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PubMed2005

In vivo optical coherence tomography imaging of the pancreatic and biliary ductal system.

BACKGROUND: In vivo optical coherence tomography (OCT) imaging has not yet been applied to the pancreatic ductal system. The aim of this study was to obtain in vivo OCT images of dog pancreatic and biliary ducts, and to correlate the images with histology. METHODS: Images of dog pancreatic and biliary ducts were obtained by using an in vivo OCT probe introduced through the respective papillary orifices. Each duct was imaged in multiple locations, and the site of imaging was marked with injected India ink. After imaging, the dogs were euthanized, and the pancreaticobiliary system was harvested. Histologic cross sections were correlated with in vivo OCT images by measuring the structures seen on in vivo OCT images and correlating them with structures seen on corresponding histology slides that contained India ink. OBSERVATIONS: Eighteen pairs of in vivo OCT images and histology slides from the bile duct and the pancreatic duct were obtained from 5 dogs. The entire duct wall could be visualized. A low reflective in vivo OCT layer corresponding to the epithelium could be discerned on many images. The bile duct showed a more complex architecture and had greater variations within the reflective OCT layers, possibly because of greater cellularity within the lamina propria. Nuclei within cells could not be identified, and structures adjacent to the ducts could not be imaged. CONCLUSIONS: In vivo OCT is capable of imaging the pancreaticobiliary ductal system and of identifying the epithelial layer. Because of limited depth of imaging (320-845 micron), OCT is unlikely to serve the purpose of tumor staging.

Animals↗
PubMed2007

A new approach to assess function after sciatic nerve lesion in the mouse - adaptation of the sciatic static index.

Among the numerous ways of assessing regeneration after peripheral nerve lesions, the analysis of gait is one of the most important, because it shows the recovery of function, which is the ultimate goal of the repair machinery. The sciatic function index was introduced as a method to assess reinnervation after an experimental sciatic nerve lesion, and was adapted to the mouse model. The sciatic static index (SSI), is more simple and practical to perform, and is not so influenced by gait's velocity, but this method has not yet been adapted to the mouse model of sciatic lesion. We used 63 male Swiss mice (Mus musculus) to develop a formula to the sciatic static index in mice (SSIm). The animals were divided on three groups (control, transection and crush). They were evaluated at the preoperative and 7th, 14th, 21st, 28th, 35th and 42nd days postoperative by the ink track method (SFI), and by the acquisition of photographs of the plantar aspects of the injured and uninjured hind paws. The parameters evaluated were the 1-5 toe spread (TS), the 2-4 toe spread (ITS) and the distance between the tip of the third toe and the most posterior aspect of the paw (PL), on both methods. After verifying the temporal pattern of function, correlation and reproducibility of the measurements, we performed a multiple regression analysis using SFI values as dependent variable, and the TS, ITS and PL measured with the photo method as independent variables, and found the formula of the SSI for mice (SSIm). The three groups (control, transection and crush) had a characteristic pattern of dysfunction. The parameters measured in the ink and photo method had variable but significant correlations between them (P<0.000), but photo method of measurement showed a better reproducibility. The correlation between SFI and SSIm showed a high correlation coefficient (r=0.892, P<0.000), and demonstrates that SSIm can be used as an alternative method to assess the functional status relative of sciatic nerve activity in mice.

Animals↗
PubMed2006

Apical leakage of root-end placed SuperEBA, MTA, and Geristore restorations in human teeth previously stored in 10% formalin.

