Laccase Activity and Azo Dye Decolorization Potential of Podoscypha elegans

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.

Caffeic acid phenethyl ester protects against photothrombotic cortical ischemic injury in mice

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.

Enhancement of quantum efficiency of a dye-sensitized electrochemical cell by using triturated zinc oxide mixed with two organic dyes, azure C and rose bengal

For efficiently utilising solar energy, when suitable nanoparticles are being engineered, triturated zinc oxide an eco-friendly, easily available, low-cost material has been used as an agent for solar energy conversion. Two organic dyes Azure C and Rose bengal having absorption bands in two different spectral regions at 545 nm and 610 nm respectively, were chosen in order to overcome the band absorption limits of each dye and utilise the broad spectrum of solar radiation. The material was mixed with these two dyes in a specially devised electrochemical cell and photovoltage with significant efficiency was generated. The energy conversion efficiency of the cell using three different potencies 6C, 30C and 200C of triturated zinc oxide with the same concentration of two dyes (0.5x10-5 M) in all cases are 0.39%, 0.43% and 0.35% respectively. The efficiency is only 0.15% for the mixed dye under similar conditions.

Photodynamic therapy with bengal rose and derivatives against leishmania amazonensis

Introduction: The treatment of cutaneous leishmaniasis [CL] is based primarily on the use of pentavalent antimonials, which may lead to many side effects limiting their use. Photodynamic therapy [PDT] is an alternative for the treatment of CL, and some xanthene dyes have the potential for use in PDT

Methods: The xanthenes rose bengal B [RB] and its derivatives rose bengal methyl ester [RBMET], and butyl ester [RBBUT] were analyzed for leishmanicidal activity against promastigotes and intracellular amastigotes of Leishmania amazonensis. Cytotoxicity was assessed in J774.A1 macrophages

Results: RB derivates RBMET [IC50 9.83 microM], and RBBUT [IC50 45.08 microM] showed leishmanicidal activity, however, were toxic to J774.A1 macrophages, resulting in low selectivity index

Conclusion: The RBMET and RBBUT showed to be effective against the L. amazonensis and the low selectivity index presented may not be a limitation for their use in PDT to CL treatment

Isolation and characterization of Brucella abortus isolates from wildlife species in South Korea

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.

Role of the Cerebral Cortex on Vestibular Compensation Following Unilateral Labyrinthectomy in Rats / 대한평형의학회지

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.

Seguimiento clínico, serológico y mediante la reacción de polimerasa en cadena de una familia con brucelosis / Clinical, serological and polymerase chain reaction follow-up of a family with brucellosis

Introduction: Human brucellosis diagnosis is based on isolation of Brucella spp. from blood or tissue cultures with a positivity rate of 40-70% and serology techniques are used as complementary tools; recently molecular biology diagnostic techniques have been developed intending to optimize the etiological confirmation. Aim: The main objective of this work was to compare the polymerase chain reaction (PCR), against serological diagnostic tests during the clinical follow-up of a family presenting brucellosis. Methods: Seven family members who lived in the urban area of Mexico City, were monitored using the Rose Bengal test, the agglutination test as well as agglutination with 2 mecapto ethanol, blood cultures and serum PCR for a period of 27 months. The suspected source of infection was fresh goat cheese from a known endemic zone. Results: Brucella melitensis was isolated from the blood cultures of two patients. All of the patients were positive in serological and PCR tests at the beginning of this follow-up. At the end of the study, three patients responded well to the treatment and showed negative results in the serological and PCR tests. While two patients with diabetes mellitus type 2, showed positive results in the serological and PCR tests as well as persistent symptoms. Conclusion: Clinical follow-up of patients with brucellosis is of great importance, to properly evaluate the given treatment. In this sense the PCR is a great supporting tool in diagnostic testing.

Introducción: El diagnóstico de brucelosis humana es difícil pues los cultivos de sangre y tejidos tienen un rendimiento limitado (40-70%) y usualmente se recurre a la serología como recurso complementario; últimamente se han desarrollado técnicas de biología molecular que intentan optimizar la confirmación etiológica. Objetivo: Comparar la reacción de la polimerasa en cadena (RPC) con las pruebas de diagnóstico serológicas en el seguimiento clínico de una familia con brucelosis. Métodos: Siete integrantes de una familia con brucelosis que habitaban la zona urbana de Ciudad de México fueron monitoreados mediante aglutinación con antígeno Rosa de Bengala, prueba de aglutinación, aglutinación en presencia de 2 mercapto-etanol, hemocultivos y RPC en suero durante 27 meses. La probable fuente de infección de los pacientes fue el consumo de queso fresco de cabra originario de una zona endémica. Resultados: Brucella melitensis se obtuvo del hemocultivo de dos pacientes. Todos los pacientes fueron positivos a las pruebas serológicas y al RPC al inicio del seguimiento. Tres pacientes respondieron bien al tratamiento y mostraron resultados negativos en serología y RPC al final del estudio. Mientras que en dos pacientes con diabetes mellitus tipo 2 la sintomatología fue persistente, serología positiva y RPC positivos al finalizar el estudio. Conclusión: El seguimiento clínico de pacientes con brucelosis es muy importante para valorar el tratamiento, en este sentido la RPC es una herramienta que puede apoyar a otras pruebas de diagnóstico.

