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[Direct detection in diarrheal stools of Escherichia coli adhesion factors CFA/I, CFA/II and E8775 by the india ink immune reaction].

An India ink immune reaction was used for the direct detection, in diarrhoeal stools, of Escherichia coli possessing the CFA/I, CFA/II and E8775 fimbriae. With this method, a presumptive diagnosis of enterotoxigenic E. coli (ETEC) can rapidly be made. Staining required the bivalency of rabbit anti-fimbriae IgG; the F(ab')2 fragment was necessary. The reaction was impossible with purified Fab fragment. A comparative study of this technique and detection by culture of ETEC strains showed good correlation when the India ink immune reaction was performed at the beginning of infection.

Antigens, Bacterial↗

Effects of the humic substances of de-inking paper sludge on the antagonism between two compost bacteria and Pythium ultimum.

We investigated the in vitro influence of humic substances (HS) extracted from de-inking paper sludge compost on the inhibition of Pythium ultimum by two compost bacteria, Rhizobium radiobacter (Agrobacterium radiobacter) and Pseudomonas aeruginosa. When low concentrations (5 or 50 mg l(-1)) of HS were added to the culture medium, fungal inhibition by R. radiobacter significantly increased (P<0.01) by 2-3%. In contrast, these low levels of HS had no effect on P. ultimum inhibition by P. aeruginosa. The Fe, chelated by HS, was in part responsible for the decrease of P. ultimum inhibition by the bacteria when increasing amounts of HS were added in the culture medium. The addition of 500 mg l(-1) of humic acids isolated from de-inking paper sludge compost or from fossil origin completely eliminated the inhibition of P. ultimum by R. radiobacter. This Fe effect also stimulated growth of R. radiobacter and reduced its siderophore production in a minimal medium supplemented with HS as sole source of Fe. The results showed that HS influence microbial antagonism when added to a culture medium. However, this effect varies with different factors such as the type of bacteria, concentration of HS, molecular weight and Fe content.

Bacteria↗

Multivariate chemometrics for the forensic discrimination of blue ball-point pen inks based on their Vis spectra.

Fifty blue ball-point pen inks of five different brands were examined on the basis of the Vis spectrum of their ethanolic solutions with a view to achieving good discrimination between them. Samples were dissolved in absolute ethanol and their absorbance values in the range of 400-750 nm, after appropriate transformations, were used as variables in the multivariate statistical techniques of cluster analysis (CA), principal component analysis (PCA) and discriminant analysis (DA). These techniques were used successively so that an effective and meaningful discriminant model was calculated in the final step. The initial 351 variables (log10 transformed ink absorption values at each wavelength) were subjected to a K-means CA over the objects (samples) and only 20 variables were retained. Principal component analysis was used to detect any outliers (four samples were removed) and the remaining samples were re-subjected to PCA to decide how many variables to enter into DA and whether original variables or components should be used. It was found that the first three principal components (in accordance with the Kaiser criterion) were good descriptors of the 20 original variables (96.97% of the data variance was explained) and their use as latent variables in DA lead to low average variable redundancy (33.6%) in the discriminant model. The calculated model had a Wilks' lambda of 8.98 x 10(-5) and was statistically significant at the P = 0.05 level. The post hoc classification of the training dataset was 100% correct. From the DA results and the component loadings it was found that discrimination was achieved on the basis of differences in the shape of the absorption bands as well as their relative intensities. The method was therefore deemed appropriate for supporting exclusionary forensic purposes.

Journal Article↗

Experimental glaucoma model in the rat induced by laser trabecular photocoagulation after an intracameral injection of India ink.

