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Changes in composition of ballpoint pen inks on aging in darkness.

A method for comparison of the relative age of ink entries written by the same ballpoint pen on documents stored in darkness is presented. Inks were extracted from the document and analyzed by HPLC (high performance liquid chromatography). On aging, changes in the chemical composition of the inks were noted. These changes were similar to those observed when inks were exposed to light or heat. The aging was followed by using ternary diagrams constructed for dyes generally present in blue-colored inks--Crystal Violet, Methyl Violet, and Tetramethyl Para Rosaniline. The procedure is applicable for relative dating of ink entries in diaries, notebooks, etc., where often several ink entries are written by the same ink. However, a prolonged exposure of the document to daylight and/or artificial light (light from fluorescent tubes) as well as to extensive heat will render the whole procedure inapplicable. An example of the use of the proposed method in casework is given.

Chromatography, High Pressure Liquid↗

Immune-indian ink method for detection of hepatitis A associated antigen and antibody.

TIndian-ink grains coated with commercial gamma globulin (immune-Indian-ink) were agglutinated by 3 percent of sera from healthy volunteer blood donors; by 4 percent of those from hospital staff in contact with patients suffering from hepatitis; and by 10 percent of those from patients with viral diseases other than hepatitis, In contrast, the rate of positive reactions was 86 percent in the case of sera taken from patients in the acute phase of an illness diagnosed as hepatitis A on the basis of epidemiological and clinical data. Investigation of serum samples taken serially from patients positive in the acute phase of illness revealed that the immune-Indian-ink agglutinating factor does not persist for long in majority of cases. Two months after discharge from the hospital it was present in 18 percent of the patients only. The reaction proved negative when a limited number of cases diagnosed as hepatitis B were investigated. The immune-Indian-ink agglutinating factor was inhibited by all but one of 36 sera taken in the convalescent phase from patients with a diagnosis of hepatitis A. Some sera displaying agglutination with immune-Indian-ink gave a reaction with uncoated Indian-ink, too. Efforts to free the sera from non-specific agglutinating factor by starch-block electrophoresis have led to partial success. Fractionation on Sephadex G-200 columns suggested that in molecular weight (or particle size) the immune-Indian-ink agglutinating factor is smaller than HBsAg and larger than the non-specific agglutinating factor. On the basis of these results it is assumed that the immune-tindian-ink reaction is suitable for detecting an antigen tentatively called IH chi Ag and its antibody (IH chi Ab) specific to hepatitis A.

Acute Disease↗

Evoked ink release in Aplysia produces inhibition of the siphon withdrawal reflex in neighboring conspecifics.

Aplysia californica exhibit a dramatic defensive reaction, the release of a cloud of dark purple ink, in response to noxious stimuli. Although the neural control of this behavior has been studied rather extensively, the functional significance of the inking response is not well understood. We have found that ink released by animals that are subjected to noxious stimuli rapidly induces inhibition of the tail-elicited siphon withdrawal reflex in neighboring Aplysia. Further experiments indicated that the inhibitor is the ink itself, and not some other substance released by the donor animals. Finally, we examined whether ink-induced inhibition of siphon withdrawal might be a secondary consequence of an elevated competing response such as increased locomotion. We found that locomotion is not affected by the concentrations of ink we employed, indicating that the ink probably modulates the withdrawal reflex directly. Because the neural circuits responsible for both tail-elicited siphon withdrawal and the inking response have already been partly delineated, one can now bring the neurobiological advantages of Aplysia to bear on the ethologically important issue of signaling between conspecifics.

Animal Communication↗

Analysis of Indian blue ballpoint pen inks tagged with rare-earth thenoyltrifluoroacetonates by inductively coupled plasma-mass spectrometry and instrumental neutron activation analysis.

Characterization and assessment of inks on sensitive documents for absolute/relative age determination is the challenging forensic problem in spite of practical difficulties. Tagging of ballpoint pen ink with suitable taggant(s) is a unique method to come out with definitive inferences on the detection of forgery in documents written with ballpoint pens. Selection of a proper taggant primarily depends on sensitivity of analytical determination and their absence in normal varieties of ink used for document writing. Rare-earth elements, from all technical considerations can be potential taggant(s) for inks. To ensure more compatibility with ink, 13 rare-earth thenoyltrifluoroacetonate chelates were prepared and characterized. The ballpoint pen inks were tagged with rare-earth thenoyltrifluoroacetonate chelates individually at about 1-100 ppm level depending on sensitivity of element under suitable optimized experimental conditions and instrumental sensitivity. Aliquots of such tagged ink having varying amounts of taggants were analyzed by ICP-MS and INAA. Satisfactory recoveries and a good linear relationship of intensity (signal) against concentrations/amounts were observed. Under the optimized experimental conditions, the detection limits were worked out. This study of tagging metal ions in combination with ICP-MS and NAA as an analytical tool can allow to draw various combination options based on different rare-earth chelates as suitable materials for tagging of ballpoint pen inks for absolute/relative age determination to aid in document related crime examination. The advantages and limitations of proposed analytical techniques are discussed.

