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Corrales F.J.,Research Center Medica Aplicada | Mora M.I.,Research Center Medica Aplicada | Pello-Palma J.,University of Oviedo | Moreno J.F.,Sexing Technologies
Journal of Proteome Research | Year: 2013

The bovine endometrium recognizes early embryos and reacts differently depending on the developmental potential of the embryo. However, it is unknown whether the endometrium can distinguish embryonic sex. Our objective was to analyze sexual dimorphism in the uterus in response to male and female embryos. Differentially expressed (DE) proteins, different levels of hexoses, and other embryotrophic differences were analyzed in uterine fluid (UF). Proteomic analysis of day-8 UF recovered from heifers after the transfer of day-5 male or female embryos identified 23 DE proteins. Regulated proteasome/immunoproteasome protein subunits indicated differences in antigen processing between UF carrying male embryos (male-UF) or female embryos (female-UF). Several enzymes involved in glycolysis/gluconeogenesis and antioxidative/antistress responses were up-regulated in female-UF. Fructose concentration was increased in female-UF versus male-UF, while glucose levels were similar. In vitro cultures with molecules isolated from male-UF were found to improve male embryo development compared to female embryos cultured with molecules isolated from female-UF. We postulated that, in vivo, male embryos induce changes in the endometrium to help ensure their survival. In contrast, female embryos do not appear to induce these changes. © 2013 American Chemical Society.


Gosalvez J.,Autonomous University of Madrid | Ramirez M.A.,Autonomous University of Madrid | Lopez-Fernandez C.,Autonomous University of Madrid | Crespo F.,Centro Militar Of Cria Caballar Of Avila | And 3 more authors.
Theriogenology | Year: 2011

This study examined the dynamic response of Spermatozoa DNA Fragmentation after sex selection in bulls using a MoFlo® SX (Beckman Coulter, Miami FL) spermatozoa sorter. The dynamic response of spermatozoa DNA fragmentation refers to the changing values of SDF, i.e., rate of SDF (rSDF), when analyzed periodically over a set incubation time at 37 °C. A dynamic assessment of SDF using non-sorted and sex-sorted spermatozoa samples during 72 h of incubation at 37 °C was performed. Results showed a reduced DNA longevity in sex-sorted frozen-thawed spermatozoa, with spermatozoa DNA damage appearing between 24 h and 48 h. The baseline SDF level was higher in conventional frozen-thawed than in sex-sorted frozen-thawed spermatozoa samples; while the reverse occurred for the rSDF. The afore-mentioned result produced a crossover point between both dynamic tendencies of SDF for sex-sorted versus conventional samples. We defined this crossover point as the Crossover Positioning Time (CPT) or the time (in hours) where both curves crossover after a period of spermatozoa incubation at 37 °C. The point at which the CPT occurs could be used as an indicator of the rSDF for individual bulls after X- and Y-chromosome bearing spermatozoa selection. CPT values produced a window of SDF ranging between 24 h and 48 h in the present experiment. It is proposed that higher values for CPT are indicative of bulls presenting chromatin that is more resistant to the external stressors affecting spermatozoa DNA after spermatozoa sorting. © 2011 Elsevier Inc.


Gosalvez J.,Autonomous University of Madrid | Ramirez M.A.,Autonomous University of Madrid | Lopez-Fernandez C.,Autonomous University of Madrid | Crespo F.,Centro Militar Of Cria Caballar Of Avila | And 3 more authors.
Theriogenology | Year: 2011