The purpose of this study was to determine if storage of extracted teeth in 10% formalin affects microleakage of MTA, Geristore, or SuperEBA root-end fillings. There were 130 freshly extracted single-rooted teeth collected, immediately placed in phosphate buffered saline (PBS) and stored for less than 24 hours. Teeth were divided and either placed in 10% formalin for a 4-weeks immersion or immediately prepared. Preparation for all teeth consisted of canal instrumentation, obturation, apical resection, retrograde preparation, and placement of a root-end filling material. After 72 hours, the apical portions were immersed in India ink under vacuum pressure for 30 minutes, and then stored in ink for 1 week. Negative controls and positive controls performed as expected. In general, there was significantly less dye leakage of root-end restorations in teeth stored in 10% formalin when compared to the freshly extracted teeth (p < 0.0001). No difference in dye leakage was observed between Geristore restorations placed in teeth stored in formalin as compared to fresh teeth (p = 0.892). Less dye leakage was noted in teeth restored with Geristore as compared to MTA and SuperEBA, regardless of storage medium (p < 0.0001). No difference was observed between MTA and SuperEBA root-end restorations (p = 0.157). The results of this study provide evidence that storage of teeth in 10% formalin over a 4-week period may significantly influence dye leakage as compared to leakage in freshly extracted teeth.

Aluminum Compounds↗
PubMed2006

Cytostatic lung perfusion results in heterogeneous spatial regional blood flow and drug distribution: evaluation of different cytostatic lung perfusion techniques in a porcine model.

OBJECTIVES: Comparison of doxorubicin uptake, leakage and spatial regional blood flow, and drug distribution was made for antegrade, retrograde, combined antegrade and retrograde isolated lung perfusion, and pulmonary artery infusion by endovascular inflow occlusion (blood flow occlusion), as opposed to intravenous administration in a porcine model. METHODS: White pigs underwent single-pass lung perfusion with doxorubicin (320 mug/mL), labeled 99mTc-microspheres, and Indian ink. Visual assessment of the ink distribution and perfusion scintigraphy of the perfused lung was performed. 99mTc activity and doxorubicin levels were measured by gamma counting and high-performance liquid chromatography on 15 tissue samples from each perfused lung at predetermined localizations. RESULTS: Overall doxorubicin uptake in the perfused lung was significantly higher (P = .001) and the plasma concentration was significantly lower (P < .0001) after all isolated lung perfusion techniques, compared with intravenous administration, without differences between them. Pulmonary artery infusion (blood flow occlusion) showed an equally high doxorubicin uptake in the perfused lung but a higher systemic leakage than surgical isolated lung perfusion (P < .0001). The geometric coefficients of variation of the doxorubicin lung tissue levels were 175%, 279%, 226%, and 151% for antegrade, retrograde, combined antegrade and retrograde isolated lung perfusion, and pulmonary artery infusion by endovascular inflow occlusion (blood flow occlusion), respectively, compared with 51% for intravenous administration (P = .09). 99mTc activity measurements of the samples paralleled the doxorubicin level measurements, indicating a trend to a more heterogeneous spatial regional blood flow and drug distribution after isolated lung perfusion and blood flow occlusion compared with intravenous administration. CONCLUSIONS: Cytostatic lung perfusion results in a high overall doxorubicin uptake, which is, however, heterogeneously distributed within the perfused lung.

Animals↗
PubMed2005

Interferon beta increases c-Myc proteolysis in mouse monocyte/macrophage leukemia cells.

Growth inhibitory activity of interferons (IFNs) has been attributed to several events. These include rapid induction of cyclin-dependent kinase inhibitors, such as those in the Cip/Kip and Ink 4 families and down-regulation of c-myc mRNA and c-Myc transcriptional activity. Here, we report an additional mechanism, involving regulation of Myc protein levels, through which type 1 IFN may halt proliferation of cells. This was discovered using a cell line which constitutively expresses c-myc from a retrovirus vector and which was reported to have undergone deletion of genes encoding the Ink 4 tumor suppressors p15 and p16. IFNbeta caused a reduction in the steady state level of c-Myc protein by increasing degradation through the 26S proteasome. Our data, as well as that of others, indicate that multiple levels of c-Myc expression can be affected by IFN treatment and this contributes to rapid growth arrest in the G1 phase of the cell cycle.

Animals↗
PubMed2005

Cloning and biochemical characterization of APIT, a new l-amino acid oxidase from Aplysia punctata.

