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PubMed1999

Microvasculature of crotaline snake pit organs: possible function as a heat exchange mechanism.

The infrared sensory membranes of the pit organs of pit vipers have an extremely rich capillary vasculature, which has been noted passim in the literature, but never illustrated or studied in detail. We rendered the pit vasculature visible in various ways, namely, by microinjection of India ink, by a combination of ink and succinate dehydrogenase staining, and by making resin casts for scanning electron microscope study. We also used transmission electron microscopy for identifying the types (arterioles, venules, capillaries) of blood vessels. Then we compared the pit vasculature with that of the retina and the dermis. Good visualization of the vasculature was obtained with both ink and resin injection. Arterioles, venules, and capillaries could be distinguished with all methods used. The monolayer vasculature was denser in the pit membrane than in the retina or skin. Each loop of the network enclosed a small number of infrared receptors so that all receptors were in contact with a capillary on at least one side. The forward-looking areas of the pit had a denser network than side-looking areas. Since infrared rays cause nerve impulses by raising the temperature of individual receptors, the capillary network functions not only as a supplier of energy but also as a cooling mechanism to reduce afterimages. Thus the denser network in the forward-looking areas causes these areas to be more sensitive and have better image resolution than the rest of the membrane.

Agkistrodon↗
PubMed1994

The relationship between microscopic margins of resection and the risk of local recurrence in patients with breast cancer treated with breast-conserving surgery and radiation therapy.

BACKGROUND: The relationships among the involvement of tumor at the final margins of resection, the presence of an extensive intraductal component (EIC), and the risk of local recurrence are important considerations in patients treated with conservative surgery and radiation therapy for early stage breast cancer but have not been defined adequately. METHODS: Between 1982 and 1985, 885 patients were treated for clinical Stage I or II invasive breast cancer. The study population was limited to 181 patients with an infiltrating ductal carcinoma who received a radiation dose to the surgical site of 60 Gy or greater, whose final microscopic margins of resection were evaluable, and who had at least 5 years of follow-up. A positive margin was defined as tumor present at the inked margin of resection, a close margin as tumor within 1 mm of the inked margin, and a negative margin as no tumor within 1 mm of the inked margin. A focally positive margin was defined as tumor at the margin in three or fewer low-power fields. In 157 patients (87%), the tumor was evaluable for the presence or absence of an EIC. The median follow-up was 86 months. RESULTS: In 12 of 181 patients (7%), a recurrence developed at or near the primary site (true recurrence/marginal miss [TR/MM]) within 5 years. The 5-year rate of TR/MM (with 95% confidence intervals) among patients with negative, close, focally positive, and more than focally positive margins was 0% (0-4%), 4% (0-20%), 6% (1-17%) and 21% (10-37%), respectively. Patients with positive margins also were more likely to have a distant failure within 5 years (14%, 8%, 25%, and 32% in the four groups, respectively). However, patients with positive margins more often had positive axillary lymph nodes than patients with negative or close margins (59% vs. 38%, P < 0.02). The 5-year rate of TR/MM was 20% for patients with an EIC-positive tumor and 7% for patients with an EIC-negative tumor. However, among the 127 patients with an EIC-negative tumor, the 5-year rate of TR/MM was less than 10% in all margin groups. Among the 30 patients with an EIC-positive tumor, the 5-year rate of TR/MM was 0% when margins were negative or close but 50% when margins were more than focally positive. CONCLUSIONS: These results provide support for the use of breast-conserving surgery and breast irradiation in all patients with uninvolved margins, whether the tumor is EIC-positive or EIC-negative. This study suggests that breast-conserving therapy (including a radiation boost to the primary site) also may be a reasonable option for some patients with an EIC-negative tumor and margin involvement.

Breast↗
PubMed1986

Localized vascular regression during limb morphogenesis in the chicken embryo: II. Morphological changes in the vasculature.

