Impact of reducing iodinated intravenous contrast volume in brain CT on image diagnostic quality.

PURPOSE: To investigate whether reducing the volume of intravenous iodinated contrast material injected during brain computed tomography (CT) provides reliable and accurate imaging without compromising diagnostic accuracy. METHODS: This prospective study enrolled patients undergoing enhanced brain CT at a single tertiary hospital. Subjects who agreed to participate received a reduced dose of 60 ml contrast. The images were compared to an age and gender-matched control group who received the conventional 80 cc dose. Neuroradiologists assessed image quality and interpretation using a 5-point Likert scale with six specific domains. Based on ICC, inter-rater reliability was high at 0.873. Multiple linear regression predicted overall diagnostic accuracy based on contrast dose, age, and gender. Visual Grading Characteristics (VGC) analysis was also performed to quantify regional brain enhancement differences between the two contrast groups. RESULTS: The study included 47 patients in the 60 cc group and 55 in the 80 cc control group. The results showed the 80 cc group had significantly higher enhancement ratings compared to 60 cc for all six structures assessed. The differences between groups ranged from -0.241 to -0.433 (p < 0.001) on the 5-point scale.The VGC analysis confirmed significantly greater brain parenchymal enhancement in the 80 cc group compared to the 60 cc group. CONCLUSION: The findings indicate that reducing the intravenous iodinated contrast material volume during brain CT from 80 cc to 60 cc leads to a statistically significant reduction in image quality and diagnostic accuracy. Further research with larger cohorts is needed to confirm these findings and assess the clinical impact of these differences.

Evaluation of Artificial Intelligence-Powered Identification of Large-Vessel Occlusions in a Comprehensive Stroke Center.

BACKGROUND AND PURPOSE: Artificial intelligence algorithms have the potential to become an important diagnostic tool to optimize stroke workflow. Viz LVO is a medical product leveraging a convolutional neural network designed to detect large-vessel occlusions on CTA scans and notify the treatment team within minutes via a dedicated mobile application. We aimed to evaluate the detection accuracy of the Viz LVO in real clinical practice at a comprehensive stroke center. MATERIALS AND METHODS: Viz LVO was installed for this study in a comprehensive stroke center. All consecutive head and neck CTAs performed from January 2018 to March 2019 were scanned by the algorithm for detection of large-vessel occlusions. The system results were compared with the formal reports of senior neuroradiologists used as ground truth for the presence of a large-vessel occlusion. RESULTS: A total of 1167 CTAs were included in the study. Of these, 404 were stroke protocols. Seventy-five (6.4%) patients had a large-vessel occlusion as ground truth; 61 were detected by the system. Sensitivity was 0.81, negative predictive value was 0.99, and accuracy was 0.94. In the stroke protocol subgroup, 72 (17.8%) of 404 patients had a large-vessel occlusion, with 59 identified by the system, showing a sensitivity of 0.82, negative predictive value of 0.96, and accuracy of 0.89. CONCLUSIONS: Our experience evaluating Viz LVO shows that the system has the potential for early identification of patients with stroke with large-vessel occlusions, hopefully improving future management and stroke care.

Localized effects of microwave radiation on the intact eye lens in culture conditions.

A novel experimental system was used to investigate the localized effects of microwave radiation on bovine eye lenses in culture for over 2 weeks. Using this setup, we found clear evidence that this radiation has a significant impact on the eye lens. At the macroscopic level, it is demonstrated that exposure to a few mW at 1 GHz for over 36 h affects the optical function of the lens. Most importantly, self-recovery occurs if the exposure is interrupted. At the microscopic level, close examination of the lens indicates that the interaction mechanism is completely different from the mechanism-causing cataract via temperature increase. Contrary to the latter's effect, that is particularly pronounced in the vicinity of the sutures and it is assumed to be a result of local friction between the edges of the fibers consisting the lens. Even if macroscopically the lens has recovered from the irradiation, microscopically the indicators of radiation impact remain.

