EZH2 mutations at diagnosis in follicular lymphoma: a promising biomarker to guide frontline treatment.

EZH2 is mutated in nearly 25% of follicular lymphoma (FL) cases. Little is known about how EZH2 affects patients' response to therapy. In this context, the aim of this study was to retrospectively analyze the frequency of mutations in EZH2 at diagnosis in tissue and ctDNA in patients with FL and to assess the patients' outcomes after receiving immunochemotherapy, depending on the EZH2 mutation status. Among the 154 patients included in the study, 27% had mutated EZH2 (46% with high-grade and 26% with low-grade FL). Of the mutated tissue samples, the mutation in ctDNA was identified in 44% of cases. EZH2 mutation in ctDNA was not identified in any patient unmutated in the tissue.Unmutated patients who received R-CHOP had significantly more relapses than patients who received R-Bendamustine (16/49 vs. 2/23, p = 0.040). Furthermore, our results show that patients with mutated EZH2 treated with R-CHOP vs. those treated with R-Bendamustine present a lower incidence of relapse (10% vs. 42% p = 0.09 at 4 years), a higher PFS (92% vs. 40% p = 0.039 at 4 years), and higher OS (100% vs. 78% p = 0.039 at 4 years). Based on these data, RCHOP could be a more suitable regimen for mutated patients, and R-bendamustine for unmutated patients. These findings could mean the first-time identification of a useful biomarker to guide upfront therapy in FL.

Modified quasispecies model: the analysis of a periodic therapy.

We propose a modified mathematical model of the quasispecies type to analyze an unstable tumor progression evolution. In our study, we consider a heterogeneous population with different individuals, generated by the accumulation of successive mutations. Our model's main feature is that it allows for variable growth rates for each subpopulation and takes into account mutations from nonconsecutive types of mutants. Bifurcations and linear stability of the steady states are analyzed. We focus on two equilibria; one of them implies the coexistence of anomalous growth and genetically unstable cells. The other one yields the dominance of the anomalous growth population and the extinction of the malignant cells. However, linear stability analysis of the second equilibrium is inconclusive and suggests a suitable environment for the study of periodic therapy. This is carried out by introducing a small perturbation modeling the effect of a periodic medical treatment. As a result, a Zero-Hopf periodic orbit is identified, showing a cyclic behavior among the populations, with a strong dominance of the parental anomalous growth cell population.

Comparison of different sucrose-based extenders for stallion sperm vitrification in straws.

Vitrification of sperm is based on high-speed freezing by direct exposure to liquid nitrogen using non-permeable cryoprotectants, mainly disaccharides; yet, the concentration of cryoprotectants has a species-specific effect on the sperm cell. The aim of this study was to assess different sucrose concentrations for stallion sperm vitrification. Semen samples (n = 9) were collected from three stallions, centrifuged and resuspended to a concentration of 50 × 106  sperm/ml in a base extender (INRA96 + 1% of bovine serum albumin) with three different sucrose concentrations (Molar): 20 mM (S1), 100 mM (S2), or 200 mM (S3). Then, sperm were filled in covered 0.25 ml straws and directly plunged into liquid nitrogen. For warming, 0.25 ml straw was pulled out the covering straw and immersed in 3 ml of INRA96 at 43°C, with gentle pipetting to accelerate the melting. Total (TM, %) and progressive sperm motility (PM, %) were analysed using computer-assisted sperm analysis. Plasma (PMI, %) and acrosome membrane integrity (AIS, %) were assessed under epifluorescence microscopy. Post-warmed sperm parameters were compared between treatments by ANOVA. S2 showed significantly higher values in comparison with S1 and S3 for TM (S2 = 54.7 ± 5.5a ; S1 = 29.1 ± 3.3b ; S3 = 28.6 ± 3.0b ; p < 0.001) and PM (S2 = 31.3 ± 3.8a ; S1 = 18.5 ± 2.6b ; S3 = 17.7 ± 2.9b ; p < 0.01), respectively. No significant differences were found among treatments for PMI (S2 = 70.3 ± 5.2; S1 = 67.4 ± 4.3; S3 = 70.0 ± 3.7) neither for AIS (S2 = 57.1 ± 3.9; S1 = 53.9 ± 4.2; S3 = 57.0 ± 7.9). In conclusion, a concentration of 100 mM sucrose is recommended for stallion sperm vitrification in straws.