This study examined the static response of Spermatozoa DNA Fragmentation (SDF) after sex selection in bulls using a MoFlo® SX (Beckman Coulter, Miami FL) spermatozoa sorter to produce three different subpopulations: 1) Spermatozoa bearing X- chromosomes with a purity of 95%, 2) Spermatozoa bearing Y-chromosomes with a purity of 95%, and 3) non-viable spermatozoa. The static response of SDF refers to the baseline values observed for DNA damage when analyzed pre- and post sex-sorting. Results showed that while the baseline level SDF in pre-sorted bull spermatozoa samples ranged from 5.3% to 11% with an average of 7.9% ± 2.1%, the level of SDF obtained in X- and Y-chromosome sorted samples was much lower (3.1% ± 1.9%) and statistical differences were obtained after comparing both groups (P < 0.01). Spermatozoa containing a fragmented DNA molecule tend to be accumulated in the non-viable subpopulation. The baseline SDF level in X- and Y-chromosome sorted subpopulations is reduced, by 63% on average when compared to the values obtained in the neat semen sample. Different bulls exhibit unique SDF reduction efficiencies via the X- and Y-chromosome sex selection process. © 2011 Elsevier Inc.


Gonzalez-Marin C.,Sexing Technologies | Gosalvez J.,Autonomous University of Madrid | Roy R.,Autonomous University of Madrid
International Journal of Molecular Sciences | Year: 2012

Concentration, motility and morphology are parameters commonly used to determine the fertilization potential of an ejaculate. These parameters give a general view on the quality of sperm but do not provide information about one of the most important components of the reproductive outcome: DNA. Either single or double DNA strand breaks can set the difference between fertile and infertile males. Sperm DNA fragmentation can be caused by intrinsic factors like abortive apoptosis, deficiencies in recombination, protamine imbalances or oxidative stress. Damage can also occur due to extrinsic factors such as storage temperatures, extenders, handling conditions, time after ejaculation, infections and reaction to medicines or post-testicular oxidative stress, among others. Two singular characteristics differentiate sperm from somatic cells: Protamination and absence of DNA repair. DNA repair in sperm is terminated as transcription and translation stops post-spermiogenesis, so these cells have no mechanism to repair the damage occurred during their transit through the epididymis and post-ejaculation. Oocytes and early embryos have been shown to repair sperm DNA damage, so the effect of sperm DNA fragmentation depends on the combined effects of sperm chromatin damage and the capacity of the oocyte to repair it. In this contribution we review some of these issues. © 2012 by the authors; licensee MDPI, Basel, Switzerland.