Long-term lens organ culture system with a method for monitoring lens optical quality.

The Scan Tox System is a method for monitoring lens optical quality (focus or lack of focus) in culture conditions, which mimic conditions inside the eye. The ocular lens is an ideal organ for long-term culture experiments because it has no direct blood supply and no connection to the nervous system. The Scan Tox System makes it possible to keep lenses for long-term studies of up to a few weeks. The use of cultured lenses, mainly bovine, replaces the need for testing the effects of potentially damaging agents on live animals. This optical monitoring apparatus uses a computer-operated scanning laser beam, a video-camera system and a video frame analyzer to record the focal length and transmittance of the cultured lens. The scanner is designed to measure the focal length at points across the diameter of the lens. The lens container permits the lens to be exposed to a vertical laser beam from below. The laser source projects its light onto a plain mirror, which is mounted at 45 degrees C on a carriage assembly. The mirror reflects the laser beam directly up through the test lens. The mirror carriage is connected to a positioning motor, which moves the laser beam across the lens. The camera sees the cross section of the beams and, by examining the image at each position of the mirror, Scan Tox software is able to measure the quality of the lens by calculating the back vertex distance for each beam position. The cultured lenses continue to maintain their original refractive function. When foreign substances are introduced to a cultured lens, the Scan Tox System measures the resulting optical response. This provides a very sensitive means to follow early damage to the eye lens. Because the lens is maintained in an intact state in solutions that are similar to those inside the eye, the lens retains its normal recuperative powers. So in addition to measuring early damage, this system allows measurement of recovery from damage.

UV-A-related alterations of young and adult lens water-insoluble alpha-crystallin, plasma membranous and cytoskeletal proteins.

The damaging effects of UV-A irradiation on lens water-insoluble alpha-crystallin, plasma membranous and cytoskeletal proteins derived from bovine lenses were studied. Young and adult bovine lenses were kept viable for 2 months in organ culture. After 24 h of incubation they were irradiated, and analyses of the proteins by one-dimensional and two-dimensional gel electrophoresis followed by Western blotting were carried out at several time intervals. RNA isolation, PCR and Northern blotting were also performed. We identified age-related changes in water-insoluble alpha-crystallin, the major membrane protein MP26 and the cytoskeletal proteins vimentin, phakinin and actin between control and UV-irradiated lenses. It appeared that adult lenses are more susceptible to UV light than young lenses, and protein modification occurred more frequently in adult lenses. UV-A irradiation affects not only the cytoskeletal structure, as deduced by the abnormal arrangement of actin in the fiber cells, but also leads to degradation of actin mRNA. Furthermore, analysis of the expression of hsp25 and hsp70 revealed some alteration in the protein pattern of adult lenses. We suggest that degradation of the cytoskeletal proteins following irradiation is due to, at least in part, the decreased protective ability of heat shock proteins upon aging.

In vitro filament-like formation upon interaction between lens alpha-crystallin and betaL-crystallin promoted by stress.

