Evaluation of the Putative Efficacy of a Methanolic Extract of Ocimum Basilicum in Preventing Disruption of Structural Proteins in an in Vitro System of Selenite-induced Cataractogenesis.

Purpose: To evaluate whether a methanolic extract of Ocimum basilicum (OB) leaves prevented lenticular protein alterations in an in-vitro model of selenite-induced cataractogenesis.Materials and Methods: Transparent lenses extirpated from Wistar rats were divided into three groups: control; selenite only; treated. Control lenses were cultured in Dulbecco's modified Eagle's medium (DMEM) alone, selenite only lenses were cultured in DMEM containing sodium selenite only (100 µM selenite/ml DMEM) and treated lenses were cultured in DMEM containing sodium selenite and the methanolic extract of OB leaves (200 µg of extract/ml DMEM); all lenses were cultured for 24 h and then processed. The parameters assessed in lenticular homogenates were lenticular protein sulfhydryl and carbonyl content, calcium level, insoluble to soluble protein ratio, sodium dodecyl sulphate-polyacrylamide gel electrophoretic (SDS-PAGE) patterns of lenticular proteins, and mRNA transcript and protein levels of αA-crystallin and ßB1-crystallins.Results: Selenite only lenses exhibited alterations in all parameters assessed. Treated lenses exhibited values for these parameters that were comparable to those noted in normal control lenses.Conclusions: The methanolic extract of OB leaves prevented alterations in lenticular protein sulfhydryl and carbonyl content, calcium level, insoluble to soluble protein ratio, SDS-PAGE patterns of lenticular proteins, and expression of αA-crystallin and ßB1-crystallin gene and proteins in cultured selenite-challenged lenses. OB may be further evaluated as a promising agent for the prevention of cataract.

Multi-Targeting Andrographolide, a Novel NF-κB Inhibitor, as a Potential Therapeutic Agent for Stroke.

A key focus in the field of drug discovery has been motivated by the neuroprotection of natural compounds. Cerebral ischemia is a multifaceted pathological process with a series of mechanisms, and a perspective for the development of neuroprotectants from traditional herbal medicine or natural products is a promising treatment for this disease. Natural compounds with the effects of anti-oxidation, anti-inflammation, anti-apoptosis, and neurofunctional regulation exhibit therapeutic effects on experimental ischemic brain injury. Conferring to the pharmacological mechanisms underlying neuroprotection, a study found that androgapholide, a diterpene lactone compound, exhibits varying degrees of neuroprotective activities in both in vitro and in vivo experimental models of stroke. The neuroprotective mechanisms of andrographolide are suggested as: (I) increasing nuclear factor E2-related factor 2-heme oxygenase (Nrf2-HO-1) expression through p38-mitogen activated protein kinase (MAPK) regulation, (II) inducing cerebral endothelial cells (CEC) apoptosis and caspase-3 activation, (III) down regulating Bax, inducible nitric oxide synthase (iNOS), and (IV) inhibiting hydroxyl radical (OH-) formation, and activating transcription factor NF-κB signaling pathways. Recently, several researchers have also been trying to unveil the principal mechanisms involved in the neuroprotective effects of andrographolide. Therefore, this review aims to summarize an overview on the neuroprotective effects of andrographolide and exemplifies the essential mechanisms involved. This paper can provide information that andrographolide drug discovery may be a promising strategy for the development of a novel class of neuroprotective drug.

The neuroprotective effects of Tao-Ren-Cheng-Qi Tang against embolic stroke in rats.

