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Emission of ink aerosol from ink-jet recorders.

As a follow-up of two patients found to be allergic to EKG*ink, a study was made, using spectrophotometric and light microscopic methods, of the factors influencing the emission of ink aerosol from ink-jet recorders used in electrocardiographs. Ink aerosol concentrations were only detectable by spectrophotometry under extreme conditions. However, the light microscopic method was sufficiently sensitive. At the slow paper speeds normally used (10-25 mm/sec) and under normal working conditions, the concentration of ink aerosol near the recorder was less than 0.1 microgram/m3; but aerosol emission increased exponentially with increasing ink pressure, i.e. increasing paper speed. At the highest paper speed studied (200 mm/sec), the aerosol concentration 36 cm from the ink nozzles was about 65 micrograms/m3. An increase in the distance from the ink nozzles to the chart paper resulted in a decrease in ink aerosol emissions. Aerosol dissipation was decreased by local exhaust ventilation of the ink-jet recorder. However, a protective shield designed to catch emitted ink aerosol only reduced the aerosol concentration at the measuring point at slow paper speeds. At higher paper speeds, the concentration increased. The average ink aerosol concentration near modern electrocardiographs is low, but some EKG operations may result in high peak concentrations. The formation of ink aerosol could be considerably reduced by changing the rheologic properties of the ink.

Aerosols

Inking in Aplysia californica. III. Two different synaptic conductance mechanisms for triggering central program for inking.

1. In the previous two papers we described the motor elements which mediate all-or-none inking behavior in Aplysia (4) and the contribution of specific membrane properties of the ink-gland motor cells to the central program for inking (5). In the present paper we show that the central program for inking (a characteristic accelerating burst of action potentials in the motor cells) can be triggered in two different ways: 1) by means of a suprathreshold stimulus delivered to a single site on the animal; and 2) by means of two subthreshold stimuli closely spaced in time, delivered either to the same or to different sites on the animal. 2. The single suprathreshold stimulus triggers the central program primarily by means of a complex increased-conductance EPSP. The two subthreshold stimuli trigger the program by means of a complex decreased-conductance EPSP which is capable of augmenting subsequent afferent synaptic input. 3. The synaptic augmentation produced by the decreased-conductance EPSP is attributable to two properties of the EPSP: 1) it increases the input resistance of the ink-gland motor cells so that the synaptic current from afferent input onto the cells produces larger, more effective EPSP's; and 2) it increases the electrotonic coupling among the motor cells, thereby producing increased positive feedback among them. Thus, the decreased-conductance EPSP provides a novel mechanism for spatial and temporal summation.

Animals

Higher susceptibility of mast-cell-deficient W/WV mutant mice to brain thromboembolism and mortality caused by intravenous injection of India ink.

(WB X C57BL/6)F1-W/WV mice possess a genetic defect in multipotential hematopoietic stem cells; the mice are anemic and lack mast cells. The authors injected diluted India ink intravenously into W/WV mice and congenic normal +/+ mice and searched for genetically determined differences in the development of complications of the injection. In both W/WV and +/+ mice, intravenous ink resulted in thrombocytopenia and markedly prolonged bleeding times, as well as prolonged partial thromboplastin and prothrombin times and reduced fibrinogen concentrations. These effects were similar in W/WV and +/+ mice, although the reduction in platelet counts was greater in W/WV mice. In addition, the mortality associated with ink injection was significantly higher in W/WV mice than in congenic +/+ mice. Most W/WV mice which died first exhibited paralysis, and examination under the dissection microscope revealed that ink injection resulted in significantly more cerebral thromboemboli in W/WV mice than in +/+ controls. Bone marrow transplantation from +/+ mice corrected both the mast cell deficiency and the anemia of W/WV mice and protected the W/WV recipients from the adverse consequences of ink injection. By contrast, +/+ mice rendered as anemic as W/WV mice by breeding did not exhibit increased morbidity and mortality after ink injection. (WC X C57BL/6)F1-Sl/Sld mice, which are anemic and lack mast cells because of a genetic defect different from that of W/WV mice, also exhibited increased morbidity and mortality after intravenous ink. Finally, mixture of ink with commercial heparin prior to intravenous injection markedly reduced the incidence of cerebral thromboembolism and death in W/WV mice. Taken together, these findings suggest that the increased morbidity and mortality exhibited by W/WV and Sl/Sld mice that received injected ink might be related to their mast cell deficiency rather than to their anemia. But measurement of the histamine content of the blood and various tissues of WBB6F1-+/+ mice injected with ink, and examination of their tissues in 1-mu sections, indicated that intravenous ink did not cause substantial mast cell degranulation. As a result, the possibility that mast cells protect +/+ mice from the adverse effects of intravenous ink by a mechanism other than degranulation and release of heparin, or that the differences in the response of W/WV or Sl/Sld mice and their +/+ littermates are due to defects other than their lack of mast cells, cannot be excluded.