BEVERLY, Mass. and TORONTO, Feb. 22, 2017 (GLOBE NEWSWIRE) -- Hamilton Thorne Ltd. (TSX-V:HTL), a leading provider of precision instruments, consumables, software and services to the Assisted Reproductive Technologies (ART) and developmental biology research markets, today reported preliminary selected unaudited financial results for the fourth quarter and year-ended December 31, 2016. Preliminary unaudited results show that 2016 revenues increased 16% to over $10.5 million, led by the contribution from its newly acquired Embryotech Laboratories business, as well as strong growth in sales of LYKOS® clinical laser systems and increased revenues from after sale services.  Fourth quarter sales increased 37% to over $3.6 million, with net income and EBITDA expected to be over $500,000 and $750,000 respectively, a record quarter for the Company. For the year, including the effect of approximately $275,000 of one-time expenses relating to the Embryotech acquisition, the Company is expecting to report net income of approximately $650,000, and EBITDA of approximately $1.2 million. David Wolf, President and Chief Executive Officer of Hamilton Thorne Ltd. commented, “2016 was a transformational year for us as we completed a significant expansion of both our financial profile and business capabilities when we acquired Embryotech Laboratories in September.  I am pleased to report that the integration of the businesses is progressing smoothly and we are actively working on several growth initiatives brought about by marketing synergies between the Hamilton Thorne and Embryotech businesses.   Building on our internal growth initiatives and the acquisitions of the Oosight product line in 2015 and Embryotech in 2016, we remain committed to our strategy of growth through new product innovation, organic growth and acquisition.” Mr. Wolf added, “Based on continued foreign exchange headwinds as well as changes in distributor relationships in certain of our markets, we saw some challenges in our instrument business, particularly in the fourth quarter.  Gross profit margins for the year improved to over 65% as, despite some significant pricing actions to reflect currency issues, we increased sales of high-margin software, consumables and services with recurring revenues.  In order to accelerate our sales growth and get closer to our customers, we have begun to put more investment into direct sales resources in both our instrument and services businesses.” The financial information contained in this news release is based on management's estimates and is subject to adjustment. The Company expects to release its completed audited financial statements for the year ended December 31, 2016 on or about April 26, 2017. Expansion of Board of Directors The Company also announced that David Sable, portfolio manager for New York-based Special Situations Life Sciences Fund has joined the Company’s board of directors.  Dr. Sable is a reproductive endocrinologist by training and an expert in the field of human assisted reproduction.  Dr. Sable also serves on the board of directors of RESOLVE: The National Infertility Society and on the advisory board of OvaScience. All amounts are in US dollars, unless specified otherwise, and results, with the exception of EBITDA, are expressed in accordance with the International Financial Reporting Standards ("IFRS"). About Hamilton Thorne Ltd. (www.hamiltonthorne.com)   Hamilton Thorne is a leading world-wide provider of precision instruments, consumables, software and services that reduce cost, increase productivity, improve results and enable breakthroughs in Assisted Reproductive Technologies (ART) and developmental biology research markets. Hamilton Thorne's laser products attach to standard inverted microscopes and operate as micro-surgical devices, enabling a wide array of scientific applications and In Vitro Fertilization (IVF) procedures. Its image analysis systems are designed to bring quality, efficiency and reliability to studies of reproductive cells in the human fertility, animal sciences and reproductive toxicology fields.  Hamilton Thorne’s standardized toxicology assays and quality control testing services help to improve outcomes in human IVF clinics.  Hamilton Thorne’s growing worldwide customer base consists of pharmaceutical companies, biotechnology companies, fertility clinics, university research centers, animal breeding companies, and other commercial and academic research establishments, including Harvard, MIT, Yale, McGill, Oxford, Cambridge, the Smithsonian Institution, Charles River Labs, Covance, ABS Global, Sexing Technologies, Merck, Cook Medical, Novartis, Pfizer, and Dow Chemical. Neither the Toronto Venture Exchange, nor its regulation services provider (as that term is defined in the policies of the exchange), accepts responsibility for the adequacy or accuracy of this release. The Company has included earnings before interest, income taxes, depreciation and amortization, (“EBITDA”) as a non-IFRS measure, which is used by management as a measure of financial performance. See section entitled “Use of Non-IFRS Measures” in the Company’s Management Discussion and Analysis for the periods covered for further information. Certain information in this press release may contain forward-looking statements. This information is based on current expectations that are subject to significant risks and uncertainties that are difficult to predict. Actual results might differ materially from results suggested in any forward-looking statements. The Company assumes no obligation to update the forward-looking statements, or to update the reasons why actual results could differ from those reflected in the forward-looking statements unless and until required by securities laws applicable to the Company. Additional information identifying risks and uncertainties is contained in filings by the Company with the Canadian securities regulators, which filings are available at www.sedar.com.