Carbon↗

Melanogenesis in the ink gland of Sepia officinalis.

Among the various melanin-producing systems, the ink gland of the cuttlefish (Sepia officinalis) has traditionally been regarded as a most convenient model system for the studies of melanogenesis. The ink gland is a highly specialized organ with immature cells in the inner portion, from where the cells gradually mature, migrate towards the outer portion of the gland and become competent to produce melanin giving rise to particulate melanosomes. When cell maturation is complete, melanin is secreted into the lumen of the gland, accumulated into the ink sac and ejected on demand. Biochemical studies carried out over the past two decades have shown that the ink gland contains a variety of melanogenic enzymes, including tyrosinase, a peculiar dopachrome rearranging enzyme (which catalyses the rearrangement of dopachrome to 5,6-dihydroxyindole) and a peroxidase (presumably involved in the later stages of melanin biosynthesis). These enzymes are functionally interactive in close subcellular compartments of ink gland cells and appear to act in a concerted fashion during the process of melanogenesis in the mature portion of the gland. More recent studies have revealed that ink production and ejection are affected and modulated by the N-methyl-D-aspartate (NMDA)-nitric oxide (NO)-cyclic GMP (cGMP) signalling pathway. Glutamate NMDA receptor and NO synthase, the enzyme responsible for the synthesis of NO, have been detected by biochemical and immunohistochemical techniques in immature ink gland cells. Stimulation of NMDA receptors caused a marked elevation of cGMP levels, activation of tyrosinase and increased melanin synthesis in the mature portion of the gland, via the NO-guanylyl cyclase interaction. This signalling is also present in different regions of the nervous system in Sepia and in certain neural pathways controlling contraction of the ink sac sphincters and wall muscle in the ejection mechanism. Overall, these and other findings allowed elaboration of an improved model of melanin formation in Sepia, which underscores the complex interplay of melanogenic enzymes and regulatory factors, highlighting both the similarities and the differences with melanogenesis in mammals.

Animals↗

Differentiating writing inks using direct analysis in real time mass spectrometry.

Writing ink analysis is used in establishing document authenticity and the sources and relative ages of written entries. Most analytical methods require removing samples or visibly altering the document. Nondestructive, in situ analysis of writing inks on paper without visible alteration is possible using mass spectrometry with a new ion source called Direct Analysis in Real Time. Forty-three different black and blue ballpoint, black fluid, and black gel inks were examined. Both dyes and persistent but thermally labile components of the inks contribute to the mass spectra, principally as protonated molecules [M+H](+). Numerous ink components were identified from the spectra. The spectra were placed in a searchable library, which was then challenged with two spectra from each of the 43 inks. The best match for each of the challenge spectra was correct for all but one ink, which matched with a very similar ink by the same manufacturer.

Journal Article↗

Aplysia ink release: central locus for selective sensitivity to long-duration stimuli.

1. A behavioral and electrophysiological analysis of defensive ink release in Aplysia californica was performed to examine the response of this behavior and its underlying neural circuit to various-duration noxious stimuli. 2. Three separate behavioral protocols were employed using electrical shocks to the head as noxious stimuli to elicit ink release. Ink release was found to be selectively responsive to longer duration stimuli, and to increase in a steeply graded fashion as duration is increased. 3. Intracellular stimulation of ink motor neurons revealed that ink release is a linear function of motor neuron spike train duration, indicating that the selective sensitivity of the behavior to long-duration stimuli is not due to a nonlinearity in the glandular secretory process. 4. In contrast, electrophysiological examination of ink motor neuron activity in response to sustained head shock revealed an accelerating spike train. During the later part of the spike train, compound excitatory synaptic potentials show a positive shift in reversal potential. 5. Our results suggest a central locus for the mechanisms that determine sensitivity of inking behavior to stimulus duration. 6. In contrast to ink release, defensive gill withdrawal was found to be extremely sensitive to short-duration stimuli.

