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PubMed1995

Morphology of pancreatic microcirculation in the monkey: light and scanning electron microscopic study.

The morphology of the microcirculation of the pancreas in 20 monkeys (Macaca mulatta) was studied by scanning electron microscopy (SEM) of vascular corrosion casts and light microscopy (LM) of China ink-injected/cleared tissues. The principal results were that 1) insulo-acinar portal vessels were found between the endocrine and exocrine parts in the pancreas. The blood flows from the endocrine to the exocrine part. 2) Depending on the different microvascular arrangement, there were two patterns of microcirculation in the islet: in 66% of islets the direction of microcirculation was from cortex to core, and in 44% from core to cortex. 3) Islets could be categorized in three classes on the basis of size: the small islets (40-100 microns in diameter), the intermediate islets (101-240 microns in diameter), and the large islets (241-340 microns in diameter). 4) Insulo-insular portal routes were observed in the pancreas of the monkey. Some intermediate or large islets were connected to an adjacent small islet by one or two, occasionally more, efferent vessels. These small islets received no arterial branch and were entirely supplied by the portal vessels--the efferent vessels of intermediate or large islets. The authors suggest this new pattern to be termed the insulo-insular portal system. 5) A single centrally located intralobular artery as the exclusive vessel supplied each pancreatic lobule of the monkey, there being no anastomosis between the intralobular arteries and any of their branches. This anatomic feature might be the morphological basis of the pancreatic microcirculatory disturbance and microvascular impairment occurring during acute pancreatitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗
PubMed1995

Blood supply of subcutaneous tissue in the leg and its clinical application.

The vascular anatomy of the subcutaneous or adipose layer of the leg is described. The major arteries of the distal thigh and leg were injected with either a colored latex or an India ink and gelatin mixture to demonstrate the principal sources of blood supply to the skin and the superficial tissues of the leg. Microdissection and the technique of Spalteholz for dehydrating and clearing the tissues (rendering them transparent) facilitated the study of the direct and indirect arterial branches which form the three principal networks: the deep fascia, the subcutaneous or adipose layer, and the skin or the dermoepidermic layer. The results are of practical importance since the fascio-subcutaneous tissue layer can be used as a flap for covering soft tissue defects of the leg.

Adipose Tissue↗
PubMed2006

Imaging by sensitized oxygenations of photochromic anthracene films: examination of effects that improve performance and reversibility.

The aliphatic anthracene compound 1 and the oligomeric anthracene 2 were synthesized. Thin films of 1 and 2 mixed with the sensitizers tetraphenylporphyrin (TPP) and methylene blue (MB) were irradiated with visible light in air. Upon formation of singlet oxygen, the anthracene units were converted quantitatively to the corresponding endoperoxides. Heating of the irradiated samples afforded the parent anthracenes with high yields. Here, we demonstrate that the kinetics and reversibility of this reaction strongly depend on the microenvironment of the anthracene groups in the two compounds. The photooxidation of thin films of 1 is accompanied by interesting changes in the morphology of the film and allows the first application of 1 as a nondestructive negative-tone photoresist for lithography and as an oxidizing ink. The morphology of 2 remained unchanged after photooxidation as a result of the stabilizing oligomer backbone. This stabilizing effect significantly improves the photochromic performance of 2. The reversibility of the photooxidation is very high (>90 %) for oligomeric films of 2 after several cycles of irradiation and heating. Decomposition of the anthracene and a loss of the activity of the sensitizer diminish slightly the performance of the monomeric species.

Journal Article↗
PubMed1983

The islands of Calleja complex of rat basal forebrain. III. Histochemical evidence for a striatopallidal system.