The purple ink of the sea hare Aplysia punctata contains a 60 kDa protein with tumoricidal activity. This A. punctata ink toxin (APIT) kills tumor cells within 6--8h in an apoptosis independent manner by the production of high amounts of hydrogen peroxide which induce a necrotic form of oxidative stress. Here, we describe the biochemical features of APIT associated with its anti-tumor activity. APIT is a weakly glycosylated FAD-binding L-amino acid oxidase that catalyzes the oxidative deamination of L-lysine and L-arginine and thereby produces hydrogen peroxide (H(2)O(2)), ammonia (NH(4)(+)) and the corresponding alpha-keto acids. The tumoricidal effect is completely abrogated in the absence of the amino acids L-lysine and L-arginine. The enzyme is stable at temperatures from 0 to 50 degrees C. Similar to other FAD-binding enzymes, it is resistant against tryptic digest. Even digest with proteinase K fails to degrade the enzyme. Cloning of the APIT gene and subsequent sequencing revealed a FAD-binding domain followed by a so-called GG-motif, which is typical for L-amino acid oxidases. Strongest homology exists to escapin, aplysianin A precursor, the cyplasins L and S and achacin.

Amino Acid Sequence↗
PubMed2005

Laparoscopic radical prostatectomy: evaluation of specimen pathologic features to critically assess and modify surgical technique.

OBJECTIVES: To evaluate laparoscopic radical prostatectomy (LRP) specimens for pathologic stage and the malignant and benign surgical margin status to determine areas for improvement in surgical technique. LRP is gaining significant attention as a viable alternative to retropubic radical prostatectomy. METHODS: A total of 110 consecutive LRP specimens were retrospectively reviewed. A single experienced genitourinary pathologist examined all slides. In addition, exposure of benign glands at the inked margin was reported. Oncologic data were assessed by standard pathologic examination for stage, grade, and margin status. The pathologic data from year 1 (group 1, 52 patients) were compared with the data from subsequent years (group 2, 58 patients). RESULTS: The overall positive surgical margin rate was 18.2% (13.5% for pT2 disease). Benign parenchymal exposure was identified in 16 patients (14.5%). Six of these patients had concomitant negative surgical margins for cancer. The positive margin rate for patients with Stage pT2 disease was 22.2% in group 1 and 5.9% in group 2. The positive margin rate was 34.8%, 13.6%, and 13.8% for bilateral, unilateral, and non-nerve-sparing procedures, respectively. CONCLUSIONS: LRP offers comparable surgical margins to those of reported open prostatectomy series. Careful pathologic review provides feedback so that technique modifications can be implemented and the positive margin rates improved further. A low, but significant, positive margin or benign parenchymal exposure rate in pT2 disease indicates room for improvement in the surgical technique. The presence of benign glands at the inked margin is not routinely reported but does provide an additional parameter of surgical adequacy.

Adult↗
PubMed1997

C57BL/6 strain is most susceptible to cerebral ischemia following bilateral common carotid occlusion among seven mouse strains: selective neuronal death in the murine transient forebrain ischemia.

Rats and gerbils have been used widely to investigate the molecular mechanism of selective neuronal death following transient global ischemia. Recently, the availability of transgenic mice has enabled us to examine the involvement of specific gene products in various pathophysiological conditions. However, there has been only limited information about the experimental model of cerebral ischemia in mice, particularly in regard to selective neuronal death. We examined whether bilateral carotid occlusion produced global forebrain ischemia in seven common mouse strains including C57BL/6, ICR, BALB/c, C3H, CBA, ddY and DBA/2, based on neurological signs, histological findings and cortical microcirculatory as well as India ink perfusion patterns. The C57BL/6 strain was found to be the most susceptible among seven strains. All C57BL/6 mice died within 6 h after permanent bilateral carotid occlusion. After transient bilateral carotid occlusion for 20 min, more than 90% of C57BL/6 mice showed typical neurological signs such as torsion of the neck and rolling fits, and developed selective neuronal death in the hippocampus and caudoputamen. Hypothermia prevented the neuronal death. Visualization of brain vasculature by India ink perfusion indicated that the susceptibility of the mice after bilateral carotid occlusion depended mainly on the degree of anastomosis between carotid and basilar arteries. Our results showed the feasibility of investigating selective neuronal death in transgenic mice with simple temporary occlusion of both common carotid arteries, when those from the C57BL/6 strain or inbred transgenic mice from other strains with the C57BL/6 strain in a back-cross manner are used.

