Analyze Diet

Topic:Genomics

Genomics in horses involves the study and analysis of the horse genome to understand genetic variations and their implications for equine health, performance, and breeding. This field encompasses the identification and mapping of genes associated with specific traits, diseases, and conditions in horses. Techniques such as whole-genome sequencing and genome-wide association studies (GWAS) are employed to explore genetic diversity and inheritance patterns among different horse breeds. Genomics provides insights into hereditary disorders, informs selective breeding practices, and aids in the development of personalized veterinary care. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings of genomic research in equine science.
Liposome-mediated uptake of exogenous DNA by equine spermatozoa and applications in sperm-mediated gene transfer.
Equine veterinary journal    December 18, 2007   Volume 40, Issue 1 76-82 doi: 10.2746/042516407X235786
Ball BA, Sabeur K, Allen WR.Sperm-mediated gene transfer has been reported as a method for production of transgenic animals in a variety of species, and this technique represents a possible method for production of transgenic equids. Objective: To evaluate the uptake of exogenous DNA (enhanced green fluorescent protein; pEGFP) by equine spermatozoa and to assess the ability of transfected spermatozoa to introduce this transgene into early equine embryos. Methods: To evaluate incorporation of pEGFP into equine spermatozoa, washed spermatozoa were incubated with 32P-pEGFP, with or without lipofection. Spermatozoa were also...
Review of genetic aspects of radiological alterations in the navicular bone of the horse.
DTW. Deutsche tierarztliche Wochenschrift    December 15, 2007   Volume 114, Issue 11 404-411 
Diesterbeck U, Distl O.Navicular disease or podotrochlosis has long been known to cause forelimb lameness in horses. It had been proposed that the development of podotrochlosis has similarities to the human osteoarthritis (OA) complex. Alterations of the navicular bone can be made visible early in life only on the basis of radiographs. Reports on the prevalences of navicular disease indicate that radiological alterations in the navicular bone are present in different warmblood populations at frequencies of between 14.9% and 87.6%. Genetic factors play an important role in the development of the radiological signs. E...
Loss of expression of protein kinase a regulatory subunit 1alpha in pigmented epithelioid melanocytoma but not in melanoma or other melanocytic lesions.
The American journal of surgical pathology    December 7, 2007   Volume 31, Issue 11 1764-1775 doi: 10.1097/PAS.0b013e318057faa7
Zembowicz A, Knoepp SM, Bei T, Stergiopoulos S, Eng C, Mihm MC, Stratakis CA.Pigmented epithelioid melanocytoma (PEM) is a recently described entity comprising most cases previously described as "animal-type melanoma" and epithelioid blue nevus (EBN) occurring in patients with the multiple neoplasia syndrome Carney complex (CNC). Mutations of the protein kinase A regulatory subunit type 1alpha (R1alpha) (coded by the PRKAR1A gene) are found in more than half of CNC patients. In this study, we investigated whether PEM and EBN are related at the molecular level, and whether changes in the PRKAR1A gene status and the expression of the R1alpha protein may be involved in th...
The horse genome project–sequence based insights into male reproductive mechanisms.
Reproduction in domestic animals = Zuchthygiene    December 6, 2007   Volume 42 Suppl 2 45-50 doi: 10.1111/j.1439-0531.2007.00897.x
Leeb T.The growing knowledge on physiology, cell biology and biochemistry of the reproductive organs has provided many insights into molecular mechanisms that are required for successful reproduction. Research directed at the investigation of reproduction physiology in domestic animals was hampered in the past by a lack of species-specific genomic information. The genome sequences of dog, cattle and horse have become publicly available in 2005, 2006 and 2007 respectively. Although the gene content of mammalian genomes is generally very similar, genes involved in reproduction tend to be less conserved...
Allelic heterogeneity at the equine KIT locus in dominant white (W) horses.
PLoS genetics    November 14, 2007   Volume 3, Issue 11 e195 doi: 10.1371/journal.pgen.0030195
Haase B, Brooks SA, Schlumbaum A, Azor PJ, Bailey E, Alaeddine F, Mevissen M, Burger D, Poncet PA, Rieder S, Leeb T.White coat color has been a highly valued trait in horses for at least 2,000 years. Dominant white (W) is one of several known depigmentation phenotypes in horses. It shows considerable phenotypic variation, ranging from approximately 50% depigmented areas up to a completely white coat. In the horse, the four depigmentation phenotypes roan, sabino, tobiano, and dominant white were independently mapped to a chromosomal region on ECA 3 harboring the KIT gene. KIT plays an important role in melanoblast survival during embryonic development. We determined the sequence and genomic organization of t...
