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Morphometry and subpopulation structure of Holstein bull spermatozoa： variations in ejaculates and cryopreservation straws

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Asian Journal of Andrology 2016年第6期18卷 851-857页

作者： Anthony Valverde Hector Arenan Maria Sancho Jesus contell Jesus Yaniz Alejandro Fernandez caries soler Technological Institute of costa Rica San Carlos Campus S Xenetica Fontao Fontao-Esperante 27210 Lugo Spain R+D Department Proiser R+D Scientific Park University of Valencia 46980 Paterna Spain Department of Animal Production and Food Science Politechnical School of Huesca University of Zaragoza 22071 Huesca Spain Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Spain

Sperm quality is evaluated for the calculation of sperm dosage in artificial reproductive programs. The most common parameter used is motility, but morphology has a higher potential as a predictor of genetic quality. Morphometry calculations from cASA-Morph technology improve morphological evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vagina, and several ejaculates were studied. After general evaluation, samples were diluted, packaged in 0.25 ml straws, and stored in liquid nitrogen. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a cASA-Morph system for eight morphometric parameters. In addition to the ＂classical＂ statistical approach, based on variance analysis （revealing differences between animals, ejaculates, and straws）, principal component （Pc） analysis showed that the variables were grouped into Pc1, related to size, and Pc2 to shape. Subpopulation structure analysis showed four groups, namely, big, small, short, and narrow from their dominant characteristics, representing 31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering an ejaculate sperm population as divided into subpopulations reflecting quantifiable parameters generated by cASA-Morph systems technology opens a new view on sperm function. This is the first study applying this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive programs.

关键词： cryopreservation Holstein bull sperm morphometry subpopulations

Morphometric comparison by the ISAS cASA-DNAf system of two techniques for the evaluation of DNA fragmentation in human spermatozoa

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Asian Journal of Andrology 2016年第6期18卷 835-839页

作者： Sara Sadeghi Almudena Garcia-Molina Ferran celma Anthony Valverde Sogol Fereidounfar caries soler Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Valencia Spain R+D Department Proiser R+D Scientific Park Universitat de Valencia 46980 Paterna Valencia Spain Technological Institute of costa Rica San Carlos Campus School of Agronomy 223-21001 Alajuela Costa Rica Department of Botany Kharazmi University Tehran Iran

DNA fragmentation has been shown to be one of the causes of male infertility, particularly related to repeated abortions, and different methods have been developed to analyze it. In the present study, two commercial kits based on the ScD technique （Halosperm~ and SDFA） were evaluated by the use of the DNA fragmentation module of the ISAS vl cASA system. Seven semen samples from volunteers were analyzed. To compare the results between techniques, the Kruskal-Wallis test was used. Data were used for calculation of Principal components （two Pcs were obtained）, and subsequent subpopulations were identified using the Halo, Halo/core Ratio, and Pc data. Results from both kits were significantly different （P 〈 0.001）. In each case, four subpopulations were obtained, independently of the classification method used. The distribution of subpopulations differed depending on the kit used. From the Pc data, a discriminant analysis matrix was obtained and a good a posterioriclassification was obtained （97.1% for Halosperm and 96.6% for SDFA）. The present results are the first approach on morphometric evaluation of DNA fragmentation from the ScD technique. This approach could be used for the future definition of a classification matrix surpassing the current subjective evaluation of this important sperm factor.

关键词： DNA fragmentation Halosperm morphometry principal component analysis SDFA subpopulation

A functional tacrolimus-releasing nerve wrap for enhancing nerve regeneration following surgical nerve repair

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Neural Regeneration Research 2025年第1期20卷 291-304页

作者： Simeon c.Daeschler Katelyn J.W.So Konstantin Feinberg Marina Manoraj Jenny cheung Jennifer Zhang Kaveh Mirmoeini JPaul Santerre Tessa Gordon Gregory HBorschel SickKids Research Institute Neuroscience and Mental Health ProgramTorontoONCanada Institute of Biomaterials and Biomedical Engineering University of TorontoTorontoONCanada Indiana University School of Medicine IndianapolisINUSA Division of Plastic and Reconstructive Surgery the Hospital for Sick ChildrenTorontoONCanada Translational Biology and Engineering Program Ted Rogers Centre for Heart ResearchInstitute of Biomedical EngineeringFaculty of DentistryUniversity of TorontoTorontoONCanada