BACKGROUND: Combinations of the traditional Chinese and Western medicines have been used to treat numerous diseases throughout the world, and there is a growing body of evidence showing that some of the herbs used in traditional Chinese medicine elicit significant pharmacological effects. The aim of this study was to demonstrate the neuroprotective effects of Tao-Ren-Cheng-Qi Tang (TRCQT) in combination with aspirin following middle cerebral artery occlusion (MCAO)-induced embolic stroke in rats. METHODS: A blood clot was embolized into the middle cerebral artery of rats to induce focal ischemic brain injury. After 24 h of MCAO occlusion, the rats were arbitrarily separated into five groups and subjected to different oral treatment processes with TRCQT and aspirin for 30 days before being evaluated in terms of their neurological behavior using a four-point system. The rats were sacrificed at 30 days after drug treatment and the infarct volumes were measured using a 2,3,5-triphenyltetrazolium chloride staining method. Tumor necrosis factor-α (TNF-α), c-Jun N-terminal kinases (JNK), activated caspase-3 and Bax were detected by western blot analysis. The apoptotic cells were identified by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. ROS generation was also measured by electron spin resonance spectrometry. RESULTS: Rats treated with TRCQT alone or in combination with aspirin showed a significantly reduced infarct volume (P < 0.001) and improved neurological outcome compared with those treated with distilled water. Rats treated with TRCQT alone (P = 0.021) or in combination with aspirin (P = 0.02) also showed significantly reduced MCAO-induced expression levels of TNF-α and pJNK (P < 0.001) in their ischemic regions. Rats treated with TRCQT alone or in combination with aspirin showed decreased apoptosis by a reduction in the number of TUNEL positive cells, which inhibited the expression of activated caspase-3 (P = 0.038) and Bax (P = 0.004; P = 0.003). TRCQT also led to a significant concentration-dependent reduction in the formation of hydroxyl radicals (P < 0.001). CONCLUSIONS: TRCQT reduced brain infarct volume and improved neurological outcomes by reducing apoptosis, attenuating the expression of TNF-α and p-JNK, and reducing the formation of hydroxyl radicals in MCAO-induced embolic stroke of rats.

Hinokitiol Exerts Anticancer Activity through Downregulation of MMPs 9/2 and Enhancement of Catalase and SOD Enzymes: In Vivo Augmentation of Lung Histoarchitecture.

Melanoma is extremely resistant to chemotherapy and the death rate is increasing hastily worldwide. Extracellular matrix promotes the migration and invasion of tumor cells through the production of matrix metalloproteinase (MMP)-2 and -9. Evidence has shown that natural dietary antioxidants are capable of inhibiting cancer cell growth. Our recent studies showed that hinokitiol, a natural bioactive compound, inhibited vascular smooth muscle cell proliferation and platelets aggregation. The present study is to investigate the anticancer efficacy of hinokitiol against B16-F10 melanoma cells via modulating tumor invasion factors MMPs, antioxidant enzymes in vitro. An in vivo mice model of histological investigation was performed to study the patterns of elastic and collagen fibers. Hinokitiol inhibited the expression and activity of MMPs-2 and -9 in B16-F10 melanoma cells, as measured by western blotting and gelatin zymography, respectively. An observed increase in protein expression of MMPs 2/9 in melanoma cells was significantly inhibited by hinokitiol. Notably, hinokitiol (1-5 µM) increased the activities of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD) from the reduction in melanoma cells. Also, hinokitiol (2-10 µM) concentration dependently reduced in vitro Fenton reaction induced hydroxyl radical (OH·) formation. An in vivo study showed that hinokitiol treatment increased elastic fibers (EF), collagens dispersion, and improved alveolar alterations in the lungs of B16/F10 injected mice. Overall, our findings propose that hinokitiol may be a potent anticancer candidate through down regulation of MMPs 9/2, reduction of OH· production and enhancement of antioxidant enzymes SOD and CAT.

Keratitis due to Fusarium langsethiae: clinical profile, molecular identification, and susceptibility to antifungals.

We report a case of keratitis due to Fusarium langsethiae in a 56-year-old man. The patient presented with pain and tearing of 10 days duration in the right eye, which had sustained a paddy stalk injury. On examination, a hypopyon corneal ulcer was noted in the right eye. Multiple scrapings were obtained from the affected part of the cornea. A lactophenol cotton blue wet mount and a Gram-stained smear of scrapings were made. Scrapings were also inoculated on various culture media, including Sabouraud dextrose agar (SDA). A fungal etiology was sought by conventional microbiological techniques and polymerase chain reaction. In vitro susceptibility testing was performed by an agar dilution method. Direct microscopy of corneal scrapings revealed septate hyphae, leading to initiation of intensive topical therapy with natamycin (5 %). However, the keratitis progressed, necessitating therapeutic penetrating keratoplasty. White, powdery-like colonies, with abundant aerial mycelium, were recovered on SDA from corneal scrape material. Based on macroscopic and microscopic morphological features, the isolated fungus was initially identified as a Fusarium species. Sequence analysis of the 28S rRNA region of the fungal genome led to a specific identification of F. langsethiae. Antifungal susceptibility testing results suggested that the strain isolated was susceptible to voriconazole, ketoconazole, and itraconazole. To our knowledge, this is the first reported case of keratitis due to F. langsethiae; attention is drawn to the unique characteristics of the fungal isolate, difficulties in identification and non-responsiveness to medical therapy.