Equine umbilical cord blood contains a population of stem cells that express Oct4 and differentiate into mesodermal and endodermal cell types.
Journal of cellular physiology    October 12, 2007   Volume 215, Issue 2 329-336 doi: 10.1002/jcp.21312
Reed SA, Johnson SE.Mesenchymal stem cells (MSCs) offer promise as therapeutic aids in the repair of tendon, ligament, and bone damage suffered by sport horses. The objective of the study was to identify and characterize stem-like cells from newborn foal umbilical cord blood (UCB). UCB was collected and MSC isolated using human reagents. The cells exhibit a fibroblast-like morphology and express the stem cell markers Oct4, SSEA-1, Tra1-60 and Tra1-81. Culture of the cells in tissue-specific differentiation media leads to the formation of cell types characteristic of mesodermal and endodermal origins. Chondrogenic...
Molecular characterization and chromosomal assignment of equine cartilage derived retinoic acid sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA).
Gene    October 5, 2007   Volume 407, Issue 1-2 98-104 doi: 10.1016/j.gene.2007.09.022
Berg LC, Mata X, Thomsen PD.Cartilage-derived retinoic acid sensitive protein (CD-RAP) also known as melanoma inhibitory activity (MIA) has already been established as a marker for chondrocyte differentiation and a number of cancerous conditions in humans. Studies have also shown that CD-RAP/MIA is a potential marker of joint disease. The objective of this study was to characterize the equine CD-RAP/MIA gene and thus make it available as a marker in cartilage research and clinical studies. Gene analysis revealed that the equine gene (GenBank accession no. EF679787) consists of four exons and three introns, and the homolo...
Sequence analysis of the equine SLC26A2 gene locus on chromosome 14q15–>q21.
Cytogenetic and genome research    September 29, 2007   Volume 118, Issue 1 55-62 doi: 10.1159/000106441
Hansen M, Knorr C, Hall AJ, Broad TE, Brenig B.The solute carrier family 26, member 2 (SLC26A2) gene belongs to a family of multifunctional anion exchangers. Mutations in the human SLC26A2 gene are associated with autosomal recessively inherited chondrodysplasias. Hence, we postulate that the equine SLC26A2 could be a candidate gene for conformational traits in horses. An equine BAC clone harboring the SLC26A2 gene was isolated. The complete 142,625 bp insert sequence of this clone was determined by transposon sequencing. Together with the SLC26A2 gene the BAC clone contains four genes, i.e. the macrophage colony stimulating factor 1 recep...
Genome-wide search for markers associated with osteochondrosis in Hanoverian warmblood horses.
Mammalian genome : official journal of the International Mammalian Genome Society    September 29, 2007   Volume 18, Issue 10 739-747 doi: 10.1007/s00335-007-9058-9
Dierks C, Löhring K, Lampe V, Wittwer C, Drögemüller C, Distl O.A genome-wide scan was performed to detect quantitative trait loci (QTLs) for osteochondrosis (OC) and osteochondrosis dissecans (OCD) in horses. The marker set comprised 260 microsatellites. We collected data from 211 Hanoverian warmblood horses consisting of 14 paternal half-sib families. Traits used were OC (fetlock and/or hock joints affected), OCD (fetlock and/or hock joints affected), fetlock OC, fetlock OCD, hock OC, and hock OCD. The first genome scan included 172 microsatellite markers. In a second step 88 additional markers were chosen to refine putative QTLs found in the first scan....
A new view on dam lines in Polish Arabian horses based on mtDNA analysis.
Genetics, selection, evolution : GSE    September 27, 2007   Volume 39, Issue 5 609-619 doi: 10.1186/1297-9686-39-5-609
Głazewska I, Wysocka A, Gralak B, Sell J.Polish Arabian horses are one of the oldest and the most important Arab populations in the world. The Polish Arabian Stud Book and the Genealogical Charts by Skorkowski are the main sources of information on the ancestors of Polish Arabs. Both publications were viewed as credible sources of information until the 1990s when the data regarding one of the dam lines was questioned. The aim of the current study was to check the accuracy of the pedigree data of Polish dam lines using mtDNA analysis. The analyses of a 458 bp mtDNA D-loop fragment from representatives of 15 Polish Arabian dam lines re...
