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Preparation and characterisation of rose Bengal-loaded surface-modified albumin nanoparticles.

Surface-modified albumin nanoparticles were prepared from two poly(ethylene glycol)-human serum albumin conjugates: poly(thioetheramido acid)-poly(ethylene glycol) copolymer-grafted HSA (HSA-PTAAC-PEG) and methoxy poly(ethylene glycol)-grafted HSA (HSA-mPEG). Rose bengal (RB) was used as a model drug for encapsulation into the nanoparticles either during the particle production or by adsorption post particle preparation. The drug incorporation and release was affected by the different production methods and the different polymer compositions. When RB was loaded in HSA and HSA/HSA-PTAAC-PEG nanoparticles, up to 5% (w/w) drug content was achieved. The drug loading in HSA-mPEG nanoparticles was much lower and the results from the microcalorimetry study indicated that the low loading efficiency was due to less drug-protein binding sites available in the HSA-mPEG molecule as compared to the HSA molecule. The release of RB from the albumin nanoparticles was very slow in PBS and dramatically accelerated in the presence of trypsin. Compared with unmodified nanoparticles, the slower release of RB from the surface-modified HSA nanoparticles in the presence of the enzyme suggested that the existence of a steric hydrophilic barrier on the surface of the nanoparticles made digestion of the nanoparticles more difficult.

Administration, Oral↗

A simple luminescence method for detecting lipid peroxidation and antioxidant activity in vitro.

The excited carbonyl species formed during lipid peroxidation may transfer their energy to dye molecules containing heavy atoms such as rose bengal to produce excited singlet state dye molecules. The excited dye then emits photons to return to its ground state. The aim of the present study was to investigate the relationship between methemoglobin (metHb)-induced lipid peroxidation, rose bengal luminescence, and O2 consumption in a system containing 0-850 mu M phospholipid and 1 mu M metHb and 1 mu M rose bengal in Tris-HCl buffer, pH 7.4. The rapid rise in rose bengal luminescence was accompanied by a rapid rise in O2 consumption. Further, the luminescence of rose bengal could be quenched by alpha -tocopherol or probucol present in the phospholipid liposomes, with the latter being strikingly more effective than the former. This simple luminescence method is a useful practical approach to detect lipid peroxidation and determined antioxidant effects of water insoluble compounds in vitro.

Antioxidants↗

Laser in situ keratomileusis-induced (presumed) neurotrophic epitheliopathy.

OBJECTIVE: To evaluate tear production, corneal topography, accuracy of refractive correction, and best spectacle-corrected visual acuity in eyes that had moderate to severe rose bengal staining develop on the flap compared with eyes with little or no staining on the flap, the first few months after laser in situ keratomileusis (LASIK). None of the eyes in this study had significant preoperative dry eye disease. DESIGN: Retrospective case control study. PARTICIPANTS: Individual eyes of 19 consecutive patients with moderate to severe punctate epithelial erosions and rose bengal staining on the flap 1 to 3 months after LASIK were compared with eyes of 19 concurrent patients who did not have punctate epithelial erosions or more than trace staining on the flap develop. METHODS: Nonparametric statistical analyses were used to compare tear secretion, corneal topographic irregularity, spherical equivalent, and visual acuity 3 and 6 months after surgery. Some eyes in both groups also had analysis of tear secretion 1 month after surgery. MAIN OUTCOME MEASURES: Schirmer's test without anesthesia, the topographic corneal irregularity measurement (CIM), the difference between attempted and achieved spherical equivalent, and the loss of best spectacle-corrected visual acuity. RESULTS: There was no difference in tear production 1, 3, or 6 months after LASIK in patients who had punctate epithelial erosions and rose bengal staining on the flap develop and those who did not. There was no significant difference in the CIM or mean accuracy of the refractive correction in the two groups, but some patients had a transient decrease in best spectacle-corrected visual acuity. Flap rose bengal staining resolved by 6 months after LASIK in most affected patients. CONCLUSIONS: LASIK-induced rose bengal staining in patients without preexisting dry eye is likely neurotrophic epitheliopathy, because there is no difference in mean tear production between patients who have significant punctate epithelial erosions and rose bengal staining develop on the flap and those who do not. The signs and symptoms of LASIK-induced (presumed) neurotrophic epitheliopathy tend to resolve approximately 6 months after surgery. This disorder tends to be more common and severe in patients with pre-existing dry eye disease.