PURPOSE: To determine whether alpha-crystallin is capable of forming filament-like structures with other members of the crystallin family. METHODS: Water-soluble crystallins were isolated from calf lenses and fractionated into alpha-, betaH-, betaL-, and gamma-crystallins according to standard procedures. Chaperone-like activity of alpha-crystallin was determined in control and UV-A-irradiated lenses by the heat-induced aggregation assay of betaL-crystallin. Protein samples from this assay were analyzed by electron microscopy. In vitro filament formation was examined by transmission immunoelectron microscopy using specific antibodies directed against the crystallins. Involvement of intermediate filament constituents was excluded by the results of Western blot analysis, which were all negative. Moreover, the in vitro amyloid fibril interaction test using thioflavin T (ThT) was also performed. RESULTS: At the supramolecular level heating at 60 degrees C has no effect on the morphologic appearance of alpha-crystallin as observed by transmission electron microscopy. Moreover alpha-crystallin obtained from UV-A-irradiated lenses shows a virtually identical shape. However, heating in the presence of betaL-crystallin results in the formation of filament-like alphabeta-hybrids as demonstrated by immunoelectron microscopy using specific antibodies directed either against alpha- or betaL-crystallin. Parallel experiments with alpha-crystallin derived from UV-A-irradiated lenses showed even more pronounced filamentous structures, compared with the controls. Nonetheless, we were able to show that the UV-light treatment affected the chaperone-like capacity of alpha-crystallin, as revealed by a diminished ability to inhibit in vitro denaturation of betaL-crystallin. To exclude the presence of cytoskeletal contamination in the crystallin preparations, vimentin antibodies were also tested. These latter experiments were negative. The filamentous nature of the hybrids was further confirmed by the results obtained with the ThT assay earlier applied for the detection of amyloid fibrils. CONCLUSIONS: Crystallin hybrids have previously been detected in the water-soluble lens crystallin fraction. Our findings indicate that such endogenous hybrids, formerly called "rods," may result from stress-induced interaction between alpha-crystallin and other lens constituents such as betaL-crystallin. Because the hybrid formation is enhanced when alpha-crystallin from UV-A-irradiated lenses is used as one of the two components of the hybrid, one can only speculate that this formation may be one of the factors leading to UV-A cataract.

Analysis of UVA-related alterations upon aging of eye lens proteins by mini two-dimensional polyacrylamide gel electrophoresis.

This study is a first approach to identify UVA-related alterations in situ of bovine eye lens proteins from the water-soluble and urea-soluble fractions upon aging. The fractions were obtained from irradiated long-term organ culture lenses and analyzed by mini two-dimensional gel electrophoresis. This micropreparative method followed by computer analysis allows high resolution and separation of microgram quantities of proteins and to detect spots which arose as a consequence of irradiation. To facilitate the analysis we first separated the water-soluble fraction into the major crystallin classes by gel filtration. Moreover, we immunoblotted the gel of the urea-soluble fraction with a specific antibody against the intermediate filament protein vimentin. Upon irradiation of young and adult lenses, alphaA-crystallin and vimentin showed obvious modifications. During aging the susceptibility to irradiation increased when vimentin started to degrade, whereas deamidation of alphaA-crystallin seems to occur.

Effect of UV-A light on the chaperone-like properties of young and old lens alpha-crystallin.

PURPOSE: To study the damaging effect of UV-A irradiation on the chaperone-like properties of alpha-crystallin and the subsequent recovery process of young and old bovine lenses. METHODS: Young and old bovine lenses were kept in organ culture. After 24 hours of incubation they were irradiated with UV-A at 365 nm, and optical quality measurements were performed during the experiments (192 hours). alpha-Crystallin and alpha1-, alphaA2-, alphaB1-, and alphaB2-crystallin subunits were analyzed, separated by gel filtration and cation exchange chromatography, respectively, after different culture times. Protein patterns were obtained after two-dimensional (2-D) gel electrophoresis. Chaperone-like activity was determined on the basis of insulin B-chain and betaL-crystallin aggregation assays. Aggregation of alpha-crystallin was analyzed, tryptophan fluorescence measurements were performed, and alpha-crystallin mRNA levels were determined. RESULTS: The water-soluble alpha-crystallin obtained from old lenses compared with young lenses after UV irradiation had decreased chaperone activity, a higher molecular weight, and increased loss of tryptophan fluorescence. Moreover, alpha-crystallin mRNA virtually disappeared, whereas extra spots on the 2-D protein pattern appeared, possibly because of deamidation. CONCLUSIONS: alpha-Crystallin obtained from old lenses is more affected by irradiation than alpha-crystallin derived from young lenses. Moreover, it appeared that alphaB-crystallin from UV-treated old lenses compared with control lenses was less susceptible to UV-A than alphaA-crystallin. It may well be that alphaB-crystallin protects alphaA-crystallin in vivo.