A simple and reproducible rodent glaucoma model is required to elucidate the pathophysiology of damage to the optic nerve. We developed chronically elevated intraocular pressure (IOP) unilaterally in rats by injecting india ink into the anterior chamber of one eye using a 30-gauge needle. One week later, trapped carbon particles in the chamber angle formed a black band along the corneal limbus in the injected eyes. We performed direct laser photocoagulation without a gonio lens, and selectively burned the trabecular meshwork. Intraocular pressure was measured every week and laser photocoagulation was repeated until mean IOP in the experimental eyes rose above 25 mmHg. Unilateral IOP elevation was attained in all rats within 4 weeks. Twelve weeks after ink injection, we sacrificed the rats and excised the eyes for histologic analysis. The anterior chamber angle showed peripheral anterior synechia caused by laser photocoagulation, and carbon particles were engulfed by macrophages that infiltrated the ciliary cleft. In the optic nerve head, a remarkable decrease in the nerve fiber layer and cavernous degeneration were observed, suggesting glaucomatous optic nerve damage. This experimental rodent model should facilitate the study of the complex mechanisms involved in glaucoma.

Animals↗

Molecular cloning of a peroxidase mRNA specifically expressed in the ink gland of Sepia officinalis.

A full-length cDNA encoding a peroxidase has been assembled from two different overlapping cDNA clones isolated from the cDNA library of Sepia officinalis ink gland and 5' RACE procedure. The cDNA of 2898 bp contains an open reading frame of 2778 bp corresponding to a protein of 926 amino acids. The protein shows 76% identity with a peroxidase isolated from the light organ of the squid Euprymna scolopes and approximately 30% identity with mammalian peroxidases. Northern blot analysis of mRNA from various tissues of Sepia officinalis reveals that peroxidase is exclusively expressed in the ink gland. These results represent the first characterization of a melanogenic peroxidase whose possible involvement in the biosynthesis of melanin is discussed.

Amino Acid Sequence↗

A drop-on-demand ink-jet printer for combinatorial libraries and functionally graded ceramics.

A printer has been designed and built for the preparation of combinatorial libraries of ceramics and for solid freeforming of functionally graded ceramics with three-dimensionally programmable spatial variation in composition. Several ceramic suspensions (as inks) can be subjected to micromixing behind the nozzle and printed at precise positions. Both mixing and positioning are computer-controlled. The machine consists of an XY table to control the geometry, a set of electromagnetic valves that manage the mixing, a combined electromagnetic valve and sapphire nozzle that form the print head, and a computer that controls the whole system. The mixing valves can eject as little as 1 mg/s ink into the mixing chamber. The printer has been controlled, run, calibrated and tested; the composition and geometry of printed mixtures can be controlled precisely. This method for the controlled mixing of powders facilitates the advance of combinatorial methods within the materials sciences.

Journal Article↗

The effect of preparation procedures on the morphology of melanin from the ink sac of Sepia officinalis.

The structure of melanin extracted from the ink sac of the cuttlefish Sepia officinalis was examined for different methods of isolation and purification of the pigment. Scanning electron microscopy (SEM) images of Sepia eumelanin prepared by different procedures establish that multi-microm-sized aggregates reported by previous workers are generated by their sample preparation, and that the dominant constituents of Sepia melanin are approximately 150 nm spherical granules. Brunauer-Emmett-Teller (BET) measurements reveal that Sepia eumelanin from Sigma (prepared by spray drying the pigment) has a surface area of 14.3 m2/g. Pigment extracted directly from the fresh ink sac and then freeze-dried has a surface area of 21.5 m2/g, while CO2-supercritically dried has a surface area of 37.5 m2/g. This is consistent with SEM images showing that the process of freeze-drying produces aggregates, but to a lesser extent than spray drying. Supercritical drying of the sample produces suspensions of the individual approximately 150 nm granule, which is more reflective of the natural pigment. Brunauer-Emmett-Teller surface area analysis and Barrett-Joyner-Halenda (BJH) pore volume analysis indicate that the surface of the granules is not smooth and the interior of the granules is not porous, but rather the aggregates of granules are porous. Ultra-high resolution SEM and atomic force microscopy (AFM) images show the granules are easily deformed and are comprised of smaller constituents. De-aggregation of the granules by sonication and ultra-filtration reveal a range of structures depending on the pore size of the membrane used. The implications of these results on quantifying photochemical properties and kinetic reaction rate constants of melanin are discussed.

