Asian Journal of Biochemistry, Genetics and Molecular Biology最新文献

{"title":"Biological, Serological and Molecular Characterization of Cucumber mosaic virus (CMV) Infecting Cucurbita moschata (Duchene ex. Poir) (Summer Squash) in Itu and Uyo Local Governments Areas, Akwa Ibom State, Nigeria","authors":"Daniel A. Usanga, A. Owolabi, O. I. Eyong","doi":"10.9734/ajbgmb/2024/v16i6379","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i6379","url":null,"abstract":"Cucurbita moschata is an edible member of the family Cucurbitaceae. The plant is commonly observed showing typical virus-like symptoms during the growing season in Akwa Ibom State, Nigeria. A total of 7 leaf samples showing typical virus-like symptoms were collected from C. moschata in different locations in Itu and Uyo Local Government, Akwa Ibom State. All the virus isolates used in this study were sap transmissible with a wide host range. The virus isolates irrespective of the place of collection were transmitted in a fore-gut manner by A. spiraecola. Aphis citricida did not transmit the viruses. All the virus isolates reacted positively to cucumovirus polyclonal antibody in antigen coated plate-ELISA (ACP-ELISA). Gel electrophoresis of the cDNA of the virus isolates produced 500 base pairs typical of Cucumber mosaic virus. The gene sequence analysis revealed that all the isolates had sequence homology of 95% and above with Cucumber mosaic virus (CMV) and are therefore considered strains of CMV. The results also showed that CMV isolates investigated in this study shared common ancestry, though belonging to different clusters. The results also indicated that CMV could be widespread and therefore a threat to C. moschata production in Itu and Uyo Local Government Areas of Akwa Ibom State. This is the first report of CMV infecting C. moschata in Akwa Ibom State, Nigeria. Further work should be carried out to cover the entire state.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140671978","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of Aqueous Extract of Irvingia wombolu Seeds on Lipid Profile and Atherogenic Indices in Hyperlipidemic Wistar Rats","authors":"Joy O. Uba, O. E. Ezim","doi":"10.9734/ajbgmb/2024/v16i6378","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i6378","url":null,"abstract":"The effect of aqueous extract of Irvingia wombolu seeds on lipid profile and atherogenic indices in hyperlipidemic wistar rats was evaluated. Forty five (45) wistar rats were grouped into five groups of nine rats each. The animals were allowed seven days acclimatization period. Group one was the control group and it received normal rat chow and water throughout the study. Groups 2 to 5 were given high fat diet for 14 days after which they were fed with normal rat chow till the end of the study. At the end of the 14 days, group 2 was not treated while group 3-5 were treated with 250, 500 and 1000mg/kg body weight aqueous extract of Irvingia wombolu seed respectively for 28 days. The lipid profile of animals was assayed three times: first after 14 days induction period (phase 1) i.e day 0 of treatment, second was taken 14 days after treatment (phase 2), third was taken 28 days after treatment (phase 3).The study lasted for 49 days and distilled water was used as a vehicle for the extract.In phase 1, there was a significant decrease (p<0.05) in HDL (high density lipoprotein) level in all groups (2-5) compared to the control.there was a significant increase (p<0.05) in LDL ( low density lipoprotein), Total cholesterol, Triglyceride, VLDL (very low density lipoprotein), Non HDL, Cardic risk factor, Atherogenic coefficient, Atherogenic index of plasma levels in all groups (2-5) compared to the control. All doses of aqueous extract of Irvingia wombolu seed were able to increase HDL level and decrease other lipid profile parameters and atherogenic indices in hyperlipidemic rats. Thus, aqueous extract of Irvingia wombolu seed has antidyslipidemic potential.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140708330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adaptation and Growth Performance of Tea Plants (Camelia sinensis (L) O. Kuntze) in Obudu Hills, Cross River State","authors":"Idu, J. I., Edu, N. E., Egbaji, C. I., Nsude, L.","doi":"10.9734/ajbgmb/2024/v16i6377","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i6377","url":null,"abstract":"Young seedlings of five tea clones under cultivation (318, 236, 68, BB35, and 143) and five from the germplasm), were collected from the Kakara Highland tea Plantations, Mambila Plateau, Taraba State. Seedlings were planted in fifty sample boxes to enable the seedlings to be established first before transplanting to the field. After the establishment, they were transported to Obudu Hills for investigation. The experiment was laid out in a randomized complete block design (RBCD) with 3 replications. Data collected on morphological traits were subjected to an analysis of variance (ANOVA) test. Multivariate analysis such as principal component analysis (PCA), hierarchical cluster analysis and correlation analysis were used to estimate genetic diversity between the tea plant clones. The result showed that the tea clones survived the first two months after transplanting. However, subsequently, the percentage of survival was reduced. Results from the morphological studies revealed that the accessions differed significantly (p>0.001) in all evaluated traits except for leaf width and dry biomass.  