Equivalent seminal characteristics in human and stallion at first and second ejaculated fractions.

Sperm quality was assessed in normozoospermic human (n = 10) and Spanish breed stallion (n = 10) after sperm fractionation during ejaculation. The first ejaculated fraction was separated from the second. A third sample was reconstituted using equivalent proportion of both fractions (RAW). Fraction 1, Fraction 2 and RAW semen were incubated for 30 min at 37°C to homogenise the impact of iatrogenic damage between both species. Sperm concentration, motility and sperm DNA damage were assessed in each fraction and RAW semen. The results showed two important facts: (i) spermatozoa confined at Fraction 1 exhibit superior parameters than those included at Fraction 2 in both species, and (ii) there is a certain level of concordance between species in the proportion of benefit observed when Fraction 1 is compared to RAW semen. Altogether, these results call into question whether the standard practice of whole ejaculate collection can be considered the best strategy when using male gametes for artificial insemination. In fact, the reconstituted RAW semen exhibits poorer semen characteristics than those found in Fraction 1.

Reduction in the incidence of pressure ulcers upon implementation of a reminder system for health-care providers.

PURPOSE: To measure the clinical impact of the introduction of a reminder system for healthcare professionals to alert patients who are at risk for pressure ulcers (PU). METHODS: This was a pre- and post-test study of patients who were discharged from 6 medical-surgical units of the University Hospital of Fuenlabrada in 2009 and 2010. Beginning in January 2010, implementation of an on-screen list of reminders was automatically updated daily on the units' computers including patient arrival date, last assessment of ulceration risk and location of any PU. The cumulative incidence of PU was measured for patients discharged in 2009 (group A: healthcare professionals were not exposed to on-screen reminder) and 2010 (group B: healthcare professionals were exposed to on-screen reminder list). The relative risk (RR) was estimated. The study was completed with a stratified analysis and binary logistic regression. RESULTS: In group A, there were 84 cases of PU among 9263 patients discharged (0.9%); whereas in group B, there were 59 cases among 9220 patients discharged (0.6%). The RR of PU for group B/group A was 0.706 (p=0.038). In the logistic regression analysis, after adjusting for study variables, the odds ratio of PU B/A was 0.558. CONCLUSION: A list of on-screen reminders at the beginning of a healthcare professional's shift to inform them of patients at risk for developing a PU was effective at reducing the incidence of these clinical burdens.

Effects of different strategies of mineral supplementation (marine algae alone or combined with rumen boluses) in organic dairy systems.

This study was designed to evaluate the effect of marine algae supplementation alone or in combination with a regular mineral supplement (rumen boluses) to improve the mineral status in organic dairy cattle and their effect on the milk mineral composition, milk production, composition (% of fat and protein) and quality (SCC). Thirty-two Holstein Friesian lactating cows were randomly selected and assigned to the algae (A), boluses (B), algae+boluses (AB) and control group (C). For the algae groups (A, AB), a supplement composed of Sea Lettuce (80%), Japanese Wireweed (17.5%) and Furbelows (2.5%) was formulated to be given to the cows at the rate of 100 g/animal per day (A1) for the length of 4 weeks. In the second half of the experiment (weeks 5-8), the algae mixture was reformulated and the proportion of Furbelows was increased from 2.5% to 5.0% with a subsequent decrease of Lettuce to 77.5% (A2). In the boluses group (B), each cow received 2 boluses after calving. Blood (serum) and milk samples were collected at 2 and 4 week intervals, respectively, and analysed for trace element concentrations by ICP-MS. Information related to the milk composition and SCC during a 305-day lactation for each animal were obtained from the Dairy Records Management System. The supplementation with algae, boluses or the combination of both treatments showed a statistically significant effect on the iodine (algae), selenium (boluses) and cobalt (algae+boluses) status of the animals. In milk, treatments had a statistical significant increase on iodine, and a tendency to increase selenium concentrations. The assayed algae mixture combined with another source of selenium could be an effective tool to improve the mineral status in serum and milk.

Differences in preservation of canine chilled semen using simple sperm washing, single-layer centrifugation and modified swim-up preparation techniques.