News Article | November 28, 2016
Site: www.marketwired.com

BEVERLY, MA and TORONTO, ON--(Marketwired - Nov 28, 2016) - Hamilton Thorne Ltd. (TSX VENTURE: HTL), a leading provider of precision instruments, consumables, software and services to the Assisted Reproductive Technologies (ART) and developmental biology research markets today reported operational and financial results for the quarter and nine months ended September 30, 2016. Sales increased 18% to $2.45 million for the quarter ended September 30, 2016 versus the prior year. Sales for the first nine months of 2016 increased to $6.86 million, up 7%. Net income and EBITDA for the quarter and nine months declined versus the prior year periods due to planned investments in research and development and sales and marketing resources, as well as significant increases in general and administrative expenses in the second and third quarter relating to the Company's acquisition program. David Wolf, President and Chief Executive Officer stated, "We continue to see good growth in our instrument business as a result of the significant investments in sales and support resources and market development funding made over the past 6-12 months. Sales into our core in vitro fertilization (IVF) clinic market led our growth, driven by a substantial increase in sales of our clinical laser systems. Margins were up slightly at 64.5% for the quarter and flat at 63.9% for the nine-months ended September 30, 2016, due to product mix and the impact of higher Embryotech margins." Mr. Wolf continued, "We were pleased to complete the acquisition of the Embryotech Laboratories business in the third quarter. Embryotech is the acknowledged US leader in providing quality control services and testing assays to the ART community. In addition to acquiring a great brand, we will be strengthening the financial performance of Hamilton Thorne. Not only will this acquisition substantially increase our scale, we anticipate that it will diversify our revenue base from a predominantly one-time sale of instruments model to a projected 40% of recurring revenues from the sales of services and consumables. We expect that our revenue, profitability, and cash flow will be positively impacted in future quarters as the results from our combined operations begin to be realized." The Company reported that operating expenses were generally in line with expectations, reflecting its increased investment in R&D, staffing, sales and marketing, significant expenses related to its acquisition program, and increases in non-cash amortization and expense resulting from stock option grants. All amounts are in US dollars, unless specified otherwise, and results, with the exception of EBITDA, are expressed in accordance with the International Financial Reporting Standards ("IFRS"). Cash generated by operations was $686,981 for the nine-months ended September 30, 2016, compared to $711,176 in the comparable period of the prior year, a decrease of $24,195, primarily due to the decrease in net income and increases in acquisition expenses. In September 2016, the Company closed on a new credit facility consisting of a $5.5 million five-year term loan at 4.5% interest rate and a $2.5 million revolving line of credit at prime, maturing in September 2018 ($1.5 million of which was drawn at closing and outstanding at September 30, 2016), for net new borrowings of $3.5 million, which was used to fund the Embryotech acquisition. After utilizing existing cash of $2.6 million for the acquisition, the Company ended the quarter with $1.9 million of cash on hand. The Financial Statements and accompanying Management Discussion and Analysis for the periods are available on www.sedar.com and the Hamilton Thorne website. Hamilton Thorne is a leading world-wide provider of precision instruments, consumables, software and services that reduce cost, increase productivity, improve results and enable breakthroughs in Assisted Reproductive Technologies (ART) and developmental biology research markets. Hamilton Thorne's laser products attach to standard inverted microscopes and operate as micro-surgical devices, enabling a wide array of scientific applications and In Vitro Fertilization (IVF) procedures. Its image analysis systems are designed to bring quality, efficiency and reliability to studies of reproductive cells in the human fertility, animal sciences and reproductive toxicology fields. Hamilton Thorne's standardized toxicology assays and quality control testing services help to improve outcomes in human IVF clinics. Hamilton Thorne's growing worldwide customer base consists of over 1,000 pharmaceutical companies, biotechnology companies, fertility clinics, university research centers, animal breeding companies, and other commercial and academic research establishments, including Harvard, MIT, Yale, McGill, Oxford, Cambridge, the Smithsonian Institution, Charles River Labs, Covance, ABS Global, Sexing Technologies, Merck, Cook Medical, Novartis, Pfizer, and Dow Chemical. Neither the Toronto Venture Exchange, nor its regulation services provider (as that term is defined in the policies of the exchange), accepts responsibility for the adequacy or accuracy of this release. The Company has included earnings before interest, income taxes, depreciation and amortization, ("EBITDA") as a non-IFRS measure, which is used by management as a measure of financial performance. See section entitled "Use of Non-IFRS Measures" in the Company's Management Discussion and Analysis for the periods covered for further information. Certain information in this press release may contain forward-looking statements. This information is based on current expectations that are subject to significant risks and uncertainties that are difficult to predict including the risk that the Company may not be able to obtain the necessary regulatory approvals, as applicable. Actual results might differ materially from results suggested in any forward-looking statements. The Company assumes no obligation to update the forward-looking statements, or to update the reasons why actual results could differ from those reflected in the forward-looking statements unless and until required by securities laws applicable to the Company. Additional information identifying risks and uncertainties is contained in filings by the Company with the Canadian securities regulators, which filings are available at www.sedar.com


