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PubMed1989

A search for lymphatic drainage of the monkey orbit.

Colloid solutions of technetium Tc 99m and india ink injected into the retrobulbar space of the cynomolgus monkey outside the extraocular muscle cone were removed from the orbit by the lymphatic vessels of the conjunctiva and eyelids and were then concentrated within the lymph nodes that drained the conjunctival and eyelid areas. Colloid solutions injected into the retrobulbar space inside the extraocular muscle cone did not reach the conjunctiva and did not collect in any lymph nodes over a 24-hour period. Within the orbit, the injected colloids spread along the planes of the connective-tissue septa. No lymphatic vessels were identified within the orbits posterior to the conjunctiva. Small amounts of india ink left the posterior orbit and ultimately entered the contralateral orbit. This posterior pathway did not lead to lymphatic vessels or lymph nodes and therefore does not appear to represent a prelymphatic pathway.

Animals↗

PubMed1977

Methylphenidate hydrochloride effects on psychological tests in acute schizophrenic and nonpsychotic patients.

Actively psychotic schizophrenic and nonpsychotic psychiatric inpatients received intravenous methylphenidate hydrochlroide (0.5 mg/kg). Each patient was rated for level of psychosis and talkativeness, and each received the Holtzman projective ink blot tests and the Kent-Rosanoff word-association tests before, during, and after methylphenidate infusion. Minnesota Multiphasic Personality Inventory Goldberg Index scores were also obtained as a general measure of psychosis. For the entire patient group, methylphenidate infusion was followed by a significant increase in talkativeness and psychosis ratings and in pathological responses to the Holtzman ink blot test. There was a significant decrease in common word associations. Since neither of the psychological tests allow more than one response per item, it appears that methylphenidate truly effects pathological thought processes and decreases common word associations as such, rather than merely making these processes more evident by increasing verbalization.

Acute Disease↗

PubMed1997

Primitive interventricular septum, its primordium, and its contribution in the definitive interventricular septum: in vivo labelling study in the chick embryo heart.

BACKGROUND: Because the studies on the embryological development of the primitive interventricular septum have been done with postmortem material, we do not know the site within the cardiac tube and the developmental stage at which the primordium appears and its anatomical manifestation in the mature heart. Consequently, we do not know its real contribution to the constitution of the definitive interventricular septum. METHODS: With this purpose, we selected an adequate biological model, the chick embryo heart and the in vivo labelling technique. We placed a label of gelatin India ink in the ventral fusion line of both cardiac primordia at the level of the interventricular grooves in the straight tube heart (stage 9+HH), and we traced the ink up to the mature heart (stage 36HH). We made histological sections of some hearts, of the zone where the label was found to investigate the first morphological manifestation of the primitive interventricular septum. We also made microdissections and scanning electron microscopic studies. RESULTS: The label placed at stage 9+HH in the ventral fusion line of both cardiac primordia, at the level of the interventricular grooves, was found at stage 14HH in the greater curvature of the looped heart, opposite the left interventricular groove. This label at stage 17HH was found in the apical trabecular region of the first cardiac septum (8-shaped septum) and in the mature heart (stage 36HH) in the definitive interventricular septum at the limit between the basal and the medial third of the definitive interventricular septum. CONCLUSIONS: Firstly, the primordium of the primitive interventricular septum appears at stage 9+HH, in the ventral fusion line of both cardiac primordia at the level of the interventricular grooves. Secondly, its first morphological manifestation takes place at stage 17HH, and it forms the apical trabeculated region of the first cardiac septum (8-shaped septum). Thirdly, the primitive interventricular septum gives origin to the middle and apical third of the definitive interventricular septum.

Animals↗

PubMed1996

The arterial anatomy of larynx transplantation: microsurgical revascularization of the larynx.

Advances in immunosuppression and selective reinnervation may soon make laryngeal transplantation a potential therapy for patients undergoing total laryngectomy. Successful transplantation requires a clear delineation of those vessels necessary to completely revascularize the larynx. Our hypothesis is that the arterial inflow provided by a single superior thyroid artery is sufficient to revascularize the entire larynx. To test this hypothesis, 8 cadavers were studied via either barium latex injection (n = 4) to assess contralateral tissue perfusion or India ink (n = 4), to determine the degree of mucosal perfusion. Following injection via a single superior thyroid artery, all larynges demonstrated either complete, bilateral tissue perfusion evidenced by x-ray visualization of the barium latex injected specimen or bilateral mucosal staining with India ink. We conclude that bilateral perfusion of the entire larynx transplant, including laryngeal and epiglottic mucosa, would occur after revascularization of a single superior thyroid artery. These findings suggest that reliable revascularization of a larynx transplant is technically possible using modern microsurgical techniques.