This study compared the efficacy of simple sperm washing (SW), single-layer centrifugation (SLC) and modified swim-up (SU) techniques in the preparation of dog spermatozoa for cooling. Eighteen ejaculates, collected from three dogs (six per dog), were pooled (three ejaculates per pool) and divided into three aliquots: (1) one aliquot was washed and cooled at 5°C for 72h, considered as control (SW-control), (2) the second aliquot was selected by SLC through Androcoll-C and subsequently cooled in the same way as the SW-control samples (SLC-AC) and (3) the last aliquot was selected by a modified SU method with Androcoll-C and cooled as mentioned above (SU-AC). Assessment of sperm motility, sperm morphology, sperm membrane integrity and acrosome integrity were performed on aliquots of fresh semen and chilled-rewarmed samples. Sperm membrane integrity and progressive motility were significantly (PPP>0.05). The recovery rates were not significantly (P>0.05) different between SW-control, SLC-AC and SU-AC samples. Our results confirm that SU-AC may be a successful method for the preparation of dog spermatozoa for cooling.

Colloidal centrifugation of stallion semen results in a reduced rate of sperm DNA fragmentation.

Stallion spermatozoa recovered and examined immediately after colloidal centrifugation resulted in a higher straight-line velocity (VSL) than sperm processed using direct conventional centrifugation (p = 0.000), but there was no differences in the progressive motility or sperm DNA fragmentation (SDF) as determined by the sperm chromatin dispersion assay. However, when centrifuged spermatozoa were incubated at 37 °C for 24 h to determine the rate of SDF (r-SDF), a lower r-SDF (p = 0.0011) was observed in those sperm recovered after colloidal separation (0.5 ± 0.1%/h) compared to direct (1.2 ± 0.4%/h) or no centrifugation (r-SDF = 1.2 ± 0.3%/h). These results confirm that colloidal separation of stallion spermatozoa results in prolonged sperm DNA longevity, but these differences were only apparent following a period of incubation and dynamic assessment. Consequently, we strongly recommend the use of the dynamic form of the SDF assay for evaluating centrifugation and/or other ex vivo procedures, as a single basal assessment of SDF may inadvertently result in a false-positive evaluation of DNA quality.

Outcome of ST-elevation myocardial infarction versus non-ST-elevation acute coronary syndrome treated with titanium-nitride-oxide-coated versus everolimus-eluting stents: insights from the BASE-ACS trial.

AIM: The BASE-ACS trial demonstrated an outcome of titanium-nitride-oxide-coated bioactive stents (BAS) that was statistically non-inferior to that of everolimus-eluting stents (EES) at 12-month follow-up, in patients presenting with acute coronary syndrome (ACS) who underwent early percutaneous coronary intervention (PCI). We explored a post-hoc analysis of the 12-month outcome of the BASE-ACS trial in the subgroup of patients with ST-elevation myocardial infarction (STEMI) versus non-ST-elevation ACS (non-STEACS). METHODS: A total of 827 patients with ACS (321 STEMI) were randomly assigned to receive either BAS or EES. The primary endpoint was a composite of cardiac death, non-fatal myocardial infarction (MI) and ischemia-driven target lesion revascularization (TLR) at 12-month follow-up. RESULTS: The 12-month cumulative incidence of the primary endpoint was similar between the two subgroups (9% versus 9.5%, in STEMI versus non-STEACS patients respectively, P=0.90). The 12-month rate of cardiac death was significantly higher in the STEMI subgroup as compared with the non-STEACS subgroup (2.8 versus 0.6%, respectively, P=0.01). However, the rates of non-fatal MI, ischemia-driven TLR, definite stent thrombosis, and non-cardiac death were all statistically matched between the two subgroups (P>0.05 for all). CONCLUSION: In the current post-hoc analysis of the BASE-ACS trial based on the infarction type, the 12-month outcome of patients who underwent early PCI for ACS was slightly worse in the setting of STEMI as compared with non-STEACS, as reflected by a significantly higher rate of cardiac death.

[Translated article] Metal on metal total hip arthroplasty: Correlation between inclination of the acetabular and metal ion levels.