Cornea↗

Reactive oxygen species-induced apoptosis and necrosis in bovine corneal endothelial cells.

PURPOSE: The loss of corneal endothelial cells associated with aging and possibly other causes has been speculated to be related to exposure to reactive oxygen species (ROS). The current study was conducted to investigate, by use of photosensitizers, the underlying mechanisms involved in the death of bovine corneal endothelial cells (BCENs) caused by ROS. METHODS: BCEN cells in primary culture were treated with a photosensitizer (riboflavin or rose bengal) with light exposure. The patterns of cell damage and death were assessed using an acridine orange-ethidium bromide differential staining method, TdT-mediated dUTP nick-end labeling (TUNEL) assay, and transmission electron microscopy. The cytotoxicity was assayed by mitochondrial function using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) testing. Antioxidants, including catalase, L-histidine, salicylic acid, and superoxide dismutase, were used to determine the types of ROS involved. Activation of nuclear factor (NF)-kappaB was examined by fluorescent immunocytochemistry with anti-p65 antibody. RESULTS: Light-irradiated riboflavin or rose bengal resulted in a significant decrease in viability of BCEN cells. Chromosomal condensation and fragmentation were observed in apoptotic cells, and membrane lysis and damage of cell ultrastructures were observed in necrotic cells. Riboflavin induced apoptosis at 30 minutes and thereafter and induced necrosis after 2 hours. Rose bengal was shown to cause similar effects within half the time required for the effects of riboflavin. Catalase and salicylic acid were found to provide protection for BCENs from cytotoxic effects of riboflavin, and L-histidine was found to protect BCENs from cytotoxicity induced by rose bengal. Kinetic studies using immunocytochemistry showed that NF-kappaB was translocated into the nucleus within 15 minutes and 30 minutes after treatment with rose bengal and riboflavin, respectively. CONCLUSIONS: The cytotoxic effects of photo-irradiated riboflavin and rose bengal are shown to be mediated by two distinct but parallel pathways, one leading to apoptosis and the other to necrosis. Possible involvement of NF-kappaB in cell death is suggested. These findings provide potential leads for future investigation into the molecular mechanisms of loss of corneal endothelial cells related to aging, oxidative stress, and possibly other similar causes.

Acridine Orange↗

A study on the ability of copper complexes to act as active oxygen species scavengers.

The ability of two low-molecular-weight copper complexes to influence the hemolysis of human erythrocytes caused by active oxygen species-generating systems was studied. Cu(II) (glycine)2 and Cu(II) (tyrosine)2 did not inhibit hemolysis due to O-2 and H2O2 generated by xanthine oxidase plus acetaldehyde but rather has a prooxidant effect. The same copper complexes as well as Cu(II) strongly inhibited the hemolysis caused by the 1O2-generating system (Rose Bengal + light). It was found that except for 1O2 the other active oxygen species (O-2, H2O2 and OH.) did not participate in the Rose Bengal + light-induced hemolysis. Thus we examined whether the inhibitory effect of copper complexes was due to 1O2 quenching. Cu(II) (glycine)2 inhibited the Rose Bengal + light-induced oxidation of compounds known to react chemically with 1O2 and its effects were analogous to the effects of physical 1O2 quenchers, e. g. NaN3 and NiCl2. The oxygen consumption upon NADH-photooxidation in the presence of Rose Bengal was inhibited competitively by Cu(II) (glycine)2 but when concentration of Rose Bengal or light intensity were varied the extent of Cu(II) (glycine)2-caused inhibition was not changed. It is concluded that the effects of Cu(II) (glycine)2 and possibly of Cu(II) (tyrosine)2 are due to quenching of 1O2 but quenching of the excited state of the dye could not be excluded.