Hinokitiol, a tropolone derivative, inhibits mouse melanoma (B16-F10) cell migration and in vivo tumor formation.

Invasion and metastasis are the major causes of treatment failure in patients with cancer. Hinokitiol, a natural bioactive compound found in Chamacyparis taiwanensis, has been used in hair tonics, cosmetics, and food as an antimicrobial agent. In this study, we investigated the effects and possible mechanisms of action of hinokitiol on migration by the metastatic melanoma cell line, B16-F10, in which matrix metalloproteinase-1 (MMP-1) is found to be highly- expressed. Treatment with hinokitiol revealed a concentration-dependent inhibition of migration of B16-F10 melanoma cells. Hinokitiol appeared to achieve this effect by reducing the expression of MMP-1 and by suppressing the phosphorylation of mitogen- activated protein kinase (MAPK) signaling molecules such as extracellular signal-regulated kinase (ERK) 1/2, p38 MAPK and c-Jun N-terminal kinases (JNK). On the other hand, hinokitiol treatment reversed IκB-α degradation and inhibited the phosphorylation of p65 nuclear factor kappa B (NF-κB) and cJun in B16-F10 cells. In addition, hinokitiol suppressed the translocation of p65 NF-κB from the cytosol to the nucleus, suggesting reduced NF-κB activation. Consistent with these in vitro findings, our in vivo study demonstrated that hinokitiol treatment significantly reduced the total number of mouse lung metastatic nodules and improved histological alterations in B16-F10 injected C57BL/6 mice. These findings suggest that treatment of B16-F10 cells with hinokitiol significantly inhibits metastasis, possibly by blocking MMP-1 activation, MAPK signaling pathways and inhibition of the transcription factors, NF-κB and c-Jun, involved in cancer cell migration. These results may accelerate the development of novel therapeutic agents for the treatment of malignant cancers.

Astaxanthin, a Carotenoid, Stimulates Immune Responses by Enhancing IFN-γ and IL-2 Secretion in Primary Cultured Lymphocytes in Vitro and ex Vivo.

Astaxanthin, a potent antioxidant carotenoid, plays a major role in modulating the immune response. In this study, we examined the immunomodulatory effects of astaxanthin on cytokine production in primary cultured lymphocytes both in vitro and ex vivo. Direct administration of astaxanthin (70-300 nM) did not produce cytotoxicity in lipopolysaccharide (LPS, 100 µg/ mL)- or concanavalin A (Con A, 10 µg/ mL)-activated lymphocytes, whereas astaxanthin alone at 300 nM induced proliferation of splenic lymphocytes (p < 0.05) in vitro. Although astaxanthin, alone or with Con A, had no apparent effect on interferon (INF-γ) and interleukin (IL-2) production in primary cultured lymphocytes, it enhanced LPS-induced INF-γ production. In an ex vivo experiment, oral administration of astaxanthin (0.28, 1.4 and 7 mg/kg/day) for 14 days did not cause alterations in the body or spleen weights of mice and also was not toxic to lymphocyte cells derived from the mice. Moreover, treatment with astaxanthin significantly increased LPS-induced lymphocyte proliferation ex vivo but not Con A-stimulated lymphocyte proliferation ex vivo. Enzyme linked immunosorbent assay (ELISA) analysis revealed that administration of astaxanthin significantly enhanced INF-γ production in response to both LPS and Con A stimulation, whereas IL-2 production increased only in response to Con A stimulation. Also, astaxanthin treatment alone significantly increased IL-2 production in lymphocytes derived from mice, but did not significantly change production of INF-γ. These findings suggest that astaxanthin modulates lymphocytic immune responses in vitro, and that it partly exerts its ex vivo immunomodulatory effects by increasing INF-γ and IL-2 production without inducing cytotoxicity.

Ex vivo and in vivo studies of CME-1, a novel polysaccharide purified from the mycelia of Cordyceps sinensis that inhibits human platelet activation by activating adenylate cyclase/cyclic AMP.

