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PubMed1996

Apical and coronal seal of roots obturated with a dentine bonding agent and resin.

The efficacy of sealing root canal systems with a dentine bonding agent and resin cement was compared with that of a glass ionomer sealer. Scanning electron microscopic examination of the bonding agent-dentine interface was also performed. The root canals of 50 single rooted teeth with mature apices were prepared chemomechanically and the smear layer removed with 17% REDTA and 5.25% NaOCl. Specimens were divided into two groups of 24 teeth each. One group had the dentine conditioned with a 10:3 citric acid-ferric chloride solution and obturated with the dentine bonding agent and resin, radiopaque C & B Metabond. The other group was obturated with the glass ionomer sealer, Ketac-Endo and a single cone of gutta-percha. After immersion in Indian ink for 90 h, the teeth were cleared and the quality of the apical and coronal seal was assessed using India ink dye penetration. There was a significantly better seal in both the apical and coronal directions when using the dentine bonding agent and resin obturation material. Scanning electron microscopic examination of the demineralized dentine and the C & B Metabond interface revealed the presence of the characteristic hybrid layer along with microtags of resin penetrating deep into the dentine tubules.

Boron Compounds↗
PubMed1994

The nasal route of cerebrospinal fluid drainage in man. A light-microscope study.

The drainage routes from the subarachnoid space to the nasal mucosa were investigated in autopsy material. Indian ink, applied post-mortem to the olfactory groove, promptly filled the perineurial spaces around the olfactory nerve branches in the dura, the lamina cribrosa and the submucosal tissue in the nose. In a case of recent subarachnoid haemorrhage, the perineurial spaces even around the most distal olfactory nerve branches were congested with blood and there was an abundant accumulation of red corpuscles in the apical part of the nasal mucosa. Iron-containing pigment was found in the perineurial spaces of proximal and distal olfactory nerve branches as well as in the nasal mucosal stroma in cases with older haemorrhagic lesions. The findings show that the perineurial spaces provide an efficient drainage route from the subarachnoid space to the nasal mucosa in cases with haemorrhagic cerebral lesions. A complementary drainage route for the cerebrospinal fluid was indicated by the presence of indian ink, red corpuscles and iron pigment in arachnoid villi, which penetrated the lamina cribrosa and ended in the nasal submucosal tissue. Iron in the deep cervical lymph nodes should not be taken as evidence of transport from the CNS, since iron pigment was also found in cases without intracranial haemorrhage.

Adult↗
PubMed2006

Optical coherence tomographic pattern of fluorescein angiographic leakage site in acute central serous chorioretinopathy.

PURPOSE: To study the optical coherence tomography (OCT) pattern of the fluorescein angiographic leakage in acute central serous chorioretinopathy (CSCR). METHODS: This is a non-interventional pilot case study. OCT line scan was performed over the fluorescein angiographic leak site in eyes clinically diagnosed acute CSCR. Clinical fundus photograph, site of leakage on fundus fluorescein angiography and corresponding OCT were analysed. RESULTS: The mean age of 10 consecutive patients was 38.8 +/- 6.9 years. Six patients were male and the mean duration of symptom was 7 days. Six eyes (60%) showed a characteristic dipping pattern of neurosensory retina with intervening hyper-reflective echoes suggestive of fibrin over the leakage site. All these eyes had ink-blot leak and subretinal fibrin. CONCLUSIONS: Ink-blot leak in acute CSCR with subretinal fibrin generates a dipping morphological pattern on OCT.

Acute Disease↗
PubMed2006

Expression and significance of nerve growth factor receptor p75 in rats' cathartic colonic wall.

