Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.

Correlation of Serum C-Reactive Protein with Disease Severity in Tuberculosis Patients

Purpose: To study the factors influencing sputum smear conversion including Serum C-Reactive Protein (CRP) and its correlation with disease severity in tuberculosis patients. Method: Levels of Serum-CRP concentrations were deter-mined in 60 patients with pulmonary tuberculosis, 30 healthy volunteers and patients in follow-up after completion of antitubercular treatment (DOTS therapy). Results: Serum-CRP levels were found to be significantly higher in smear-positive group as compared with the follow-up patients and smear-negative control group. The values were 43.65 ± 23.68, 9.88 ± 5.23 and 4.04 ± 3.85 mg/L respectively (P Conclusion: Serum-CRP levels are significantly correlated with disease severity in patients with active pulmonary tuberculosis. Thus these findings from the present study would certainly add new criteria for early diagnosis of TB, which may lead to development of new strategies to treat TB.

Protecting future antimalarials from the trap of resistance:Lessons from artemisinin-based combination therapy (ACT) failures

Having faced increased clinical treatment failures with dihydroartemisinin-piperaquine(DHA-PPQ),Cambodia swapped the first line artemisinin-based combination therapy(ACT)from DHA-PPQ to artesunate-mefloquine given that parasites resistant to piperaquine are susceptible to mefloquine.However,triple mutants have now emerged,suggesting that drug rotations may not be adequate to keep resistance at bay.There is,therefore,an urgent need for alternative treatment strategies to tackle resistance and prevent its spread.A proper understanding of all contributors to artemisinin resistance may help us identify novel strategies to keep artemisinins effective until new drugs become available for their replacement.This review highlights the role of the key players in artemisinin resistance,the current strategies to deal with it and suggests ways of protecting future antimalarial drugs from bowing to resistance as their predecessors did.

Assessment of anti-diabetic potential of aqueous and ethanol extracts of Strophanthus hispidus stem bark in Wistar rats

OBJECTIVE Diabetes mellitus is a chronic disease,for which there is no known cure except in a very specific situation.The available orthodox drugs used for its management are having disadvantages.This study investigated the anti-diabetic potential of aqueous and ethanol extracts of Strophanthus hispidus(SH) stem bark in Wistar rats.METHODS Glucose concentrations,serum α-amylase and lipid profile parameters of normal and diabetic rats were monitored for 12 weeks.Diabetes mellitus was induced by intraperitoneal injection of streptozotocin(55 mg · kg^(-1)).30 Male rats were randomly selected into six groups of five rats per group.Two groups served as normal and diabetic controls,respectively,and received distilled water for 12 weeks.Another two groups(normal rats) were treated orally with the aid of a gavage,250 mg·kg^(-1) of aqueous and ethanol extracts of SH respectively for 12 weeks whereas the other two groups(diabetic rats) received the respective dose of aqueous and ethanol extracts for the same period.RESULTS The results revealed significant(P<0.05) progressive decrease in the fasting blood glucose concentration on the 2nd-12th weeks in normal rats,whereas the diabetic treated rats,showed reduction(P<0.05) in the fasting blood glucose concentration on the 4th-12th weeks.Total cholesterol,LDL and TG levels were lowered significantly(P<0.05) by the extracts in normal and diabetic treated rats,whereas HDL levels(17.6±0.50) mg·dL^(-1)(aqueous) and(21.4±1.28) mg·dL^(-1)(ethanol)were elevated in diabetic treated rats.The activity of serum α-amylase was also elevated by the extract in diabetic treated rats.CONCLUSION These findings indicated that SH showed hypoglycemic,antihyperlipidemic and anti-diabetic activities.These findings may be connected with the presence and amount of phytochemicals in the plant.

Prevalence of Polycystic Ovary Syndrome in Nigerian Women with Infertility: A Prospective Study of the Three Assessment Criteria

