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ROSE: decision trees, automatic learning and their applications in cardiac medicine.

Computerized information systems, especially decision support systems, have acquired an increasingly important role in medical applications, particularly in those where important decisions must be made effectively and reliably. But the possibility of using computers in medical decision making is limited by many difficulties, including the complexity of conventional computer languages, methodologies, and tools. Thus a conceptual simple decision making model with the possibility of automating learning should be used. In this paper, we introduce a cardiological knowledge-based system based on the decision tree approach supporting the mitral valve prolapse determination. Prolapse is defined as the displacement of a bodily part from its normal position. The term mitral valve prolapse (PMV), therefore, implies that the mitral leaflets are displaced relative to some structure, generally taken to be the mitral annulus. The implications of the PMV are: disturbed normal laminar blood flow, turbulence of the blood flow, injury of the chordae tendinae, the possibility of thrombus's composition, bacterial endocarditis, and, finally, hemodynamic changes defined as mitral insufficiency and mitral regurgitation. Uncertainty persists about how it should be diagnosed and about its clinical importance. It is our deep belief that the echocardiography enables properly trained expert armed with proper criteria to evaluate PMV almost 100%. But, unfortunately, there are some problems concerned with the use of echocardiography. With this in mind, we have decided to start a research project aimed at finding new criteria and enabling the general practitioner to evaluate the PMV using conventional methods and to select potential patients from the general population. To empower doctors to perform needed activities, we have developed a computer tool called ROSE (computeRized prOlaps Syndrome dEtermination) based on algorithms of automatic learning. This tool supports the definition of new criteria and the selection of potential PMV-patients. The ROSE is based on concepts of decision trees and automatic learning. The decisions and learning process can be presented with an easily visualized two dimensional model; thus decision trees are straightforward to build and interpret. Decision trees use different object attributes to classify different subsets of objects. (Their great advantage is that they don't use a fixed number of predetermined attributes.) In the decision tree approach, the members of a set of objects are classified as either positive or negative instances (in our case patients with PMV Syndrome or without it). Candidate attributes that may possibly describe the concept are then outlined. A decision tree construction tool uses outlined attributes to formulate the appropriate decision tree that identifies all positive instances of the underlying concept according to objects with known classification. (In our case the classification is done with the echo examination). The first set of objects used for the tree generation is usually called the training set. This decision tree characterization next becomes a basis: 1) forecasting whether an object previously unseen is a positive or negative instance of the concept being modeled, and 2) the hierarchical representation of the most important attributes of the concept being investigated. Our main interest and idea is to discover symptoms, syndromes, and illnesses related to PMV that can be distinguished by general practitioners in their everyday job and which should help them to identify possible PMV candidate patients. To this end, we constructed a computerized tool called ROSE. According to the principles presented above, we first taught ROSE using the sample of 400 examined volunteers. ROSE, considering that the clinical PMV diagnosis is practically not researched, is relatively successful. (abstract truncated)

Algorithms↗

Encircling photothrombotic therapy for choroidal Greene melanoma using rose bengal.

The photosensitizing dye rose bengal in combination with an argon green laser (514.5 nm) operated at low power was evaluated in 49 rabbit eyes for treatment of experimental choroidal Greene melanoma by circumferential occlusion of the choroidal vasculature. The effects of no treatment, laser alone, and rose bengal alone were observed in 16 control eyes, all of which showed rapid tumor growth. Immediately following rose bengal injection, 3 minutes of continuous irradiation at 20.4 W/cm2 (500-microns spot, 40 mW) applied in three to four circumferential revolutions around the base of tumor nodules, without direct tumor irradiation, produced peripheral vascular occlusion and consequent tumor inhibition. Similar therapy at higher laser intensity (30.6 W/cm2) and with multiple retreatment sessions (28.0 to 30.6 W/cm2) resulted in increased tumor-inhibiting effect. Low-dose rose bengal phototherapy did not appear to directly damage ocular tissues adjacent to treatment areas; however, when multiple irradiation sessions were given within a short interval, an increased incidence of retinal detachment was observed.