OBJECTIVE: To investigate the expression of nerve growth factor receptor p75 in a normal and cathartic colon and its significance in the formation of the cathartic colon in rats. METHODS: Sixty Sprague-Dawley rats were equally divided into normal control group, rhubarb group and phenolphthalein group. A model of the cathartic colon was constructed by gastric infusion with rhubarb or phenolphthalein in rats. The first dose of rhubarb and phenolphthalein was both 200 mg/kg/d and was increased by 200 mg/kg/d with each passing day. The last dose of rhubarb and phenolphthalein was 3200 mg/kg/d and 4200 mg/kg/d, respectively. The transit function of colon was measured by the Chinese ink expulsion test; the p75 in colon wall was determined by the immunohistochemical method. RESULTS: The transit speed was much slower in the cathartic colon group than that in the control group. The imprinted Chinese ink length and the ratio of imprinted length/total colon length in the rhubarb-induced cathartic colon was significantly shorter than that of the control group (77.38 +/- 8.42 vs 94.25 +/- 7.07 cm, P < 0.01). Those in the phenolphthalein-induced group (83.38 +/- 9.75 cm) were also significantly shorter than those of the control group but to a lesser degree (P < 0.05). p75 was abundantly expressed in the submucosal nerve plexus and weakly expressed in the myenteric plexus. The expression of p75 was much higher in the rhubarb-induced group. The expression was strongly positive in the submucosal nerve plexus, significantly higher than that in the controls (P < 0.01). In the myenteric plexus, p75 was also highly expressed (P < 0.05). In the case of the phenolphthalein-induced group, the expression of p75 was positive in the submucosal nerve plexus but was positive in the myenteric plexus of three rats only. The remaining rats were negative or weakly positive. This was not significantly different from that of the control group. CONCLUSIONS: The abnormal expression of p75 in cathartic colon probably has some effect on the degeneration or apoptosis of neuronal cells in the enteric nerve plexus, with a subsequent pathological change of the enteric nervous system, and thus leads to abnormalities in colonic dynamics. The kind of lesion is probably associated with long-term use of irritative cathartics.

Animals↗
PubMedDate not supplied

Regional perfusion of the equine carpus for antibiotic delivery.

Regional perfusion of carpal tissues by forced intramedullary administration of fluids was evaluated in 10 horses. Results of subtraction radiography after perfusion with a contrast medium demonstrated that perfusate was delivered to the carpal tissues by the venous system. Perfused India ink was distributed uniformly in the antebrachiocarpal and middle carpal synovial membranes. Histologically, the ink was within the venules of the synovial villi. Immediately after perfusion with gentamicin sulfate (1 g), the gentamicin concentrations in the synovial fluid and synovial membrane of the antebrachiocarpal joint were 349 +/- 240 micrograms/mL and 358 +/- 264 micrograms/g, respectively. When gentamicin concentrations in the synovial fluid of the antebrachiocarpal joint and serum were measured 0, 0.5, 1, 4, 8, 12, and 24 hours after carpal perfusion, the mean peak gentamicin concentration in the synovial fluid was 589 +/- 429 micrograms/mL. At hour 24, the mean gentamicin concentration in the synovial fluid was 4.8 +/- 2.0 micrograms/mL. The resulting peak gentamicin concentration in the serum was 23.7 +/- 14.5 micrograms/mL immediately after the perfusion; it decreased below the desired trough level of 1 micrograms/mL between hours 4 and 8.

Animals↗
PubMed2006

Equatorial ring-like channels in the Cryptococcus neoformans polysaccharide capsule.

Under certain conditions, India ink particles can penetrate the capsule of the opportunistic pathogen Cryptococcus neoformans. India ink penetration gave two distinct patterns, one as a ring in the middle of the capsule, and another as a double spot located at opposite poles of the cells. These spots were perpendicularly orientated to the bud. This pattern suggests the existence of a localized structure deep in the capsule that can accumulate large insoluble particles. Although the mechanisms responsible for the assemblage and maintenance of ring-like channels are not understood, their existence deep within the capsule implies a new level of complexity for this enigmatic structure.

Animals↗
PubMed1988

Attempts to demonstrate a polysaccharide capsule in Neisseria gonorrhoeae.