Axonal regeneration following surgical nerve repair is slow and often incomplete,resulting in poor functional recovery which sometimes contributes to lifelong ***,there are no FDA-approved therapies available to promote nerve *** accelerates axonal regeneration,but systemic side effects presently outweigh its potential benefits for peripheral nerve *** authors describe herein a biodegradable polyurethane-based drug delivery system for the sustained local release of tacrolimus at the nerve repair site,with suitable properties for scalable production and clinical application,aiming to promote nerve regeneration and functional recovery with minimal systemic drug *** is encapsulated into co-axially electrospun polycarbonate-urethane nanofibers to generate an implantable nerve wrap that releases therapeutic doses of bioactive tacrolimus over 31 *** and drug loading are adjustable for applications in small and large caliber nerves,and the wrap degrades within 120 days into biocompatible *** released from the nerve wrap promotes axon elongation in vitro and accelerates nerve regeneration and functional recovery in preclinical nerve repair models while off-target systemic drug exposure is reduced by 80%compared with systemic *** its surgical suitability and preclinical efficacy and safety,this system may provide a readily translatable approach to support axonal regeneration and recovery in patients undergoing nerve surgery.

关键词： biodegradable local drug delivery nerve injury nerve regeneration nerve wrap tacrolimus

Relationship of sperm plasma membrane andacrosomal integrities with sperm morphometry inBos taurus

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Asian Journal of Andrology 2020年第6期22卷 578-582页

作者： Inmaculada Palacin Pilar Santolaria carlos Alquezar-Baeta carles soler Miguel A Silvestre Jesus Yaniz BIOFITER Research Group Environmental Sciences Institute(IUCA)Department of Animal Production and Food SciencesUniversity of ZaragozaHuesca 22071Spain Department of computer Science University of ZaragozaZaragoza 50018Spain Department of cellular Biology Functional Biology and Physical AnthropologyUniversity of ValenciaBurjassot 46100Spain

To date,sperm morphometric studies have assessed whole sperm populations without considering sperm *** aim of this study was to evaluate the relati on ship of sperm membrane and acrosomal integrity with sperm morphometry in liquid semen samples collected from *** this end,sperm morphometry was performed on cryopreserved semen samples from 16 bulls by a combination of fluorescent dyes,including Hoechst 33343,carboxyfluorescein diacetate,and propidium *** allowed discrimination of different subpopulations on the basis of sperm membrane and acrosomal integrity and analysis of the morphometries of the sperm head,nucleus,and acrosome using a specific plug-in module created on *** integrity was related to sperm morphometry as the heads of spermatozoa with a damaged acrosome were significantly smaller than those with a normal acrosome(Pcase of spermatozoa with an intact acrosome,those with a damaged plasma membrane had a larger sperm head and acrosome than spermatozoa with an intact plasma membrane(Pcant differences in the sperm head size were observed between sperm subpopulations without an acrosome or in the nuclear sperm morphometry of the different *** was a positive correlation between the sperm motility values of the samples and the morphometric parameters for in tact *** correlations were particularly strong for the morphometric parameters of the sperm *** conclude that there are clear differences in the sperm morphometry depending on the status of the sperm membrane and acrosome and this should be considered when performing this kind of analysis.

关键词： Bos taurus fluoresce nee microscopy image analysis sperm morphometry sperm quality

Sperm kinematic, head morphometric and kinetic- morphometric subpopulations in the blue fox （Alopex lagopus）

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Asian Journal of Andrology 2017年第2期19卷 154-159页

作者： caries soler Jesus contell Lorena Bori Maria Sancho Almudena Garcia-Molina Anthony Valverde Jan Segarvall Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Spain R+D Department Proiser R+D Scientific Park University of Valencia 46980 Paterna Spain Technological Institute of costa Rica San Carlos Campus School of Agronomy 223-21001 Alajuela Costa Rica Finnish Fur Breeders Association ProFur 65101 Vaasa Finland