Due to their low friction index, hip arthroplasties with metal-metal (M-M) friction torque have been an attractive option in young patients with high functional demand, currently they have suffered a decrease in their use due to the complications of some models and physiological reactions adverse reactions related to the elevation of metal ions in the blood. Our objective is to review the patients with M-M pair operated in our centre, correlating the ion level with the position of the acetabular component and with the size of the head. MATERIAL AND METHODS: Retrospective review of 166 M-M hip prostheses operated between 2002 and 2011. Sixty five ruled out for different causes (death, loss of follow-up, no current ion control, no radiography or others), leaving a sample of 101 patients to be analysed. Follow-up time, cup inclination angle, blood ion level, Harris Hip Score (HHS), and complications were recorded. RESULTS: One hundred and one patients (25 women and 76 men), 55 years of average age (between 26 and 70), of these 8 were surface prostheses and 93 total prostheses. The mean follow-up time was 10 years (between 5 and 17 years). The mean head diameter was 46.25 (between 38 and 56). The mean inclination of the butts was 45.7° (between 26° and 71°). The correlation force between the verticality of the cup and the increase in ions is moderate r=0.31 for Cr and slight r=0.25 for Co. The correlation force between head size and ion increase is weak and inverse r=-0.14 for Cr and r=0.1 for Co. Five patients (4.9%) required revision (2 [1%] due to increased ions with pseudotumor). The mean time to revision was 6.5 years in which the ions increased. The mean HHS was 94.01 (between 55.8 and 100). In the review of patients, we found 3 with a significant increase in ions who had not followed controls, all 3 had an HHS of 100. The angles of the acetabular components were 69°, 60° and 48° and the diameter of the head was 48.42 and 48mm, respectively. DISCUSSION AND CONCLUSIONS: M-M prostheses have been a valid option in patients with high functional demand. A bi-annual analytical follow-up is recommended, since in our case we have detected 3 patients with HHS 100 who presented unacceptable elevation of cobalt >20µm/l (according to SECCA) of the ions and 4 with very abnormal elevation of cobalt ≥10µm/l (according to SECCA), all of them with cup orientation angles >50°. With our review we can conclude that there is a moderate correlation between the verticality of the acetabular component and the increase in blood ions and that the follow-up of this patient with angles >50° is essential.

Metal on metal total hip arthroplasty: Correlation between inclination of the acetabular and metal ion levels. / Artroplastias metal-metal en cadera: correlación entre el aumento de iones y el ángulo del componente acetabular.

Due to their low friction index, hip arthroplasties with metal-metal (M-M) friction torque have been an attractive option in young patients with high functional demand, currently they have suffered a decrease in their use due to the complications of some models and physiological reactions adverse reactions related to the elevation of metal ions in the blood. Our objective is to review the patients with M-M pair operated in our center, correlating the ion level with the position of the acetabular component and with the size of the head. MATERIAL AND METHODS: Retrospective review of 166 M-M hip prostheses operated between 2002 and 2011. Sixty five ruled out for different causes (death, loss of follow-up, no current ion control, no radiography or others), leaving a sample of 101 patients to be analyzed. Follow-up time, cup inclination angle, blood ion level, Harris Hip Score (HHS), and complications were recorded. RESULTS: One hundred and one patients (25 women and 76 men), 55years of average age (between 26 and 70), of these 8 were surface prostheses and 93 total prostheses. The mean follow-up time was 10years (between 5 and 17years). The mean head diameters was 46.25 (between 38 and 56). The mean inclination of the butts was 45.7° (between 26° and 71°). The correlation force between the verticality of the cup and the increase in ions is moderate r=0.31 for Cr and slight r=0.25 for Co. The correlation force between head size and ion increase is weak and inverse r=-0.14 for Cr and r=0.1 for Co. Five patients (4.9%) required revision (2 [1%] due to increased ions with pseudotumor). The mean time to revision was 6.5years in which the ions increased. The mean HHS was 94.01 (between 55.8 and 100). In the review of patients, we found 3 with a significant increase in ions who had not followed controls, all 3 had an HHS of 100. The angles of the acetabular components were 69°, 60° and 48° and the diameter of the head was 48.42 and 48mm, respectively. DISCUSSION AND CONCLUSIONS: M-M prostheses have been a valid option in patients with high functional demand. A bi-annual analytical follow-up is recommended, since in our case we have detected 3patients with HHS100 who presented unacceptable elevation of cobalt >20µm/L (according to SECCA) of the ions and 4 with very abnormal elevation of cobalt ≥10µm/L (according to SECCA), all of them with cup orientation angles >50°. With our review we can conclude that there is a moderate correlation between the verticality of the acetabular component and the increase in blood ions and that the follow-up of this patient with angles >50° is essential.