Adult↗

PubMed1976

Renal vasculature of the trout demonstrated by scanning electron microscopy, compared with canine glomerular vessels.

In order to gain additional information regarding renal circulatory patterns, we have used both ink and resin injections to study the arterial supply to the mesonephric kidney of trout. Arterial injections through the dorsal aorta with ink were made for histological preparations in which the length, termination and relationship of glomerular vessels were examined. Similar injections with methyl methacrylate were made in preparation of corrosion casts to provide us with gross replicas of the aortic branches to the kidney as well as casts of glomerular structure for scanning electron microscopy. The sequence of vessels through which arterial blood passed to the renal corpuscle and ultimately to the uriniferous tubules was traced. Each afferent arteriole was found to terminate in three to six branches which formed anastomosing circuits of capillaries; these vessels reunited at the hilum to form a single efferent arteriole. The efferent arterioles in trun traveled a short distance to peritubular capillary beds and sinusoids. Morphological evidence was found for preglomerular sphincter-like action only. The glomerular vessels were found to be similar to, although less complex than, those of the outer and mid-cortical regions of the dog kidney.

Animals↗

PubMed1986

Localized vascular regression during limb morphogenesis in the chicken embryo. I. Spatial and temporal changes in the vascular pattern.

Morphogenesis of the vertebrate limb bud depends upon reciprocal interactions between epithelial and mesenchymal tissues. A characteristic limb vascular pattern is essential for normal limb outgrowth. The vascular pattern in the distal portion of the wing bud was examined by ink injection and compared to the sites of cartilage differentiation, as indicated by [35S]-incorporation. During development, avascular areas arose in three distinct locations within the vascularized mesoderm. These areas corresponded to the distal skeletal elements, referred to as digits 2, 3, and 4. Incorporation of radioactive sulfate was high in the avascular areas and low in the adjacent vascular tissue. Examination of autoradiographs of ink-injected limbs suggested that the appearance of avascular regions preceded the accumulation of sulfated cartilage matrix. These results indicate that remodeling of the limb vasculature is related to the formation of the skeletal pattern.

Animals↗

PubMed1991

Endocytotic activity of the free floating cells and epithelial cells in the endolymphatic sac: an electron microscopic study.

The fine structure and its functional properties of both free floating cells and epithelial cells in the endolymphatic sac after injections of India ink particles or polystyrene latex beads, 0.24 micron in diameter, into the endolymphatic space, were studied using light and electron microscopy. Twenty-four to 48 hours after injections, these foreign materials had accumulated in the lumen of the endolymphatic sac and a large number of them were ingested into free floating cells, most of which appear to be macrophages. Granular leucocytes taking up the foreign materials into the cytoplasm were also recognized in the sac lumen. A few of these leucocytes passed through the epithelium and migrated to the subepithelial connective tissue, while the others were degenerated and phagocytosed by the free floating cells in the lumen of the sac. In addition to the free floating cells, ink particles, latex beads, and degenerated leucocytes were endocytosed into some of the epithelial cells. As we reported previously, the epithelial cells were clearly classified into two types; type-1 epithelial cells (cytoorganelle-rich cells) and type-2 epithelial cells (filament-rich cells). The foreign materials and degenerated cells were taken up mainly into the type-1 epithelial cells, while the type-2 epithelial cells did not show so marked endocytotic activity in comparison with the type-1 epithelial cells. Thus, it becomes clear that the endolymphatic sac plays an important role in the endocytotic activity for foreign materials and waste products, and both the free floating cells and the type-1 epithelial cells of the sac have strong endocytotic activity.

Animals↗

PubMed2001

JNK phosphorylates the HSF1 transcriptional activation domain: role of JNK in the regulation of the heat shock response.