Animals↗

Hydrogen peroxide produced by Aplysia ink toxin kills tumor cells independent of apoptosis via peroxiredoxin I sensitive pathways.

Marine snails of the genus Aplysia possess numerous bioactive substances. We have purified a 60 kDa protein, APIT (Aplysia punctata ink toxin), from the defensive ink of A. punctata that triggers cell death with profound tumor specificity. Tumor cell death induced by APIT is independent of apoptosis but is characterized by the rapid loss of metabolic activity, membrane permeabilization, and shrinkage of nuclei. Proteome analysis of APIT-treated tumor cells indicated a modification of peroxiredoxin I, a cytoplasmic peroxidase involved in the detoxification of peroxides. Interestingly, knockdown of peroxiredoxin I expression by RNA interference sensitized cells for APIT-induced cell death. APIT induced the death of tumor cells via the enzymatic production of H2O2 and catalase completely blocked APITs' activity. Our data suggest that H2O2 induced stress and the modulation of peroxiredoxins might be a promising approach for tumor therapy.

Animals↗

An intelligence ink for oxygen.

A generic ink formulation is described, comprising semiconductor photocatalyst particles, a brightly-coloured redox dye, a mild reducing agent, a polymer and a solvent, that creates an irreversible, reusable, UV-light-activated, colorimetric indicator or intelligence ink for oxygen.

Journal Article↗

A calcium/calmodulin-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate in the CNS of the cuttlefish Sepia officinalis: localization in specific neural pathways controlling the inking system.

Chemical, biochemical, and immunohistochemical evidence is reported demonstrating the presence in the brain of the cuttlefish Sepia officinalis of a Ca2+-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate, occurring in neurons and fibers functionally related to the inking system. Nitric oxide synthase activity was concentrated for the most part in the cytosolic fraction and was masked by other citrulline-forming enzyme(s). The labile nitric oxide synthase could be partially purified by ammonium sulfate precipitation of tissue extracts, followed by affinity chromatography on 2',5'-ADP-agarose and calmodulin-agarose. The resulting activity, immunolabeled at 150 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis by antibodies to rat neuronal nitric oxide synthase, depended on NADPH and tetrahydro-L-biopterin, and was inhibited by N(G)-nitro-L-arginine. NMDAR2/3 subunit-immunoreactive proteins migrating at 170 kDa could also be detected in brain extracts, along with glutamate (whole brain: 0.32 +/- 0.03 micromol of glutamate/mg of protein; optic lobes: 0.22 +/- 0.04; vertical complex: 0.65 +/- 0.06; basal lobes: 0.58 +/- 0.04; brachial lobe: 0.77 +/- 0.06; pedal lobe: 1.04 +/- 0.08; palliovisceral lobe: 0.86 +/- 0.05). Incubation of intact brains with 1.5 mM glutamate or NMDA or the nitric oxide donor 2-(N,N-diethylamino)diazenolate-2-oxide caused a fivefold rise in the levels of cyclic GMP, indicating operation of the glutamate-nitric oxide-cyclic GMP signaling pathway. Immunohistochemical mapping of Sepia CNS showed specific localization of nitric oxide synthase-like and NMDAR2/3-like immunoreactivities in the lateroventral palliovisceral lobe, the visceral lobe, and the pallial and visceral nerves, as well as in the sphincters and wall of the ink sac.

Animals↗

Phlegmonous gastritis after Indian ink marking.

A report is presented on a case of phlegmonous gastritis following gastroscopic Indian ink injection into the mucosa of the stomach in a 60-year-old woman patient wih early gastric carcinoma. The patient was discharged from hospital on the 11th day after Billroth II resection, in good condition. The necessity for disinfection and sterilization of the instruments employed is emphasized; the continued use of Indian ink marking is recommended.