Estimation of genetic parameters and prediction of breeding values for multivariate threshold and continuous data in a simulated horse population using Gibbs sampling and residual maximum likelihood.
Journal of animal breeding and genetics = Zeitschrift fur Tierzuchtung und Zuchtungsbiologie    September 18, 2007   Volume 124, Issue 5 308-319 doi: 10.1111/j.1439-0388.2007.00666.x
Stock KF, Hoeschele I, Distl O.Simulated horse data were used to compare multivariate estimation of genetic parameters and prediction of breeding values (BV) for categorical, continuous and molecular genetic data using linear animal models via residual maximum likelihood (REML) and best linear unbiased prediction (BLUP) and mixed linear-threshold animal models via Gibbs sampling (GS). Simulation included additive genetic values, residuals and fixed effects for one continuous trait, liabilities of four binary traits, and quantitative trait locus (QTL) effects and genetic markers with different recombination rates and polymor...
Characterization of the equine glycogen debranching enzyme gene (AGL): Genomic and cDNA structure, localization, polymorphism and expression.
Gene    August 23, 2007   Volume 404, Issue 1-2 1-9 doi: 10.1016/j.gene.2007.07.034
Herszberg B, Mata X, Giulotto E, Decaunes P, Piras FM, Chowdhary BP, Chaffaux S, Guérin G.Glycogen debranching enzyme (AGL) is a multifunctional enzyme acting in the glycogen degradation pathway. In humans, the AGL activity deficiency causes a type III glycogen storage disease (Cori-Forbes disease). One particularity of AGL gene expression lies in the multiple alternative splicing in its 5' region. The AGL gene was localized on ECA5q14-q15. The sequence of the equine cDNA was determined to be 7.5 kb in length with an open reading frame of 4602 bp. The gene is 69 kb long and contains 35 exons. The equine AGL gene has an ubiquitous expression and presents five tissue-dependent cDNA v...
Karyotypic relationships among Equus grevyi, Equus burchelli and domestic horse defined using horse chromosome arm-specific probes.
Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology    August 23, 2007   Volume 15, Issue 6 807-813 doi: 10.1007/s10577-007-1164-8
Musilova P, Kubickova S, Zrnova E, Horin P, Vahala J, Rubes J.Using laser microdissection we prepared a set of horse chromosome arm-specific probes. Most of the probes were generated from horse chromosomes, some of them were derived from Equus zebra hartmannae. The set of probes were hybridized onto E. grevyi chromosomes in order to establish a genome-wide chromosomal correspondence between this zebra and horse. The use of arm-specific probes provided us with more information on the mutual arrangement of the genomes than we could obtain by means of whole-chromosome paints generated by flow sorting, even if we used reciprocal painting with probe sets from...
High altitude adaptation and phylogenetic analysis of Tibetan horse based on the mitochondrial genome.
Journal of genetics and genomics = Yi chuan xue bao    August 21, 2007   Volume 34, Issue 8 720-729 doi: 10.1016/S1673-8527(07)60081-2
Xu S, Luosang J, Hua S, He J, Ciren A, Wang W, Tong X, Liang Y, Wang J, Zheng X.To investigate genetic mechanisms of high altitude adaptations of animals living in the Tibetan Plateau, three mitochondrial genomes (mt-genome) of Tibetan horses living in Naqu (4,500 m) of Tibetan, Zhongdian (3,300 m) and Deqin (3,100 m) of Yunnan province were sequenced. The structures and lengths of these three mt-genomes are similar to the Cheju horse, which is related to Tibetan horses, but little shorter than the Swedish horse. The pair-wise identity of these three horses on nucleotide level is more than 99.3%. When the gene encoding the mitochondrial protein of Tibetan horses was analy...
Sequence analysis of the equid herpesvirus 2 chemokine receptor homologues E1, ORF74 and E6 demonstrates high sequence divergence between field isolates.