The role of c-Jun NH2-terminal kinase (JNK) signaling cascade in the stress-inducible phosphorylation of heat shock factor 1 (HSF1) was investigated using known agonists and antagonists of JNK. We showed that treatment of HeLa cells with MG132, a proteasome inhibitor and known INK activator, caused the transcriptional activation domain of HSF1 to be targeted and phosphorylated by JNK2 in vivo. Dose-response and time course studies of the effects of heat shock and anisomycin treatment showed a close correlation of the activation of JNK and hyperphosphorylation of HSF1. SB203580 inhibited INK at the 100 microM concentration and significantly reduced the amount of hyperphosphorylated HSF1 upon heat shock or anisomycin treatment. SB203580 and dominant-negative JNK suppress hsp70 promoter-driven reporter gene expression selectively at 45 degrees C but not at 42 degrees C heat stress, suggesting that JNK would be preferentially associated with the protective heat shock response against severe heat stress. The possibility that JNK-mediated phosphorylation of HSF1 may selectively stabilize the HSF1 protein and confers protection to cells under conditions of severe stress is discussed.

Animals↗

PubMed1993

Examples of evolution of microstructure in ceramics and composites.

In this article we describe a number of studies involving the direct observation of microstructural evolution. In general these investigations were carried out to establish the mechanistic paths involved. The materials studied range from fibers being evaluated for use in high-temperature ceramic composites to energetic materials used as propellants. In particular we discuss the room temperature imaging of materials difficult to image by conventional means and the use of the chamber atmosphere to influence microstructural evolution. Imaging of hydroxyapatite formed by chemical means is briefly described as an example of a difficult microstructure. Microstructural evolution during calcium aluminate cement hydration relies on the chamber atmosphere to control moisture loss from the hydrating specimens. In some instances microstructural evolution with heating occurred independently of the chamber atmosphere. Grain growth in PZT films formed by sol-gel processes depends strongly on temperature but does not appear to depend on the chamber atmosphere. This is also the case for the combustion of nitroamine propellants in that their combustion does not depend on access to an external source of oxygen. In other studies, the chamber atmosphere played an indirect role in determining microstructure. However, the mechanistic path driving microstructural evolution in copper-based inks used as conductive paths on electronic substrates is atmosphere dependent. These inks are formulated from copper powder, glass, and an organic binder, and the interaction of the binder with an oxidizing atmosphere allows it to be burned out before significant interaction occurs between the copper powder and the glass. Finally, the microstructural variations during the oxidation of structural composites at high temperature were used to allow assessments of their likely failure mechanisms.

Animals↗

PubMed2002

Magnetite tattoos.

BACKGROUND AND OBJECTIVES: Tattoo removal is a significant problem. The extraction of magnetite (Fe(3)O(4)) ink tattoos by a magnetic field was investigated, with and without Q-switched laser treatment. STUDY DESIGN/MATERIALS AND METHODS: Magnetite particles (1.4 microm) were used to make mature, black skin tattoos on hairless albino rats. A Q-switched ruby laser (QSRL) 3.5 J/cm(2), 6.5-mm spot size, 40-nanosecond pulse width was used for treatment. Permanent magnets (1.4 T, 6-mm diameter) were tested to extract the magnetite particles, alone and after QSRL. Lightening of treated tattoos was measured from digital photographs, and the amount and distribution of magnetite in skin biopsies was scored blindly. RESULTS: External application of magnets on mature magnetite tattoos without prior QSRL treatment, did not significantly extract, lighten, darken, or change their histologic appearance. A magnetic field applied immediately after QSRL treatment extracted some ink when epidermal injury was present, and caused significant redistribution of magnetite into the upper dermis with vertical banding along magnetic field lines. When applied for 3 weeks following QSRL, magnets caused darkening of tattoos. CONCLUSIONS: Magnetite skin tattoos can be manipulated by external magnets, especially after Q-switched laser treatment. Magnetically-extractable tattoos may be feasible.

Albinism↗

PubMed1994

Prefabrication of a vascularized nerve graft by vessel implantation: preliminary report of an experimental model.

Regeneration through vascularized nerve grafts (VNG) seems to be better than nonvascularized nerve grafts (NVNG), especially in hostile beds. We report on an experimental technique of prefabrication of VNG by direct vessel implantation. An arteriovenous fistula was created in the groin region with autologous vein grafts in the Wistar rat model, and implanted into the sciatic nerve. Five weeks later the sciatic VNG was elevated on the prefabricated pedicle. The flap was free-transferred orthotopically over a silicone sheet to impede plasmatic imbibition. Flap viability at 3 days was complete. India ink injection of the AV fistula resulted in capillary ink filling within the nerve and surrounding tissues. Histologic sections of the flap were examined, revealing its neovascularity. In an ongoing study, the regeneration through this prefabricated VNG is being compared to native VNG.

