In this study, we aimed to investigate the neuroprotective effects of caffeic acid phenethyl ester (CAPE), an active component of propolis purified from honeybee hives, on photothrombotic cortical ischemic injury in mice. Permanent focal ischemia was achieved in the medial frontal and somatosensory cortices of anesthetized male C57BL/6 mice by irradiation of the skull with cold light laser in combination with systemic administration of rose bengal. The animals were treated with CAPE (0.5–5 mg/kg, i.p.) twice 1 and 6 h after ischemic insult. CAPE significantly reduced the infarct size as well as the expression of tumor necrosis factor-α, hypoxiainducible factor-1α, monocyte chemoattractant protein-1, interleukin-1α, and indoleamine 2,3-dioxygenase in the cerebral cortex ipsilateral to the photothrombosis. Moreover, it induced an increase in heme oxygenase-1 immunoreactivity and interleukin-10 expression. These results suggest that CAPE exerts a remarkable neuroprotective effect on ischemic brain injury via its anti-inflammatory properties, thereby providing a benefit to the therapy of cerebral infarction.
Animals ; Brain Injuries ; Brain Ischemia ; Cerebral Cortex ; Cerebral Infarction ; Chemokine CCL2 ; Heme Oxygenase-1 ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; Interleukin-10 ; Ischemia ; Male ; Mice* ; Necrosis ; Neuroprotective Agents ; Propolis ; Rose Bengal ; Skull ; Urticaria

Azo dyes containing effluents from different industries pose threats to the environment. Though there are physico-chemical methods to treat such effluents, bioremediation is considered to be the best eco-compatible technique. In this communication, we discuss the decolorization potentiality of five azo dyes by Podoscypha elegans (G. Mey.) Pat., a macro-fungus, found growing on the leaf-litter layer of Bethuadahari Wildlife Sanctuary in West Bengal, India. The fungus exhibited high laccase and very low manganese peroxidase activities under different culture conditions. Decolorization of five high-molecular weight azo dyes, viz., Orange G, Congo Red, Direct Blue 15, Rose Bengal and Direct Yellow 27 by the fungus was found to be positive in all cases. Maximum and minimum mean decolorization percentages were recorded in Rose Bengal (70.41%) and Direct Blue 15 (24.8%), respectively. This is the first record of lignolytic study and dye decolorization by P. elegans.
Azo Compounds ; Biodegradation, Environmental ; Citrus sinensis ; Congo Red ; Fungi ; India ; Laccase* ; Manganese ; Peroxidase ; Rose Bengal

A total of 782 blood and 465 tissue samples from 1,039 wild animals and 127 dairy goats were collected from January 2011 to December 2013 in 10 provinces of South Korea and tested for the presence of brucellosis. The Rose Bengal test revealed that 8.0% (52/650) of the serum samples were seropositive, while 4.2% (33/782) of the serum samples were positive for Brucella antibodies by competitive enzyme-linked immunosorbent assay. Of the 650 sera examined, only 16 (2.5%) were positive by both serological tests. Direct polymerase chain reaction (PCR) assay using B4/B5 primers for Brucella abortus (BCSP31) revealed the prevalence of Brucella to be 26.5% (129/487) in blood samples and 21% (98/465) in tissue samples while, 16S rRNA PCR detected Brucella DNA in 6.8% (33/487) and 2.6% (12/465) in blood and tissue samples, respectively. Of PCR-positive samples, only 6.2% (30/487) of blood samples and 2.4% (11/465) of tissue samples were found to be positive by both BCSP31 and 16S rRNA PCRs. However, Brucella strains were isolated by blood culture from only two out of 487 blood samples (0.4%). This characterization and identification of pathogenic Brucella isolates is the first to clearly indicate that the organisms were Brucella abortus biovar 1.
Animals ; Animals, Wild ; Antibodies ; Brucella abortus* ; Brucella* ; Brucellosis ; DNA ; Enzyme-Linked Immunosorbent Assay ; Goats ; Korea* ; Polymerase Chain Reaction ; Prevalence ; Rose Bengal ; Serologic Tests