INTRODUCTION: CME-1, a novel water-soluble polysaccharide, was purified from the mycelia of Cordyceps sinensis, and its chemical structure was characterized to contain mannose and galactose in a ratio of 4:6 (27.6 kDa). CME-1 was originally observed to exert a potent inhibitory effect on tumor migration and a cytoprotective effect against oxidative stress. Activation of platelets caused by arterial thrombosis is relevant to various cardiovascular diseases (CVDs). However, no data are available concerning the effects of CME-1 on platelet activation. Hence, the purpose of this study was to examine the ex vivo and in vivo antithrombotic effects of CME-1 and its possible mechanisms in platelet activation. METHODS: The aggregometry, immunoblotting, flow cytometric analysis and platelet functional analysis were used in this study. RESULTS: CME-1 (2.3-7.6 µM) exhibited highly potent activity in inhibiting human platelet aggregation when stimulated by collagen, thrombin, and arachidonic acid but not by U46619. CME-1 inhibited platelet activation accompanied by inhibiting Akt, mitogen-activated protein kinases (MAPKs), thromboxane B2 (TxB2) and hydroxyl radical (OH(●)) formation. However, CME-1 interrupted neither FITC-triflavin nor FITC-collagen binding to platelets. CME-1 markedly increased cyclic AMP levels, but not cyclic GMP levels, and stimulated vasodilator-stimulated phosphoprotein (VASP) phosphorylation. SQ22536, an inhibitor of adenylate cyclase, but not ODQ, an inhibitor of guanylate cyclase, obviously reversed the CME-1-mediated effects on platelet aggregation and vasodilator-stimulated phosphoprotein (VASP), Akt, p38 MAPK phosphorylation, and TxB2 formation. CME-1 substantially prolonged the closure time of whole blood and the occlusion time of platelet plug formation. CONCLUSION: This study demonstrates for the first time that CME-1 exhibits highly potent antiplatelet activity that may initially activate adenylate cyclase/cyclic AMP and, subsequently, inhibit intracellular signals (such as Akt and MAPKs), ultimately inhibiting platelet activation. This novel role of CME-1 indicates that CME-1 exhibits high potential for application in treating and preventing CVDs.

Amarogentin, a secoiridoid glycoside, abrogates platelet activation through PLC γ 2-PKC and MAPK pathways.

Amarogentin, an active principle of Gentiana lutea, possess antitumorigenic, antidiabetic, and antioxidative properties. Activation of platelets is associated with intravascular thrombosis and cardiovascular diseases. The present study examined the effects of amarogentin on platelet activation. Amarogentin treatment (15~60 µM) inhibited platelet aggregation induced by collagen, but not thrombin, arachidonic acid, and U46619. Amarogentin inhibited collagen-induced phosphorylation of phospholipase C (PLC) γ2, protein kinase C (PKC), and mitogen-activated protein kinases (MAPKs). It also inhibits in vivo thrombus formation in mice. In addition, neither the guanylate cyclase inhibitor ODQ nor the adenylate cyclase inhibitor SQ22536 affected the amarogentin-mediated inhibition of platelet aggregation, which suggests that amarogentin does not regulate the levels of cyclic AMP and cyclic GMP. In conclusion, amarogentin prevents platelet activation through the inhibition of PLC γ2-PKC cascade and MAPK pathway. Our findings suggest that amarogentin may offer therapeutic potential for preventing or treating thromboembolic disorders.

Role of a Janus kinase 2-dependent signaling pathway in platelet activation.

INTRODUCTION: Janus kinases (JAKs) are intracellular non-receptor tyrosine kinases that transduce cytokine-mediated signals through a pathway mediated by JAK and the signal transducer and activator of transcription (STAT) proteins. The JAK-STAT pathway is involved in immune response, inflammation, and tumorigenesis. Platelets are anuclear blood cells that play a central role in hemostasis. METHODS: The aggregometry, immunoblotting, and platelet functional analysis used in this study. RESULTS: We found that the JAK2 inhibitor AG490 (25 and 50µM) attenuated collagen-induced platelet aggregation and calcium mobilization in a concentration-dependent manner. In the presence of AG490, the phosphorylation of PLCγ2, protein kinase C (PKC), Akt or JNK in collagen-activated aggregation of human platelets was also inhibited. In addition, we found that various inhibitors, such as the PLCγ2 inhibitor U73122, the PKC inhibitor Ro318220, the phospoinositide 3-kinase inhibitor LY294002, the p38 mitogen-activated protein kinase inhibitor SB203580, the ERK inhibitor PD98059, and the JNK inhibitor SP600125, had no effects on collagen-induced JAK2 activity. However, U73122, Ro318220 and SP600125 significantly diminished collagen-induced STAT3 phosphorylation. These findings suggest that PLCγ2-PKC and JNK are involved in JAK2-STAT3 signaling in collagen-activated platelets. CONCLUSION: Our results demonstrate that the JAK2-STAT3 pathway is involved in collagen-induced platelet activation through the activation of JAK2-JNK/PKC-STAT3 signaling. The inhibition of JAK2 may represent a potential therapeutic strategy for the preventing or treating thromboembolic disorders.