Aggression↗

Identification of surgical biopsy borders by use of india ink.

Separation of surgical biopsy borders from artifactual borders created during trimming of biopsy specimens is necessary to avoid misinterpretation of histologic borders. Misinterpretation of a contaminated trimming border as a surgical border may lead to additional surgery and excessive removal of normal tissue. Likewise, a neoplasm may regrow locally or metastasize if a surgical border infiltrated with neoplastic cells is falsely assumed to be an artifactual trimming border. The use of India ink for distinguishing between surgical biopsy borders and artifactual borders was evaluated. Ten normal tissue specimens from 8 types of tissue (skin, small intestine, urinary bladder, bone, muscle, lung, large intestine, and uterus) were obtained from freshly euthanatized dogs. The specimens were painted with India ink and examined for adherence of the ink to the cut surface of the specimen. Adherence of the ink was observed in all specimens with the exception of the cut surface of the lung. Twenty-five biopsy specimens from dogs with clinical cases of disease were similarly painted with India ink and evaluated. Twenty-two were identified as neoplastic and 3 as inflammatory lesions. Wedges of tissue were obtained from the center of the biopsy specimens to purposely create borders that contained neoplastic tissue. These positive controls were painted with India ink to evaluate the effect of the ink on the histologic appearance of the neoplastic cells. Distortion or alteration of the cellular architecture was not observed in any of the normal specimens, specimens from dogs with clinical cases, or positive controls. The use of India ink for delineation of biopsy borders is a simple technique that presents few technical difficulties.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Cosmetic tattoo ink darkening. A complication of Q-switched and pulsed-laser treatment.

BACKGROUND: High-energy, short-pulse lasers, eg, Q-switched lasers, emitting visible and near-infrared light have recently been developed for removing tattoos, with little risk of scarring. The mechanisms of action, and possible adverse effects other than scarring and hypopigmentation, are not fully understood. OBSERVATIONS: We describe five cases of pulsed-laser-induced, immediate, irreversible darkening of cosmetic, white, flesh (skin-color), and pink-red colored tattoos. Irreversible ink darkening can be an insidious complication, because immediate whitening of the skin temporarily obscures the subsequently impressive color change. Among these cases, irreversible ink darkening occurred with Q-switched ruby (694 nm), Q-switched neodymium (Nd):YAG (1064 nm/532 nm), and pulsed green dye (510 nm) lasers. Attempts to remove the darkened ink with further laser treatment failed in two cases, and surgical excision was necessary. In the other three cases, subsequent laser treatments successfully removed the darkened ink. The red cosmetic tattoo ink used in one of the cases was placed in agar in vitro and was converted to a black compound immediately on Q-switched ruby laser exposure. Ferric oxide, a brown-red ingredient commonly used in cosmetic tattoos, was similarly tested and blackened in vitro by Q-switched ruby laser exposures. CONCLUSIONS: Although most tattoos are not darkened by laser treatment, short-pulsed lasers over a wide spectrum can cause immediate darkening of some tattoo inks. Patients should be warned of the potential for irreversible cosmetic tattoo darkening, and test-site exposures should be performed prior to treatment. In some cases, subsequent laser treatments may remove the blackened ink. The mechanism probably involves, at least for some tattoos, reduction of ferric oxide (Fe2O3, "rust") to ferrous oxide (FeO, jet black), but the chemical reaction that is involved remains unknown.

Adult↗

Pollution-preventing anionic lithographic inks.

Lithographic printing presses use aliphatic and aromatic solvents for cleaning various surfaces, which are coated with the ink. The conventional printing inks also contain volatile solvents. During printing and cleaning operations, volatile organic compounds (VOCs) present in the inks and cleaning solvents are lost to the atmosphere by evaporation and these losses are quantifiable and alarmingly high. A new type of ink based on castor oil, which completely eliminates emissions of VOCs, has been developed. Synthesis of resins and the kinetics of washing of the ink, similar to the recently announced pollution-preventing ink [development of a VOC-free lithographic printing system. TAGA Proc., 324], with water at a slightly elevated pH are studied. A correlation between the mass-transfer coefficient characterizing the washing of ink and the experimental process variables is confirmed. New data are also reported on flake-formation dynamics.

Air Pollutants, Occupational↗

Dermatitis from aziridine hardener in printing ink.