The regression of blood vessels in the distal wing bud of chicken embryos from stages 19 to 31 was examined by light and electron microscopy. The vessels were double-labelled by an injection of Monastral blue B (MB) to label the regressing endothelial cells, followed 6-48 hours later with an injection of India ink which marked the lumens of patent vessels. Prior to stage 26 the vessels contained only India ink since the endothelial cells were not phagocytic at this stage. Vessels at stage 26 or later were often double-labelled, with MB sequestered in the endothelial cell cytoplasm and India ink in the vessel lumens. After stage 27 cells not associated with lumens, but labelled with MB, were observed in areas undergoing vascular regression. Ultrastructural changes in the endothelial cells as the vessels regressed included formation of luminal and abluminal processes, long complex junctions, and vacuoles containing MB. In many involuting vessels the endothelial cells appeared normal even though the lumens were collapsed. Occasionally, isolated pyknotic cells were observed in regions that had been previously vascularized. At stage 31 cells in the developing cartilage had vacuoles containing MB. Our study suggests that blood vessels may disappear from the prechondrogenic zone of the distal wing bud by several mechanisms. These could include a type of cell death that does not elicit a cellular infiltrate, migration of the endothelial cells away from vascularized regions, and/or transdifferentiation into cells that resembled chondrocytes.

Animals↗
PubMed2003

Cell and organ printing 1: protein and cell printers.

We have developed several devices for positioning organic molecules, molecular aggregates, cells, and single-cell organisms onto solid supports. These printers can create stable, functional protein arrays using an inexpensive technology. The cell printer allows us to create cell libraries as well as cellular assemblies that mimic their respective position in organs. The printers are derived from commercially available ink-jet printers that are modified to dispense protein or cell solutions instead of ink. We describe here the modifications to the print heads, and the printer hardware and software that enabled us to adapt the ink-jet printers for the manufacture of cell and protein arrays. The printers have the advantage of being fully automated and computer controlled, and allow for the high-throughput manufacture of protein and cell arrays.

Animals↗
PubMed2003

Indentation testing of human cartilage: sensitivity to articular surface degeneration.

OBJECTIVE: To determine, for clinical indentation testing of human articular cartilage, the effects of aging and degeneration on indentation stiffness and traditional indices of cartilage degeneration; the relationship between indentation stiffness and indices of degeneration; and the sensitivity and specificity of indentation stiffness to cartilage degeneration. METHODS: Osteochondral cores from femoral condyles of cadaveric human donors were harvested. Samples were distributed into experimental groups based on donor age (young [20-39 years], middle [40-59 years], and old [>/=60 years]), and a macroscopic articular surface appearance that was either normal or mildly degenerate, without deep erosion. Samples were analyzed for indentation stiffness, cartilage thickness, India ink staining (quantitated as the reflected light score), and Mankin-Shapiro histopathology score. RESULTS: Indentation stiffness, India ink staining, and the histopathology score each varied markedly between normal-sample and degenerate-sample groups but varied relatively little between normal samples obtained from different age groups. A decrease in indentation stiffness (softening) correlated with a decrease in the reflectance score and an increase in the overall histopathology score, especially the surface irregularity component of the histopathology score. Receiver operating characteristic analysis suggested that the indentation testing could accurately detect cartilage degeneration as indicated by macroscopic appearance, India ink staining, and histopathology score. CONCLUSION: The indentation stiffness of the normal to mildly degenerate samples tested in this study was sensitive to mild degeneration at the articular surface and was insensitive to changes associated with normal aging or to slight variations in cartilage thickness. This suggests that indentation testing may be a useful clinical tool for the evaluation of early-stage degenerative changes in articular cartilage.

Adult↗
PubMed2005

Analysis of atypical urine cytology in a tertiary care center.