Three principal components were identified based on eigenvalues; the first principal component contributed 44.18% of the total variation with an eigenvalue of 3.977, the second principal component accounted for 24.36% with total eigenvalue of 2.193 while PC3 accounted for 15.345% of the total variation with eigenvalue of 1.381.  Cluster analysis showed that the evaluated tea clones were grouped into two main clusters containing sub-clusters. The high genetic diversity identified in Camellia sinensis clone in the study will serve as a guide in breeding, genetic improvement, utilization and conservation of this important nutritional and medicinal plant species.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140708974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Neutrophil-lymphocyte and Platelet-lymphocyte Ratios of Pregnant Women Living with HIV/AIDS","authors":"Lebechukwu, B.C., Mbachu, N.A., Eleje, G.U., Orji, E. C., Obiegbu, N.P., Umennadi, R.O","doi":"10.9734/ajbgmb/2024/v16i5376","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i5376","url":null,"abstract":"Background: Chronic inflammation, a factor for high mortality, persists throughout the pregnancy period of women living with human immunodeficiency Virus (HIV). \u0000Aim: To determine whether pregnancy alone or the combined effects of HIV and pregnancy are the sources of the inflammation, this study evaluated the predictive variables for inflammation, lymphocyte ratio (NLR), and Platelet-Lymphocyte Ratio (PLR). \u0000Materials and Methods: A population of 128 women, aged 15-49 years were grouped into 4; Group A- 32 HIV positive and pregnant, Group B- 32 HIV negative and pregnant, Group C- 32 HIV positive not pregnant, Group D- 32 HIV negative not pregnant (control group). Differential white blood cell and platelet count methods were used to determine the neutrophil (N), Lymphocyte (L) and platelet (P) counts. The ratio calculation method was used to derive the NLR and PLR. \u0000Results: NLR was significantly higher for group A (2.62±0.41) compared to groups B (2.25 ± 0.18); C (0.95 ± 0.07); D (1.68 ± 0.19) at p ≤ 0.00. However, the NLR of groups A and B (p ≤ 0.05) were not significantly different, PLR was significantly higher for group A (9.86±1.12) compared to groups B (6.65 ± 1.12); C (5.35 ± 0.47); D (6.34 ± 0.43). There was no significant difference in blood pressure across the groups, significant difference was however observed in their Body Mass Index. \u0000Conclusion: Hence, Inflammation may have been caused by pregnancy alone.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140730189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of Benzo (a) Pyrene on Thyroid Glands and their Associated Hormones of Male Mice Mus musculus","authors":"Anita Singh Purvia, V. Shrivastava","doi":"10.9734/ajbgmb/2024/v16i5375","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i5375","url":null,"abstract":"Benzo [a] pyrene is the main representative of polycyclic aromatic hydrocarbons, and has been repeatedly found in the air, surface water, soil, and sediments. Human exposure to Benzo [a] pyrene is therefore common. Benzo (a) pyrane (BaP) which is mainly presented in newspaper ink and inhibits the function of the thyroid and other endocrine glands of the human system, due to several disorders that occur in the human body. Healthy mature male mice Mus musculus having weight of 125 ± 5 gm were used for the experiments. The effects of a Benzo (a) pyrene (0.65mg/25g body weight/twice in a week) during experimental periods of 60 and 90 days in male mice Mus musculus showed significant difference in body weights compared to control. We assessed the hormonal estimation of thyroid treated with Benzo (a) pyrene by using an appropriate ELISA assay kit and histopathological changes were observed under a light microscope (magnifications 100X) in the thyroid tissue of male mice Mus musculus exposed to BaP after 60 and 90 days. During the study, we observed that in treated mice thyroid hormone concertation was affected the structure of the thyroid follicles was disturbed and epithelial cells were necrotic compared with the control mice which indicates that the BaP may lead to changes in the thyroid morphology as well as secreted hormone concentration in exposed treated male mice.