Adult↗

Effects of xanthene dyes in cattle diets for insect control: diet digestibility and fecal excretion.

The effects of rose bengal, erythrosin B and fluorescein on ruminant digestion were evaluated by an in vitro rumen technique. Rose bengal reduced in vitro dry matter disappearance (IVDMD) values starting at .5 mM in the nutrient media and erythrosin B reduced IVDMD values starting at .05 mM in the nutrient media. Maximum reduction in IVDMD values was observed at 3.0 mM of rose bengal and erythrosin B. Fluorescein depressed digestion but not as severely as rose bengal or erythrosin B. Dry matter digestibility (DMD) values for steers given erythrosin B at a daily dosage of 6.5 mg/kg body weight were higher (P less than .05) than DMD values for control steers. DMD values were higher for steers given 16.3 or 26.1 mg erythrosin B/kg body weight daily than for the control steers, but the differences were not significant. There were no significant differences among animals given the different dosages in digestible energy. Recovery of erythrosin B from feces of treated steers varied with time, indicating that steers fed erythrosin B at a dosage of 6.5 mg/kg body weight might excrete feces during some periods of the day which would not control face fly development; however, administration of erythrosin B at a dosage of 16.3 mg/kg body weight would provide enough erythrosin B in feces to control face fly development throughout a 24-h period.

Animals↗

Effect of NADP+ and its analogs on the Rose Bengal-sensitized photoinactivation of D-erythrulose reductase from beef liver.

Upon addition of NADP+, the rose bengal-sensitized photoinactivation of D-erythrulose reductase from beef liver is prevented to a remarkable extent. Adenosine 2',5'-diphosphate (2',5'-ADP) also has a protective effect, but to a lesser extent. On the other hand, 2'-AMP markedly enhances the photoinactivation. Other nucleotides which have no 2'-phosphoryl group, such as NAD+, 3'-AMP, 5'-AMP, ADP, and NMN, are ineffective. Further, only 2'-AMP derivatives (NADP+, 2',5'-ADP, and 2'-AMP) among these nucleotides were found to be potent competitive inhibitors of the enzyme with small Ki's (6--13 muM). Photooxidation of some methionine residues in the enzyme is prevented by the addition of NADP+ and accelerated in the presence of 2'-AMP. Photooxidation products(s) of 2'-AMP derivatives have no effect upon the enzymatic activity. Although NADP+ and 2'-AMP induce detectable conformational changes of the enzyme, the changes are not characteristic to the compounds. Based on these observations, we present a possible action mechanism of 2'-AMP derivatives on the photoinactivation of D-erythrulose reductase.

Adenosine Monophosphate↗

Staining characteristics and antiviral activity of sulforhodamine B and lissamine green B.