Abdominal Muscles↗

PubMed2002

Identification of loci associated with putative recurrence genes in transitional cell carcinoma of the urinary bladder.

Following an earlier study linking monosomy 9 with recurrence of transitional cell carcinomas (TCCs) of the urinary bladder, 109 primary and recurrent TCCs (from 47 patients) were examined to explore genetic alterations at chromosome 9 associated with recurrence. Patient DNA was microdissected and extracted from archival tissue sections and analysed for loss of heterozygosity (LOH) at three regions on chromosome 9 where tumour suppressor genes (TSGs) are known to reside (INK 4A, DBC1, and TSC1). Patients were categorized into two groups, non-recurrent TCC (NR, n=18) and recurrent TCC (REC, n=29). It was noted that 12% of NR tumours, compared with 54% of REC primary tumours (p=0.01), had LOH at all informative markers spanning the TSC1 region. The risk of recurrence was significantly higher in patients with deleted TSC1 than in those who retained the TSC1 region (p=0.035). Levels of LOH at DBC1 or INK 4A were not significantly different in NR tumours than in REC primary tumours and recurrence-free survival was not affected by loss of either of these genes. Loss of all informative markers spanning chromosome 9 was observed in 0% of NR tumours compared with 25% of REC primary tumours (p=0.04). The probability of recurrence was also significantly increased in patients who had LOH at all informative markers spanning chromosome 9 (p=0.016), confirming earlier fluorescence in situ hybridization results. This study provides further evidence that recurrence in bladder cancer is a distinct event, with underlying molecular causes. It also identifies the TSC1 locus as a candidate for a TSG, which drives recurrence in a proportion of TCC patients. Loss of all informative markers, including those residing in the TSC1 region, spanning chromosome 9 was also linked to recurrence.

Carcinoma, Transitional Cell↗

PubMed2005

Peptide mass fingerprinting by matrix-assisted laser desorption ionization mass spectrometry of proteins detected by immunostaining on nitrocellulose.

We have developed an approach that allows peptide mass mapping by matrix-assisted laser desorption ionization-mass spectrometry of proteins visualized on a nitrocellulose membrane by immunochemical detection. Proteins are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), electroblotted onto a nitrocellulose membrane and after blocking with a nonprotein-containing polymer such as polyvinylpyrrolidone 40 (PVP-40) or Tween 20, the proteins are stained with fount India ink. After incubation with primary and, if required, secondary peroxidase-coupled antibodies, immunochemically reactive proteins can be visualized using conventional enhanced chemiluminescence detection and assigned to the India ink-stained membrane by simple superposition. The proteins of interest are excised, submitted to "on-membrane" cleavage and the peptides are analyzed by mass spectrometry. Protein-based blocking reagents normally used in standard immunodetection protocols, such as skimmed milk, can be employed. We have obtained high-quality mass spectra of bovine serum albumin (BSA) detected on an immunoblot with an estimated amount of 100 fmol applied onto the gel, indicating the sensitivity of the present method. In addition, the approach is demonstrated with two other commercially available proteins, a serum protein, the successful identification of a tyrosine phosphorylated protein from total rat liver homogenate and serine phosphorylated proteins from an EcR 293 nuclear extract separated by two-dimensional (2-D) SDS-PAGE.

Animals↗

PubMed2001

Turnover of rat brain perivascular cells.

Brain perivascular spaces harbor a population of cells which exhibit high phagocytic capacity. Therefore, these cells can be labeled by intraventricular injection of tracers. Such perivascular cells at the interface between blood and brain are believed to belong to the monocyte/macrophage lineage and to be involved in antigen presentation. Currently, it is unclear whether these cells undergo a continuous turnover by entering and leaving the bloodstream. Using bone-marrow-chimeric animals, migration of donor macrophages into brain perivascular spaces has been reported. On the other hand, following intracerebral injection of india ink into nontransplanted animals, ink-labeled perivascular cells were still found 2 years after injection, suggesting a high stability of this cell pool. Thus, the turnover of perivascular cells observed in chimeras might be a result of bone marrow transplantation rather than a physiological occurrence. To address this issue, we monitored de novo invasion of macrophages into perivascular spaces of apparently healthy adult rats by applying techniques other than bone marrow transplantation, (i) consecutive injections of different tracers and (ii) ex vivo isolation of macrophages from the blood, cell labeling, and reinjection into the same animal to avoid MHC mismatch. Both approaches revealed vivid de novo invasion of macrophages into perivascular spaces, but not into brain parenchyma, rendering untenable the concept of perivascular cells forming a stable population of macrophages in the brain. Thus, brain perivascular spaces are under permanent immune surveillance of blood borne macrophages in normal adult rats.