Background: Polycystic ovarian syndrome (PCOS) is a frequent diagnosis in oligomenorrheic and infertile Nigerian women. However, to date there is a paucity of data on the prevalence of PCOS in Nigerian women. The objective of this study was to investigate the prevalence of PCOS in a cross-section of women attending Infertility Clinics in Benin City, Nigeria using the three assessment criteria namely: the 1990 National Institutes of Health (NIH), the 2003 Rotterdam and 2006 Androgen Excess Society (AES) criteria. Method: Four hundred and twenty-one consecutive infertile premenopausal women aged 18 - 45 years were recruited and evaluated with a proforma that elicited information about their maternal and reproductive health history. Blood samples were analyzed for hormone levels using standard immunoassay procedures, while trans-vaginal ultrasound scan was carried out to determine the presence of ultrasonic features of PCOS. The control group comprised of eumenorrheics (n = 180). Results: An estimated prevalence of biochemical hyperandrogenism (BHA) was as high as 20.9% (88 women), while 3.6% (15 women) presented with clinical hyperandrogenism (CHA). Also the prevalence of polycystic ovaries (PCO) was 13.8%. The prevalence of PCOS based on NIH, Rotterdam and AES criteria was 16.9% (71 women), 27.6% (116 women) and 20.7% (87 women) respectively. However, women with PCOS were significantly younger and had higher total testosterone levels (p = 0.001) when compared to controls. Conclusion: The prevalence of PCOS is as high in the population under study as in other prevalence studies. The hormonal investigations were clinically useful in assessing the prevalence rates. However, the recruitment criteria, together with the regional and racial factors may have contributed to the estimates obtained, and the high incidence of biochemical hyperandrogenism in this region.

Glycation Induced Physicochemical Changes in Low-Density Lipoprotein and Its Role in Promoting Cholesterol Accumulation in Macrophages along with Antiglycation Effect of Aminoguanidine

The present study aimed at investigating physicochemical changes in modified LDL by sugars specifically fructose due to recent reports on its involvement in cardiovascular diseases and also glucose and their role in subsequent in vitro accumulation of cholesterol in macrophages. Antiglycation action of aminoguanidine was also investigated. LDL isolated from human blood was incubated with fructose or glucose and aminoguanidine where indicated. The physicochemical changes in modified LDL were detected by electrophoretic, spectroscopic and chemical analysis. Accumulation of cholesterol and its inhibiton in human monocyte-derived macrophages incubated with modified LDL was determined by HPLC. Results showed increased relative electrophoretic mobility, hyperchromicity at 280 nm, development of AGE fluorescence, decrease in free amino groups and increased carbonyl content in glycated LDL as compared to native LDL. Also total cholesterol accumulated in macrophages was more for glycated LDL as compared to native LDL. The magnitude of changes was more prominent in case of fructose as compared to glucose. Aminoguanidine showed remarkable restriction of glycation-induced alterations in LDL and also in accumulation of cholesterol in macrophages. The study thus proclaims that LDL-AGEs formed by fructose may contribute to accelerated initiation of diabetes induced atherosclerosis via foam cells generation and aminoguanidine may have therapeutic potential against it.

FgGyp8 as a putative FgRab1 GAP is required for growth and pathogenesis by regulating FgSnc1-mediated secretory vesicles fusion in Fusarium graminearum

Fusarium graminearum is an important plant pathogenic fungus that causes disease and yield reduction in many cereal crops, such as wheat and barley. Gyp8 stimulates GTP hydrolysis on Ypt1 in yeast. However, the functions of Gyp8 in plant pathogenic fungi are still unknown. In this study, we investigated the roles of Fg Gyp8 in F. graminearum by genetic and pathological analyses. Through gene knockout and phenotypic analyses, we found that Fg Gyp8 is required for vegetative growth in F. graminearum. The conidiation, conidial size and number of septa per conidium of ΔFggyp8 mutant are significantly reduced when compared to the wild type PH-1. Furthermore, Fg Gyp8 is crucial for pathogenicity on wheat coleoptiles and wheat heads. Fg Gyp8 contains a conserved TBC domain. Domain deletion analysis showed that the TBC domain, C-and N-terminal regions of Fg Gyp8 are all important for its biological functions in F. graminearum. Moreover, we showed that Fg Gyp8 catalyzes the hydrolysis of the GTP on Fg Rab1 to GDP in vitro, indicating that Fg Gyp8 is a GTPase-activating protein(GAP) for Fg Rab1. In addition, we demonstrated that Fg Gyp8 is required for Fg Snc1-mediated fusion of secretory vesicles with the plasma membrane in F. graminearum. Finally, we showed that Fg Gyp8 has functional redundancy with another Fg Rab1 GAP, Fg Gyp1, in F. graminearum. Taken together, we conclude that Fg Gyp8 is required for vegetative growth, conidiogenesis, pathogenicity and acts as a GAP for Fg Rab1 in F. graminearum.