PURPOSE: Fluorescein and rose bengal are dyes used routinely in the examination of the ocular surface. As part of an ongoing search for a superior ophthalmic dye with optimal specificity and sensitivity and a lack of interference with subsequent viral cultures, and as part of studies that use chemical dyes to understand better the pathophysiology of ocular surface disorders, the staining characteristics and antiviral activity of sulforhodamine B and lissamine green B were investigated. METHODS: Staining of rabbit corneal epithelial cell cultures by sulforhodamine B and lissamine green B was compared to that of fluorescein and rose bengal. Diffusion of each dye through a collagen gel was measured. Uptake of lissamine green B by herpes simplex virus type 1 (HSV-1)-infected Vero cell cultures was compared at several times postinfection. The effect of sulforhodamine B and lissamine green B on HSV-1 plaque formation in Vero cells was determined. The cellular toxicity of sulforhodamine B and lissamine green B in vitro was examined by a quantitative 14C-amino acid uptake assay and by a qualitative cell viability assay. Finally, the effect of sulforhodamine B and lissamine green B on viral replication was compared in vivo with that of rose bengal in a rabbit model of herpetic epithelial keratitis. RESULTS: Rose bengal vividly stained cell monolayers of explant cultures of rabbit corneal epithelium. By light microscopy, sulforhodamine B and lissamine green B, like fluorescein, did not stain the epithelial cells, but did stain the corneal explant stroma. Pretreatment of epithelial cells with 0.25% trypsin for 5 minutes failed to induce dye uptake; however, pretreatment with 0.5% Triton X-100 for 5 minutes resulted in nuclear staining by lissamine green B, but not sulforhodamine B. When added to a collagen gel, the relative diffusion rate was fluorescein > lissamine green B > sulforhodamine B > rose bengal. By spectrophotometric analysis, HSV-1-infected and uninfected Vero cells bound equivalent amounts of lissamine green B until late in infection, when infected cells took up more dye (P < 0.001). A direct neutralization assay showed that 0.06% lissamine green B or 0.5% sulforhodamine B reduced HSV-1 plaque formation in Vero cells by greater than 50%, when present at the time of viral adsorption. By a quantitative 14C-amino acid uptake assay, lissamine green B was toxic to Vero cells in a dose-dependent manner, whereas sulforhodamine B was relatively nontoxic at the concentrations tested. By a cell viability assay, however, neither dye showed significant cellular toxicity. In a rabbit model of herpetic epithelial keratitis, rose bengal significantly reduced viral replication and recovery, whereas sulforhodamine B and lissamine green B had no effect. CONCLUSIONS: Neither sulforhodamine B nor lissamine green B stain healthy, normal cells. Lissamine green B stains membrane-damaged epithelial cells, but sulforhodamine B does not. Both sulforhodamine B and lissamine green B stain corneal stroma. Lissamine green B inhibits HSV-1 plaque formation at low concentrations of dye in vitro, which correlates with suppression of cellular metabolism as demonstrated by a 14C-amino acid uptake assay, but does not affect cell viability. Neither sulforhodamine B nor lissamine green B inhibit viral replication or recovery in vivo.

Animals↗

Comparison of tear function tests and impression cytology with the ocular findings in acne rosacea.

PURPOSE: The aim of this study was to determine the correlation between tear function tests and impression cytology results and the clinical findings in ocular rosacea. METHODS: Patients with a histopathologic and clinical diagnosis of acne rosacea were studied. Ocular examination consisted of best-corrected visual acuity measurement, slitlamp examination, tear break-up time (BUT), basal Schirmer test, and rose Bengal staining. Impression cytology was done two days later. Ocular findings and symptoms were checked and scored. Age-matched controls were assessed using the same parameters. RESULTS: The most frequent symptoms were itching (80%), redness (68%), burning (64%), and photophobia (60%). The most frequent clinical findings were meibomitis (92%), telangiectasis (88%), blepharitis (84%), superficial punctate keratopathy (72%), and hyperemia (60%). Mean tear BUT was 9.6+/-3.1 (SD) seconds (range 5-17 seconds). Mean Schirmer test measured 11.4+/-1.6 mm (SD) (range 8-15 mm). Mean rose Bengal staining scored 1.8+/-0.1. Mean tear BUT, Schirmer test and rose Bengal staining scores were lower in the rosacea group than the control group (p<0.003, p=0.04, p< or =0.038, respectively). Staining was pathologic in 18 patients (36%) and the highest staining score was 4. No significant difference was found between the stages of the nasal and temporal conjunctival impression cytologies in each eye, and there was no relationship between impression cytology stages and rose Bengal scores (p>0.05). A significant relation was observed between the stages of impression cytology and the severity of meibomitis (Fisher's test, chi2=9.625, p=0.001). The Schirmer test gave lower results in patients with severe blepharitis (Mann-Whitney U Test, U=180.5, p=0.034). CONCLUSIONS: Early diagnosis prevents serious ocular complications and chronic dry eye in rosacea. We suggest that in addition to tear function tests, rose Bengal staining and impression cytology can be successfully used in the early diagnosis of dry eye and in monitoring medical treatment in ocular rosacea. Meibomian glands play an important role in the pathogenesis of the ocular disease.