Animals↗

Inhibition of vesicular glutamate uptake by Rose Bengal-related compounds: structure-activity relationship.

Synaptic vesicular accumulation of glutamate is a vital initial step in glutamate transmission. We have previously shown that Rose Bengal, a polyhalogenated fluorescein analog, is a potent inhibitor of glutamate uptake into synaptic vesicles. Here, we report the structural features of Rose Bengal required for this inhibition. Various Rose Bengal-related compounds, with systematic structural variations, were tested. Results indicate that the four iodo groups and the phenyl group attached to the xanthene moiety are critical for potent inhibitory activity. Replacement of these groups with two iodo groups and an alkyl group, respectively, results in substantial reduction in potency. Of further interest in creating high potency is the critical nature of the oxygen atom which links the two benzene rings of xanthene. Thus, the phenyl group and multiple iodo groups, as well as the bridging oxygen of xanthene, are crucial elements of Rose Bengal required for its potent inhibitory action.

Animals↗

Effect of sodium orthovanadate on the hepatobiliary clearance of rose bengal in streptozotocin-induced diabetic rats.

Sodium orthovanadate is known to promote glucose uptake in muscle and adipose tissues and has been suggested as a possible oral hypoglycemic agent. In addition, insulin-dependent diabetes has been shown to alter the hepatobiliary clearance of several drugs in rats. This study has determined whether orthovanadate, like insulin, can reverse diabetes-induced changes in the biliary excretion of endogenous bile acids and in the hepatobiliary clearance of rose bengal. Six groups of male Sprague-Dawley rats were used; normal, insulin-treated normal, vanadate-treated normal, diabetic, insulin-treated diabetic, and vanadate-treated diabetic. Diabetes was induced by injection of streptozotocin (45 mg/kg, i.v.). One week later, insulin (2-4 U/day, s.c.) and sodium orthovanadate (877 +/- 82 mumol/kg/day, p.o.) treatments were initiated. After 4 weeks, the clearance and biliary excretion of rose bengal (60 mumol/kg, i.v.) were determined for 3 hr. Bile flow rate, rose bengal excretion, and excretion of endogenous bile acids were unchanged in the two treated normal groups and in the insulin-treated diabetic rats. These parameters were increased in untreated diabetic and vanadate-treated diabetic rats as compared with normal. Pharmacokinetic analyses indicated that total and biliary clearances of rose bengal were increased in diabetic rats and that orthovanadate did not reverse these changes. However, liver weight and serum glucose concentrations were reduced by orthovanadate treatment. These data indicate that the oral insulinomimetic chemical sodium orthovanadate effectively reversed some, but not all, of the diabetes-induced alterations of hepatic function.

Animals↗

Cellular stress by light and rose bengal in human lymphocytes.

Human lymphocyte cultures were supplemented with 10(-8)-10(-4) M Rose Bengal and irradiated with fluorescent light (Philips 40-W daylight-fluorescent lamp, 380-550 nm) and chromosomal aberrations and catalase and superoxide dismutase activities in the cells were determined. Chromosomal lesions and both enzymatic activities increased additively or synergistically in human lymphocytes after Rose Bengal supplementation and fluorescent light irradiation. Chromosomal lesions (expressed as chromosomal aberrations/cells) were (a) 0.12 +/- 0.03, (b) 0.18 +/- 0.12, (c) 1.58 +/- 0.11 and (d) 3.20 +/- 0.17 in the following conditions: (a) control; (b) after 3 h light irradiation, (c) supplemented with 10(-5) M Rose Bengal and (d) with dye treatment and light irradiation. Superoxide dismutase activity was: (a) 16.5 +/- 1.5; (b) 19.5 +/- 1.2; (c) 29.2 +/- 1.5 and (d) 35.4 +/- 2.1 U/mg protein and catalase activity was: (a) 10.3 +/- 0.5, (b) 12.8 +/- 0.7, (c) 22.4 +/- 0.5 and (d) 27.6 +/- 1.1 U/mg protein in the same experimental conditions. These findings suggest that Rose Bengal supplementation plus fluorescent light irradiation of human lymphocytes lead to the synthesis of superoxide dismutase and catalase in a manner similar to the heat-shock response. A threshold of chromosomal damage (about 2 chromosomal aberrations/cell) is apparently required to activate oxidative stress genes.