The Correlation of Routine Tear Function Tests and Conjunctival Impression Cytology in Dry Eye Syndrome

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 Schirmer's > RBS, and the specificity was Schirmer's > TBUT > RBS in decreasing order when CIC was considered the gold standard.

Tratamiento del ojo seco severo con suero autólogo / Treatment of severe dry eye by autologous serum eyedrops

Objetivo: evaluar la eficacia de la aplicación de suero autólogo 20% en gotas tópicas en el tratamiento de pacientes con ojo seco severo que acudieron a Centro Visión (Lima, Perú) durante el año 2010. Material y Métodos: Se incluyeron un total de 32 ojos (16 pacientes) con diagnóstico de ojo seco severo que fueron tratados con suero autólogo 20% por 30 d¡as. La evaluación de los síntomas se realizó con el cuestionario OSDI, y el estudio de los signos através de las pruebas de tiempo de ruptura lagrimal (BUT), coloración con fluoresceína, rosa de Bengala y verde lisamina. Los hallazgos se compararon antes y después de 15 y 30 días de tratamiento. Resultados: con la terapia de suero autólogo mejoraron los s¡ntomas registrados por la escala OSDI de 87.31 (pretratamiento), a 24.68 (15 d¡as) y 20.69 (30 d¡as postratamiento). En promedio, la prueba del BUT mejoró de 3.31 (pretratamiento) a 5.94 (15 dias) y 6.91 (30 días postratamiento). La coloración de fluoresce¡na mejoró de 5.94 (pretratamiento) a 2.53 (15 días) y 2.28 (30 días postratamiento). La coloración con rosa de Bengala y/o verde de lisamina mejoró de 5.69 (pretratamiento) a 1.84 (15 d¡as) y 1.66 (30 d¡as postratamiento). Todos los resultados fueron estadísticamente significativos. Conclusiones: el suero autólogo fue efectivo para tratar el ojo seco severo, mejorando los síntomas subjetivos, la estabilidad del filme lagrimal y la puntuación de los colorantes vitales en la superficie ocular.

Objective: to evaluate the effectiveness of 20% autologous serum eyedrops in the treatment of severe dry eye patients, who attended Centro Visi¢n (Lima, Per£) in 2010. Material and Metods: thirty-two eyes of 16 severe dry eye patients were enrolled and treated with 20% autologous serum for 30 days. The subjective symptoms were evaluated with the OSDI score. The objective signs were assessed with tear film breakup time (BUT), fluorescein and rose Bengal staining scores. The findings were compared before, 15 days and 30 days after treatment. Results: the subjective symptoms OSDI scores showed improvement from 87.31 (pretreatment) to 24.68 (15 days post) and 20.69 (30 days post). As an average, BUT improved from 3.31 (pretreatment) to 5.94 (15 days post) and 6.91 (30 days). Fluorescein staining scores improved from 5.94 (pretreatment) to 2.53 (15 days post) and 2.28 (30 days post). Rose Bengal and/or lissamine green staining scores improved from 5.69 (pretreatment) to 1.84 (15 days post) and 1.66 (30 days post). All results were statistically significant. Conclusion: autologous serum eyedrops were found effective in the treatment of severe dry eye disease, improving subjective symptoms, the tear film stability and ocular surface vital staining scores.

Dry Eye — A Clinical Study in a Hospital Based Population.

Objectives: To determine the prevalence of dry eye based on dry eye symptoms attending the Department of Ophthalmology and Medicine, Tripura Medical College. Materials and Methods: This study was conducted at Tripura Medical College, Hapania, Agartala, from 1st December 2009 to 31st December 2010. Complete history and ocular examinations was recorded on a pre designed proforma in the Department of Ophthalmology. The diagnosis was made from history and objective dry eye test in the following sequence: tear meniscus height, tear break up time test,fluorescein staining, schirmer test, and rose Bengal staining. If 2 or more of the above test were positive, the patient was deemed to be suffering from dry eye. All patients were sent to the Dept. of Medicine for systemic examination and to rule out any systemic diseases. Results: In this study, total number of patients that presented with dry eye symptoms were 762 and among them dry eye was diagnosed in 403 patients. The Prevalence rate was 3.10% and it was highest in the age of more than 70 years of age. Females had higher prevalence (1.94%) than males (1.15%). Conclusion: The prevalence of dry eye is less in Tripura at around 3.10%. This may be due to the geographical and climatic implication with a high degree of humidity reigning in this region.