Catalyzed degradation of disperse dyes by calcium alginate-pectin entrapped bitter gourd (Momordica charantia) peroxidase

Calcium-alginate pectin entrapped bitter gourd peroxidase （BGP） has been employed for the treatment of disperse dyes： Disperse Brown 1 （DB 1） and Disperse Red 17 （DR 17）. Peroxidase alone was unable to decolorize DR 17 and DB 1. However, the investigated dyes were decolorized maximally by BGP in the presence of 0.2 mmol/L redox mediator, violuric acid （VA）. A slow decrease in percent decolorization was observed when VA concentration was higher than 0.2 mmol/L which could likely be due to the high reactivity of its aminoxyl radical （ N–O . ） intermediate, that might undergo chemical reactions with aromatic amino acid side chains of the enzyme thereby inactivating it. Maximum decolorization of the dyes was observed at pH 3.0 and 40°C within 2 hr of incubation. Immobilized peroxidase decolorized 98% DR 17 and 71% DB 1 using 35 U of BGP in batch process in 90 min. Immobilized enzyme decolorized 85% DR 17 and 51% DB 1 whereas soluble enzyme decolorized DR 17 to 48% and DB 1 to 30% at 60°C. UV-visible spectral analysis was used to evaluate the degradation of these dyes and their toxicity was tested by Allium cepa test. The generally observed higher stability of the bioaffinity bound enzymes against various forms of inactivation may be related to the specific and strong binding of enzyme with bioaffinity support which prevents the unfolding/denaturation of enzyme. Thus entrapped peroxidase was found to be effective in the decolorization of the investigated dyes.

Validation of an Optimized Spectrophotometric Method for the Selective Determination of Labetalol Hydrochloride

A simple sensitive and economical method for the determination of labetalol hydrochloride has been proposed, based on the reaction of labetalol with sodium nitroprusside and hydroxylamine hydrochloride in sodium dihydrogen phosphate-sodium hydroxide buffer solution of pH 12. The green-blue color produced due to the formation of a nitroso derivative has been measured at 695 nm. The Beer＇s range was obeyed in the concentration range of 2-51 μg·mL^-1 with molar absorptivity of 0.48 × 10^4 L·mol^-1·cm^-1. Rigorous statistical analyses were performed for the validation of the method. A detailed investigation of the selectivity of the method has been done to find it to be highly selective for the determination of labetalol hydrochloride in the presence of its acidic degradation product and common excipients of formulations. The proposed method was successfully applied to the determination of labetalol hydrochloride in the laboratory prepared dosage forms. Comparison of the means of the proposed procedure with a reference method using point as well as interval hypotheses showed no statistically significant difference. The developed method was extended to investigate its applicability to biological samples.

Anti-inflammatory and bronchodilatory constituents of leaf extracts of Anacardium occidentale L. in animal models

Objective： Anocardium occidentale L. leaf is useful in the treatment of inflammation and asthma, but the bioactive constituents responsible for these activities have not been characterized. Therefore, this study was aimed at identifying the bioactive constituent（s） ofA. occidentale ethanolic leaf extract （AOEL） and its solvent-soluble portions, and evaluating their effects on histamine-induced paw edema and bronchocon- striction. Methods： The bronchodilatory effect was determined by measuring the percentage protection provided by plant extracts in the histamine-induced bronchoconstriction model in guinea pigs. The anti- inflammatory effect of the extracts on histamine-induced paw edema in rats was determined by measur- ing the increase in paw diameter, after which the percent edema inhibition was calculated. The extracts were analyzed using gas chromatography-mass spectrometry to identify the bioactive constituents. Column chromatography and Fourier transform infrared spectroscopy were used respectively to isolate and characterize the constituents. The bronchodilatory and anti-inflammatory activities of the isolated bioactive constituent were evaluated. Results： Histamine induced bronchoconstriction in the guinea pigs and edema in the rat paw. AOEL, hexane-soluble portion of AOEL, ethyl acetate-soluble portion of AOEL, and chloroform-soluble portion of AOEL significantly increased bronchodilatory and anti-inflammatory activities （P 〈 0.05）. Oleamide （9-octadecenamide） was identified as the most abundant compound in the extracts and was isolated. Oleamide significantly increased bronchodilatory and anti-inflammatory activities by 32.97% and 98.41%, respectively （P 〈 0.05）. Conclusion： These results indicate that oleamide is one of the bioactive constituents responsible for the bronchodilatory and anti-inflammatory activity of A. occidentale leaf, and can therefore be employed in the management of bronchoconstriction and inflammation.