The presence or absence of a polysaccharide capsule on the human pathogen Neisseria (N.) gonorrhoeae is still a topic of controversy. For this reason we compared the results obtained by light microscopy (dry India ink-Fuchsin stain) and electron microscopy (Alcian blue-lanthanum nitrate stain) of encapsulated strains of N. meningitidis and Streptococcus (S.) pneumoniae and of non-encapsulated strains of S. pneumoniae and Escherichia (E.) coli with those obtained using the same methods on strains of pilliated and non-pilliated N. gonorrhoeae. After staining with India ink-Fuchsin no capsules could be demonstrated on any of the N. gonorrhoeae strains studied. If present the capsules on these cells are too delicate to be identified by light microscopy. After treatment with Alcian blue-lanthanum nitrate sections of cells of N. meningitidis and S. pneumoniae generally showed the presence of a capsular layer. Sections of cells of the non-encapsulated strain of S. pneumoniae which possess C (common)-polysaccharide also showed surface associated capsule-like material. Similarly the surface of the cells of the E. coli strain showed material which appeared to be tufts of pili and/or M (mucoid)-antigen. In experiments where the N. gonorrhoeae cells were harvested as early as after six hours of growth a capsule-like material was demonstrated on cells of all strains studied.

Culture Media↗
PubMed1984

Absorption of the cerebrospinal fluid and intracranial compliance in an amphibian, Rana pipiens.

Experiments have been carried out to investigate the absorption of the cerebrospinal fluid (c.s.f.) and the intracranial compliance in an amphibian, Rana pipiens, using infusions into the c.s.f. system through glass micropipettes. Resistance to absorption of the c.s.f. was estimated by the constant rate infusion technique. Mean absorption resistance for infusions of artificial c.s.f. into the lateral ventricles and into the cerebral subarachnoid space were 15.48 and 16.52 mmH2O min microliter-1 respectively. This difference was not significant and it is concluded that the pores in the posterior tela situated in the roof of the fourth ventricle do not offer any resistance to the flow of c.s.f. out of the ventricles. The resistance to drainage of the c.s.f. in this amphibian is higher than that found for mammals. Mean resting c.s.f. pressure, estimated from the intercept of the regression line with the pressure axis at zero infusion rate was 18.0 mmH2O. Absorption resistance was measured from the cerebral subarachnoid space before and after injection of 4 microliter Indian ink solution. There was a 3-fold increase in resistance following ink injection. Two-way analysis of variance showed the difference to be significant (P less than 0.01) suggesting that the outflow sites can become partially blocked by particulate matter. During a continuous 3 h infusion of artificial c.s.f. containing [14C]dextran or [125I]-labelled human serum albumin (RISA) into the lateral ventricles, the mean percentage uptakes into the systemic circulation after the first 0.5 h of a 3 h period were 74.1 and 61.9% respectively. The difference is not significant. The rapid and high uptake into blood suggests there is a direct communication between c.s.f. and blood in amphibians. During continuous infusion of RISA into the lateral ventricles, simultaneous blood samples were taken from the femoral artery and the internal dorsal vertebral vein. Radioactivity was found to be 13.2% higher in venous samples. This suggests that at least some c.s.f. drainage takes place directly into the spinal venous system. Intracranial compliance was investigated by recording the peak pressure in response to a series of bolus injections of artificial c.s.f. into one lateral ventricle. Compliance was estimated to be 0.11, 0.10 and 0.09 microliter mmH2O-1 for injection rates of 12.74, 16.62 and 25.10 microliters min-1 respectively. The difference between these values is not significant. The results suggest that for injection volumes over 5 microliters the c.s.f. system behaves elastically.

Absorption↗
PubMed1987

A system of canals in semilunar menisci.

Semilunar menisci of young calves, and infants and young children were examined with the scanning electron microscope and the light microscope. Canal-like structures were noted opening onto the surface of the menisci. After filling the joints with Indian ink, ink was noted within canal-like structures deep in the substance of the tissue. The canals may play a part in the transport of fluid within the meniscus, or may carry nutrients from the synovial fluid or the blood vessels to the avascular sections of the meniscus.