OBJECTIVE: The cerebral cortex can modulate vestibular functions through direct control of neuronal activities in the vestibular nuclei. The purpose of this study was to investigate the effect of unilateral cortical lesion or cortical stimulation on static vestibular symptoms and vestibular nuclear activities at the acute stage of vestibular compensation following unilateral labyrinthectomy (UL) in rats. METHODS: The photothrombic ischemic injury using rose bengal was induced in the primary motor cortex or primary sensory cortex, and electrical stimulation was applied to the primary motor cortex, primary sensory cortex, or sencondary sensory cortex, respectively, in unilateral labyrinthectomized rats. Static vestibular symptoms including ocular movement and postural deficits, and expression of c-Fos protein in the medial vestibular nucleus (MVN) were measured. RESULTS: Lesion of the motor cortex produced a marked postural deficit with paralytic weakness in the hindlimb contralateral to UL. Number of spontaneous nystagmus in animals receiving cortical lesion was significantly increased 2, 6, and 12 hours after UL compared with animals being UL only. Lesion of the primary motor cortex or stimulation of the S2 sensory cortex decreased expression of c-Fos protein in MVN following UL compared with UL only group. Electrical stimulation of S2 sensory areas caused significant reduction of static vestibular symptoms and decreased expression of c-Fos protein in MVN 24 hours following UL. CONCLUSION: The present results suggest that cerebral cortex involves in recovery of static vestibular symptoms during vestibular compensation following UL.
Animals ; Cerebral Cortex ; Compensation and Redress ; Electric Stimulation ; Hindlimb ; Motor Cortex ; Neurons ; Rats ; Rose Bengal ; Vestibular Nuclei

PURPOSE: To establish the strength of the association between routine tear function tests and conjunctival impression cytology (CIC) and to determine whether they simulate the morphological and cytological changes that occur on the ocular surface in dry eye. What are the sensitivity, specificity and positive predictive values of these tests when CIC is considered the gold standard? METHODS: The tear film profile included tear film break up time (TBUT), Schirmer's-1, Rose Bengal scores (RBS), and impression cytology. CIC samples were obtained from the inferior bulbar conjunctiva and stained with periodic acid-Schiff and counter stained with hematoxylin and eosin. RESULTS: The mean Schirmer's value was 11.66 +/- 5.90 in patients and 17.17 +/- 2.97 in controls (p < 0.001). The mean TBUT in participants was 8.88 +/- 3.54 and 13.53 +/- 2.12 in controls (p < 0.001). Patients had a mean goblet cell density (GCD) of 490 +/- 213, while the value for controls was 1,462 +/- 661 (p < 0.001). Abnormal CIC was observed in 46.7% cases of dry eye and in 32.8% of controls. The correlation coefficient (L) for Schirmer's was 0.2 and 0.24 for participants and controls, respectively, while TBUT values were 0.26 and 0.38, RBS were 0.5 and 0.5, and GCD was 0.8 and 0.6 in cases and controls, respectively. CONCLUSIONS: GCD, RBS, and TBUT were better predictors of morphological and cytological changes in the conjunctiva than Schirmer's in dry eye syndrome. The sensitivity of tear function tests in diagnosing dry eye was TBUT > Schirmer's > RBS, and the specificity was Schirmer's > TBUT > RBS in decreasing order when CIC was considered the gold standard.
Adolescent ; Adult ; Case-Control Studies ; Child ; Conjunctiva/pathology ; Diagnostic Techniques, Ophthalmological/*standards ; Dry Eye Syndromes/*diagnosis/*pathology ; Eosine Yellowish-(YS)/diagnostic use ; Female ; Goblet Cells/pathology ; Hematoxylin/diagnostic use ; Humans ; Male ; Middle Aged ; Ophthalmology/*standards ; Periodic Acid-Schiff Reaction/standards ; Predictive Value of Tests ; Rose Bengal/diagnostic use ; Sensitivity and Specificity ; *Tears ; Young Adult