Effects of UV-A radiation on lens epithelial NaK-ATPase in organ culture.

PURPOSE: To investigate the mechanisms involved in the damage caused by UV-A irradiation at 365 nm on the eye lens. METHODS: Bovine lenses obtained from animals 1 to 5 years of age were placed in specially designed organ culture chambers for preincubation. Twenty-four hours later, the lenses were irradiated by 33 J/cm2 UV-A at 365 nm. During irradiation, the lenses were oriented in the culture so that the anterior surface faced the incident UV-A radiation source. After irradiation, lens optical quality was monitored throughout the 8 days of the culture period, and lens samples were taken for analysis of NaK-ATPase activity. RESULTS: Lens optics and NaK-ATPase activity were affected by irradiation of 33 J/cm2. The effects on lens epithelial NaK-ATPase activity were stronger at the equators than at the center. The damage to the activity at the center was reversible, as the lens optically recovered from the LW-A damage. CONCLUSIONS: Lens NaK-ATPase activity can recover from damage caused by UV-A at 365 nm. When the lenses received irradiation of 33 J/cm2, NaK-ATPase activity recovered from the damage during the culture period only at the center and not at the equators of the epithelium.

Transglutaminase involvement in UV-A damage to the eye lens.

Solar radiation is believed to be one of the major environmental factors involved in lens cataractogenesis. The purpose of the study was to investigate the mechanisms by which UV-A at 365 nm causes damage to the eye lens. Bovine lenses were placed in special culture cells for pre-incubation of 24 hr. The lenses were positioned so that the anterior surface faced the incident UV-A radiation source and were maintained in the cells during irradiation. After irradiation, lens optical quality was monitored throughout the culture period and lens epithelium, cortex and nuclear samples were taken for biochemical analysis. Transglutaminase activity in the lens was affected by the radiation. The activity of transglutaminase in lens epithelium cortex and nucleus increased as a result of the irradiation and then declined towards control levels during the culture period, as the lens recovered from the UV-A damage. Specific lens proteins, alpha B and beta B1 crystallins (the enzyme substrates) were analyzed by SDS polyacrylamide gel electrophoresis and immunoblotting with specific antibodies. Seventy-two hours after irradiation of 44.8 J cm-2 UV-A, alpha B crystallins were affected as was shown by the appearance of aggregation and degradation products. Some protein changes seem to be reversible. It appears that transglutaminase may be involved in the mechanism by which UV-A causes damage to the eye lens.

Long-term organ culture system to study the effects of UV-A irradiation on lens transglutaminase.

The purpose of the study was to investigate the mechanisms by which UV-A at 365 nm causes damage to the eye lens. Bovine lenses were placed in special culture cells and were oriented so that the anterior surface faced the incident UV-A radiation source. The lenses were maintained in the cells during irradiation. After 5 days in culture, samples of lens epithelium, cortex, and nucleus were taken for biochemical analysis. Transglutaminase activity in the lens was affected by the radiation and showed increased activity in the lens cortex and nucleus. Specific lens protein, betaB1 crystallin (one of the enzyme substrates), was analyzed by SDS polyacrylamide gel electrophoresis and immunobloting with specific antibodies. The crystallin was affected as was shown by the appearance of degradation products. There is correlative information but no clear proof that transglutaminase is involved in the mechanisms by which UV-A causes damage to the eye lens.

Recovery of lens optics and epithelial enzymes after ultraviolet A radiation.