Mycoflora of Soybeans Used for Meju Fermentation

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.

Rose Bengal staining - diagnostic aid for potentially malignant and malignant disorders: A pilot study.

Background: The word "Cancer" itself is sufficient to cause a fear in the minds of people. Early detection of oral potentially malignant and malignant disorders is still a diagnostic challenge for most of the clinicians. In the present study, we have evaluated the efficacy of Rose Bengal staining in the detection of oral premalignant and malignant lesions. Aim: The aim of the study was to evaluate the efficacy of the Rose Bengal staining for the identification of dysplastic areas clinically, and then correlating the findings with the histological grading of dysplasia. Methods and Materials: A total of 20 patients were selected for the study. Patients had been evaluated clinically and histopathologically along with the grading of dysplasia, and Rose Bengal stain was painted over the lesional mucosa with the patient's consent. Incisional biopsies were taken from the stained area of the Rose Bengal dye, and studied histopathologically. Grading of dysplasia and intensity of the Rose Bengal staining were found to be correlated. Chi square test was performed and a statistical significance of P<0.001 was observed. Statistical significance was defined as P<0.001. Results and Conclusion: Grading of dysplasia and the intensity of Rose Bengal staining were directly proportional to each other according to this study. P value was found to be significant. Thus Rose Bengal stain can be used as a diagnostic aid in the detection of oral potentially malignant and malignant disorders.

Seroprevalence study on brucellosis in native and imported camels

Seroprevalence study was carried out on 470 camels [123 native camels in contact with other farm animals in Assiut and El-Wadi El Gadid Governorates and 347 imported camels from Sudan, through Daraw, Aswan Governorate] these camels with different ages and sexes were examined for detection of brucella antibodies. A high incidence of seroreactive cases was observed among native camels [2.43%, 12.19%, 4.87% and 2.43%] while it was [0.57%; 0.86%; 0.29% and 0.29%] among imported camels using Rose Bengal plate Test, Buffered Acidified plate Antigen Test, Tube Agglutination Test and Rivanol test, respectively. Quantitatively, four serum samples were true seropositive - without any clinical signs - with a titer ranged from 1/25 to 1/400 using Rivanol test, while 3 serum samples were seroreactors with a titer of 1/40, 1/80 and 1/320 using TAT. Three [1.07%] of positive camels were adult over 4 years old and the remaining 1[0.52%] was young under 4 years old. It can be concluded that brucella seroprevelance in native camels was significantly different [P<0.05] than in imported ones, however no significant differences could be recorded based on the sex of the age. The study proved that Rose Bengal plate Test followed by Tube Agglutination. Test agreed with Rivanol test in the rapid diagnosis of brucellosis in camels

Time Point Expression of Apoptosis Regulatory Proteins in a Photochemically-Induced Focal Cerebral Ischemic Rat Brain / 전남의대학술지

Apoptosis after global or focal cerebral ischemia plays a crucial role in mediating cell death. In this study, we observed the time point expression of physiologic events involving apoptosis regulatory proteins after photochemically-induced focal cerebral ischemia in Sprague-Dawley rats. Protein expression was evaluated at days 1, 3, and 7 by Western blot. Bcl-2, Bax, caspase-3, and phosphorylated Akt (pAkt) activity markedly increased in the ischemic hemisphere in a time-dependent manner, not affected. The expression of Bcl-2, Bax, and caspase-3 was dramatically changed around day 3, whereas changes in pAkt expression occurred at day 1. Differential elevation of these apoptosis regulatory proteins at various time points indicates that different modes of cell death occur in photochemically-induced focal cerebral ischemia in a rat brain.

Stem Cell Dynamics in an Experimental Model of Stroke / 전남의대학술지

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.

Brucellosis in sheep and goat of Bogra and Mymensingh districts of Bangladesh / 대한수의학회지

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.

Matrix Metalloproteinase Inhibitors Attenuate Neuroinflammation Following Focal Cerebral Ischemia in Mice

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.

Activation of Matrix Metalloproteinases-9 after Photothrombotic Spinal Cord Injury Model in Rats

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.