Antioxidant defense system induced by cysteinestabilized peptide fraction of aqueous extract of Morinda lucida leaf in selected tissues of Plasmodium berghei-infected mice

OBJECTIVE： This study evaluated the responses of some antioxidant parameters in selected tissues of Plasmodium berghei-infected mice treated with cysteine-stabilized peptide fraction（CSPF） of aqueous extract of Morinda lucida leaf.METHODS：Fifty-six mice were randomly divided into seven groups.Group A（normal control）was uninfected and received 5%dimethyl sulfoxide（DMSO）.Mice in Groups B（negative control）,C,D,E and F were inoculated with P.berghei NK65 and were administered with 5%DMSO and 15.63,31.25,61.5 and 125 mg/kg body weight of CSPF respectively.Group G animals,were also inoculated with P.berghei NK65,and received 20 mg/kg body weight of chloroquine.The administration lasted for three days,after which malondialdehyde（MDA）concentration and various antioxidant parameters in selected tissues of mice were determined on days 4 and 8 postinoculation.RESULTS：The results revealed that MDA concentration was significantly increased（P〈0.05）in the tissues of the negative control and chloroquine-treated groups.The increased MDA concentration was reduced by CSPF in a dose-dependent manner,which was significant（P〈0.05）at higher doses.The activities of superoxide dismutase,catalase,glutathione peroxidase,glutathione reductase and glutathioneS-transferase and the concentration of reduced glutathione were significantly reduced（P〈0.05）in the tissues of the negative control animals compared to the normal controls.This observed reduction in the negative control animals was reverted in a dose-dependent manner in infected animals given CSPF,even to the range of the normal controls at highest dose,as did chloroquine.CONCLUSION：The results suggest that CSPF of M.lucida leaf extract may induce the antioxidant defense system in vivo against Plasmodium species infection.

Interplay of transport vesicles during plant-fungal pathogen interaction

Vesicle trafficking is an essential cellular process upon which many physiological processes of eukaryotic cells rely.It is usually the‘language’of communication among the components of the endomembrane system within a cell,between cells and between a cell and its external environment.Generally,cells have the potential to internalize membrane-bound vesicles from external sources by endocytosis.Plants constantly interact with both mutualistic and pathogenic microbes.A large part of this interaction involves the exchange of transport vesicles between the plant cells and the microbes.Usually,in a pathogenic interaction,the pathogen releases vesicles containing bioactive molecules that can modulate the host immunity when absorbed by the host cells.In response to this attack,the host cells similarly mobilize some vesicles containing pathogenesis-related compounds to the pathogen infection site to destroy the pathogen,prevent it from penetrating the host cell or annul its influence.In fact,vesicle trafficking is involved in nearly all the strategies of phytopathogen attack subsequent plant immune responses.However,this field of plant-pathogen interaction is still at its infancy when narrowed down to plant-fungal pathogen interaction in relation to exchange of transport vesicles.Herein,we summarized some recent and novel findings unveiling the involvement of transport vesicles as a crosstalk in plant-fungal phytopathogen interaction,discussed their significance and identified some knowledge gaps to direct future research in the field.The roles of vesicles trafficking in the development of both organisms are also established.

FgRab5 and FgRab7 are essential for endosomes biogenesis and nonredundantly recruit the retromer complex to the endosomes in Fusarium graminearum

The retromer complex,composed of the cargo-selective complex(CSC)Vps35-Vps29-Vps26 in complex with the sorting nexin dimer Vps5-Vps17,mediates the sorting and retrograde transport of cargo proteins from the endosomes to the trans-Golgi network in eukaryotic cells.Rab proteins belong to the Ras superfamily of small GTPases and regulate many trafficking events including vesicle formation,budding,transport,tethering,docking and fusion with target membranes.Herein,we investigated the potential functional relationship between the retromer complex and the 11 Rab proteins that exist in Fusarium graminearum using genetic and high-resolution laser confocal microscopic approaches.We found that only FgRab5(FgRab5A and FgRab5B)and FgRab7 associate with the retromer complex.Both FgVps35-GFP and FgVps17-GFP are mis-localized and appear diffused in the cytoplasm ofΔFgrab5A,ΔFgrab5B andΔFgrab7 mutants as compared to their punctate localization within the endosomes of the wild-type.FgRab7 and FgRab5B were found to co-localize with the retromer on endosomal membranes.Most strikingly,we found that these three Rab GTPases are indispensable for endosome biogenesis as both early and late endosomes could not be detected in the cells of the mutants after FM4-64 staining of the cells,while they were very clearly seen in the wild-type PH-1.Furthermore,FgRab7 was found to recruit FgVps35 but not FgVps17 to the endosomal membranes,whereas FgRab5B recruits both FgVps35 and FgVps17 to the membranes.Thus,we conclude that the Rab proteins FgRab5A,FgRab5B and FgRab7 play critical roles in the biogenesis of endosomes and in regulating retromer-mediated trafficking in F.graminearum.