The Journal of general virology    August 19, 2007   Volume 88, Issue Pt 9 2450-2462 doi: 10.1099/vir.0.82942-0
Sharp EL, Farrell HE, Borchers K, Holmes EC, Davis-Poynter NJ.Equid herpesvirus 2 (EHV-2), in common with other members of the subfamily Gammaherpesvirinae, encodes homologues of cellular seven-transmembrane receptors (7TMR), namely open reading frames (ORFs) E1, 74 and E6, which each show some similarity to cellular chemokine receptors. Whereas ORF74 and E6 are members of gammaherpesvirus-conserved 7TMR gene families, E1 is currently unique to EHV-2. To investigate their genetic variability, EHV-2 7TMRs from a panel of equine gammaherpesvirus isolates were sequenced. A region of gB was sequenced to provide comparative sequence data. Phylogenetic analysi...
The diversity of coliphages and coliforms in horse feces reveals a complex pattern of ecological interactions.
Applied and environmental microbiology    August 17, 2007   Volume 73, Issue 19 5975-5981 doi: 10.1128/AEM.01145-07
Golomidova A, Kulikov E, Isaeva A, Manykin A, Letarov A.The diversity of coliphages and indigenous coliform strains (ICSs) simultaneously present in horse feces was investigated by culture-based and molecular methods. The richness of coliforms (as estimated by the Chao1 method) is about 1,000 individual ICSs distinguishable by genomic fingerprinting present in a single sample of feces. This unexpectedly high value indicates that some factor limits the competition of coliform bacteria in the horse gut microbial system. In contrast, the diversity of phages active against any selected ICS is generally limited to one to three viral genotypes present in...
Mapping quantitative trait loci for osteochondrosis in fetlock and hock joints and palmar/plantar osseus fragments in fetlock joints of South German Coldblood horses.
Animal genetics    June 9, 2007   Volume 38, Issue 4 350-357 doi: 10.1111/j.1365-2052.2007.01610.x
Wittwer C, Löhring K, Drögemüller C, Hamann H, Rosenberger E, Distl O.The aim of this study was to identify quantitative trait loci (QTL) for osteochondrosis (OC) and palmar/plantar osseous fragments (POF) in fetlock joints in a whole-genome scan of 219 South German Coldblood horses. Symptoms of OC and POF were checked by radiography in 117 South German Coldblood horses at a mean age of 17 months. The radiographic examination comprised the fetlock and hock joints of all limbs. The genome scan included 157 polymorphic microsatellite markers. All microsatellite markers were equally spaced over the 31 autosomes and the X chromosome, with an average distance of 17.7...
The mechanisms determining the nucleolar-organizing regions inactivation of domestic horse chromosomes.
Journal of animal breeding and genetics = Zeitschrift fur Tierzuchtung und Zuchtungsbiologie    June 7, 2007   Volume 124, Issue 3 163-171 doi: 10.1111/j.1439-0388.2007.00642.x
Slota E, Wnuk M, Bugno M, Pienkowska-Schelling A, Schelling C, Bratus A, Kotylak Z.Cytogenetic investigations of the nucleolar-organizing regions (NORs) show that there is variation in the transcriptional activity of rDNA in many organisms. As a consequence, genetic polymorphism of these regions has been detected. The aim of the present study was to evaluate the hypothetic genetic mechanisms determining the NORs polymorphism of the domestic horse chromosomes. Molecular cytogenetic analyses were carried out on Hucul horses and the following techniques were used: fluorescence in situ hybridization (FISH), telomere primed in situ synthesis (PRINS), in situ nick-translation with...
Molecular characterization of the equine herpesvirus 1 strains RacL11 and Kentucky D.
The Journal of veterinary medical science    June 7, 2007   Volume 69, Issue 5 573-576 doi: 10.1292/jvms.69.573
Ghanem YM, Ibrahim el-SM, Yamada S, Matsumura T, Osterrieder N, Yamaguchi T, Fukushi H.The pathogenicities of RacL11 and Kentucky D strains of equine herpesvirus 1 in the hamster infection model are different from those of Ab4p and the Japanese isolates. Virus genome restriction fragment length polymorphism analysis and sequence comparison of an intergenic region, glycoproteins and tegument genes showed higher conservation but with some strain-specific differences. These results indicate that point nucleotide differences in RacL11 and Kentucky D might be responsible for their pathogenicity in rodent models.