PURPOSE: To establish the mechanism by which ultraviolet A (UVA) radiation causes irreversible damage to the eye lens. METHODS: The authors irradiated 223 bovine lenses in organ culture with 22.4, 33.6, and 44.8 J/cm2 of UVA radiation (365 nm) and studied biochemical and optical properties of the lenses in long-term culture conditions. Each lens tested was placed in a specially designed cell. The lenses were oriented so that the anterior surface faced the incident UVA radiation source, and they were maintained in their cells during irradiation. After irradiation, lens optical quality was monitored throughout the culture period, and lens samples were taken for enzyme analysis. RESULTS: Full recovery of lens optical damage and activity of the enzymes hexokinase, catalase, and glucose-6-phosphate dehydrogenase in lens epithelium was observed after 8 days in culture after irradiation with 22.4J/cm2. After irradiation with 33.6J/cm2, partial recovery of optical damage was found, and there was between 80% to 90% recovery of the enzyme activity. No recovery of optical and enzyme activity was found after 44.8J/cm2 irradiation. CONCLUSIONS: Irradiation between 22.4J/cm2 to 33.6J/cm2 of UVA at 365 nm is the minimal level of irradiation that causes irreversible damage to lens enzymes and optics.

Influence of 17 beta-estradiol and progesterone on rat ocular lens.

The effect of 17 beta-estradiol and progesterone on ocular lens in rats and untreated controls was studied. In the treated lenses, the activity of hexokinase and glucose-6-phosphate dehydrogenase remained unchanged. The activity of aldolase was increased in 18- and 20-month-old lenses as compared to controls. Aldose reductase activity was decreased at the age of 20 months (p < 0.001). Structural lens proteins studies by SDS polyacrylamide gel electrophoresis and immunodecoration with specific antibodies for crystallines alpha A + alpha B and beta + gamma suggest some protective effect in treated animals.

Long-term organ culture system to study the effect of UV-radiation on lens enzymes.

Environmental factors such as solar radiation and drug treatment are potential cataractogenic agents. It is suggested that their damaging effects accumulate with age. The purpose of the study was to isolate the effect of one factor (UV-radiation) and find out the mechanism by which UV radiation causes damage to the eye lens. We irradiated bovine lenses with UV-A (365 nm) radiation for 50, 75, 90, 100, and 120 min and followed the optical changes of the lenses in a long-term organ culture. Enzyme activities were analyzed in lens epithelium after five days of incubation in organ culture. The enzymes analyzed were ATPase, which belongs to the transport mechanism in lens epithelium cells, hexokinase, the key enzyme of the glycolysis pathway, G6PD, which provides NADPH to the glutathione system and catalase, which protects the cells from H(2)O(2). Optical damage was observed even for the minimal radiation. The same amount of radiation also affected ATPase and hexokinase activities. G6PD and catalase were affected only in lenses which received radiation for 90 min, We can conclude that enzymes involved in the transport mechanism and metabolism are more sensitive to UV-A (365 nm) radiation than enzymes involved in the defense mechanism against oxidation.

DL-propranolol inhibits lens hexokinase activity and affects lens optics.

A clinico-biochemical study indicated that the beta-blocker DL-propranolol may affect human lens epithelial hexokinase (HK) activity. In that study five key enzymes were analysed in 192 freshly excised human lens epithelia obtained during cataract surgery. In a large number of patients the epithelial HK was found to be inactive. Medical records of these patients showed widespread use of the drug DL-propranolol. In vitro experiments demonstrated a direct inhibitory effect of the drug on human lens HK activity. Lens refractive function was monitored during long term bovine lens culture experiments in which the potential cataractogenic agent was added to the culture media. DL-propranolol in a concentration of 0.1 mM reduced HK activity in bovine lens epithelium after 72 hr in organ culture and disrupted lens light focusing ability after 250 hr of incubation. Kinetic studies of HK inhibition suggested a competitive inhibitory effect of the drug on the enzyme.

Enzyme activities and crystallin profiles of clear and cataractous lenses of the RCS rat.