Animals

Flow and distribution of India ink in microvessels of the frog.

Velocities of an India ink front and of RBCs moving ahead of it were studied by intravital microscopy in capillaries of frog mesentery and skeletal muscle. Measurements were made during microperfusion of single vessels or groups of connected vessels in mesentery, and following intravenous ink injection in both tissues. The presence of ink did not appear to interfere with microvascular flow or vasomotion during the period of observation. On the average, the ink spearhead moved only slightly faster than the RBCs. There was substantial variation in relative velocities of RBCs in the same vessel and in the relative velocities of ink front and RBCs. The time course of ink filling showed substantial heterogeneity of flow in mesentery, more nearly uniform flow in skeletal muscle. Comparison of the measured velocity ratios of ink to RBCs with published observations on relative velocities of RBC to blood suggest that the advancing, apparently parabolic front of ink moves at less than twice the mean blood velocity. This is due in small part to diffusive dispersion of the ink particles in the laminar flow gradient, but more largely to stochastic dispersion of the front by interaction with RBCs and by displacement at branches.

Animals

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

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

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

[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

Filling of microcirculation in skeletal muscles during timed India ink perfusion.

Lower leg muscles of anesthetized rabbits were perfused in situ with heparinized India ink at flows and pressures comparable to normal resting levels of 4 ml.min-1.100 g-1). Paired cross sections were counterstained with eosin to show ink-containing microvessels and reacted for alkaline phosphate to show all vessels. The fraction of microvessels filled with ink (Fi) increased progressively with perfusion time. At 3.5 s, mean Fi for the muscles studied fell between 0.12 and 0.19. At 60-90 s, the following levels were reached: medial gastrocnemius 0.74, lateral gastrocnemius 0.76, tibialis anterior 0.59, and soleus 0.80. The number of open capillaries and their distribution of flow velocities can be inferred from such data only if the perfusion rate is known or by recourse to a specified anatomic model. The time course of ink appearance shows best agreement with 60-80% of the vessels open and accessible to ink, with microvascular transit times ranging from less than 3 to greater than 30 s. If microvascular path lengths are assumed to be uniform, the range of velocities must be four times to one-fourth the mean, with 15-30% of the microvessels perfused at velocities equal to or greater than the mean. Alternatively, if microvascular velocity is assumed uniform, flow path lengths must vary from one-fourth to four times the average. Anatomic measurements of other suggest that less than one-half the variability in ink transit is attributable to differences in microvascular length. Thus both length and velocity must vary among alternate arteriovenous pathways.

Animals

India ink staining of proteins on nylon and hydrophobic membranes.

India ink was found to be an acceptable stain for proteins blotted or dotted onto positively charged nylon or hydrophobic membranes. The hydrophobic membrane, Immobilon, was an outstanding matrix for binding proteins and displayed low levels of background staining. The least amount of protein detected by india ink staining was between 1.0 and 10 ng. India ink staining of proteins on nylon membranes is an easy, inexpensive, and quick method for the unequivocal detection of both standards and unknowns in the same blot. However, inks, ink concentrations, fixing conditions, staining times, pH, washing conditions, and membrane lots all need to be controlled to achieve maximum sensitivity for protein detection following india ink staining.

Blotting, Western