BACKGROUND: Voided urine cytology continues to play a paramount role in the surveillance of transitional cell neoplasms and as a screening modality in certain high-risk situations. Although a significant number of samples are diagnosed as atypical, there is little known about the outcome of these patients. In addition, the significance of transitional cell fragments in voided urine samples is uncertain. The objective of the current study was to evaluate retrospectively a large series of voided urine specimens that were reported as atypical. Standard cytomorphologic parameters were used with the aim of refining the atypical category. METHODS: The authors studied 201 consecutive voided urine samples from 1995 that were evaluated by liquid-based cytology for the following features: specimen cellularity, the presence and number of cell clusters, and the architecture of the cell fragments. In addition, cells were examined for the following cytologic features: high nuclear-to-cytoplasmic ratios, nuclear membrane irregularities, hyperchromasia, India-ink nuclei, and chromatin pattern. Cytoplasmic features were evaluated but were limited significantly by poor preservation. Only specimens with a biopsy or cystoscopy and/or from patients who were followed for > 5 years were analyzed further. RESULTS: In total, 23.4% of specimens showed transitional cell carcinoma (TCC) on biopsy, including 20 Grade 1 TCCs and 21 Grade 2 TCCs. Specimen cellularity, cluster numbers, nuclear membrane abnormalities, hyperchromasia, and India-ink nuclei were associated significantly with TCC. Twenty-six specimens (18.9%) were associated with renal calculi. CONCLUSIONS: The atypical category contained a significant proportion of low-grade TCCs. Transitional cell clusters in voided urine are relevant clinically. The clinical picture, including the previous history of TCC and the presence of urinary calculi, provides valuable information when evaluating voided urine cytology. These features and careful attention to India-ink nuclei and nuclear membrane abnormalities could help make the "atypical" category a more meaningful group.

Adolescent↗
PubMed1984

Demonstration of M cells in the specialized follicle-associated epithelium overlying isolated lymphoid follicles in the gut.

Within the epithelium that overlies the dome regions of Peyer's patches, exist specialized surface epithelial cells (M) which function to take up macromolecules from the gut lumen. These cells may be of great importance in processing antigenic material in the gut. The predominant lymphoid structures of the small intestine are isolated lymphoid follicles, by virtue of their frequency. These follicles are difficult to study because they are not grossly visible. In the present study, three guinea pigs drank India ink mixed into their water for 1, 3, and 5 months. Two hours prior to sacrifice, animals were given an intraintestinal injection of ferritin or India ink. Using a hand lens, the Peyer's patches and isolated follicles were clearly identified among the villi of the intestine. Light microscopy revealed ink in the surface epithelium covering the isolated follicles and within the substance of the follicles. Transmission electron microscopy demonstrated M cells over isolated follicles and Peyer's patches. These cells had lighter staining cytoplasm, while the mitochrondria stained darker with prominent cristae, and the microvilli were shorter. Therefore, M cells do exist within isolated follicles and structurally appear the same as those found in Peyer's patches. This implicates the isolated follicles in the overall antigen processing role of gut-associated lymphoid tissues. The present method facilitates identification of isolated lymphoid follicles which will allow functional studies to be performed on these structures.

Animals↗
PubMed1987

Transactivation of several genes of two native Serratia prophages after superinfection by phage kappa.

Serratia marcescens HY bacteria must be lysogenic with either prophage y or psi to make it possible for phage kappa to form plaques unless they carry a so-called ink mutation. Genes in y and psi termed any and anp were identified that after infection of ink+ cells are necessary for an effective propagation of these phages as well as of coinfecting kappa phage. When kappa infects y and/or psi-lysogenic cells it transactivates the respective prophage genes by means of two early genes termed tay and tap. It appears that on infection of nonlysogenic ink+ cells kappa damps its own development, provided the regulatory region of the responsible gene is undermethylated. After kappa infection duly to achieve the special methylation of this region seems to be the function of any and anp. There are some more genes in y and psi prophage under the control of tay and tap, concerning in both cases a Dam methylation (recognition sequence GATC) of kappa DNA, a recombination proneness under restricting conditions of kappa DNA not modified by the modification enzyme of HY, and the kappa plaque size. By hybridization studies a region of homology common to y and psi was demonstrated which from its size might comprise all the transactivated genes. The view is supported by genetic data indicating an affinity among the any and anp genes. Investigation of various any mutants were indicative of DNA inversions in this region of the y genome. Surprisingly some of the any mutants had become sensitive in their plaque forming ability to an inhibitory activity exerted by prophage psi. Mutants of psi unable to interfere but still able to lysogenize were isolated. A model is presented accounting for the formation of pleiotropic and nonpleiotropic mutations with Any phenotype and their reversion types. Possible functions of the y genes and their counterparts in psi are discussed.