Adult↗

Comparison of fluorescein and rose bengal staining.

The authors have recently reported that rose bengal is not a vital dye, and stains whenever the cultured cells are not covered by such components as albumin and mucin, and such a tear substitute as carboxycellulose. In this report, using cultured cells as well as normal rabbit corneas, they characterize and correlate the staining differences between rose bengal and fluorescein with the differences in their chemical structures. Fluorescein differs from rose bengal in its lack of intrinsic toxicity, photodynamic action, and ability to be blocked by the above-mentioned substances. Fluorescein staining is increased by rapid stromal diffusion and hence can manifest whenever there is disruption of cell-cell junctions. In contrast, rose bengal staining ensues whenever there is deficiency of preocular tear film protection. These experimental data may help interpret the clinical staining properties of these two dyes and enhance the understanding of the pathogenesis of various ocular surface disorders.

Animals↗

Lid imbrication syndrome. Diagnosis with rose bengal staining.

BACKGROUND: Lid imbrication syndrome is an abnormality of lid apposition in which the upper lid overlies the lower lid. Patients often complain of irritation, tearing, and foreign body sensation. The condition may be difficult to diagnose. METHOD: Twenty-one patients with suspected lid imbrication syndrome were compared with 21 age-matched controls. All patients were given 0.5% topical rose bengal. RESULTS: The diagnosis of lid imbrication syndrome was confirmed by the presence of rose bengal staining of the tarsal conjunctiva of the upper lid margin. The amount of rose bengal staining correlated with the severity of lid imbrication. Eighteen of 21 patients had received a diagnosis previously of dry eye syndrome. Four patients had persistent epithelial defects. CONCLUSIONS: Rose bengal staining of the superior lid margin tarsal conjunctiva offers an extremely reliable aid for diagnosing lid imbrication syndrome. The condition commonly presents as a dry eye or persistent epithelial defect. Treatment ranges from viscous tear substitutes to horizontal lid shortening of the upper lid.

Administration, Topical↗

Correlation of goblet cell density and mucosal epithelial membrane mucin expression with rose bengal staining in patients with ocular irritation.

PURPOSE: This study was designed to compare goblet cell densities and mucosal epithelial membrane mucin (MEM) expression in impression cytology specimens obtained from control subjects and patients with one of the following clinically defined diseases: aqueous tear deficiency (ATD) associated with Sjögren syndrome, ATD not associated with Sjögren syndrome, inflammatory Meibomian gland disease associated with rosacea, and Meibomian gland atrophy. These data were correlated with ocular surface rose Bengal staining scores, Schirmer scores, and HLA-DR antigen staining of conjunctival epithelial cells. METHODS: Goblet cell density and MEM expression were studied by impression imprints with immunohistochemical staining using an anti-mucosal epithelial membrane mucin antibody in the temporal and inferior bulbar and inferior tarsal conjunctiva of study subjects. RESULTS: Goblet cell density adjacent to the temporal limbus was significantly reduced at 3 mm posterior to the temporal limbus in both aqueous tear deficiency groups compared with the other groups and in patients with Sjögren syndrome compared with all other groups. In the inferior tarsus, goblet cell density was significantly reduced in patients with non-Sjögren syndrome ATD as compared with all other groups, except those with inflammatory Meibomian gland disease. Mucosal epithelial membrane mucin expression in the bulbar and tarsal conjunctiva was absent in a greater percentage of patients with Sjögren syndrome compared with all other groups. Total ocular surface rose Bengal staining scores were significantly higher in patients with Sjögren syndrome as compared with all other groups and in patients with non-Sjögren syndrome ATD as compared with control groups. Rose Bengal staining scores and Schirmer I test results (without anesthesia) were inversely correlated with bulbar, but not tarsal, conjunctival goblet cell densities, and with the absence of bulbar conjunctival MEM expression. CONCLUSIONS: These results suggest that reduced goblet cell density and mucosal epithelial cell mucin expression could explain increased rose Bengal staining in patients with aqueous tear deficiency. In addition, MEM may be regarded as a marker for normal differentiation of ocular surface epithelia, with its absence signifying the development of squamous metaplasia.