The characteristics of the islands of Calleja complex (ICC) in the basal forebrain of the rat were studied with immunohistochemistry, histofluorescence, acetylcholinesterase staining, India ink vascular perfusions, electron microscopy, and steroid autoradiography. The ICC contains clusters of granule cells and associated medium-sized and large cells in the surrounding neuropil of the olfactory tubercle and septum-nucleus accumbens interface. The ICCs were found to contain monoamine fibers (dopamine and norepinephrine), neuroactive peptide fibers (leu-enkephalin, met-enkephalin, substance P, cholecystokinin, luteinizing hormone-releasing hormone), acetylcholinesterase-containing somata and dendrites, and medium-sized and large cells that concentrate [3H] estradiol. The specific overlap and combination of putative neurotransmitters in separate compartments of the ICC suggest that these structures contain striatum- and pallidumlike components. Striatumlike regions are defined as the zone in the rim regions of the ICC and are innervated predominantly by dopamine and cholecystokinin inputs. Pallidumlike regions are defined as the synaptic zone near the medium-sized and large cells of the cap and core regions of the ICC and they are innervated predominantly by enkephalin, substance P, and gamma aminobutyric acid inputs. The morphology, connections, and neurotransmitter relationships of the ICC, therefore, resemble classical striatopallidal systems. The additional presence of substances involved in the reproductive neuroendocrine systems (luteinizing hormone-releasing hormone, estradiol-binding cells, especially in the medial ICC, suggest that some ICC are involved in an endocrine corticostriatopallidal system. These endocrine systems resemble other neocortically and allocortically originating corticostriatopallidal systems in terms of their cell types, connections, and neurotransmitter systems. A functional role for the ICC in extrapyramidal motor systems is proposed.

Acetylcholinesterase↗
PubMed1993

Transient neovascularisation of the frog retina during optic nerve regeneration.

In conditions such as diabetic retinopathy, degenerative events in the retina are associated with neovascularisation. It is well established that a proportion of retinal ganglion cells die during optic nerve regeneration in the frog. The present study has determined whether neovascularisation takes place during this regenerative process. To do so, the pattern of blood vessels overlying the retinal ganglion cell layer was analysed in the frog Litoria (Hyla) moorei. We examined normal animals and those undergoing optic nerve regeneration following nerve crush. Blood vessels were visualised by perfusion with Indian ink and retinae were prepared as wholeamounts. In normal animals, the vascular tree was found to lie superficial to the nerve fibre layer and was more complex in regions overlying the area centralis and visual streak. After nerve crush, abnormal blood vessels transiently formed between the existing branches of the vascular tree. The new vessels were concentrated as an annulus centred on the optic nerve head and over the area centralis in midtemporal retina. The neovascularisation became most extensive between 6 and 10 weeks postcrush and disappeared by 12 weeks. The spatiotemporal sequence of neovascularisation suggests that it is triggered by accumulations of degenerating material formed as a proportion of the ganglion cells die during optic nerve regeneration.

Animals↗
PubMed1984

Maternal discrimination of pup sex in rats.

In recent studies (Moore, 1981, 1982; Moore & Morelli, 1979), it has been found that maternal female rats spend more time licking the anogenital region of male than female offspring. This discrimination of pup sex has important implications for the development of a number of sex differences in behavior later in life, and the present study was designed to re-examine Moore's observations while allowing each pup to be individually identified. Litters of Sprague-Dawley rats were delivered by Caesarean section, and the footpads of each animal were tattooed with India ink to permit identification throughout testing. Each litter was then fostered to a maternal female and remained with her except when observations of maternal licking were made--on the fourth, seventh, and tenth days after delivery. On each of these days, the foster mother was placed in a test chamber for 30 min, and each pup was then presented individually for a 10-min session. During this time the number and duration of anogenital licking episodes were recorded. We found, as did Moore, that adult females do distinguish between their male and female pups, spending significantly more time licking the anogenital region of the males in their litters on all but the first day of testing. The possibility that the attentional bias toward male young has important consequences for behavior later in life and contributes to behavioral differences between the sexes (Beach, 1979; Moore, 1982) makes the identification of individual animals throughout development important in the analysis of these effects.

Animals↗
PubMed2003

Morphogenesis of blood vessels in the head muscles of avian embryo: spatial, temporal, and VEGF expression analyses.