Pleurotus ostreatus, an edible mushroom, enhances glucose 6-phosphate dehydrogenase, ascorbate peroxidase and reduces xanthine dehydrogenase in major organs of aged rats.

CONTEXT: Aging is now considered to be associated with an elevation in oxidative damage to macromolecules and enhanced levels of inflammation. Therefore, inhibition of age-related oxidative stress by natural supplement is an important study. OBJECTIVES: To investigate whether the treatment with Pleurotus ostreatus (Jacq.: Fr) Kumm, (Pleurotaceae) can ameliorate oxidative damage in aged rats. MATERIALS AND METHODS: Male Wistar rats were divided into three groups of six each: group 1, normal young rats; group 2, normal aged untreated rats; group 3, normal aged rats treated with P. ostreatus (200 mg/kg body wt administered intraperitoneally for 21 days). On the 22nd day, rats were sacrificed by decapitation; the liver, kidneys, heart and brain were removed from each rat for the biochemical and isozyme analyses of the antioxidant enzymes glucose 6-phosphate dehydrogenase (G6PDH), ascorbate peroxidase (Apx) and xanthine dehydrogenase (XDH). RESULTS: An elevated activity of XDH was observed in the liver (G2:13.72 ± 4.1 versus G1: 7.57 ± 1.15; p < 0.05), kidneys (G2:101.48 ± 12.3 versus G1: 31.15 ± 1.71; p < 0.01), heart (G2: 63.21 ± 3.96 versus G1: 37.3 ± 2.70; p < 0.01) and brain (G2: 39.02 ± 3.96 versus G1: 19.84 ± 1.22; p < 0.001). The activities of G6PDH and Apx were lowered in major organs of aged untreated rats. However, treatment of P. ostreatus to aged rats resulted in decreased XDH and increased G6PDH and Apx activities in liver, kidneys, heart and brain. Interestingly, analyses of isozyme pattern of these enzymes are support the results obtained from the spectrophotometric determinations. CONCLUSION: These results suggest that an extract of P. ostreatus can protect the age-related oxidative damage in major organs of Wistar rats by enhancing the antioxidant enzymes G6PDH and Apx and by reducing XDH.

Putative free radical-scavenging activity of an extract of Cineraria maritima in preventing selenite-induced cataractogenesis in Wistar rat pups.

PURPOSE: To investigate the possible free radical-scavenging activity of an extract of Cineraria maritima on selenite-induced cataractous lenses in Wistar rat pups. METHODS: In the present study, Wistar rat pups were divided into three experimental groups. On P10, Group I (control) rat pups received an intraperitoneal injection of 0.89% saline. Rats in groups II (selenite-challenged, untreated) and III (selenite-challenged, C. maritima treated) received a subcutaneous injection of sodium selenite (19 µmol/kg bodyweight); Group III rat pups also received an intraperitoneal injection of the extract of C. maritima (350 mg/kg bodyweight) once daily P9-14. Both eyes of each pup were examined from P16 until P30. Cytochemical localization of nitroblue tetrazolium salts and generation of superoxide, hydroxyl, and nitric oxide levels were measured. The expression of the inducible nitric oxide synthase gene was evaluated with reverse transcription-PCR. Immunoblot analysis was also performed to confirm the differential expression of the inducible nitric oxide synthase protein. RESULTS: Subcutaneous injection of sodium selenite led to severe oxidative damage in the lenticular tissues, shown by increased formation of formazan crystals, elevated generation of superoxide, hydroxyl, and nitric oxide radicals, and elevated inducible nitric oxide synthase gene and protein expression that possibly contributed to the opacification of the lens and thus cataract formation. When rat pups were treated with intraperitoneal administration of the extract of C. maritima, the generation of free radicals as well as the messenger ribonucleic acid and protein expression of inducible nitric oxide synthase were maintained at near normal levels. CONCLUSIONS: The data generated by this study suggest that an ethanolic extract of C. maritima possibly prevents cataractogenesis in a rat model by minimizing free radical generation.