Abscess↗

Transdermal transport of India ink by electromagnetic electroporation in Guinea pigs: an ultrastructural study.

Transdermic administration by electroporation has developed over recent years for applying drugs in a variety of pathological processes. However, mechanisms are still not finally settled. India ink was applied to the backs of guinea pigs and for the transdermic transport short, high-voltage pulses (TDES, Dencort Dell) were administrated. Punch biopsies (4 mm) immediately taken after 24, 48, 72, 96 and at 26 days were studied by light and electronic microscopy. The ultrastructural characteristics and image pigment particles were reported. Particles of India ink were observed in the stratum corneum and in the epidermic keratinocytes of samples studied immediately after treatment. Particles were also seen in the epidermic and folicular keratinocytes, and in the papillary and reticular dermis (among collagen fibers, vessel walls, and macrophages) in all the subsequent biopsies; but not in the controls, which were conducted with electromagnetic waves alone. No tissue alterations were observed. The efficacy and noninvasive nature of electroporation for the transdermic administration of macromolecules is confirmed.

Administration, Cutaneous↗

Successful removal by ruby laser of darkened ink after ruby laser treatment of mismatched tattoos for acne scars.

Cosmetic tattoos are becoming increasingly popular. Elimination of cosmetic tattoos is sought because of misplacement or migration of tattoo pigment, allergic reactions to the various pigments or dissatisfaction of the customer for various reasons. Removal of unwanted pigment is a domain of laser surgery using various Q-switched laser systems, such as the ruby, alexandrite, pulsed dye and Nd:YAG lasers. Dark colours are easily removed by these lasers, whereas red, pink and skin-toned pigment may turn black if exposed to Q-switched laser light. This ink-blackening occurs because Q-switched lasers heat up the pigments, which consist of ferric oxide, and reduce them into ferrous oxide, which is black. Laser-induced black ink is not always readily removed. A successful ruby laser-removal of laser-induced blackened cosmetic tattoos for acne scar camouflage is reported. The advantageous outcome in this case contrasts with other published cases where laser-darkened pigment had to be removed by other measures, or was permanent. Test site treatment can limit the problem to some degree but, in addition to test-treating, some kind of 'tattoo identification card' could help to prevent problems in this field and allow 'in vitro' test treatment.

Acne Vulgaris↗

Modified India ink preparation for Cryptococcus neoformans in cerebrospinal fluid specimens.

A novel modified India ink technique for the diagnosis of Cryptococcus neoformans in cerebrospinal fluid specimens is described. It employs 2% chromium mercury and India ink. This technique allows a clear identification of some external and internal structures of the organism. Three layers from the outer capsule that have previously been discerned only by electron microscopy are distinguished.

Carbon↗

Changes of tumor microcirculation after transcatheter arterial chemoembolization: first pass perfusion MR imaging and Chinese ink casting in a rabbit model.