Adult↗

Photodynamic therapy of subretinal neovascularization in the monkey eye.

Experimental subretinal neovascularization in the monkey eye was treated by photodynamic therapy with rose bengal. Following intravenous injection of rose bengal, the subretinal vessels were irradiated with filtered light. Successful treatment was achieved, provided the subretinal vessels were irradiated during the period in which the dye was present in and around the subretinal vessels but had already cleared from the retinal vasculature. The successfully treated lesions demonstrated replacement of the leaking and pooling subretinal vessels with a non-leaky scar. Morphologic evaluation revealed immediate destruction of the subretinal plexus, with minimal damage to the overlying retina. The destroyed subretinal tuft was replaced by a scar containing mainly fibroblasts embedded in collagen fibers. Our results suggest that photodynamic therapy is potentially useful for destruction of subretinal vessels without damaging the overlying retina.

Animals↗

Hepatic radiation injury in the rat.

The whole livers of rats were exposed intraoperatively to graded doses (0 to 75 Gy) of 137Cs gamma radiation. At various times (0 to 155 days) after liver irradiation, minimally invasive, nondestructive tests (rose bengal retention and plasma alkaline phosphatase, glutamic-oxaloacetic acid transaminase, glutamic-pyruvic transaminase) were performed on at least half the surviving animals in each dose group to assess developing liver injury. Liver histology was done on animals sacrificed 96 to 100 days after irradiation. Radiation damage to the stomach killed approximately 50% of the animals 30 to 60 days after exposure to doses of 25 Gy or higher. These deaths were significantly reduced when care was taken to shield the stomach during irradiation. Stomach injury did not, however, appreciably affect liver function as measured by rose bengal retention. Whole-liver irradiation to 15 Gy resulted in reduced liver size and minimal histological changes, but did not result in increased rose bengal retention or plasma alkaline phosphatase concentration. The next highest dose group studied (25 Gy) showed significant histological abnormalities and liver injury as measured by increased rose bengal retention and liver enzymes. The latent period for development of hepatic injury, as measured by increased rose bengal retention, was 35 to 42 days and was relatively invariant over the 25- to 75-Gy dose range. Hepatic vein lesions and cellular necrosis were the most prominent histological lesions observed in 25-Gy-irradiated liver.

Alanine Transaminase↗

A new model for photochemically induced thrombosis in the inner ear microcirculation and the use of hearing loss as a measure for microcirculatory disorders.

A new photochemical method was employed to cause disorders in the inner ear's microcirculation, using the rat as an animal model. Hearing loss was used as a measure for establishing the altered microcirculation. Under pentobarbital anesthesia, the middle ear was opened by a ventral approach. The lateral wall of the cochlea was then illuminated with a filtered xenon lamp (wavelength 540 nm) while rose bengal was infused intravenously. Photoactivated rose bengal produces oxygen radicals and oxygen singlets, which subsequently damage the vascular epithelium to cause the adhesion and aggregation of platelets in the small vessels. Disintegration of the inner ear hair cells at the irradiated site became evident 24 h after the illumination. These findings further suggest that the photochemical occlusion in the inner ear's microcirculation led to ischemic damage of the stria vascularis and the hair cells in the inner ear. When the action potential (AP) of the cochlea was measured with an electrocochleogram a gradual decrease occurred after the illumination. When acetylsalicylic acid was injected intravenously before treatment, the time required to completely suppress the AP was prolonged in a dose-dependent manner. Findings indicate that our method causes a photochemically induced occlusion in the inner ear's microcirculation and is therefore potentially useful for evaluating the various effects of drugs on the ear.

Action Potentials↗

Membrane potential fluctuations and transient inward currents induced by reactive oxygen intermediates in isolated rabbit ventricular cells.