Bacteriophages↗
PubMed2002

Management of old landfills by utilizing forest and energy industry waste flows.

The lack of landfill capacity, forthcoming EU waste disposal and landfill management legislation and the use of non-renewable and energy intensive natural resources for the end-treatment of old landfills increase pressures to develop new landfill management methods. This paper considers a method for the end-management of old landfills in Finland, which is based on the utilization of forest and paper industry waste flows, wastes from paper recycling (de-inking) and wastes from forest industry energy production. Fibre clay wastes from paper mills, de-inking sludges from de-inking of recovered waste paper and incineration ash from forest industry power plants serve to substitute the use of natural clay for the building of landfill structures for closed landfills. Arguably, this method is preferable to existing practices of natural clay use for landfill building, because it (1) substitutes non-renewable natural clay, (2) consumes less energy and generates less CO2 emissions than the use of natural clay, and (3) eliminates considerable amounts of wastes from paper production, paper consumption and from forest industry energy production. Some difficulties in the application of the method are considered and the waste flow utilization is incorporated into a local forest industry recycling network.

Aluminum Silicates↗
PubMed1984

In vivo effects of lipopolysaccharide on lymphoid and non-lymphoid cells in the mouse spleen. Reduction of T-lymphocytes and phagocytosis in the inner parts of the periarteriolar lymphocyte sheath.

In the present study the effects of lipopolysaccharide (LPS) on the cellular composition and phagocytosis of India ink in the inner parts of the periarteriolar lymphocyte sheaths (PALS) are described. Staining for B-, T-lymphocytes, and reticulin fibers in the spleen of normal and LPS-injected mice shows that the B-dependent follicular area is increased in size after LPS administration. However, the number of T-lymphocytes in the inner PALS is reduced markedly and a relatively high number of B-lymphocytes can be found in this area. The significance of this phenomenon is discussed. In untreated mouse spleen, carbon particles become localized in strongly acid-phosphatase (AP)-positive macrophages of the red pulp, marginal zone and white pulp 24 h after an intravenous injection of India ink. All these macrophages contain numerous carbon particles. After LPS pretreatment, the phagocytosis of carbon particles in the inner PALS is dramatically diminished, although many strongly AP-positive macrophages can be found in this area. The phagocytosis of carbon particles in the other compartments of the spleen did not change. It appears that injection of 2 micrograms LPS or more is sufficient to induce this phenomenon which is most significant when LPS is injected 24 or 48 h before exposure to India ink.

Acid Phosphatase↗
PubMed1993

Calcium and potassium currents in porcine granulosa cells maintained in follicular or monolayer tissue culture.

We studied membrane currents in granulosa cells (GC), immediately after collection or after variable culture time in the everted-follicle wall or in the monolayer. GC in both systems express an inward calcium current (ICa) with T-type kinetics and voltage dependence. GC in the everted-follicle culture express an outward potassium current (IK) kinetics, which remains unchanged during three days in culture. IK has delayed-rectifier kinetics, but is insensitive to TEA, 4-AP and apamine. GC in monolayer culture develop a new, inactivating delayed-rectifier potassium current (InK), which progressively dominates as cells advance from day one to day three in culture. A similar InK was recorded in large luteal cells. A possible link between luteinization and the appearance of InK is hypothesized.

Animals↗
PubMed1993

Perivascular cells act as scavengers in the cerebral perivascular spaces and remain distinct from pericytes, microglia and macrophages.