Genome-wide search for microsatellite markers associated with radiologic alterations in the navicular bone of Hanoverian warmblood horses.
Mammalian genome : official journal of the International Mammalian Genome Society    June 6, 2007   Volume 18, Issue 5 373-381 doi: 10.1007/s00335-007-9021-9
Diesterbeck US, Hertsch B, Distl O.The aim of this study was to identify quantitative trait loci (QTLs) for pathologic changes in the navicular bone in Hanoverian warmblood horses. Seventeen paternal half-sib groups comprising 192 individuals were analyzed in a whole-genome scan. These families included 144 progeny and grandchildren, which were randomly chosen from the Hanoverian warmblood. Three different traits were considered: deformed canales sesamoidales and radiographic changes in the contour and in the structure of the navicular bone. The genome scan included in total 214 highly polymorphic microsatellite markers. The pu...
DNA archive for companion animals.
The Veterinary record    June 5, 2007   Volume 160, Issue 22 776 doi: 10.1136/vr.160.22.776
Clements DN, Carter S, Innes J, Kennedy L, Bennett D, Day M.No abstract available
Identification and characterization of equine granzyme B.
Veterinary immunology and immunopathology    June 3, 2007   Volume 118, Issue 3-4 239-251 doi: 10.1016/j.vetimm.2007.05.002
Piuko K, Bravo IG, Müller M.In the present study we describe the isolation and characterization of putative equine granzyme B for which we propose the designation 'eqGrzmB'. Sequence analysis revealed characteristic features of a GrzmB protease such as the presence of a signal (leader-) peptide and an activation di-peptide. The isolated eqGrzmB is functionally active when expressed in human or in insect cells. Furthermore, exchange of any of three putative active site amino acids, which are highly conserved along granzyme B enzymes, led to a complete loss of enzymatic activity in the newly identified eqGrzmB. Phylogeneti...
Evaluation of Compass as a comparative mapping tool for ESTs using horse radiation hybrid maps.
Animal genetics    June 2, 2007   Volume 38, Issue 3 294-302 doi: 10.1111/j.1365-2052.2007.01603.x
Coleman SJ, Gong G, Gaile DP, Chowdhary BP, Bailey E, Liu L, MacLeod JN.Loci for 9322 equine expressed sequence tags (ESTs) were predicted using the Comparative Mapping by Annotation and Sequence Similarity (Compass) strategy in order to evaluate the programme's ability to make accurate locus predictions in species with comparative gene maps. Using human genome sequence information from Build 35 (May 2004) and published marker information from the radiation hybrid (RH) maps for equine chromosomes (ECA) 17 and X, 162 ESTs were predicted to locations on ECA17 and 328 ESTs to locations on ECAX by selection of the 'top blast hit'. The locations of 30 ESTs were assesse...
Equine lysozyme: the molecular basis of folding, self-assembly and innate amyloid toxicity.
FEBS letters    May 21, 2007   Volume 581, Issue 14 2587-2592 doi: 10.1016/j.febslet.2007.05.023
Morozova-Roche LA.Calcium-binding equine lysozyme (EL) combines the structural and folding properties of c-type lysozymes and alpha-lactalbumins, connecting these two most studied subfamilies. The structural insight into its native and partially folded states is particularly illuminating in revealing the general principles of protein folding, amyloid formation and its inhibition. Among lysozymes EL forms one of the most stable molten globules and shows the most uncooperative refolding kinetics. Its partially-folded states serve as precursors for calcium-dependent self-assembly into ring-shaped and linear amyloi...
Heterospecific nuclear-transferred embryos derived from equine fibroblast cells and enucleated bovine oocytes.
Reproduction in domestic animals = Zuchthygiene    May 18, 2007   Volume 42, Issue 3 243-247 doi: 10.1111/j.1439-0531.2006.00759.x
Zhou H, Liu C, Wang W.This study was conducted to reconstruct heterogeneous embryos using equine skin fibroblast cells as donor karyoplasts and the bovine oocytes as recipient cytoplast for investigating the reprogramming of equine somatic cell nuclear in bovine oocyte cytoplasm and the developmental potential of the reconstructed embryos. Adult horse skin fibroblast cells serum-starved were used as donor somatic cells. Bovine oocytes matured in vitro were employed as recipient cytoplasts. The fusion of fibroblast cells into recipient cytoplasm was induced by electofusion. The fused eggs were activated by inomycin ...