Adult↗

Laser intensity and wavelength dependence of Rose-Bengal-photosensitized inhibition of red blood cell acetylcholinesterase.

The intensity and wavelength-dependence of Rose-Bengal-mediated photoinhibition of red blood cell acetylcholinesterase has been studied. Irradiation of dye-membrane suspensions with 308 nm laser excitation resulted in enzyme inhibition almost 50% greater than that obtained with 514 nm laser excitation. Sodium azide and argon purging greatly decreased the photosensitized enzyme inhibition at both wavelengths. Although Rose Bengal photosensitized enzyme inhibition more efficiently upon excitation into Sn (308 nm) than into S1 (514 nm), Stern-Volmer analysis of sodium azide quenching data gave similar quenching efficiencies at both wavelengths. Irradiation of dye-membrane suspensions with increasing intensities (Nd:YAG, 532 nm, 40 ps pulse duration) resulted in a decrease in enzyme inhibition. Saturation of the Rose Bengal fluorescence intensity and light transmission occurred with nearly the same intensity-dependence, suggesting that ground-state depletion occurs at the higher intensities. Our results demonstrate that excitation of a sensitizer into higher-lying excited singlet states can result in enhanced sensitizing efficiency. However, attempts to populate such states in Rose Bengal by sequential two-photon absorption using high intensities resulted only in ground-state depletion.

Cholinesterase Inhibitors↗

Site-directed photochemical disruption of the actin cytoskeleton by actin-binding Rose Bengal-conjugates.

The in situ light-induced, non-enzymatic digestion of cytoskeletal actin by a xanthene dye conjugated to heavy meromyosin, anti-actin antibodies and/or anti-myosin antibodies is reported. The dye Rose Bengal was conjugated to either anti-actin antibodies, anti-myosin antibodies or heavy meromyosin. Under our experimental conditions, visible light induced the non-enzymatic breakdown of cytoskeletal actin when mammalian tissue culture cells were probed either with Rose Bengal-conjugated anti-actin and/or anti-myosin antibodies. Similar results were obtained when tissue culture cells were probed with Rose Bengal-conjugated heavy meromyosin before irradiation with visible light. The in situ photochemical reaction depended on the presence of actin-binding Rose Bengal-conjugates.

Actins↗

Toxic effect of rose bengal dye on the living human corneal epithelium.

PURPOSE: To investigate whether rose bengal dye at a concentration of 1% has a toxic in vivo effect on the human corneal epithelium. MATERIAL AND METHODS: 80 patients with presumed keratoconjunctivitis sicca (KCS) were examined with break-up time, Schirmer 1 test, and staining with fluorescein sodium and rose bengal. The corneal surface was observed in the slit lamp and photographed by means of non-contact photomicrography. RESULTS: After the application of rose bengal 1% in eyes preinstilled with fluorescein sodium 1% myriads of green dots appeared on the corneal surface in all 32 patients with KCS according to Copenhagen criteria, in 17/18 with low Schirmer 1 test values only, and in 21 of the remaining 30 in whom the KCS diagnosis wasn't verified. CONCLUSIONS: The appearance of green microdots after the application of rose bengal was probably due to the interruption of intercellular junctions and the penetration of fluorescein stained fluid below the superficial cells. Whether this phenomenon reveals an unhealthy corneal surface and thus has a clinical significance has to be further investigated.

Adult↗

Photoinactivation of herpes simplex virus by rose bengal and fluorescein. In vitro and in vivo studies.