Adult skeletal muscle is a highly vascularized tissue, but the development of intramuscular endothelial networks has not been well studied. In quail embryos, QH1-positive angioblasts are present and moving throughout myogenic head mesoderm before the onset of primary myotube formation. On day 5 of incubation, concurrent with early myotube formation and aggregation, angioblasts establish a transient vascular plexus surrounding the myogenic condensations. Between days 5 and 9, the intramuscular vessels form an irregular network of endothelial cords and patent channels and only later are the parallel arrays of capillaries characteristic of adult muscles established. Microinjections using India ink, QH1, and Mercox resin reveal that these intramuscular capillaries are typically not connected to systemic vessels of the head until day 10, which is near the end of primary myogenesis and corresponds to the onset of muscular function. Morphometric analyses performed during primary myogenesis stages show a decrease in muscle cell density but no significant changes in intramuscular vascular density between days 5 and 9. This finding was surprising, as it is generally assumed that muscle growth requires elevated oxygen and nutrient levels. Moreover, there are no significant morphometric differences in vascular supply to embryonic fast and slow muscles. Endothelial tissue density is similar in slow muscles (oculorotatory, e.g., lateral rectus), fast muscles (mandibular depressor), and mixed muscles, in which the fiber types can be interspersed (jaw adductors) or segregated (branchiomandibular). Vascular endothelial growth factor (VEGF) protein is abundant in myotubes but not endothelial cells within both fast and slow head muscles at days 7 and 9. However, in some mixed muscles, only a minority of myotubes, which do not correspond to one specific fiber type, express VEGF. These results document a dynamic set of intramuscular and perimuscular angiogenic reorganizations during avian head myogenesis. Thus far, no vasculogenic distinctions between fast and slow muscles have been observed, although muscle heterogeneity in VEGF expression is evident.

Animals↗
PubMed1998

Maturation of phagosomes is accompanied by specific patterns of small GTPases.

In this study we purified phagosomes of the ciliated protozoan Tetrahymena thermophila to analyze aspects of the maturation pathway of phagocytotic vesicles. Phagosomes were labeled with magnetic microparticles and then purified in high amounts with the help of a permanent magnet. By combining a pulse-chase labeling protocol with the magnetic separation procedure we were able to isolate phagosomes of defined ages, which represent distinct stages of their maturation pathway. GTP-overlay assays showed that a set of small GTPases of the ras superfamily is associated with these phagosomes. Phagosomes isolated at different stages of maturation revealed a change in the pattern of the small GTPases. Some small GTPases identified by the GTPase overlay assays could be aligned to India ink stained protein spots in two-dimensional gels of isolated phagosomes. The results presented here are a first step to identify the members of small GTPases associated with phagosomes during their maturation pathway. Microsequencing of pooled polypeptides by mass-spectrometric techniques will identify the primary structure of these small GTPases.

Animals↗
PubMed2005

Integration of a carbon microelectrode with a microfabricated palladium decoupler for use in microchip capillary electrophoresis/electrochemistry.

A method to integrate a carbon microelectrode with a microfabricated palladium decoupler for use in microchip capillary electrophoresis (CE) is detailed. As opposed to previous studies with decouplers for microchip CE, the working electrode material, which is made by micromolding of a carbon ink, is different from the decoupling electrode material (palladium). The manner in which the working electrode is made does not add additional etching or lithographic steps to the fabrication of the glass electrode plate. The hybrid poly(dimethylsiloxane)/glass device was characterized with fluorescence microscopy and by monitoring the CE-based separation of dopamine. Hydrodynamic voltammograms exhibited diffusion-limited currents occurring at potentials above +1.0 V. It was also shown that the half-wave potential does not shift as the separation potential is changed, as is the case in nondecoupled systems. Gated injections of dopamine in a 25 mM boric acid buffer (pH 9.2) showed a linear response from 200 to 5 microM (r2 = 0.9992), with a sensitivity of 5.47 pA/microM and an estimated limit of detection of 2.3 microM (0.621 fmol, S/N = 3). This is the first report of coupling a carbon electrode with a decoupler in microchip CE.

Carbon↗
PubMed2006

Use of micromolded carbon dual electrodes with a palladium decoupler for amperometric detection in microchip electrophoresis.