Animals↗

PubMed1997

A Theoretical Model of Flotation Deinking Efficiency

The associated probabilities of each microprocess occurring in flotation deinking are employed in the development of a kinetic or population balance-type model of the overall flotation process. The overall model contains two kinetic constants: The first, k 1 , governs the overall probability of a free ink particle successfully being intercepted by and adhering to an air bubble; the second, k 2 , is a measure of the probability that a particle/bubble aggregate pair will become unstable and split to yield a "new" free ink particle. The solution to the kinetic model is presented in terms of k 1 and k 2 , which are themselves functions of system parameters such as bubble and particle physical properties (e.g., diameter, density) and fluid properties (e.g., viscosity, surface tension). From this solution, a definition of theoretical flotation efficiency is presented, as well as definitions of other system performance parameters, and selected predictions are displayed.

Journal Article↗

PubMed2004

Anatomical studies of the mouse genital tubercle.

BACKGROUND: To study the etiology of hypospadias, we propose the use of a mouse model, the embryonic mouse genital tubercle. In this study, we define the development of the mouse genital tubercle with special emphasis on urethral formation demonstrating anatomical similarities to human development. MATERIALS AND METHODS: Serial sections of genital tubercles from embryonic male and female mice ages 14 to 21 days gestation from timed pregnant animals, newborn and adult mice were immunohistochemical stained with antibodies to E-cadherin, cytokeratins 7, 10, and 14. Patency of the urethral was assessed by india ink injection via the bladder. Urethral lumen morphology was determined by the creation of plastic resin cast. Surface morphology of the genital tubercle was defined by scanning electron microscopy. RESULTS: India Ink injection into the bladder showed that the urethral lumen was patent from 14 days gestation. Plastic resin casts revealed that the male urethra was characterized by a S shaped curve, the presence of the bulbar urethral gland and a longer length than age matched females. The ontogeny of the genital tubercle development revealed two epithelial edges that subsequently touched and fused into the completed urethra. During development cytokeratin immunohistochemical staining demonstrated that the epithelial cells of the urethral lumen are of bladder origin and the surface cells of skin origin. CONCLUSION: The functional and developmental anatomy of the mouse genital tubercle provides a useful model to study normal and abnormal human urethral development.

Animals↗

PubMed1990

Structural alterations of pancreatic microvasculature in cerulein-induced pancreatitis in the rat.

Pancreatic microvasculature in cerulein-induced pancreatitis was studied at the electron and light microscopic level. Using a modified model of in situ india ink perfusion, focally a marked reduction of the number of india-ink-filled microvessels was found. The numerical density (NA) of filled microvessels was reduced by 66.7% in pancreatitis. The morphological correlate for this extreme numeric reduction might be a collapse of the lumen of pancreatic microvessels. In the presence of cerulein pancreatitis 42% of pancreatic capillaries had a collapsed lumen as shown by electron microscopy. The endothelium of capillaries in acute pancreatitis demonstrated surface blebbing, the formation of cytoplasmatic vacuoles, edema, and swollen mitochondria. Some capillaries had irregularities at the interendothelial junction, and the majority of examined vessels were surrounded by marked perivascular edema. No strict correspondence between histological signs of pancreatitis and light microscopic and/or ultrastructural microvascular alterations could be demonstrated. Our study emphasizes that alterations of microvasculature are present early in experimental edematous pancreatitis. Therefore, alterations of pancreatic microcirculation seem to be of great importance in the pathogenesis of pancreatic inflammation, and should be the subject of further studies.

Animals↗

PubMed1992

Comparative anatomy of the vitreous body in rhesus monkeys and man.

In the isolated unfixed vitreous body a structural organization can be visualized by slitlamp microscopy or by an ink-injection technique. We discuss the observations on human and rhesus monkey (Macaca mulatta) vitreous bodies using the ink-injection technique. Advantages and disadvantages of this method compared with slitlamp microscopy are described. Eventual functional implications of vitreal organization are mentioned briefly.

Animals↗