Animals↗
PubMed2001

Effects of the endothelin receptor antagonist Bosentan on ischaemia/reperfusion injury in rat skeletal muscle.

We examined the role of endothelin in ischaemia/reperfusion injury in skeletal muscle, using the endothelin receptor antagonist Bosentan. In the rat hindlimb tourniquet ischaemia model, one hindlimb was rendered ischaemic for 2 h at 36 degrees C, then blood flow was re-established for either 24 h to assess muscle survival or 1.5 h for a study of capillary perfusion. In the first set of rats, the gastrocnemius muscle was removed from the postischaemic limb and assessed for viability histochemically using the nitro blue tetrazolium stain. Tissue water content (a measure of oedema) and myeloperoxidase activity (a measure of neutrophil accumulation) were also assessed in the ischaemic muscle, the contralateral non-ischaemic muscle and the lungs. In the second set of rats, the hind limb was infused with India ink after 2-h ischaemia and 1.5-h reperfusion and the muscle was harvested, fixed and cleared. In control rats, muscle viability was 17+/-2% (S.E.M.). In rats treated with Bosentan (10 mg/kg, i.p.) 30 min before release of the tourniquet, muscle viability (48+/-7%) was significantly increased compared to the control group (P<0.01). Bosentan treatment had no significant effect on tissue water content or myeloperoxidase activity in the ischaemic muscle, the contralateral non-ischaemic muscle or the lung. Immunoreactive endothelin levels in serum increased to a peak at 90 min of reperfusion and returned to control levels by 24-h reperfusion. India ink studies demonstrated a significantly increased functional capillary density in postischaemic Bosentan-treated muscles compared with postischaemic control muscles (P<0.05). These results suggest that endothelin plays an important role in the necrosis which results from a period of ischaemia and reperfusion in skeletal muscle, by mediating a decrease in postischaemic microvascular perfusion.

Animals↗
PubMed1981

Endometriosis and the development of tuboperitoneal fistulas after tubal ligation.

The present study details gross and histologic findings of 79 previously ligated fallopian tubes from 3 groups of patients. Of 20 oviducts removed after documented sterilization failure (group I), 6 revealed a process compatible with endometriosis. Four of nine previously ligated fallopian tubes removed at the Johns Hopkins Hospital (group II) were successfully injected with India ink. In two patients histologic examination demonstrated the India ink in epithelium-lined spaces that lay beyond the muscle of the tubal wall extending from the tubal lumen to the serosal surface. Fifty oviducts were studied in twenty-five patients requesting reversal of their sterilizations (group III). A higher percentage of fistulas was demonstrated in patients with less than 4 cm of remaining proximal tubal segment. Furthermore, most of these fistulas were demonstrated in patients for whom 3 years had elapsed since the original sterilization procedure. Patients sterilzed by laparoscopic cautery methods were observed to have a higher percentage of fistula formation and histologic documentation of endometriosis at the sterilization site as compared with patients sterilized by other methods. Our observations suggest that ligation of the oviduct within 4 cm of the uterine cornu may predispose to the development of endometriosis and subsequent fistula formation in the tip of the ligated oviduct.

Endometriosis↗
PubMed2003

Effect of processing and measuring procedures on estimated sizes of bull sperm heads.