The fabrication and evaluation of micromolded dual carbon ink electrodes and their integration with a fabricated palladium decoupler for use in microchip electrophoresis is described. As opposed to previous work involving carbon-based dual electrodes with microchip electrophoresis, this approach results in electrodes that are amenable to mass production in a manner where the decoupler/electrode alignment is fixed and reproducible. In this work, electrode sizes and spacings were optimized to result in dual carbon electrodes that are 1 microm in height and separated by 100 microm. Fluorescence microscopy was used to investigate leakage around the electrode/channel interface as well as to investigate what effect the dual electrodes have on band broadening phenomena. The performance of the microelectrodes was demonstrated by the separation and selective dual electrode detection of neurotransmitters in the presence of ascorbic acid. It was also found that addition of SDS to the buffer system improved both the LODs and collection efficiencies. This approach, which is the first involving carbon-based dual electrodes with an on-chip palladium decoupler, will be useful for separating and detecting neurotransmitters that are either collected by in vivo sampling or released from cells on-chip.

Ascorbic Acid↗
PubMed2003

Oligodendrocyte precursor hypercellularity and abnormal retina development in mice overexpressing PDGF-B in myelinating tracts.

Platelet-derived growth factor (PDGF) influences the generation of neurons and glia during embryogenesis and in early postnatal life. In an attempt to determine the consequences of an overexpression of PDGF-B during the first weeks of life, we targeted transgenic expression of a human PDGF-B cDNA to myelinating tracts using the promoter region of the myelin basic protein (MBP) gene. Transgenic mRNA and protein were expressed in the brain and the expression profile of the human PDGF-B during early postnatal development closely paralleled that of the endogenous mouse MBP gene. The gross morphological appearance of transgenic brains was normal but at the cellular level several phenotypic alterations could be identified. In white matter tracts such as the corpus callosum and cerebellar medulla, there was a marked hypercellularity. The number of oligodendrocyte precursors was increased and astrocytes were more abundant. In adult mice carrying the MBP-PDGF-B transgene, however, myelination appeared normal and the amount of oligodendrocytes was similar to that of control littermates. In addition to the phenotypic alterations in the brain, investigation of eye structure revealed a striking disorganization of retinal architecture. The retina was folded with cells collected in papillar or follicular-like structures. Retinal whole mount preparations after India ink perfusion revealed capillary disorganization with large-caliber vessels supporting only a few fine branches. Our observations strengthen the notion that PDGF is an important effector molecule in postnatal CNS development.

Animals↗
PubMed2002

Distinguishing depression from dementia in later life: a pilot study employing the Emotional Stroop task.

OBJECTIVES: In later life, cognitive impairment is common in depression often making it difficult to distinguish a dementing illness from depression. We examined whether people with depression could be differentiated from those with dementia on their performance on a task that examines attentional bias to depression related material. METHODS: Twelve older adults who fulfilled DSM-IV criteria for major depression were compared with 12 people with Alzheimer's Disease (AD) and 12 age matched controls on a test of cognitive biases: the Emotional Stroop task. In this task participants were presented with words written in different coloured inks, and they had to name the colour the word was written in. Four types of material were presented-neutral, positive, and negative emotion words as well as a condition of meaningless symbols. RESULTS: People with depression and those with AD were both slower than the controls on the task generally. However, the depressed group alone showed a statistically significant and specific increase in response time when colour naming the negative emotion words. The other two groups did not demonstrate such a pattern and colour named neutral, positive and negative words equally quickly. CONCLUSIONS: The biased processing of depression related material may have a valuable role in distinguishing depression from dementia in later life. Although the Emotional Stroop in its present form is not sufficient for such a purpose. Furthermore, the demonstration that older adults with depression exhibit such biases helps provide a theoretical basis for the application of cognitive behavioural treatments with older adults.

Aged↗
PubMed2005

Anterior cingulate cortex: an fMRI analysis of conflict specificity and functional differentiation.

In this event-related functional magnetic resonance imaging (fMRI) study, we provide evidence that the role of the anterior cingulate cortex (ACC) in cognitive control may not be unitary, as the responses of different ACC subregions vary depending upon the nature of task-irrelevant information. More specifically, using the color-word Stroop task (congruent, incongruent, and neutral trial types), we examined the degree to which increases in neural activity within ACC are specific to conditions of conflict, as posited by the conflict monitoring theory (Botvinick et al. [1999]: Rev Neurosci 10:49-57; Carter et al. [1998]: Science 280:747-749). Although incongruent and congruent trials both involve two competing sources of color information (color word and ink color), only incongruent trials involve a direct conflict between task-relevant and task-irrelevant information. Although the anterior division of the ACC rostral zone exhibited conflict specific increases in neural activity (i.e., incongruent > congruent = neutral), the posterior division exhibited a more generalized pattern, increasing whenever the task-irrelevant information was color related, regardless of whether it was conflicting (i.e., incongruent and congruent > neutral). Our data thus suggest a possible functional differentiation within the ACC. As such, it is unlikely that the role of the ACC in cognitive control will be able to be accommodated by a single unifying theory.