13 of 51 workers developed dermatitis of the hands and face after handling a water-based ink containing a polyfunctional aziridine hardening agent. Improper work practices resulted in skin contamination with the ink and its ingredients. The aziridine hardener contained trimethylolpropane triacrylate (TMPTA). The incidence of dermatitis was highest among the ink mixers who handled the undiluted aziridine (6 of 8 workers affected), was lower among printers who handled ink containing 2 to 4% aziridine (7 of 22 workers affected), and was absent in workers who did not handle ink. The mean latency from first contact with the ink to the development of the rash was 3.2 months among the ink mixers, and 6.2 months among the printers. The present findings demonstrate the risk of handling aziridine hardeners when protective clothing is not properly used and when work practices result in direct skin contact. Further research should be performed to discern whether aziridine compounds themselves, free of TMPTA, can cause dermatitis.

Acrylates↗

Development of biometric DNA ink for authentication security.

Among the various types of biometric personal identification systems, DNA provides the most reliable personal identification. It is intrinsically digital and unchangeable while the person is alive, and even after his/her death. Increasing the number of DNA loci examined can enhance the power of discrimination. This report describes the development of DNA ink, which contains synthetic DNA mixed with printing inks. Single-stranded DNA fragments encoding a personalized set of short tandem repeats (STR) were synthesized. The sequence was defined as follows. First, a decimal DNA personal identification (DNA-ID) was established based on the number of STRs in the locus. Next, this DNA-ID was encrypted using a binary, 160-bit algorithm, using a hashing function to protect privacy. Since this function is irreversible, no one can recover the original information from the encrypted code. Finally, the bit series generated above is transformed into base sequences, and double-stranded DNA fragments are amplified by the polymerase chain reaction (PCR) to protect against physical attacks. Synthesized DNA was detected successfully after samples printed in DNA ink were subjected to several resistance tests used to assess the stability of printing inks. Endurance test results showed that this DNA ink would be suitable for practical use as a printing ink and was resistant to 40 hours of ultraviolet exposure, performance commensurate with that of photogravure ink.

Algorithms↗

[Gastroscopic mucosal biopsy and carbon ink injection marking for determination of resection line on the gastric wall in stomach cancer].

Gastroscopic mucosal biopsies and carbon ink injection marking were performed in 31 patients with gastric cancer before operation. The resection line of gastric wall was determined during operation according to the marked points. The method, dose, site, and opportunity of ink injection were studied. The results were: 1. None of these 31 patients had positive biopsies from cancer free areas. None of the resected specimens showed deep cancer infiltration beyond the site of carbon ink injection. This method is significant in recognizing the extent of intramucosal cancer infiltration; 2. None of the 31 patients marked by ink injection had residual cancer on the resected line whereas 8-10% of those unmarked had a positive margin. This result indicates that this method is significant in avoiding residual cancer on the resection line; and 3. Before surgery, the home-made carbon ink was satisfactory. The optimum dose for an ideal ink point was 0.05-0.1 ml/point which would give a marking of 0.5-1.5 cm in diameter on the serosa. The ink point was clearly shown on the anterior wall but less satisfactorily on the lesser curvature of the stomach. Injection performed as early as the fifth week before operation was valid.

Biopsy↗

Uptake of india ink particles and latex beads by corneal fibroblasts.

The fate of India ink particles and polystyrene latex beads injected into the corneal stroma of rabbits was studied by the naked eye, light microscopy, and electron microscopy. All the injected ink particles or latex beads were unchanged in shape, size, and number for at least 6 months. India ink particles and latex beads were endocytosed by the corneal fibroblasts within 3-4 days after injection. Numerous ink particles were packed into vacuoles, 0.5-10 micron in diameter, which occupy a large volume of the cytoplasm of the cell body and processes of fibroblasts in and near the injected area. Each latex bead, 0.72 micron in diameter, is usually enclosed in one vesicle, and a large number of vesicles are distributed throughout the cytoplasm. In corneal tissue removed 10 min after injection of India ink and cultured for 3 or 7 days, uptake of many ink particles by the fibroblasts was seen. By this experiment, the contribution of the blood-derived cells was completely excluded, and it is more distinctly shown that the corneal fibroblast has a strong endocytotic activity. The uptake and long-term storage of ink particles and latex beads by the corneal fibroblast are reactions that protect the organ without inflammation from the injury and harm by non-toxic foreign materials.

Animals↗

The uptake and long-term storage of India ink particles and latex beads by fibroblasts in the dermis and subcutis of mice, with special regard to the non-inflammatory defense reaction by fibroblasts.