Reported estimates of sperm head size within a species vary considerably, partly due to procedural effects. A simple India ink method was developed that provided good contrast without inducing artifacts. Semen from five fertile bulls was smeared on replicate slides and left unfixed or fixed in Carnoys solution, with ink added for background. Other slides were fixed, and sperm were stained by the Feulgen procedure. Sperm head area was measured four ways. These were linear measurements made with the aid of an ocular micrometer and an oil immersion objective, plus three methods of measuring sperm heads projected at magnification 5,000 x. The areas of unfixed and fixed sperm heads did not differ (41.5 microm(2) versus 41.6 microm(2), respectively, P>0.05). The Feulgen-stained head area was smaller (26.2 microm(2), P<0.05). Sperm head areas calculated from ocular micrometer measurements were slightly smaller (P<0.05) than areas measured using projection. Identical results obtained by two technicians were treated as duplicates and approximately half of the variation was biological, due to source of semen. There was an interaction (P<0.05) between the sample source and fixation procedures. Thus, preparative techniques must be carefully controlled, and experiments designed to partition possible interactions between the biological material sampled and procedures used.

Animals↗
PubMed1995

Apical sealing ability of a new glass ionomer root canal sealer.

This study compared the sealing ability of Ketac Endo with and without smear layer and Tubli Seal. Thirty upper central incisors and canines with straight canals were instrumented and randomly divided into three equal groups of 10. All teeth were obturated with laterally condensed gutta-percha. An additional group of five teeth with unobturated++ root canals served as positive controls. The sealers were Tubli Seal (group A), Ketac Endo (group B), and Ketac Endo preceded by the removal of the smear layer (group C). The teeth were immersed in India ink for 7 days, centrifuged for 5 min at 3000 rpm, cleared, and then examined under a light microscope at X 50 magnification. The mean value of ink penetration for group A was 0.14 mm, for group B 0.24 mm, and for group C 0.48 mm. No statistically significant differences were observed among groups (p > 0.05).

Acrylic Resins↗
PubMed1995

Comparison of the sealing capacity of three endodontic filling techniques.

This study compared the sealing capacity of three different endodontic gutta-percha filling techniques. Forty-seven upper central incisors with straight canals were instrumented and randomly divided into three equal groups of 14. An additional group of five teeth with unobturated canals served as positive controls. The following gutta-percha filling techniques were evaluated: Trifecta (group A), lateral condensation (group B), and a combination of SuccessFil gutta-percha and lateral condensation (group C). Tubli-Seal was used as a sealer. The teeth were radiographed in mesiodistal and bucco-lingual directions to study the quality of the filling. Afterward, the teeth were immersed in India ink and subsequently cleared. The maximal depth of penetration was measured using a light microscope. The mean value of ink penetration for group A was 1.073 mm, for group B was 0.522 mm, and for group C was 0.765 mm. No statistically significant differences were seen among groups (p < 0.05).

Dental Leakage↗
PubMed1991

Long-term sealing ability of a calcium hydroxide sealer.

A calcium hydroxide sealer (Sealapex) was compared with a zinc oxide and eugenol sealer (Tubli-Seal) over a 32-wk interval to examine solubility in an in vitro simulation. After obturation with gutta-percha and the appropriate sealer, specimens were immediately immersed in a saline solution to challenge the sealers' solubility. The solutions were changed weekly to allow for a continued dissolution of the sealers and to prevent establishment of an equilibrium between the solution and the sealers. The 2- and 32-wk specimens were removed from the solutions, immersed in India ink for 3 days, and then made transparent by a clearing process. Microscopic examination was used to determine the linear penetration of the ink for each tooth. Results revealed that Sealapex statistically had no greater dissolution (based upon linear penetration) than Tubli-Seal at both 2 and 32 wk. It is suggested that Sealapex has a sealing ability comparable to Tubli-Seal and can withstand long-term exposure to tissue fluids without significant leakage. This may allow time for the biochemical action of the calcium hydroxide to stimulate physiological calcification of the apical foramen.

Calcium Hydroxide↗
PubMed1990

An evaluation of the apical seal produced by lateral and warm lateral condensation techniques.