Adolescent↗
PubMed1990

Hepatobiliary effects of morphine are mediated in the brain.

Morphine slows hepatobiliary elimination of sulfobromophthalein in rodents, raising dye levels in plasma and liver. Earlier studies showed these effects to be independent of other opiate effects such as bile duct spasm, hypothermia or blood gas changes resulting from respiratory depression. Because opiate receptors are distributed throughout the body, within the central nervous system and at peripheral sites including the gastrointestinal tract, experiments were performed to ascertain whether central or peripheral sites mediate the hepatobiliary effects of morphine. Sulfobromophthalein was administered intravenously to mice and its levels were measured in plasma and liver. Tail-flick latency indicated centrally mediated analgesia. Inhibited intestinal transit of India ink reflected an opiate effect with a significant peripheral component. When injected into a cerebral ventricle morphine was much more potent in producing analgesia and raising sulfobromophthalein levels than when administered intravenously or intraperitoneally. An intravenous dose of naloxone that reversed morphine analgesia also prevented sulfobromophthalein elevation but did not prevent gut slowing. Naltrexone injected in a cerebral ventricle also reversed analgesia and sulfobromophthalein elevation but not intestinal slowing. The polar opiate agonist N-methylmorphine did not cause analgesia or raise sulfobromophthalein levels at peripheral intraperitoneal doses to 100 mg/kg. When given in a central ventricle at 4 x 10(-3) mg/kg, this agent produced analgesia and raised sulfobromophthalein but did not slow intestinal transit. After spinal cord transection, intravenous morphine did not retard the tail-flick response or affect sulfobromophthalein disposition, but peripherally mediated intestinal transit was slowed as it was in intact mice. These experiments demonstrate parallel opiate effects on analgesia and on BSP disposition but not on intestinal transit.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

The portal lobule in rat liver fibrosis: a re-evaluation of the liver unit.

We re-evaluated three schemes of liver organization: the classic lobule, the portal lobule, and Rappaport's liver acinus. The lobular angioarchitecture of normal rat liver and the three-dimensional structure of pseudolubules found in rat livers with fibrosis induced by swine serum were compared with the classic lobule of the pig. Normal and fibrotic rat livers and pig livers were perfused, injected with either India ink or 0.75% OsO4 through the portal and/or hepatic vein, and immersionfixed. Whole lobes and hand-cut thick sections were made transparent with a solution of benzyl benzoate and methyl salicylate. The angioarchitecture of normal rat liver differs from pig liver. In the former, terminal portal branches and central veins interdigitate, and in the latter, numerous terminal portal branches that arise from interlobular portal veins establish a vascular basket surrounding one central vein and forming classic lobule. The structure of liver acinus is never found in the pig liver. The terminal portal branch, together with the terminal hepatic artery and bile duct, are present inside each pseudolobule of fibrotic rat livers. Blood from the terminal portal branch flows through inlet venules into radiating sinusoids, and, at the periphery converges into newly formed septal and angular outlet venules; these venules terminate in fibrotic central veins located at each corner. Pseudolobules are not rugby ball-like as Rappaport's liver acini are but are polyhedron in shape. The rat pseudolobules are comparable with the portal lobule; its structure and microcirculation are the reverse of the pig classic lobule. Rat pseudolobules are different from liver acini, as shown by the following: 1) their three-dimensional shape is different; and 2) they have a reverse relationship to classic lobules while acini are defined to subdivide classic lobules. In normal and fibrotic rat livers, the liver unit is the portal lobule with a terminal portal branch as the axial branch and central veins at the periphery. The co-existence of liver acini and classic lobules is doubtful.