This work provides information on the blue fox ejaculated sperm quality needed for seminal dose calculations. Twenty semen samples, obtained by masturbation, were analyzed for kinematic and morphometric parameters by using cASA-Mot and cASA-Morph system and principal component （Pc） analysis. For motility, eight kinematic parameters were evaluated, which were reduced to Pc1, related to linear variables, and Pc2, related to oscillatory movement. The whole population was divided into three independent subpopulations： SP1, fast cells with linear movement; SP2, slow cells and nonoscillatory motility; and SP3, medium speed cells and oscillatory movement. In almost all cases, the subpopulation distribution by animal was significantly different. Head morphology analysis generated four size and four shape parameters, which were reduced to Pc 1, related to size, and Pc2, related to shape of the cells. Three morphometric subpopulations existed. SPI： large oval cells; SP2： medium size elongated cells; and SP3. small and short cells. The subpopulation distribution differed between animals. combining the kinematic and morphometric datasets produced Pc1, related to morphometric parameters, and Pc2, related to kinematics, which generated four sperm subpopulations - SPI.. high oscillatory motility, large and short heads; SP2; medium velocity with small and short heads; SP3. slow motion small and elongated cells; and SP4. high linear speed and large elongated cells. Subpopulation distribution was different in all animals. The establishment of sperm subpopulations from kinematic, morphometric, and combined variables not only improves the well-defined fox semen characteristics and offers a good conceptual basis for fertility and sperm preservation techniques in this species, but also opens the door to use this approach in other species, included humans.

关键词： integration of motility and morphology principal component analysis sperm morphometry subpopulation

Updating semen analysis:a subpopulation approach

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Asian Journal of Andrology 2020年第1期22卷 118-119页

作者： Almudena Garcia-Molina Anthony Valverde Daznia Bompart carina caldeira Alberto Vendrell carles soler Proiser R+D S.L. Scientific ParkUniversitat de ValenciaPaterna 46980ValenciaSpain Department of Functional Biology and Physical Anthropology University of ValenciaBurjassot 46100ValenciaSpain Technological Institute of costa Rica San Carlos CampusSchool of AgronomyAlajuela 223-21001Costa Rica

Dear Editor,Reproductive problems affect 8%-12%of human populations worldwide;40%-50%of all infertility cases are due to the male,and up to 2%of men have suboptimal sperm quality.

关键词： dating cases infertility

Dog sperm head morphometry： its diversity and .=volution

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Asian Journal of Andrology 2017年第2期19卷 149-153页

作者： caries soler Ana Alambiaga Maria A Marti Almudena Garcia-Molina Anthony Valverde Jesus contell Marcos campos Department of Functional Biology and Physical Anthropology University of Valencia 46100 Burjassot Spain R+D Department Proiser R+D Scientific Park University of Valencia 46980 Paterna Spain 3Technological Institute of costa Rica San Carlos Campus School of Agronomy 223-21001 Alajuela Costa Rica Department of Medicine and Animal Surgery University CardenaI-Herrera-CEU 46113 Moncada Valencia Spain Global Veterinaria (Reprovalcan) 46500 Sagunto Valencia Spain clinica Veterinaria Sangueso 46009 Valencia Spain

Dogs have been under strong artificial selection as a consequence of their relationship with man. Differences between breeds are evident that could be reflected in seminal characteristics. The present study was to evaluate differences in sperm head morphometry between seven well-defined breeds of dog： the British Bulldog, chihuahua, German Shepherd, Labrador Retriever, Spanish Mastiff, Staffordshire Terrier, and Valencian Rat Hunting dog. Semen samples were obtained by masturbation and smears stained with Diff-Quik. Morphometric analysis （cASA-Morph） produced four size and four shape parameters. Length, Ellipticity, and Elongation showed higher differences between breeds. MANOVA revealed differences among all breeds. considering the whole dataset, principal component analysis （PcA） showed that Pc1 was related to head shape and Pc2 to size. Procluster analysis showed the British Bulldog to be the most isolated breed, followed by the German Shepherd. The PcA breed by breed showed the chihuahua, Labrador Retriever, Spanish Mastiff, and Staffordshire Terrier to have Pc1 related to shape and Pc2 to size, whereas the British Bulldog, Valencia Rat Hunting dog, and German Shepherd had Pc1 related to size and Pc2 to shape. The dendrogram for cluster groupings and the distance between them showed the British Bulldog to be separated from the rest of the breeds. Future work on dog semen must take into account the large differences in the breeds＇ sperm characteristics. The results provide a base for future work on phylogenetic and evolutionary studies of dogs, based on their seminal characteristics.