Keratitis due to the wood saprobic ascomycete, Auerswaldia lignicola (Family Botryosphaeriaceae), in a carpenter in India.

Keratitis due to Auerswaldia lignicola in a 32-year-old Indian male carpenter is described. At presentation, the patient reported persistent pain and tearing (left eye) in spite of topical antimicrobial therapy for more than 3 weeks. Clinically, mycotic keratitis was suspected, and direct microscopy of corneal scrapings stained by lactophenol cotton blue and Gram stains revealed broad septate hyphae. Intensive topical antifungal therapy was then given for 15 days. The keratitis continued to progress, necessitating therapeutic penetrating keratoplasty. Following the keratoplasty, there was rapid reduction in inflammation and gradual quietening of the eye. Brown-black fungal colonies resembling Lasiodiplodia theobromae were isolated from corneal scrape and corneal button (post-surgery) material on Sabouraud glucose-neopeptone agar; however, sporulation did not occur, so the morphological identification could not be confirmed. Sequence analysis of the 18S rRNA region of extracted fungal genomic DNA yielded an identification of A. lignicola Ariyawansa, J.K. Liu & K.D. Hyde; the sequence data have been deposited in GenBank (A. lignicola strain DK/V4, accession number KC866317.1). Medical management of keratitis due to such rarely reported fungal species may be difficult, necessitating surgical procedures.

An extract of the oyster mushroom, Pleurotus ostreatus, increases catalase gene expression and reduces protein oxidation during aging in rats.

OBJECTIVE: The objective of the present study was to address the effect of mushroom Pleurotus ostreatus on the catalase (CAT) gene expression and the protein carbonyls in liver and kidney of aged (24 months old) rats. METHODS: Eighteen acclimated rats were divided into 3 groups of 6 each: group I, normal young (4 months old) rats; group II, normal aged (24 months old) untreated rats; group III, normal aged rats treated with mushroom P. ostreatus extract (200 mg/kg body weight administered intraperitoneally for 30 days). On the 31st day, rats were sacrificed by decapitation; the livers and kidneys were removed, washed free of blood, blotted dry and processed immediately. Reverse transcriptase-polymerase chain reaction (RT-PCR) and spectrophotometry were utilized for the analyses of CAT gene expression and protein carbonyl content in the tissues of livers and kidneys. RESULTS: In aged rats that had been treated with the extract of P. ostreatus (group III), the level of the transcript of CAT gene was found to be higher than that in liver (P<0.01) and kidney (P<0.05) of aged untreated (group II) rats, respectively. Treatment of aged rats with P. ostreatus extract (group III) resulted in protein carbonyl levels being significantly lower in liver (P<0.05) and kidney (P<0.01) than those observed in aged untreated (group II) rats. CONCLUSION: These results suggest that an extract of P. ostreatus can enhance the antioxidant enzyme (CAT) gene expression and could decrease the incidence of free radical-induced protein oxidation in aged rats, thereby protecting the occurrence of age-associated disorders that involve free radicals.

An extract of the pleurotus ostreatus mushroom bolsters the glutathione redox system in various organs of aged rats.

This study is to investigate the putative effect of an extract of the oyster mushroom, Pleurotus ostreatus, on reduced glutathione (GSH) and its metabolic enzymes in major organs of male albino rats. A significant (P < .05) decrease in the level of GSH was observed in liver, kidneys, heart, and brain of aged rats compared to young rats. Activities of glutathione S-transferase (GST), glutathione reductase (GR), and glucose 6-phosphate dehydrogenase (G6PDH) were significantly (P < .05) lower in the liver, kidneys, heart, and brain of aged rats. The isoform pattern of these enzymes in aged rats also revealed variations in relative concentrations, presumably due to oxidative stress during aging. Administration of the extract of P. ostreatus to aged rats resulted in a significant (P < .05) increase in the levels of GSH and elevated activities of GST, GR, and G6PDH in liver, kidney, heart, and brain tissues. An increased staining intensity of isoforms of GST and G6PDH was also noted in aged rats that had been treated with the mushroom extract compared to aged untreated rats. The results of this study may suggest that an extract of P. ostreatus, a potential antioxidant, can prevent the oxidation of GSH and protect its related enzymes during aging.