AIM: To observe the change of tumor microcirculation after transcatheter arterial chemoembolization (TACE) with bletilla microspheres by using first pass perfusion MR imaging (FP) and Chinese ink casting. METHODS: VX2 carcinoma cells were surgically implanted into the left and right lobes of liver of 30 New Zealand white rabbits, which were divided into 3 groups at random. Emulsion of lipiodol mixed with mitomycin C, and 5-FU bletilla microspheres were injected into the hepatic artery respectively, and saline was used as control agent. MR imaging was performed with turbo-flash sequence 14 d after tumor implantation and 7 d after interventional therapy. The steepest slopes (SS) of the signal intensity versus time curves were created for quantitative analysis, 7.5% Chinese ink gelatin solution was injected through ascending artery (17 cases) or portal vein (2 cases) for lesion microvessel area (MVA) measurement after the last MRI examination. The correlation between perfusion imaging and MVA was studied blindly. RESULTS: The SS values at the rim of tumor in lipiodol group (mean, 49% per second) and bletilla group (mean, 35% per second) were significantly decreased (P<0.05) as compared with control group (mean, 124% per second), no difference was found between lipiodol and bletilla groups (P>0.05). In lipiodol group, the MVAs (24 974+/-11 836 microm(2)) in the center of the tumor were significantly smaller than those of the control group (35 510+/-15 675 microm(2)) (P<0.05), while the MVAs (80 031+/-22 745 microm(2)) around the tumor were significantly increased because small and dense plexuses appeared around the tumor which correlated to intense reaction of granulation tissue. None of the vessels was seen in the tumor in bletilla group, the peripheral MVAs of the tumor were significantly smaller than those of the control group (P<0.05) and lipiodol group (P<0.05). There was a good correlation between SS and MVAs in control group (r(s), 0.985, P<0.0001) and bletilla group (r(s), 0.743, P<0.05), the correlation was not significant in lipiodol group (r(s), 0.527, P>0.05). CONCLUSION: TACE with bletilla microspheres may enhance its anti-tumor effect by inhibiting the angiogenesis, and FP-MRI provides useful information to assess the TACE effect by depicting tumor vascularization and perfusion.

Animals↗

Study on cephalopod's ink for anti-retroviral activity.

Aqueous extracts of ink from four cephalopods, adult and young Sepiella inermis and Loligo duvaucelli were tested against Moloney murine leukaemia virus reverse transcriptase (MMLV RT). Ink from young cephalopods, S. intermis and L. duvaucelli showed strong inhibition of MMLV RT.

Animals↗

Dating of ballpoint pen ink.

In this paper we describe a case in which a cash book, dated of the year 2000, was sent to the Forensic Science Division, Document Laboratory, Zurich Canton Police in March 2003. The questioned document was a list of 29 pages containing a consecutive handwritten numbering and dated entries (payments) made of blue ballpoint pen ink. By definition, a cash book has to be written by hand and the entries have to be made daily. The questioned document was suspected to have been written within a short period (e.g., a few hours) and backdated. The document lab was asked to determine the date of the entries of the questioned list. On one hand, we were asked, if the cash book had been kept on account consecutively during the period of one year, and on the other hand, the judges were interested in knowing, if the document could have been written in 2003 and back dated to 2000. To answer these questions, the document was examined for latent elements by electrostatic detection device (ESDA). The relative dating of the entries was performed by the quantification of the ballpoint pen ink dyes and their degradation products using HPLC. Results show that it is possible to determine the relative age of entries written by ballpoint pen within a relatively short time scale, if storage and supporting material of the different samples are the same or at least similar.

Journal Article↗

Reticulated black solar lentigo ('ink spot' lentigo).

BACKGROUND AND DESIGN: Pigmented lesions that are black and have an irregular outline are often considered suspicious for melanoma; however, these features may be seen in benign lesions. The reticulated black solar lentigo is such a lesion and is described in a clinicopathologic study of nine lesions in eight patients. RESULTS: The "ink spot" lentigo is distinguished clinically by its color and wiry or beaded, markedly irregular outline. It has a reticulated pattern and most resembles a spot of ink on the skin. In this series of patients, the lesions were limited to sun-exposed areas of the body and had a distribution pattern similar to that of solar lentigines. Although all the patients were of Celtic ancestry and had numerous solar lentigines, they usually had only one black lentigo (range, one to four; mean, 1.6; median, one). Histologic evaluation, including electron microscopy and dopa-incubated vertical sections, demonstrated lentiginous hyperplasia of the epidermis, marked hyperpigmentation of the basal layer with "skip" areas that involved the rete ridges, and a minimal increase in the number of melanocytes. CONCLUSIONS: Because of their dark color, irregular border, and limited number, reticulated black solar lentigines were of concern to patients and primary care physicians. However, the characteristic features of these lesions allow one to make the clinical diagnosis of a benign lentigo.

Adult↗