News Article | November 30, 2016
Site: www.marketwired.com

BEVERLY, MA and TORONTO, ON--(Marketwired - Nov 30, 2016) - Hamilton Thorne Ltd. (TSX VENTURE: HTL), a leading provider of precision instruments, consumables, software and services to the Assisted Reproductive Technologies (ART) and developmental biology research markets announced today announced that it will be presenting at the 9th annual LD Micro Main Event (www.ldmicro.com), on Wednesday, December 6, 2016 at 2:30 pm Pacific Time (5:30 pm Eastern Time), at the Luxe Sunset Boulevard Hotel in Los Angeles, CA. David Wolf, President and CEO of Hamilton Thorne will be presenting and will be available for one-on-one meetings with the investment community on December 6 and 7. Investors attending the conference who wish to schedule a meeting should notify their LD Micro conference contact or Hamilton Thorne at ir@hamiltonthorne.com. The LD Micro Main Event is the largest independent conference for small/microcap companies and will feature 240 presenting companies. About Hamilton Thorne Ltd. (www.hamiltonthorne.com) Hamilton Thorne is a leading world-wide provider of precision instruments, consumables, software and services that reduce cost, increase productivity, improve results and enable breakthroughs in Assisted Reproductive Technologies (ART) and developmental biology research markets. Hamilton Thorne's laser products attach to standard inverted microscopes and operate as micro-surgical devices, enabling a wide array of scientific applications and In Vitro Fertilization (IVF) procedures. Its image analysis systems are designed to bring quality, efficiency and reliability to studies of reproductive cells in the human fertility, animal sciences and reproductive toxicology fields. Hamilton Thorne's standardized toxicology assays and quality control testing services help to improve outcomes in human IVF clinics. Hamilton Thorne's growing worldwide customer base consists of pharmaceutical companies, biotechnology companies, fertility clinics, university research centers, animal breeding companies, and other commercial and academic research establishments, including Harvard, MIT, Yale, McGill, Oxford, Cambridge, the Smithsonian Institution, Charles River Labs, Covance, ABS Global, Sexing Technologies, Merck, Cook Medical, Novartis, Pfizer, and Dow Chemical. Neither the Toronto Venture Exchange, nor its regulation services provider (as that term is defined in the policies of the exchange), accepts responsibility for the adequacy or accuracy of this release. Certain information in this press release may contain forward-looking statements. This information is based on current expectations that are subject to significant risks and uncertainties that are difficult to predict. Actual results might differ materially from results suggested in any forward-looking statements. The Company assumes no obligation to update the forward-looking statements, or to update the reasons why actual results could differ from those reflected in the forward-looking statements unless and until required by securities laws applicable to the Company. Additional information identifying risks and uncertainties is contained in filings by the Company with the Canadian securities regulators, which filings are available at www.sedar.com.


Pena F.J.,University of Extremadura | Macias Garcia B.,University of Extremadura | Samper J.C.,Sexing Technologies | Aparicio I.M.,University of Extremadura | And 2 more authors.
Theriogenology | Year: 2011

We review recent developments in the technology of freezing stallion sperm, paying special attention to the molecular lesions that spermatozoa suffer during freezing and thawing, such as osmotic stress, oxidative damage, and apoptotic changes. We also discuss the applicability of colloidal centrifugation in stallion sperm cryobiology. Increased knowledge about the molecular injuries that occur during cryopreservation may lead to improved protective techniques and thus to further improvements in fertility in the current decade. © 2011 Elsevier Inc..


