electrogram were recorded in dogs during basal pentobarbital anesthesia, after 1% halothane or quinidine (2.38 +/- 0.22 micrograms/mL serum concentration [mean +/- SEM]), or both. Purkinje fibers from a second dog were superfused with blood from the intact (support) dog, and APs were recorded. In the intact dogs, 1% halothane caused no changes in the electrocardiogram or His bundle electrogram. Quinidine prolonged QRS duration, QT interval, and rate-corrected QT (P < 0.05). Ventricular conduction (HS interval) slowed, and atrial effective refractory period increased (P < 0.05). Quinidine combined with halothane widened QRS, QT, and rate-corrected QT, prolonged the HS interval, and increased the vulnerability of the atrioventricular node to conduction block. Three of 20 dogs developed torsades de pointes-type ventricular tachycardia during simultaneous quinidine and halothane administration. In cross-superfused Purkinje fibers, the AP duration to 50% repolarization was shortened, and conduction time was prolonged by 1% halothane (both P < 0.05). Quinidine decreased AP amplitude, prolonged AP duration to 90% repolarization, and slowed conduction (P < 0.05). Quinidine combined with halothane decreased AP amplitude, and prolonged both AP duration to 90% repolarization and conduction (P < 0.05). When 1% halothane and therapeutic concentrations of quinidine are administered in dogs, depressed conduction and an acquired long QT syndrome with malignant ventricular arrhythmias may develop.


Differences in pH management and pulsatile/nonpulsatile perfusion during cardiopulmonary bypass do not influence renal function. The renal effects of pulsatile (pulse pressure 18.0 +/- 1.5 mm Hg [mean +/- SEM]) or nonpulsatile perfusion (mean pulse pressure 1.9 +/- 0.4 mm Hg) during either alpha-stat (mean PaCO2 41.2 +/- 0.9 mm Hg measured at 37 degrees C) or pH-stat (mean PaCO2 60.6 +/- 1.7 mm Hg measured at 37 degrees C) pH management of hypothermic cardiopulmonary bypass (CPB) were studied in 100 patients undergoing elective coronary artery bypass surgery. Mean urine output, fractional excretion of sodium and potassium, and renal failure index all increased during the study period; however, there was no difference among the four different CPB management groups. Mean postoperative creatinine and blood urea nitrogen values decreased compared with preoperative values, again without differences among treatment groups. Three patients developed acute renal insufficiency; of these, two had received nonpulsatile perfusion and pH-stat management, and the other had been managed with pulsatile perfusion and pH-stat management. These three patients all had undergone prolonged CPB and required at least two vasoactive drugs and the use of an intraaortic balloon pump to be weaned from CPB. In patients with normal preoperative renal function undergoing hypothermic CPB, neither the mode of perfusion, pulsatile or nonpulsatile, nor the method of pH management, pH-stat or alpha-stat, influences perioperative renal function.