Perivascular cells in the rat brain are an immunophenotypically defined group of cells which can be identified by their expression of the ED2 antigen. The present study investigates the role of perivascular cells as scavengers in the perivascular spaces of the rat brain and the relationship of these cells to microglia, macrophages, pericytes and smooth muscle cells. Particulate matter (Indian ink) was injected selectively into the perivascular spaces of the left caudoputamen of 59 rats. Animals were killed by cardiac perfusion of formalin or glutaraldehyde 2 h-2 years after ink injection. Cerebral hemispheres were examined histologically and immunocytochemically using the ED2 antibody for perivascular cells, ED1 for microglia and macrophages and OX-6 directed against Ia antigen [major histocompatibility complex (MHC) class II]. ED2+ perivascular cells ingested Indian ink in the perivascular spaces and expressed MHC class II antigen. Reactive microglia and macrophages in the perivascular parenchyma expressed ED1, but no ED2+ cells were seen outside the perivascular spaces. Transmission electron microscopy distinguished perivascular cells, which ingested carbon particles, from pericytes, which did not. The results of this study suggest that perivascular cells remain distinct from pericytes, microglia and macrophages and that they play a major role as scavengers in the perivascular spaces of the rat brain. This role reflects the importance of perivascular spaces as drainage pathways for soluble and insoluble material from the brain.

Animals↗
PubMed1997

Experimental study of the hydrodynamic effects of irrigation suction drainage.

The function of irrigation suction drainage, as an additional method in the treatment of bone infection, is associated with hydrodynamic problems that can only be experimentally examined. The visualization of liquid movement in the experimental model was realized by adding a colored ink to the input drain of the drainage system. When only 2000 ml of liquid was used daily (80 drops/min), the ink penetrated the experimental cavity only through the most proximal holes in the input drain. It spread slowly and irregularly through the liquid in the experimental bottle, forming pocket deposits and leaving sediment. When 6000 ml of liquid was applied daily (240 drops/min), ink quickly penetrated the cavity through all holes in the drain, leading to turbulence and dispersing equally in the bottle. Decoloration of the model bottle was faster because the output drainage was more efficient. With an increased flow, i.e., with more liquid used in the drainage, a better result was achieved. This was manifested in a swift decoloration and rinsing of the cavity.

Drainage↗
PubMed1989

Distribution of ricin within the mammalian para-aortic lymph node. II. Comparison of the localization, after intramuscular dosage of colloidal gold-labelled ricin in vivo, with in vitro binding characteristics of the native toxin.

Previous work has shown that, following an intramuscular injection of ricin, the toxin becomes localized within histiocytes in the sinuses of lymph nodes draining the 'wound' site. When ricin labelled with colloidal gold was similarly injected, it was found within the same lymphoid cells as seen with native ricin. Biologically inert Indian ink apparently follows a similar fate, as demonstrated by the appearance of carbon particles within sinus histiocytes, as soon as 1 h after intramuscular injection. When the binding in vitro of Indian ink or ricin toxin to sections of lymph node was examined, ricin was seen to bind to the surfaces of the same sinusoidal cells and also, with a much lower frequency, to follicular lymphocytes, whereas Indian ink failed to bind. This indicated an interaction between ricin and cell membrane components. Moreover, this binding was inhibited markedly by the galactose-containing disaccharide, lactose, a target sugar specified by the lectin binding site of ricin and to a much lesser extent by the monosaccharide mannose.

Animals↗
PubMed1986

Distribution of blood-borne particles of two different sizes in rat spleens.

Polystyrene microspheres 5 micron in diameter and Indian ink, consisting of particles 0.03 micron in diameter, were injected into the splenic artery in rats. The distribution of the polystyrene microspheres and the ink particles in the spleen were examined microscopically and morphometrically. The polystyrene microspheres appeared mainly in the red pulp, and the Indian ink particles mostly in the marginal zone, which functions as an immunological filter. (No arteries opened into the lymphoid follicles.)

Animals↗
PubMed2003

Towards a new conservation method for ancient manuscripts by inactivation of iron via complexation and extraction.