Diverse fungi are present in Korean traditional meju and they are known to play an important role in fermented soybean products. To determine the origin of the fungi in meju, we examined the mycoflora of soybeans from 10 traditional meju factories. The samples were untreated or treated with sodium hypochlorite, and placed on malt extract agar (MEA), dichloran 18% glycerol agar (DG18), and dichloran rose bengal chloramphenicol agar (DRBC) medium. A total of 794 fungal strains were isolated and they were identified as 41 genera and 86 species. From sodium hypochlorite untreated soybeans, the genera, Cladosporium (55%), Eurotium (51%), Fusarium (33%), Penicillium (22%), and Aspergillus (exclusion of Eurotium) (20%), were mainly isolated, and Eurotium herbariorum (22%), Eurotium repens (18%), Cladosporium tenuissimum (18%), F. fujikuroi (18%), Aspergillus oryzae/flavus (7%), and Penicillium steckii (6%) were the predominant species. In case of sodium hypochlorite-treated soybeans, Eurotium (31%) and Cladosporium (5%) were frequently isolated, but Aspergillus (excluding Eurotium), Penicillium and Fusarium which were frequently isolated from untreated soybeans, were rarely isolated. Eurotium herbariorum (21%), Eurotium repens (8%), and Cladosporium tenuissimum (3%) were the predominant species. Of the 41 genera and 86 species isolated from soybeans, 13 genera and 33 species were also found in meju. These results suggest that the fungi on soybeans may influence the mycoflora of meju.
Agar ; Aniline Compounds ; Aspergillus ; Chloramphenicol ; Cladosporium ; Eurotium ; Fermentation ; Fungi ; Fusarium ; Glycerol ; Penicillium ; Rose Bengal ; Sodium ; Sodium Hypochlorite ; Soybeans

Brucellosis is the most important bacterial disease of livestock in Bangladesh. The present study was conducted to determine the seroprevalence of brucellosis in goat and sheep in Mymensingh and Bogra districts of Bangladesh using slow agglutination test and Rose Bengal test as screening test and indirect enzyme linked immunosorbent assay as confirmatory test. Questionnaire based data on age, gender, area, client's complaint, number of animals in herds, disease history, reproductive problems such as abnormal uterine discharge, abortion or previous abortion in sheep and goat and their reproductive diseases were recorded. A total of 200 sera samples were collected from 80 sheep and 120 goats. The prevalence of brucellosis in goat was 2.50% and 1.25% in sheep. Positive reactors were only detected in female of both goat and sheep. In this study, there existed a significant association among abortion and the prevalence of brucellosis (p < 0.01). The prevalence of brucellosis in sheep and goat in Bangladesh is not negligible, and it is therefore worth considering the adoption of preventive measures.
Abortion, Induced ; Adoption ; Agglutination Tests ; Animals ; Bangladesh ; Brucellosis ; Enzyme-Linked Immunosorbent Assay ; Female ; Goats ; Humans ; Livestock ; Mass Screening ; Prevalence ; Surveys and Questionnaires ; Reproductive History ; Rose Bengal ; Seroepidemiologic Studies ; Sheep

OBJECTIVE: Matrix metalloproteinases (MMPs), especially MMP-2 and MMP-9 have been known to play an important role in secondary inflammatory reaction after spinal cord injury (SCI). The aim of this study was to investigate the expression and activity of MMP-2 and MMP-9 and to determine their relationship with disruption of endothelial blood-barrier after photochemically induced SCI in rats. METHODS: Female Sprague-Dawley rats, weighing between 250 and 300 g (aged 8 weeks) received focal spinal cord ischemia by photothrombosis using Rose Bengal. Expressions and activities of MMP-2 and MMP-9 were assessed by Western blot and gelatin zymography at various times from 6 h to 7 days. Endothelial blood-barrier integrity was assessed indirectly using spinal cord water content. RESULTS: Zymography and Western blot analysis demonstrated rapid up-regulation of MMP-9 protein levels in spinal cord after ischemic onset. Expressions and activities of MMP-9 showed a significant increased at 6 h after the photothrombotic ischemic event, and reached a maximum level at 24 h after the insult. By contrast, activated MMP-2 was not detected at any time point in either the experimental or the control groups. When compared with the control group, a significant increase in spinal cord water content was detected in rats at 24 h after photothrombotic SCI. CONCLUSION: Early up-regulation of MMP-9 might be correlated with increased water content in the spinal cord at 24 h after SCI in rats. Results of this study suggest that MMP-9 is the key factor involved in disruption of the endothelial blood-barrier of the spinal cord and subsequent secondary damage after photothrombotic SCI in rats.
Animals ; Blotting, Western ; Female ; Gelatin ; Humans ; Matrix Metalloproteinases ; Rats ; Rats, Sprague-Dawley ; Rose Bengal ; Spinal Cord ; Spinal Cord Injuries ; Spinal Cord Ischemia ; Up-Regulation ; Water