关键词： canis familiaris cluster analysis diversity sperm morphometry

Prevalence of visual impairment and estimation of refractive errors among school children in Kakamega,Kenya

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International Journal of Ophthalmology(English edition) 2024年第5期17卷 932-939页

作者： Isabel Signes-soler Alfred Ragot Sheilah Nangena Andrew Wekesa Raúl Montalbán Llamusí Department of Optics and Optometry and Vision Sciences Universidat de ValènciaBurjassotValencia 46010Spain NGO Vision Without Borders(Visio Sense Fronteres VSF)Calpe 03710AlicanteSpain Masinde Muliro University of Science and Technology P.O Box 190-50100KakamegaKenya clinica Baviera Alicante c/Ausóy Monzó 16Alicante 03006Spain

AIM:To investigate the prevalence of visual impairment(VI)and provide an estimation of uncorrected refractive errors in school-aged children,conducted by optometry students as a community ***:The study was *** 3343 participants were included in the *** initial examination involved assessing the uncorrected distance visual acuity(UDVA)and visual acuity(VA)while using a+2.00 D *** inclusion criteria for a subsequent comprehensive cycloplegic eye examination,performed by an optometrist,were as follows:a UDVAcimal(0.20 logMAR)and/or a VA with+2.00 D≥0.8 decimal(0.96 logMAR).RESULTS:The sample had a mean age of 10.92±2.13y(range 4 to 17y),and 51.3%of the children were female(n=1715).The majority of the children(89.7%)fell within the age range of 8 to *** the ethnic groups,the highest representation was from the Luhya group(60.6%)followed by Luo(20.4%).Mean logMAR UDVA choosing the best eye for each student was 0.29±0.17(range 1.70 to 0.22).Out of the total,246 participants(7.4%)had a full eye *** estimated prevalence of myopia(defined as spherical equivalent≤-0.5 D)was found to be 1.45%of the total *** around 0.18%of the total sample had hyperopia value exceeding+1.75 *** astigmatism(cilce was 1.26%of the total *** our cases of VI,76.2%could be attributed to uncorrected refractive *** was detected in 0.66%(22/3343)of the screened *** was no statistically significant correlation observed between age or gender and refractive ***:The primary cause of VI is determined to be uncorrected refractive errors,with myopia being the most prevalent refractive error *** findings underscore the significance of early identification and correction of refractive errors in school-aged children as a means to alleviate the impact of VI.

关键词： visual impairment refractive errors myopia amblyopia sustainable development goals

The influence of canopy-layer composition on understory plant diversity in southern temperate forests

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Forest Ecosystems 2017年第2期4卷 154-166页

作者： Luciana Mestre Monica Toro-Manriquez Rosina soler Alejandro Huertas-Herrera Guillermo Martinez-Pastur Maria Vanessa Lencinas National Scientific and Technical Research council (cONIcET) Austral Center for Scientific Research (CADIC-CONICET) Houssay 200 9410 Ushuaia Tierradel Fuego Argentina

Background： Understory plants represents the largest component of biodiversity in most forest ecosystems and plays a key role in forest *** their importance, the influence of overstory-layer composition on understory plant diversity is relatively poorly understood within deciduous-evergreen broadleaved mixed *** aim of this work was to evaluate how tree overstory-layer composition influences on understory-layer diversity in three forest types（monospecific deciduous Nothofagus pumilio（Np）, monospecific evergreen Nothofagus betuloides（Nb）, and mixed ***（M） forests）, comparing also between two geographical locations（coast and mountain） to estimate differences at landscape ***： We recorded 46 plant species： 4 ferns, 12 monocots, and 30 ***-layer composition influences the herb-layer structure and diversity in two different ways： while mixed forests have greater similarity to evergreen forests in the understory structural features, deciduous and mixed were similar in terms of the specific composition of plant *** pure stands were the most diverse, meanwhile evergreen stands were least *** of exclusive species of mixed forest could represent a transition where evergreen and deciduous communities meet and ***, landscape has a major influence on the structure, diversity and richness of understory vegetation of pure and mixed forests likely associated to the magnitude and frequency of natural disturbances, where mountain forest not only had highest herb-layer diversity but also more exclusive ***： Our study suggests that mixed Nothofagus forest supports coexistence of both pure deciduous and pure evergreen understory plant species and different assemblages in coastal and mountain *** the mixture of canopy patch types within mixed stands will be important for conserving the natural patterns of understory plant composition in southern beech mixe

关键词： Landscape Understory communities Understory structure Forest structure Nothofagus conservation