Rose bengal and fluorescein are photosensitive dyes in widespread use in the evaluation of ocular surface diseases, including herpes simplex virus (HSV) keratitis. These dyes have recently been shown to penetrate living cells, and rose bengal was previously reported to possess antiviral activity. Several experiments reported herein suggest that these dyes do possess the potential for potent antiviral activity against extracellular virus, but only in the presence of light. Rose bengal is substantially more effective in vitro than fluorescein, and the effect is greater with increasing concentration of dye and duration of light exposure. Electron microscopic evaluation of treated virus showed no structural difference from untreated virus, in spite of 4- to 5-log decreases in virus titer. Intracellular virus was found to be markedly resistant to photoinactivation. In a rabbit model of acute primary HSV keratitis, daily application of topical rose bengal followed by light exposure had no therapeutic effect, although an adverse effect on culture sensitivity testing was seen.

Animals↗

Localization of rose bengal, aluminum phthalocyanine tetrasulfonate, and chlorin e6 in the rabbit eye.

PURPOSE: The localization and site of action of photosensitizers in the eye may be important for photodynamic therapy for fundus disorders but remain poorly understood for most agents. We investigated the intraocular localization of xanthene, phthalocyanine, and chlorin photosensitizers by using fluorescence microscopy and digital fundus fluorescence angiography. METHODS: Rose bengal (40 mg/kg), aluminum phthalocyanine tetrasulfonate (CASPc) (5 mg/kg), or chlorin e6 (2 mg/kg) was intravenously administered to albino rabbits. The eyes were enucleated and examined by means of fluorescence microscopy 5, 20, 60, and 120 minutes and 24 hours after dye injection. In vivo digital fundus fluorescence angiography with use of rose bengal (2-4 mg/kg), CASPc (2 mg/kg), and chlorin e6 (2 mg/kg) was performed. RESULTS: For all agents studied pathologically, there was moderate fluorescence from the choroid and retinal pigment epithelium 5 minutes after dye injection. Mild fluorescence detected from the photoreceptor outer segments at 5 minutes was increased at 20 minutes. Angiographic studies with use of rose bengal, CASPc, and chlorin e6 revealed differences in the pattern and rate of photosensitizer accumulation. CONCLUSIONS: Rose bengal, CASPc, and chlorin e6 accumulate rapidly in the choroid and retinal pigment epithelium and less rapidly in the outer retina. Differences in ocular localization of these photosensitizers were demonstrated. The significance of these findings for potential photodynamic therapy with these agents requires further investigation.

Animals↗

The cytotoxic and photodynamic inactivation of micro-organisms by Rose Bengal.

Rose Bengal was cytotoxic to the following bacteria at the concentrations given in parentheses (highest concentrations of dye in mol/l at which growth occurred on nutrient medium): Brochothrix thermosphacta and Deinococcus radiodurans (1 X 10(-6) or less); Streptococcus, Micrococcus, Staphylococcus, Bacillus, Arthrobacter and Kurthia spp. (1 X 10(-5)-1 X 10(-4], and Pseudomonas spp. and Enterobacteriaceae (5 X 10(-3)-1 X 10(-2) or greater). These organisms were killed rapidly when suspended in illuminated (170 microE/m2/s) solutions of Rose Bengal (1 X 10(-4) mol/l) providing oxygen was present. Singlet oxygen was identified as the lethal agent, because the rate of killing was increased by dissolving the dye in deuterium oxide while the organism were protected against photoinactivation by L-histidine or crocetin. Yeasts from chilled foods were killed in illuminated solutions of Rose Bengal but a light intensity of 315 microE/m2/s was needed for a death rate comparable with that of bacteria. The yeasts present in a range of chilled meat and dairy products failed to form colonies on Rose Bengal (5 X 10(-5) mol/l) media exposed continuously to modest illumination (55-80 microE/m2/s).

Bacteria↗

Rose Bengal test: diagnostic yield and use for the rapid diagnosis of human brucellosis in emergency departments in endemic areas.