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140746604","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation, Biological Characterization, and Quality Inspection of Human Umbilical Cord-derived Mesenchymal Stem Cells for Clinical Research","authors":"Xia Lin Zuo, Dongcheng Wu, Ding Yu, M. A. Sa’ad, Samantha Lo Yin Teng, Manickam Ravichandran, Ke Xiang Ding, Seng Kong Tham","doi":"10.9734/ajbgmb/2024/v16i4371","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i4371","url":null,"abstract":"Background: Human umbilical cord mesenchymal stem cells (hUCMSCs) represent a valuable and versatile cell type derived from the umbilical cord tissue of newborns. These cells exhibit diverse characteristics, including multipotent differentiation potential and low immunogenicity. hUCMSCs hold promise for clinical applications, yet concerns persist regarding their chromosomal stability. This study employs short tandem repeat (STR) analysis to assess genomic integrity. By focusing on chromosomal stability, the study aims to enhance confidence in the clinical viability of hUCMSCs, contributing to their safe and effective use in regenerative medicine. hUCMSCs were extracted from healthy newborn umbilical cords using tissue block adherence and enzyme digestion. The third generation of hUCMSCs undergoes biological characterization and quality checks, including examinations of cell morphology, viability, growth curves, surface markers, cell cycle, and multi-lineage differentiation potential. The result of the present study shows that hUCMSCs exhibit mesenchymal stem cell characteristics, with robust growth and high viability. Positive surface markers are expressed at rates exceeding 95%, while negative markers are ≤2%. Cell karyotyping analysis, STR spectrum identification, and chromosomal microarray analysis confirm genetic stability. Microbial testing shows the absence of contaminants, and immunological studies demonstrate the immunomodulatory capabilities of hUCMSCs. Specific residues, such as trypsin, were not detected. In conclusion, hUCMSCs produced under strict GLP and GMP conditions meet the quality guidelines for clinical application in various stem cell therapies. The findings support the clinical application of hUCMSCs in various medical contexts.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140229044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"On the Substrate Specificity and Some Properties of the Extracellular Oxidase from the Neonothopanus nambi Basidiomycete","authors":"Olga Mogilnaya, N. Ronzhin, E. Posokhina, Violeta Le, Yulia Zakharova, Andrey Sukhikh, Vladimir Bondar","doi":"10.9734/ajbgmb/2024/v16i4369","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i4369","url":null,"abstract":"The present study reports experimental data on substrate specificity and some properties of the extracellular enzyme with oxidase activity isolated from the mycelium of the higher fungus Neonothopanus nambi IBSO 2391 by treating the biomass with (beta) -glucosidase. Gel-filtration chromatography showed that molecular weight of the isolated enzyme was 80 kDa. Spectral analysis did not reveal any chromophore components in the enzyme. The extracellular oxidase of the basidiomycete N. nambi IBSO 2391 was active with most of the aromatic compounds chosen as model substrates. An important fact is that the enzyme exhibited catalytic activity with no hydrogen peroxide or any other mediators added to the reaction mixture. The highest activity of the enzyme was observed in reactions with veratryl alcohol and hydroquinone. In reactions with guaiacol and aromatic amines (diaminobenzidine, o-dianisidine), the level of activity of the extracellular oxidase was considerably lower – by a factor of 2.5–3.5. In reactions with resorcinol, phenol, and caffeic acid, the catalytic efficiency of the enzyme was no greater than 6% of its activity with veratryl alcohol. Kinetic parameters of enzymatic reactions were determined for the most efficiently oxidized substrates. The addition of the chelating agent of divalent metal ions (EDTA) did not affect the activity of the extracellular oxidase from the fungus N. nambi IBSO 2391, indicating the absence of divalent metal ions in the molecule of the enzyme. At the same time, addition of the SH reagent (DTT) increased catalytic efficiency of the enzyme. The study showed that the extracellular oxidase of the fungus N. nambi IBSO 2391 functions in wide ranges of temperature and pH of the reaction medium, showing the highest catalytic activity at temperatures between 22 and 35 °C and pH 6.0. Results obtained in the current study provide the basis for studying potential uses of the isolated enzyme in biomedical analytics and bioremediation.