The fate of India ink particles and polystyrene latex beads injected into the dermis and subcutis of the skin of the auricle and back in mice was observed with the naked eye, light microscopy and electron microscopy. The tattoo patterns made by injected ink particles remained essentially unchanged for life as observed with the naked eye. India ink particles and latex beads were endocytosed by fibroblasts as well as macrophages in the dermis and subcutis. Numerous ink particles or small latex beads (0.22 micron in diameter) were packed into vacuoles 0.1-10.0 micron in diameter which occupied a large volume of the cytoplasm of the cell body and processes of fibroblasts, whereas numerous particles and larger beads (0.22 and 2.0 micron) were taken up into the cell body of macrophages in the vicinity. Most fibroblasts, characterized by long cell processes and well developed rough endoplasmic reticulum, are easily distinguished from macrophages, the latter being round or oval in shape, and having many lysosomes and numerous irregularly shaped microvillous projections. It is believed that fibroblasts taking up and storing the ink particles or latex beads move poorly and are almost fixed in the connective tissue: the tattoos therefore do not change markedly. It is emphasized that the uptake and long-term storage of ink particles and latex beads by the dermal and subcutaneous fibroblasts represent a specific non-inflammatory defense mechanism that protects the living body, without immune reactions, against injuries and invasions by non-toxic foreign agencies. The histiocyte, a term proposed by KIYONO (1914) for a fixed macrophage on the basis of his studies using vital dye staining, is considered to include, in addition to true macrophages, fibroblasts showing endocytotic activities for small foreign bodies such as acid dyes, ink particles, and latex beads.

Animals↗

Anti-tumor activity of squid ink.

The anti-tumor activity of a new type of peptidoglycan isolated from squid ink was shown to have a cure rate of 64% for Meth A tumor from BALB/c mice. The ink delipidated in acetone, which contained the peptidoglycan at 0.1% (w/w), was administered to tumor-transplanted mice so as to examine the anti-tumor activity. One-fifth of the tumor-bearing mice was cured with 3 injections (1 mg/head) of the acetone delipidated squid ink or a prolongation of survival was observed in the treated animals. Heat treatment at 100 degrees C for 10 min did not affect the anti-tumor activity of the delipidated ink, its potentiality being preserved. The acetone-extractable fraction of the ink also brought about a similar cure rate for Meth A tumor. The delipidated ink enhanced the phagocytic activity of macrophages but no direct cytotoxicity was observed for the Meth A tumor cells. Hence it may be said that the anti-tumor activity of the delipidated ink was mainly due to the augmented cellular immunity in vivo.

Acetone↗

Systematic analysis of bulk blue ballpoint pen ink by FTIR spectrometry.

A classifying method on bulk blue ballpoint pen ink has been studied by Fourier transform infrared (FTIR) spectroscopy. By using this method, a total of 108 blue ink samples have been divided into two groups depending on their main component. Spectral characteristics of these inks such as frequency and absorbance are described by way of artificial intelligence of pattern recognition, and 35 subgroups from the 108 inks are distinguished by their correlation coefficient (lambda). Under heat or exposure to ultraviolet light a mode of change in the age of the inks has been obtained. This approach has provided a reliable and nondestructive method for the characterization of bulk ballpoint writing ink, and more importantly, it might be a basis for dating of the blue ballpoint pen ink.

Journal Article↗

Color separation of signature and stamp inks to facilitate handwriting examination.

The questioned documents laboratory often encounters cases where handwriting that is to be examined intersects with some interfering factor such as a rubber stamp, typewriting or background printing. In these cases, line direction, beginning and ending features of letters and other fine details of the handwriting may be lost in the "noise" of the intersecting ink. The purpose of this paper is to show several new digital photography methods that may be used to "subtract" the effect of the intersecting ink, thereby enhancing that of the handwriting ink in order to enable the document examiner to conduct a complete examination. These methods have the advantage of being fast and do not involve the use of expensive material or equipment. Several new methods are described that may be used to separate the colors of the handwriting ink from that of the intersecting ink: the analog method and several digital methods such as RGB-HSB-CMYK, L*a*b color, and color separation using the Channel Mixer function of Adobe Photoshop. Successful application of these color separation methods to specific handwriting ink/rubber stamp ink color combinations shows that the effect of the intersecting ink may indeed be minimalized if not canceled altogether. Application of the suggested methods may well make the difference between a nonconclusive handwriting examination and a full analysis of the questioned handwriting.

Journal Article↗