Animals↗
PubMed2003

Improved resolution by mounting of tissue sections for laser microdissection.

BACKGROUND: Laser microbeam microdissection has greatly facilitated the procurement of specific cell populations from tissue sections. However, the fact that a coverslip is not used means that the morphology of the tissue sections is often poor. AIMS: To develop a mounting method that greatly improves the morphological quality of tissue sections for laser microbeam microdissection purposes so that the identification of target cells can be facilitated. METHODS: Fresh frozen tissue and formalin fixed, paraffin wax embedded tissue specimens were used to test the morphological quality of mounted and unmounted tissue. The mounting solution consisted of an adhesive gum and blue ink diluted in water. Interference of the mounting solution with DNA quality was analysed by the polymerase chain reaction using 10-2000 cells isolated by microdissection from mounted and unmounted tissue. RESULTS: The mounting solution greatly improved the morphology of tissue sections for laser microdissection purposes and had no detrimental effects on the isolation and efficiency of amplification of DNA. One disadvantage was that the mounting solution reduced the cutting efficiency of the ultraviolet laser. To minimise this effect, the mounting solution should be diluted as much as possible. Furthermore, the addition of blue ink to the mounting medium restores the cutting efficiency of the laser. CONCLUSIONS: The mounting solution is easy to prepare and apply and can be combined with various staining methods without compromising the quality of the DNA extracted.

Coloring Agents↗
PubMed1993

Diagnosis of pulmonary cryptococcosis by ultrasound guided percutaneous aspiration.

BACKGROUND: Ultrasound is useful for locating thoracic lesions and guiding biopsy procedures. The use of sonographic appearances and ultrasound guided needle aspiration has led to the diagnosis of pulmonary cryptococcosis at this hospital. METHODS: Six hundred and eight patients who had ultrasound guided lung aspirations were reviewed retrospectively and nine with documented pulmonary cryptococcosis were collected. All patients had nodules or infiltrates on the chest radiograph. The needle aspirates obtained under ultrasound guidance were stained by Riu's or Papanicolaou's method or with India ink, and six were sent for culture. Five patients also underwent bronchoscopy and biopsy. RESULTS: The nine patients had 18 pulmonary lesions, of which 15 were nodules and three infiltrates. Fifteen lesions were detectable by ultrasound, which showed the nodules to be hypoechoic with eccentrically located air echoes. In eight of the nine cases cryptococci were detected after the lung aspirates had been stained with Riu's or Papanicolaou stain or with India ink. In five of the six aspirates sent for fungal culture Cryptococcus neoformans was isolated. The diagnostic yield was higher than that of bronchoscopy. None developed post-aspiration pneumothorax or any evidence of late dissemination. CONCLUSIONS: Because they tend to be subpleural pulmonary cryptococcal lesions seem to be identifiable by ultrasound. Ultrasound guided lung aspiration is an effective, rapid, and safe method for diagnosis.

Adult↗
PubMed1994

P-selectin and ICAM-1-dependent adherence reactions: role in the genesis of postischemic no-reflow.