The cellular basis of reactive oxygen intermediate-induced arrhythmias was investigated in isolated rabbit ventricular cells using the whole-cell voltage- and current-clamp techniques. Singlet oxygen and superoxide were generated by the photoactivation of rose bengal. Single ventricular cells exposed to rose bengal (10-100 nM) exhibited spontaneous membrane potential fluctuations at plateau potentials and at the level of the resting membrane potential. The voltage fluctuations induced in the resting potential occasionally triggered repetitive action potential discharges. At the resting membrane potential, the magnitude and dominant frequency of the voltage fluctuations were 1-3 mV and 1.5 Hz, respectively. At plateau potentials, the amplitude of the voltage fluctuations was about 2-5 mV, and the dominant oscillatory frequency was 2.6 Hz. In voltage-clamp experiments, transient inward currents were induced on repolarization after a depolarizing clamp step. Oscillatory currents also occurred occasionally during clamp steps to positive potentials. The peak frequencies of transient inward currents recorded at -20 and -70 mV were approximately 3.7 and 2.3 Hz, respectively, indicating that these currents may underlie the arrhythmogenic membrane potential fluctuations observed in current-clamp experiments. The rose bengal-induced transient inward currents were shown to be dependent on the magnitude and duration of the preceding voltage step. Studies of the voltage dependence of transient inward currents showed that these currents remained inward even at positive potentials (+30 mV), and replacement of extracellular sodium with lithium decreased transient inward current to approximately 10% of its initial value. Thus, the major component of oxidant stress-induced inward current appears to be electrogenic Na-Ca exchange. This oscillatory transient inward current may be responsible for the arrhythmias induced in isolated hearts exposed to reactive oxygen intermediates, and since oxidant stress has been implicated in reperfusion injury, it is possible that similar oscillatory currents may underlie reperfusion-induced arrhythmias.

Animals↗

Comparison of lacrimal and salivary gland involvement in Sjögren's syndrome.

OBJECTIVES: To determine the performance of different tear and salivary tests applied in Sjögren's syndrome (SS) and to disclose how these tests relate to common serologic tests in SS. DESIGN: In addition to the routine ocular and oral tests for diagnosing SS (Schirmer test, rose bengal score, unstimulated whole saliva flow, and parotid sialography), tear breakup time and flow rate of glandular saliva (parotid and submandibular-sublingual [SM/SL]) were evaluated in patients referred for diagnosis of SS. Patients were categorized into primary SS, secondary SS, and non-SS groups according to the revised European classification criteria for SS. SETTING: Referral center. PATIENTS: Referred sample of 80 consecutive patients. MAIN OUTCOME MEASURE: Correlation between ocular and salivary measures. RESULTS: Breakup time performed insufficiently in diagnosing SS, as opposed to the rose bengal score. In patients with primary and secondary SS, a clear correlation was noted between tear and saliva quality and secretion rate, and between the rose bengal score and parotid sialography. Increased rose bengal scores also correlated significantly with hyperglobulinemia and presence of SS-B antibodies in serum, with duration of subjective eye dryness, and with decreased tear-gland function. With regard to the oral tests, whole, parotid, and SM/SL salivary flow decreased significantly with increasing duration of oral complaints, with the stimulated SM/SL flow rate showing the strongest decrease and being more specific in diagnosing SS. Also, parotid sialography was more specific in excluding patients without SS than the commonly applied diagnostic criterion of secretion of unstimulated whole saliva. CONCLUSIONS: The rose bengal score remains the eye test of choice, as it has the highest specificity for SS. Hyperglobulinemia and especially positive serologic findings for SS-B may warrant close monitoring of the eyes, since these serum findings appear to relate to the severity of ocular surface damage. Parotid sialography and stimulated secretion of SM/SL saliva are more specific in diagnosing SS than unstimulated secretion of whole saliva.

Antibodies↗

Effect of pretreatment with exogenous organic anions on biliary excretion in rats.