Homozygosity mapping approach identifies a missense mutation in equine cyclophilin B (PPIB) associated with HERDA in the American Quarter Horse.
Genomics    May 11, 2007   Volume 90, Issue 1 93-102 doi: 10.1016/j.ygeno.2007.03.009
Tryon RC, White SD, Bannasch DL.Hereditary equine regional dermal asthenia (HERDA), a degenerative skin disease that affects the Quarter Horse breed, was localized to ECA1 by homozygosity mapping. Comparative genomics allowed the development of equine gene-specific markers which were used with a set of affected horses to detect a homozygous, identical-by-descent block spanning approximately 2.5 Mb, suggesting a recent origin for the HERDA mutation. We report a mutation in cyclophilin B (PPIB) as a novel, causal candidate gene for HERDA. A c.115G>A missense mutation in PPIB alters a glycine residue that has been conserved acr...
[Modeling the structure of supergenes controlling some polyallelic blood group systems in the pig Sus scrofa and horse Equus caballus].
Genetika    May 10, 2007   Volume 43, Issue 3 382-392 
Kniazev SP, Nikitin SV.Two polymorphic blood group systems (E and M) of the pig Sus scrofa L. and one blood group system (D) of the horse Equus caballus L. have been studied. On the basis of phenogroup analysis, models describing the formation of the complex allele spectra of these systems and reflecting the contributions of mutations and recombinations have been constructed. The complementary relationships between the antigens determined by the variants of supergenes within the systems, as ell as the probable number and relative positions of the subloci encoding individual groups of antigens in them, have been dete...
Bayesian estimation of genetic parameters for multivariate threshold and continuous phenotypes and molecular genetic data in simulated horse populations using Gibbs sampling.
BMC genetics    May 9, 2007   Volume 8 19 doi: 10.1186/1471-2156-8-19
Stock KF, Distl O, Hoeschele I.Requirements for successful implementation of multivariate animal threshold models including phenotypic and genotypic information are not known yet. Here simulated horse data were used to investigate the properties of multivariate estimators of genetic parameters for categorical, continuous and molecular genetic data in the context of important radiological health traits using mixed linear-threshold animal models via Gibbs sampling. The simulated pedigree comprised 7 generations and 40000 animals per generation. Additive genetic values, residuals and fixed effects for one continuous trait and ...
Phenotypical assays and partial sequencing of the hsp60 gene for identification of Streptococcus equi.
Current microbiology    May 4, 2007   Volume 54, Issue 5 331-334 doi: 10.1007/s00284-005-0458-3
Sá e Silva M, da Costa MM, de Avila Botton S, Barretta C, Groff AC, de Vargas AC.Strangles is an acute and contagious disease characterized by inflammation of the upper respiratory tract of horses. The etiological agent of strangles is the bacteria S. equi subsp. equi, which belongs to the Lancefield group C. Opportunistic agents from the same group are frequently isolated from horses with strangles and may induce mistaken diagnoses. Among the subspecies of S. equi, the phenotypic features are almost undistinguishable; however, the pathogenic potential is widely differentiated. The aim of this study was to characterize S. equi isolates obtained from clinical samples of str...
Time- and dose-dependent effects of roundup on human embryonic and placental cells.
Archives of environmental contamination and toxicology    May 4, 2007   Volume 53, Issue 1 126-133 doi: 10.1007/s00244-006-0154-8
Benachour N, Sipahutar H, Moslemi S, Gasnier C, Travert C, Séralini GE.Roundup is the major herbicide used worldwide, in particular on genetically modified plants that have been designed to tolerate it. We have tested the toxicity and endocrine disruption potential of Roundup (Bioforce on human embryonic 293 and placental-derived JEG3 cells, but also on normal human placenta and equine testis. The cell lines have proven to be suitable to estimate hormonal activity and toxicity of pollutants. The median lethal dose (LD(50)) of Roundup with embryonic cells is 0.3% within 1 h in serum-free medium, and it decreases to reach 0.06% (containing among other compounds 1.2...
1 56 57 58 59 60 77