Animals↗
PubMed2003

NDP gene mutations in 14 French families with Norrie disease.

Norrie disease is a rare X-inked recessive condition characterized by congenital blindness and occasionally deafness and mental retardation in males. This disease has been ascribed to mutations in the NDP gene on chromosome Xp11.1. Previous investigations of the NDP gene have identified largely sixty disease-causing sequence variants. Here, we report on ten different NDP gene allelic variants in fourteen of a series of 21 families fulfilling inclusion criteria. Two alterations were intragenic deletions and eight were nucleotide substitutions or splicing variants, six of them being hitherto unreported, namely c.112C>T (p.Arg38Cys), c.129C>G (p.His43Gln), c.133G>A (p.Val45Met), c.268C>T (p.Arg90Cys), c.382T>C (p.Cys128Arg), c.23479-1G>C (unknown). No NDP gene sequence variant was found in seven of the 21 families. This observation raises the issue of misdiagnosis, phenocopies, or existence of other X-linked or autosomal genes, the mutations of which would mimic the Norrie disease phenotype.

Abnormalities, Multiple↗
PubMed2006

Induction of p16INK4A mediated by beta-catenin in a TCF4-independent manner: implications for alterations in p16INK4A and pRb expression during trans-differentiation of endometrial carcinoma cells.

Excessive beta-catenin is considered to contribute to tumor progression by inducing transcription of cell cycle-related genes such as cyclin D1 and c-myc. In contrast, our recent studies demonstrated that beta-catenin could inhibit cell proliferation through activation of p14(ARF)/p53/p21(WAF1) pathway during trans-differentiation toward morular phenotype of endometrial carcinoma (Em Ca) cells. Here, we focused on associations with alterations in p16(INK4A) and pRb expression during this process. In clinical cases, p16(INK4A) immunoreactivity was found to frequently overlap with nuclear beta-catenin accumulation in small-sized morules and surrounding glandular carcinomas (Sur-Ca), demonstrating a significant positive correlation (r = 0.447, p < 0.0001) overall, while the immunoreactions showed stepwise decrease in enlarged morules, despite persistent accumulation of beta-catenin and p21(WAF1) in nuclei. Immunoreactivity for both total pRb and its phosphorylated form was apparently decreased in all morules as compared to Sur-Ca lesions, with a significantly positive correlation. In cell lines, transcriptional activation of p16(INK) (4A) promoter by active form beta-catenin, as well as p21(WAF1), occurred through the region from -385 to -280 bp relative to the translation start site, in a TCF4-independent manner. Moreover, cell proliferation was accompanied with phosphorylation of pRb and increased p16(INK4A) expression, while its inhibition by serum starvation caused decreased expression of total pRb but not p16(INK4A), resulting in high relative amounts of the latter. These findings indicate that induction of p16(INK4A) mediated by nuclear beta-catenin and p21(WAF1), along with loss of pRb expression, may be important for initial steps during trans-differentiation of Em Ca cells. In addition, its down-regulation is associated with progression of lesions.

Blotting, Western↗
PubMed1992

Mutagenicity studies on two triphenylmethane dyes, bromophenol blue and tetrabromophenol blue.

Bromophenol blue and tetrabromophenol blue are two triphenylmethane dyes. Triphenylmethane derivatives and their structurally related compounds, such as fluoresceins and xathenes, are widely used as industrial dyes for foods, drugs, cosmetics, textiles, printing inks or laboratory indicators. Since a number of these types of dyes have been reported to be genotoxic, safety concerns on these two dyes of interest have been raised. Consequently, a battery of genetic toxicology assays, including the Ames Salmonella/microsome assay, L5178Y TK+/- mouse lymphoma assay, mouse micronucleus test and mitotic recombination assay with yeast Saccharomyces cerevisiae strain D5, has been performed on each of the two dyes. The results of the evaluations indicate that both bromophenol blue and tetrabromophenol blue were not active and can be considered non-genotoxic for the three genetic endpoints assessed (gene mutation, chromosome aberrations and primary DNA damage). Genetic activities in some structurally related compounds of these dyes have been reported but may be attributed to the presence of mutagenic impurities rather than the compound itself.

Animals↗