The apical sealing ability of a standard lateral condensation technique was compared with that of two different warm lateral condensation techniques. Sixty extracted human teeth with single canals were instrumented and randomly placed into three groups. Four additional teeth were used as controls. Each group of 20 teeth was obturated with gutta-percha and sealer using one of the three techniques. After 7 days in India ink, the teeth were cleared and the linear extent of ink penetration was measured with a stereomicroscope. No leakage was evident in the two negative controls whereas complete leakage was present in the two positive control teeth. Statistical analysis of the results showed no significant difference in leakage between the experimental groups.

Dental Leakage↗
PubMed1989

A comparison of four root canal filling techniques.

This study compared the apical seal produced by four obturation techniques. Sixty-four extracted human teeth were prepared and obturated using lateral condensation of gutta-percha that was either unmodified or was dipped in chloroform, eucalyptol, or eucapercha paste. After storage in normal saline and 0.02% azide solution at 37 degrees C for 200 days, the teeth were immersed in India ink for 48 h. The most coronal extent of leakage of India ink into the canal was then determined. Significantly more apical leakage occurred in the eucapercha group than in the other three groups. All other comparisons were equivalent. The results suggest that modification of the gutta-percha master cone with solvent does not improve the apical seal in vitro. If modification is desired, then dipping the master cone in either eucalyptol or chloroform produces an apical seal superior to that achieved with eucapercha.

Chloroform↗
PubMed1997

Apical sealing using two thermoplasticized gutta-percha techniques compared with lateral condensation.

This study was conducted to compare the apical sealing capacity of three filling techniques, a multiphase gutta-percha obturation technique, JS Quickfill, and lateral gutta-percha condensation. A four-stage stepback preparation was executed with circumferential filing in a total of 70 recently extracted permanent maxillary incisors and canines. Three experimental groups (20 teeth in each group) and two control groups (5 teeth in each group) were formed at random. Each experimental group was obturated with a different technique. The positive control group was not obturated. In the negative control group, the whole root was covered with two full nail varnish layers. Teeth were immersed in India ink for 48 h, demineralized, cleared with methyl salicylate, and examined with a stereomicroscope. None of the specimens obturated with the multiphase gutta-percha obturation system showed any penetration of India ink beyond the end of the preparation. In the groups of teeth obturated by JS Quickfill or lateral gutta-percha condensation, leakage was detected in one specimen from each group. No statistically significant differences were seen among groups. We conclude that the newer thermoplasticized filling techniques can be used successfully to obturate root canals in large, straight roots.

Carbon↗
PubMed2003

Dynamic morphology of clear corneal cataract incisions.

OBJECTIVE: Clear corneal cataract incisions without sutures allow for rapid visual rehabilitation after phacoemulsification but might be associated with an increased risk of postoperative infection. The goal of this study was to examine in vitro dynamic changes in unhealed clear corneal cataract incisions that might adversely affect the risk of intraocular infection. DESIGN: Laboratory investigation. METHODS: Self-sealing clear corneal incisions were created in cadaveric human and rabbit eyes, and intraocular pressure was controlled with an infusion cannula. Incisions were imaged in real time using optical coherence tomography as intraocular pressure was varied. Using an artificial anterior chamber, India ink was applied to the surface of cadaveric human corneas with clear corneal incisions to detect possible flow of surface fluid along the incision. Intraocular pressure was varied by raising and lowering the infusion bottle so as to simulate the variation in intraocular pressure that would occur with successive blinks. RESULTS: Optical coherence tomography demonstrated variation of corneal wound morphology in response to changes in intraocular pressure. Higher intraocular pressures were associated with close apposition of the wound edges, with no tendency for wound leakage. At low intraocular pressures, however, wound edges tended to gape, starting at the internal aspect of the wound. One incision opened along the entire length, allowing fluid flow across the cornea. Histologic examination revealed India ink particles in all incisions for up to three fourths of the length of the wound. CONCLUSIONS: Transient reduction of intraocular pressure might result in poor wound apposition in clear corneal incisions, with the potential for fluid flow across the cornea and into the anterior chamber, with the attendant risk of endophthalmitis.

Animals↗