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140246721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Current Strategies for Management of Plant Viruses and Future Perspectives: Enhancing Crop Health, Yield and Productivity","authors":"Akshita Soni, R. P. Kushvaha, S. K. Snehi","doi":"10.9734/ajbgmb/2024/v16i4368","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i4368","url":null,"abstract":"Plant viruses pose significant threats to agricultural productivity and food security worldwide. This article explores various management strategies employed to overcome the impact of plant viruses on crops, with a focus on enhancing crop health and productivity. It underscores the need for proactive management strategies to minimize yield losses, reduce disease spread, and maintain sustainable crop production through preventive measures such as Conventional and Non- conventional approaches by highlighting the key methods such as the use of resistant cultivars, cultural practices, vector control, thermos, electro, and chemotherapy, along with CRISPR, RNAi, and nanotechnology. This review article provides an overview of the current management approaches employed for controlling plant viruses.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140258889","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of Ethanol Extract of Avocado Pulp (Persia americana mill) on Sperm Qualityand Histological Changes in Caffeine Induced Testicular Injury in Wistarrats","authors":"John N. Nkwocha, O. E. Ezim","doi":"10.9734/ajbgmb/2024/v16i3365","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i3365","url":null,"abstract":"This study evaluated the effect of ethanol extract of Persea americana pulp on caffeine induced testicular damage on male wistar rats. Fifty-four (54) mature male rats were used, they were divided into six (6) groups of nine (9) rats. Group 1 was not induced with caffeine, Group two(2) to group six(6) were induced with 200mg/kg caffeine for two weeks. Three rats were sacrificed from each group after two weeks of induction. Semen and  testes collected for total sperm count, sperm motility, sperm morphology, sperm vitality and histopathology. The rats were treated as follows for another four weeks, Group 1: no induction, no treatment. Group 2: induced withcaffeine without treatment. Group 3; 100mg/kg extract. Group 4:300mg/kg extract. Group 5:500mg/kg extract, Group 6:0.107mg/kg proviron. Three rats from each group were sacrificed two weeks after treatment and four weeks after treatment. semen and testes were collected for sperm parameters and histopathology respectively. Different doses of Persea americanapulp was able to improve the sperm quality in rats induced with caffeine in a dose and time dependent manner. Histopathological examination of the testes treated with extract showed recovery from damages caused by caffeine in dose dependent manner. Ethanol extract of Persea americanapulphas a dose ameliorating effect on caffeine induced testicular damage on male rats.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140264094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative In vitro Cytotoxicity Evaluation of Moringa Extract, Clitoria Extract and Coccinia grandis by MTT Assay Using Various Cell Lines","authors":"Manisha Chaudhari, Himanshi Trivedi, Shradhdha Sharma, Krunal Solanki, Darpesh Gohel","doi":"10.9734/ajbgmb/2024/v16i2360","DOIUrl":"https://doi.org/10.9734/ajbgmb/2024/v16i2360","url":null,"abstract":"Evaluation and development of potent anti-cancer treatments have been a global interest of research due to significant rise in cancer as well as related diseases. Many herbal extracts have shown the potential to be used in the treatment of such diseases. In order to evaluate the anticancer features of Moringa oleifera (ME), Clitoria ternatea (CE), and Coccinia grandis (CCD), this study was carried out using various cell lines i.e. L-929, Vero, L-132, Raw 264.7, and Balb 3T3 with six different concentrations of each test substance by using MTT cell viability Assay. Additionally, in-vitro studies are performed using these test items and claims are being done that these test items are playing a pivotal role either treatment of Alzheimer's diseases (AD) and Parkinson's disease (PD) or decreasing the severity and incidences of such neurodegenerative diseases. Globally, an average life expectancy rises, so does the severity of these conditions as well. Hence, this study was also aimed to determine whether any supportive changes are showed on different cell lines or not.   In contrast to normal cells, the test substances revealed possible anti-cancer characteristics when exposed to carcinoma cell lines based on the data obtained in this study. The results indicated that ME, CE and CCD are effective against cancer cells; hence based on the results of these study, it could be concluded that that the data of the present study supports the interpretation that these test items have significant effects either in treatment or slowing down the clinical signs of PD and AD under the conditions and procedures followed in the present study. Based on the results of cell viability results it could also be concluded that the test items are safer to use up to the concentrations of 500 µg/mL.","PeriodicalId":8498,"journal":{"name":"Asian Journal of Biochemistry, Genetics and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139961091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}