The aim of this study was to investigate whether matrix metalloproteinase (MMP) inhibitors attenuate neuroinflammation in an ischemic brain following photothrombotic cortical ischemia in mice. Male C57BL/6 mice were anesthetized, and Rose Bengal was systemically administered. Permanent focal ischemia was induced in the medial frontal and somatosensory cortices by irradiating the skull with cold white light. MMP inhibitors, such as doxycycline, minocycline, and batimastat, significantly reduced the cerebral infarct size, and the expressions of monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-alpha), and indoleamine 2,3-dioxygenase (IDO). However, they had no effect on the expressions of heme oxygenase-1 and neuroglobin in the ischemic cortex. These results suggest that MMP inhibitors attenuate ischemic brain injury by decreasing the expression levels of MCP-1, TNF-alpha, and IDO, thereby providing a therapeutic benefit against cerebral ischemia.
Animals ; Brain ; Brain Injuries ; Brain Ischemia ; Chemokine CCL2 ; Cold Temperature ; Doxycycline ; Globins ; Heme Oxygenase-1 ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; Ischemia ; Light ; Male ; Matrix Metalloproteinase Inhibitors ; Mice ; Minocycline ; Nerve Tissue Proteins ; Phenylalanine ; Rose Bengal ; Skull ; Thiophenes ; Tumor Necrosis Factor-alpha

We investigated the migration of endogenous neural stem cells (NSCs) toward an infarct lesion in a photo-thrombotic stroke model. The lesions produced by using rose bengal dye (20 mg/kg) with cold light in the motor cortex of Sprague-Dawley rats were also evaluated with sequential magnetic resonance imaging (MRI) from 30 minutes through 8 weeks. Migration of NSCs was identified by immunohistochemistry for nestin monoclonal antibody in the lesion cortex, subventricular zone (SVZ), and corpus callosum (CC). The contrast to noncontrast ratio (CNR) on MRI was greatest at 12 hours in DWI and decreased over time. By contrast, T1-weighted and T2-weighted images showed a constant CNR from the beginning through 8 weeks. MRI of the lesional cortex correlated with histopathologic findings, which could be divided into three stages: acute (edema and necrosis) within 24 hours, subacute (acute and chronic inflammatory cell infiltration) at 2 to 7 days, and chronic (gliofibrosis) at 2 to 4 weeks. The volume of the infarct was significantly reduced by reparative gliofibrosis. The number of nestin+ NSCs in the contralateral SVZ was similar to that of the ipsilateral SVZ in each group. However, the number of nestin+ NSCs in the ipsilateral cortex and CC increased at 12 hours to 3 days compared with the contralateral side (p<0.01) and was reduced significantly by 7 days (p<0.01). Active emigration of internal NSCs from the SVZ toward the infarct lesion may also contribute to decreased volume of the infarct lesion, but the self-repair mechanism by endogenous NSCs is insufficient to treat stroke causing extensive neuronal death. Further studies should be focused on amplification technologies of NSCs to enhance the collection of endogenous or transplanted NSCs for the treatment of stroke.
Cold Temperature ; Corpus Callosum ; Emigration and Immigration ; Immunohistochemistry ; Intermediate Filament Proteins ; Light ; Magnetic Resonance Imaging ; Models, Theoretical ; Motor Cortex ; Nerve Tissue Proteins ; Neural Stem Cells ; Neurons ; Rats, Sprague-Dawley ; Rose Bengal ; Stem Cells ; Stroke ; Transplants