Burroughs C.A.,Colorado State University | Graham J.K.,Colorado State University | Lenz R.W.,Sexing Technologies | Seidel G.E.,Colorado State University
Theriogenology | Year: 2013

The objective was to determine which characteristics of bovine ejaculates affected efficacy of sex sorting bovine sperm by flow cytometry. The effects of first versus second ejaculates, seminal plasma content, addition of BSA, and seminal plasma from different bulls during staining were all studied, as was the effect of 8-hour storage with and without seminal plasma. Semen collected by artificial vagina was centrifuged at 1000 × g for 15 minutes to separate sperm from seminal plasma; seminal plasma was clarified by 10 minutes of additional centrifugation at 2000 × g. Sperm were rediluted to 160 × 106 sperm per mL with: Tyrode's medium plus albumin, lactate, and pyruvate (TALP) containing 0%, 5%, 10%, or 20% homologous seminal plasma, TALP containing 10% heterologous seminal plasma, or TALP containing 0.3% (control), 0.6%, or 1.2% BSA. After incubation with Hoechst 33342 for 45 minutes, an equal volume of TALP containing red food dye was added, and sperm were analyzed by flow cytometry/cell sorting to determine percent of live-oriented sperm, X sort rate, percent of membrane-impaired sperm, and split (degree of separation between X- and Y-bearing sperm populations). The percent of live-oriented sperm was higher for sperm incubated with 0% seminal plasma (64%) than for sperm incubated with 5%, 10%, or 20% seminal plasma (60%, 59%, and 58%, respectively; P < 0.05). The X sort rate was higher for sperm incubated with 0% seminal plasma than sperm with 20% seminal plasma (4.26 vs. 3.61 × 103 sperm per second). When seminal plasma was exchanged between bull ejaculates, only one bull had seminal plasma that was detrimental to sperm, resulting in 31% membrane-impaired sperm compared with a range of 16% to 19% for seminal plasmas from other bulls (P < 0.05). The addition of BSA did not affect sort efficiency at the concentrations studied. Sperm from six bulls stored for 8 hours without seminal plasma had more membrane-impaired sperm (which were discarded) during sorting (28%) than with seminal plasma (19%; P < 0.01), but higher postthaw motility postsorting (63%) than with seminal plasma (52%; P < 0.05). In conclusion, the presence of seminal plasma during staining and sorting decreased sort rates and percent of live-oriented sperm, and storing sperm without seminal plasma increased postthaw motility. © 2013 Elsevier Inc.


Underwood S.L.,University of Sydney | Bathgate R.,University of Sydney | Pereira D.C.,Sexing Technologies | Castro A.,Sexing Technologies | And 3 more authors.
Theriogenology | Year: 2010

The objective of this study was to determine the in vitro fertilizing capacity of bull sperm derived from fresh or frozen samples and subjected to sex sorting and re-cryopreservation. Four sperm types were assessed for their ability to fertilize and sustain early embryo development in vitro. Semen from three Bos taurus bulls of different breeds (Jersey, Holstein and Simmental) was collected and either sorted immediately and then frozen (SF) or frozen for later sorting. Frozen sperm destined for sorting were thawed, sex-sorted, and re-frozen (FSF) or thawed, sex-sorted (FS), and used immediately for in vitro fertilization (IVF). Frozen-thawed nonsorted semen from the same ejaculate was used as a control. Oocytes from donor cows were aspirated via ovum pick-up and matured in vitro prior to IVF and culture. On average, 19.0 ± 1.7 (mean ± SEM) oocytes were aspirated per donor cow, of which 74.4 ± 2.2% were selected for maturation. The proportion of cleaved embryos (Day 3) did not differ between sperm groups (P = 0.91). Likewise, IVF with FSF sperm resulted in similar Day 7 blastocyst rates (as a percentage of total oocytes) as those of control, SF, and FS sperm (FSF, 34.5 ± 4.7; control, 32.2 ± 4.6; SF, 35.9 ± 4.8; and FS, 26.9 ± 4.1%; P = 0.23). These encouraging results show that frozen-thawed sex-sorted sperm may be re-frozen and used for in vitro embryo production with similar blastocyst production as that of nonsorted frozen-thawed and sex-sorted frozen-thawed sperm. © 2010 Elsevier Inc. All rights reserved.

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