The aim of this work was to study the efficiency of extraction of iron from model paper samples by use of different ligands (deferoxamine mesylate, the potassium-magnesium salt of phytic acid and diethylenetriaminepentaacetic acid) at varied concentrations (0.01, 0.005, and 0.001 mol L(-1)) and pH (7, 8, 9). Graphite furnace atomic absorption spectrometry (GFAAS) was used to monitor the total amount of iron in solutions of the respective ligands. Two types of model were used to investigate the behaviour of various iron species present in ancient iron-gall ink. Requirements for the optimal procedure, which could possibly be used in the conservation of ancient manuscripts, included high effectiveness of iron extraction from samples which modelled free iron ions (samples "Fe"), while iron deposited in the form of ink (samples "A") should remain without any visible change of the ink's intensity. The best results were achieved with the solution of 0.005 mol L(-1 )diethylenetriaminepentaacetic acid (pH=9), which allowed extraction of 97+/-1% of iron from "Fe" model samples and only 64+/-1% from "A" samples.

Journal Article↗
PubMed2001

Microarray sampling-platform fabrication using bubble-jet technology for a biochip system.

The fabrication of microarrays containing PCR-amplified genomic DNA extracts from mice tumors on a Zetaprobe membrane using a modified thermal ink-jet printer is described. A simple and cost-effective procedure for the fabrication of microarrays containing biological samples using a modified bubble-jet printing system is presented. Because of their mass-produced design, ink-jet printers are a much cheaper alternative to conventional spotting techniques. The usefulness of the biochip microarray platform is illustrated by the detection of human fragile histidine triad (FHIT), a tumor suppressor gene. Subcutaneous carcinomas were induced with MKN/FHIT and MKN/E4 cell lines in immunodeficient mice. Several weeks into their development, the tumors from both groups of mice were removed and subjected to DNA extraction by lysis of tissue samples. The extracted DNA samples were amplified by PCR (30 cycles) using the primers corresponding to nucleotides 2 to 18 of the FHIT sequence. The resulting solution was transferred to the individual reservoirs of a three-color cartridge from a conventional thermal ink-jet printer (HP 694C), and arrays were printed on to a Zetaprobe membrane. After spotting, these membranes were used in a hybridization assay, using fluorescent probes, and detected with a biochip.

Acid Anhydride Hydrolases↗
PubMed2006

The use of preoperative endoscopic tattooing in laparoscopic colorectal cancer surgery for endoscopically advanced tumors: a prospective comparative clinical study.

BACKGROUND: Endoscopic India ink marking techniques are often used for the intraoperative location of colonic polyps and early stage neoplasms. The aim of this study was to compare how effective this technique is compared with conventional localization methods, as well as its influence on the results of colorectal laparoscopy (LSCRC) for endoscopically advanced tumors. METHODS: From January 2003 to January 2005, 47 patients with colorectal carcinomas were included in the study. In one group, lesions were localized preoperatively by endoscopic India ink tattooing (n = 21; tattooed group, TG), while conventional methods were used in the others (n = 26; non-tattooed group, NTG). Patients' perioperative clinical and pathoanatomical data were prospectively collected. RESULTS: Both groups were comparable in age, sex distribution, American Society of Anesthesiologists (ASA) score, body mass index (BMI), technique performed, tumor size and proportion of patients who had previous abdominal surgery. Three patients presented ink spillage without clinical repercussions. Visualization of the correct resection site was higher in the TG (100% vs. 80.8%, P = 0.03). Operative time (147.3 +/- 46.2 vs. 187.0 +/- 52.7 minutes, P = 0.02) and blood loss (99.3 +/- 82.8 vs. 163.6 +/- 96.6 cc, P = 0.03) were lower in the TG. There were no differences between groups regarding peristalsis, introduction of oral intake, hospital stay or intra- and postoperative complication rates. No differences were observed amongst pathoanatomical data studied. CONCLUSIONS: Preoperative endoscopic tattooing is a safe and effective technique for intraoperative localization of advanced colorectal neoplasms, improving the operative results of LSCRC.

Aged↗