The effect of pretreatment with exogenous organic anions (rose bengal, amaranth, eosine, bromsulphthalein) on biliary excretion has been investigated in rats. Pretreatment with exogenous nonmetabolized organic anions (rose bengal, amaranth, eosine) had no significant influence on the biliary excretion of rose bengal, amaranth, eosine, bromsulphthalein and bromsulphthalein-glutathione conjugate. Pretreatment with bromsulphthalein, which is metabolized in the liver, significantly enhanced the biliary excretion of total bromsulphthalein due to stimulation of the conjugation of BSP with glutathione. Both the activity of glutathione S-transferase and the glutathione content in the liver were increased following BSP pretreatment. Pretreatment with rose bengal and eosine influenced neither the conjugation of bromsulphthalein with glutathione, nor the biliary excretion of total bromsulphthalein. These results indicate that the biliary excretion rate of exogenous organic anions cannot be increased by pretreatment with substrates of the hepatic transport system. The enhanced biliary output of total bromsulphthalein after bromsulphthalein pretreatment can be explained solely by stimulation of its conjugation with glutathione.

Animals↗

Validation of fluorescence polarization assay (FPA) and comparison with other tests used for diagnosis of B. melitensis infection in sheep.

Fluorescence polarization assay (FPA) is a new test for the serological diagnosis of Brucella spp. infection in animals. The FPA is validated for the diagnosis of B. melitensis infection in sheep. For this purpose, 166 sera originated from natural infected sheep (verified by culture) and 851 sera originated from healthy animals (reared in areas where B. melitensis was never been isolated) were tested. The optimum cut-off value that offers the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 87mP with the use of ROC analysis. The DSn and DSp of FPA using this cut-off value are calculated at 97.6 and 98.9% with a 95% confidence interval (CI) of 93.9-99.3% and 98.0-99.5%, respectively. The DSn and DSp of FPA have been assessed also using as positive reference (n=587), sera that gave positive results at least in two tests used for diagnosis of B. melitensis in sheep as Rose Bengal Test (RBT), modified Rose Bengal Test (m-RBT), complement fixation test (CFT), indirect Elisa (i-Elisa) and competition Elisa (c-Elisa) originated from animals reared in flocks infected by B. melitensis. The optimum cut-off value using the above panel of positive reference sera was the same offering a DSn of 95.9% with a 95% CI, 94.0-97.4%, since the DSp remains the same. The DSn and DSp as well as performance, accuracy and agreement of FPA's result were compared with those of other tests used. The accuracy of FPA is very high, similar with that of i-Elisa. FPA is a promising assay, which offers a DSn and accuracy better that of those of the tests currently approved for the diagnosis of B. melitensis in sheep and goats. Due to its simplicity, the sort time that results can be obtained and its accuracy it can be used and improve the laboratory testing capacity as well as the efficacy of the eradication program based on test-and-slaughter policy.

Animals↗

Tubifex tubifex Müll.--photosensitive organism.

The worm Tubifex tubifex Müll. (Tubificideae, Oligochaeta) is a suitable organism for the research of the biological effect of various pollutants. This pilot study deals with the responds of the organism to the treatments of two photosensitizers (bengal rose B, quinidine) and UVA radiation. The activity of the photosensitizers was evaluated by the comparison of the surfaces of tested worms and dark controls. The results showed that T. tubifex Müll. could be a suitable organism for the studies of phototoxicity. This species demonstrated relatively strong sensitivity to the effect of the selected photodynamically active substances.

Animals↗

[Seasonal features of the excretory function of the liver in the presence of tetracycline damage and correction of the disorders with antioxidants].

It was shown on pubertal albino rats that the intensity of excretion with the bile of radioactive Bengal rose was different at different seasons: the maximum and minimum levels were observed in winter and summer, respectively. When the liver was affected with tetracycline, this process was suppressed especially in summer. The use of antioxidants, such as tocopherol acetate in combination with sodium selenite promoted the recovery of liver excretion function in winter, spring and autumn. In summer, the recovery was only partial.

Animals↗