Use of chlorazol black E mounts of corneal scrapes for diagnosis of filamentous fungal keratitis.

PURPOSE: To determine whether chlorazol black E, a chitin-specific stain, can be used to detect fungal filaments in corneal scrapings and to compare its sensitivity as a diagnostic aid for fungal keratitis with that of gram and lactophenol cotton blue stains. DESIGN: Prospective study, laboratory investigation. METHODS: Between December 1, 2005 and July 31, 2006, corneal scrapes from 163 patients with ulcerative keratitis were used for culture and to prepare smears that were stained by lactophenol cotton blue, chlorazol black E, or gram stains. A diagnosis of fungal keratitis was established if fungal growth occurred on the inoculated areas of multiple culture plates. RESULTS: Fungi were isolated from corneal scrapes of 82 patients. Taking fungal culture positivity as the gold standard for diagnosis of fungal keratitis, direct microscopic examination of chlorazol black E mounts had a sensitivity of 82% and specificity of 98%; culture results and chlorazol black E results were identical in 89.6% of patients. Lactophenol cotton blue mounts and gram-stained smears had a sensitivity of 85%, specificity of 90% to 91%, and 88% agreement with culture results. CONCLUSIONS: Chlorazol black E can be used for detection of fungal filaments in corneal scrapings; however, it is less sensitive than lactophenol cotton blue and gram stains as a diagnostic aid for fungal keratitis.

Infectious keratitis.

PURPOSE OF REVIEW: Infectious keratitis is a medical emergency. Improper management can lead to marked loss of vision. This review identifies recent trends in the study of infectious keratitis. RECENT FINDINGS: A multicountry outbreak of Fusarium keratitis emphasizes that contact lens wear is a major risk factor for infectious keratitis. Acanthamoeba and fungal keratitis are the most expensive forms of infectious keratitis to treat. Noninvasive methods and molecular techniques have improved diagnosis of infectious keratitis. Fortified topical antibiotics and fluoroquinolones are still the mainstay of bacterial keratitis therapy. Voriconazole and new routes of administration of conventional antifungals appear promising for fungal keratitis. Antivirals and amelioration of host inflammatory response are promising for viral keratitis; the host response is also crucial in pathogenesis of Pseudomonas aeruginosa keratitis. Trauma-induced bacterial and fungal keratitis and contact lens-associated keratitis are preventable entities. SUMMARY: Improved modalities of diagnosis and treatment have improved the outcome of infectious keratitis, but therapy of acanthamoebal, fungal and P. aeruginosa keratitis is still a challenge. Effective strategies must neutralize potential risk factors and counter host response overactivity without impairing killing of infecting microorganisms. Trauma-induced bacterial and fungal keratitis can be prevented.

Protective effect of an extract of the oyster mushroom, Pleurotus ostreatus, on antioxidants of major organs of aged rats.

This study was undertaken to investigate the putative antioxidant activity of the oyster mushroom, Pleurotus ostreatus, on lipid peroxidation and antioxidant status of major organs of aged (24 month old) rats when compared to young (4 month old) rats. Elevated levels of malondialdehyde (MDA) and significantly lowered levels of reduced glutathione (GSH) and of vitamins C and E were observed in the liver, kidneys, heart and brain of aged rats, when compared to values in young rats. Quantitative analysis of the activities of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (Gpx) revealed significantly lower values in the liver, kidneys, heart and brain of aged rats. An analysis of isozyme pattern of these enzymes in aged rats also revealed variations in relative concentration, presumably due to oxidative stress. Administration of the extract of P. ostreatus to aged rats resulted in elevated levels of reduced glutathione and vitamins C and E and in increased activities of CAT, SOD and Gpx so that the values in most of these parameters did not differ significantly from those in young rats. In addition, the level of MDA was lowered on administration of mushroom extract to aged rats. These results suggest that treatment with an extract of P. ostreatus can improve the antioxidant status during ageing, therein minimizing the occurrence of age-associated disorders associated with involvement of free radicals.