Royal College of Surgeons (RCS) rats have hereditary retinal degeneration in association with posterior subcapsular opacities. Cataract formation is thought to be correlated with an increase in lipid peroxidation products in the vitreous (Zigler and Hess, 1985). In order to examine the possibility that parallel changes in enzyme activity are occurring within the lens, we analysed the activity of four key enzymes and the crystallin protein profile. We compared RCS rat lenses at three different stages of cataract formation to clear lenses of the nonpigmented RCS rat, lenses from pigmented RCS rat and to normal (Fisher) rat. Our data shows that concomitant with the appearance of the RCS cataract, the ratio of the crystallins beta 1, beta H and gamma to the total lens protein was reduced. The crystallin profile of a clear RCS lens was similar to that of a normal (Fisher) lens. No significant difference in the activity of the enzymes hexokinase and glucose-6-phosphate dehydrogenase (G6PD) was found among the lenses, however the activity of glutathione reductase and aldolase was reduced in the cataractous lenses.

Effect of hydrogen peroxide, DL-propranolol, and prednisone on bovine lens optical function in culture.

Lens refractive function was monitored during long-term bovine lens culture experiments in which low concentrations of potentially damaging agents were added to the culture media. The agents tested were the drugs DL-propranolol and prednisone, and hydrogen peroxide. A computer-driven scanning laser system was used to monitor lens focal length during culture. The system consists of a scanning helium-neon laser beam, a television video camera, and a video frame digitizer. The system first locates the optical center of the lens, defined as the position of little or no refractive deviation of the beam. The laser scans the lens in small steps (0.05 mm), while the digitizer measures focal length for each beam position. A graphic profile of lens focal variation is plotted. A concentration of 0.1 mM hydrogen peroxide produces no significant change in lens focus after 315 hr of incubation. When the concentration is increased 10-fold (1.0 mM), lens focal ability is radically disturbed after 60 hr of incubation. Lenses incubated with prednisone (0.03 mM) for up to 525 hr show no difference in focal characteristics in comparison to control lenses. Propranolol in a concentration of 0.1 mM disrupts lens focal ability after 250 hr of incubation, whereas a higher concentration (1.0 mM) produces a similar effect after only 150 hr. These results indicate that lens refractive function is a sensitive measure of lens function during culture.

Localization of denatured enzyme molecules in rat lenses.

Immunohistochemical localization of altered enzyme molecules was detected by the use of antibodies to denatured enzymes (ADE) conjugated with fluorescein. Denatured aldolase, glucose 6-phosphate dehydrogenase and superoxide dismutase are mostly located in the subcortical region and in the nucleus of the rat lens. In the nuclear fibres the enzyme is located near the membrane of the fibres. This study provides additional evidence that altered enzyme molecules accumulate in the lens, and indicates their exact localization. ADE antibody can distinguish between inactive enzyme molecules and active ones, using immunohistochemical techniques.

Glutathione reductase in human lens epithelium: FAD-induced in vitro activation.

Glutathione reductase (GR) activity was measured with and without the addition of flavin adenine dinucleotide (FAD) in freshly excised human lens epithelium following cataract surgery, and in control eye bank lens epithelium. Large individual variations in activity were found in both groups. Out of 32 epithelia obtained from cataractous lenses, 14 showed no measurable GR activity. Activity in eight of these 14 epithelia was restored after FAD was added, implying that the apo-enzyme in these 8 epithelia was functional but FAD was not available. Another group of 8 active epithelia showed a significant increase in GR activity upon the addition of FAD, suggesting a mixed population of active enzyme and available fraction of apo-enzyme which was reactivated by the addition of FAD. Unusually high GR activities were observed in epithelia of several patients. This correlated principally with daily intake of thyroxine which is known to have a direct effect on the metabolism of riboflavin. The fact that in a significant number of cataract patients epithelial GR was not active, and the observation that activity could be restored by adding in vitro FAD, demands that more attention should be given to riboflavin nutritional status and FAD synthesis in the eye.