The aim of this study was to determine whether immunoneutralization of P-selectin or intercellular adhesion molecule-1 (ICAM-1) (endothelial cell adhesion molecules involved in leukocyte rolling and firm adhesion, respectively) would attenuate the development of postischemic capillary no-reflow. Microvascular patency was assessed in vascularly isolated canine gracilis muscles by perfusion with contrast media (India ink) at the end of the experimental protocol. Computerized video imaging was used to quantitate the number of ink-containing microvessels (< 10 microns diam) per muscle fiber in histological samples obtained from isolated canine gracilis muscles subjected to 4.5 h of continuous perfusion (nonischemic control), 4 h of ischemia and 30 min of reperfusion (I-R), I-R + P-selectin monoclonal antibodies (MAbs) (MD6 or PB1.3), and I-R + ICAM-1 MAbs (CL18/6C7 or R6.5). The efficacy of a P-selectin MAb (MD3) that binds to a nonfunctional epitope was also evaluated. I-R was associated with a marked reduction in the number of patent capillaries per fiber (3.1 +/- 0.2 vs. 1.1 +/- 0.2 patent capillaries/fiber for control and I-R, respectively). Immunoneutralization with MAbs directed against functional epitopes on P-selectin (MD6 or PB1.3) significantly improved capillary perfusion (2.3 +/- 0.3 and 3.6 +/- 0.6 patent capillaries/fiber, respectively). On the other hand, MAb MD3, which binds to nonfunctional epitopes on P-selectin, failed to limit the development of postischemic no-reflow (1.0 +/- 0.2 patent capillaries/fiber). Immunoneutralization of ICAM-1 with CL18/6C7 and R6.5 increased the number of patent capillaries per fiber to 1.8 +/- 0.1 and 2.5 +/- 0.3, respectively. These data indicate that P-selectin and ICAM-1-dependent adherence reactions play an important role in the development of the no-reflow phenomenon in postischemic skeletal muscle.

Animals↗
PubMed1998

Effect of autotransplantation of microvessel fragments on experimental random-pattern flaps in the rat.

We examined whether autotransplantation of microvessel fragments (Mvf) and/or myofibroblasts (Mf) into an in vivo skin flap model might improve the survival of the ischemic flap. If so, this could improve blood perfusion, increase blood flow, and improve the survival of the flap. A skin flap was raised on the back of each rat (n = 15 in each group). In the control group, the flap was sutured to the original bed. In the other groups (1) phosphate-buffered saline; (2) autotransplanted Mvf, Mf, or Mvf plus Mf, and (3) a homogenized mixture of Mvf plus Mf was injected into the distal part of the flap. In a further group, Mvf labeled with DiI-acetylated low-density lipoprotein were autotransplanted with Mf into the distal part of the flap, and India ink was perfused through the abdominal aorta 7 days postoperatively. The transplanted Mvf plus Mf group showed better flap survival after 7 days than the other groups (p < 0.02). Labeling with DiI-acetylated low-density lipoprotein showed that transplanted Mvf sent arborizations into the nearby tissue. India ink was found in the lumina within such arborizations. Thus, autotransplanted Mvf may improve the survival of ischemic skin flaps by promoting the early formation of patent connections between Mvf and the host's microcirculatory system. This apparently requires the presence of Mf.

Animals↗
PubMed1999

Successful autotransplantation of microvessel fragments into the rat heart.

We examined whether microvessel fragments (Mvf), autologously transplanted with myofibroblasts (Mf) into the heart, could survive and form connections to the host's coronary microcirculation. Neither achievement has been reported before in the heart. Mvf and Mf were prepared from Sprague-Dawley rat epididymal fat pads. A mixture of Mvf (labeled with the fluorescent probe DiI-acetylated low-density lipoprotein, DiI-Ac-LDL) and Mf was injected into the superficial myocardium under general anesthesia. Rats (n = 5 in each group) were killed on postoperative days 7, 14, 21, 42 and 49, and India ink was perfused through the coronary arteries. Frozen sections of the injected area were examined under fluorescence and light microscopes. Some DiI-Ac-LDL-labeled Mvf survived in each group, and India ink was present in the lumina of microvessels coincident with DiI-Ac-LDL-labeled autotransplanted Mvf. Hematoxylin and eosin staining revealed mild inflammatory reactions followed by some fibrosis at the injection sites. These findings indicate that autotransplanted Mvf can survive for at least 49 days, and that patent microvascular anastomoses can form between them and the host's coronary microvessels. Possibly, autotransplantation of Mvf could lead to the development of a new collateral microcirculation, a phenomenon especially important in the ischemic heart.

Animals↗
PubMed1992

Fluorescein-angiographic patterns in patients with central serous chorioretinopathy at the initial visit.