The aim of the present study was to analyse the diagnostic yield of the rose Bengal test for the rapid diagnosis of human brucellosis in an emergency department in an area where the disease is endemic. The study included 711 patients diagnosed initially with brucellosis and 270 controls. Brucellosis patients were divided into three groups: group I, individuals with no regular exposure to or history of brucellosis; group II, individuals exposed repeatedly to Brucella infection; and group III, individuals infected with Brucella who had received appropriate treatment during the previous 12 months. Blood cultures were positive for 445 (62.6%) brucellosis patients, while the remaining 266 (37.4%) patients were diagnosed according to clinical and serological criteria. The overall sensitivity of the rose Bengal test was 92.9%. The specificities for groups I, II and III were 94.3%, 91.7% and 76.9%, respectively, with positive likelihood ratios of 16.5, 10.4 and 4.2, respectively. The diagnostic gain after the performance of the rose Bengal test was good or very good in patients with no previous exposure to Brucella or history of brucellosis, but poor in patients who were exposed repeatedly to Brucella or had a history of brucellosis and a low pre-test probability. Use of the rose Bengal test as the sole technique for the diagnosis of brucellosis in endemic areas should be considered very carefully in the context of patients who are exposed repeatedly to Brucella or have a history of brucellosis.

Adolescent↗

Pharmacokinetics and biliary excretion of rose bengal in rats with acute and chronic renal failure.

The effects of glycerol-induced acute renal failure (ARF) and surgically induced chronic renal failure (CRF) on the pharmacokinetics and biliary excretion of rose bengal have been examined in the rat. Both the pharmacokinetics and biliary excretion of rose bengal were unaltered in either ARF or CRF. The latter results in CRF contrast with those of Tse et al (1976, Int. J. Nucl. Med. Biol. 3: 134-137) who reported decreased removal of the dye from blood and reduced biliary excretion. In addition, rose bengal behaves differently from bromosulphophthalein and indocyanine green whose hepatic uptake and initial biliary excretion are known to be decreased in ARF. The results suggest that rose bengal may have a hepato-biliary transport route which differs from that of bromosulphophthalein and indocyanine green, and the findings also emphasize the selective nature of altered organic anion uptake by the liver in ARF.

Acute Kidney Injury↗

RcAP2L-RcAS1 complex modulates petal number in roses by targeting RcAGL80 promoter.

Double flower, which is one of the most important characteristics of ornamental plants, is closely related to their ornamental and commercial value. The double-flower trait in rose was mainly due to the increase in petal number caused by stamen petalization. However, the mechanism regulating petal number is not clear. In this study, the Rosa chinensis "Zhaiye Tengben Yuejihua" × R. chinensis "Old Blush" population was used for QTL detection and NGS-based BSA analysis to identify candidate genes related to petal number. It was found that RcAP2L and RcAS1 were highly expressed in double-flower rose, while RcAGL80 was highly expressed in single-flower rose. Silencing RcAP2L and RcAS1 reduced the petal number by inhibiting homeotic conversion of stamens to petals, separately. However, silencing RcAGL80 increased the petal number by promoting homeotic conversion of stamens to petals. The results of Y2H and BiFC assays showed that RcAP2L interacted with RcAS1. The dual-luciferase assay showed that RcAP2L was bound to the promoter of RcAGL80 and suppressed RcAGL80 transcription. In total, we found a new function of AS1 in specifying flower organ identity, and a new pathway for regulating the number of petals by RcAS1, RcAGL80, and RcAP2L, which provides new information for elucidating the mechanism of the formation of double flower in rose.

Rosa↗

The effect of photodynamic therapy with rose bengal on posterior capsule opacification in rabbit eyes.

We investigated the in vivo effect of photodynamic therapy (PDT) using rose bengal on the development of posterior capsule opacification (PCO). Endocapsular phacoemulsification was performed on white rabbits, which were divided into 4 groups: control group; group 1, treated with visible light only; group 2, treated with rose bengal only, and group 3, treated with PDT. In the case of the PDT group, rose bengal dissolved in sodium hyaluronate was injected into the empty capsular bag and treated with visible light. Three months after surgery, the rabbits were sacrificed and the eyeballs enucleated. The obstruction rate of visible light caused by PCO was measured with an optical powermeter. The mean obstruction rate was 30.6% in the control group, 28.3% in group 1, 19.3% in group 2, and 14.3% in group 3. Group 3 showed a statistically significant decrease in PCO compared with the control group and group 1 (p = 0.0014). Our results suggest that PDT using rose bengal effectively decreased PCO in rabbit eyes.

Animals↗