We reviewed the clinical records, color photographs and fluorescein angiograms of 106 eyes of 53 patients (44 men and 9 women) with central serous chorioretinopathy who had been followed up for 1 year or more. Their ages ranged from 20 to 59 years. One eye in each patient was involved at the initial visit. According to fluorescein-angiographic findings, the lesions were divided into smokestack pattern (13 eyes), ink blot appearance (33 eyes) and minimally enlarging spot (7 eyes). Most patients complained of central scotoma, and most patients had moderately decreased visual acuities at the initial visit, which improved after recovery. In particular, all 7 eyes with a minimally enlarging spot had a visual acuity of 0.8 or more at the initial visit; of these 6 eyes had 1.0 or more several weeks later. On fluorescein angiography, 49 eyes demonstrated 1 leaking spot and 4 eyes showed 2 spots. Of 57 leaking spots, 10 were observed in the foveola, 31 were noted in the fovea and 16 were found in the para- or perifovea. Of 13 smokestack patterns, 5 were found in the inferonasal area. The ink blot appearance and minimally enlarging spots were frequently found in the superonasal and superotemporal areas.

Adult↗
PubMed1995

Intracardiac blood flow patterns related to the yolk sac circulation of the chick embryo.

Intracardiac flow patterns during heart development were studied by injection of india ink into the yolk sac circulation of chick embryos at Hamburger-Hamilton stages 10 to 17. We injected india ink into a small venule or capillary, carefully preventing application of overpressure to the vascular system, and recorded the intracardiac route by video. From stage 12 onward, blood flow was laminar, and separate intracardiac currents were visualized. The yolk sac was divided into a left and a right half. Blood coursed through each half in concentric loops, ranging from the marginal sinus to the sinus venosus. This parallel array persisted within the heart. Bilateral to the embryo, two lateral regions arose that extended wedge-like within each half, resulting in six equally sized yolk sac regions at stage 16. The process of heart looping was not accompanied by a change in flow pattern. However, developmental changes of the yolk sac circulation were reflected in alteration of the intracardiac flow pattern. From stage 16 onward, the intracardiac flow pattern was no longer determined by the left- or right-hand side of the yolk sac but by bilateral anterior, lateral, and posterior regions of the yolk sac. Blood from the lateral regions of the yolk sac was preferentially distributed to the head. The results show that in preseptation stages a relatively stable flow pattern is present. We suggest that alterations in blood flow could influence the process of normal heart development.

Animals↗
PubMed1997

Unilateral vitelline vein ligation alters intracardiac blood flow patterns and morphogenesis in the chick embryo.

To study the role of blood flow in normal and abnormal heart development, an embryonic chicken model was developed. The effect of altered venous inflow on normal intracardiac blood flow patterns was studied by visualization of blood flow with India ink. At stage 17, India ink was injected into a capillary or small venule within a specific yolk sac region. After determination of the normal intracardiac flow pattern, the right lateral vitelline vein was ligated, and the new intracardiac flow pattern was studied. Ligation resulted in disturbance of normal intracardiac flow patterns, which was most obvious in the conotruncus. The long-term effect of these abnormal intracardiac flow patterns on the development of the heart and pharyngeal arch arteries was investigated by permanent ligation in ovo with a microclip at stage 17 and subsequent evaluation at stages 34, 37, and 45. These experiments revealed anomalies of the vascular system in 58 of the 91 ligated embryos studied. We observed intracardiac malformations consisting of subaortic ventricular septal defects (n = 52), semilunar valve anomalies (n = 19), atrioventricular anomalies (n = 7), and pharyngeal arch artery malformations (n = 32). It is concluded that abnormal intracardiac blood flow, resulting from hampered venous inflow, may result in serious intracardiac and pharyngeal arch artery malformations comparable to defects observed in embryonic chicken models subjected to neural crest ablation, cervical flexure experiments, and excessive retinoic acid treatment.

Animals↗