[Fungal infection by Cladophialophora bantiana and development of cerebral phaeohyphomycosis. A systematic review of 58 case reports]. / Infección micótica por Cladophialophora bantiana y desarrollo de feohifomicosis cerebral. Revisión sistemática de 58 informes de caso.

INTRODUCTION: Cladophialophora bantiana is a filamentous fungus, known as a dematiaceous fungus because of the presence of melanin. This fungus is of clinical importance because it is neurotropic and causes cerebral phaeohyphomycosis. MATERIAL AND METHODS: The available scientific information on the development of cerebral phaeohyphomycosis caused by Cladophialophora bantiana was analysed by selecting articles from the PubMed, Scopus and Google Scholar databases that describe case reports of fungal infection by C. bantiana in adults, taking into account the analysis of the patients' symptomatology, clinical history and neuroanatomical damage, in addition to considering the mortality of the condition. RESULTS: India and United States were the countries with most case reports, with 32 and 11 cases respectively. Moreover, in terms of neuroanatomical lesions, the majority of patients suffered mixed lesions (29%) and frontal lobe lesions (22%). In accordance with the patients' condition, the pathology has a mortality rate of 62%. CONCLUSIONS: It is concluded that cerebral phaeohyphomycosis has a high mortality rate, there is no standardised treatment and, in most cases, the fungal infection of the brain is mixed and affects several different parts of it. Furthermore, if not diagnosed and treated in time, it can lead to the patients' death.

TITLE: Infección micótica por Cladophialophora bantiana y desarrollo de feohifomicosis cerebral. Revisión sistemática de 58 informes de caso.Introducción. Cladophialophora bantiana es un hongo filamentoso, denominado hongo dematiáceo por la presencia de melanina. Este hongo tiene importancia clínica por ser neurotrópico y causar feohifomicosis cerebral. Material y métodos. Se analizó la información científica disponible sobre el desarrollo de feohifomicosis cerebral provocada por Cladophialophora bantiana, seleccionando artículos de las bases de PubMed, Scopus y Google Scholar, que describen informes de caso sobre infección micótica de C. bantiana en adultos, considerando el análisis de la sintomatología, el historial clínico y los daños neuroanatómicos de los pacientes, además de considerar la mortalidad de la patología. Resultados. La India y Estados Unidos fueron los países con más informes de caso, 32 y 11 casos, respectivamente. Asimismo, en cuanto a las lesiones neuroanatómicas, en su mayoría, los pacientes sufrieron lesiones mixtas (29%) y del lóbulo frontal (22%). De acuerdo con el estado de los pacientes, la patología tiene una mortalidad del 62%. Conclusiones. Se concluye que la feohifomicosis cerebral tiene una alta mortalidad, no existe un tratamiento estandarizado y, en la mayoría de los casos, la infección fúngica del cerebro es mixta y afecta a varias partes del cerebro; además, si no se diagnostica y trata a tiempo, puede ocasionar la muerte de los pacientes.

Telephone consultation service in orthopedics during COVID-19 pandemic.

BACKGROUND AND AIM: The current COVID-19 pandemic scenario has driven surgical departments to a transformation.The worldwide spread of the disease has led to a public health quarantine where health care professionals are at high risk of infection. In this context, telemedicine has been promoted and scaled up to reduce the risk of transmission. This study aims to demonstrate that a combined framework based on telematics and in-person clinical encounter not only ensures medical care but the safety of healthcare professionals and patients. MATERIAL AND METHOD: Descriptive observational study on the follow-up of patients during the COVID19 Pandemic, combining telephone and traditional. RESULTS: A total of 5031 telephone calls were made, differentiating between medical referrals, specialised primary care visits, and outpatient consultation.They were classified as successful, required an in-person visit, or no successful telephone contact. Furthermore, we divided them into 2 groups: resolved and unresolved.53% of all telematic visits were successful. CONCLUSIONS: Telematic medical systems are a feasible option in a orthopedics department and an interesting resource to preserve once the pandemic is resolved. Future lines of research should be opened to improve system success, analyze its cost-effectiveness ratio, and correct any legal conflicts that may exist.

ANTECEDENTES Y OBJETIVO: Durante la pandemia COVID-19 la actividad de los servicios quirúrgicos se ha visto obligada a adaptarse y transformarse. La telemedicina se está implantando como nunca antes en esta nueva situación en la que los pacientes están confinados y los profesionales sanitarios presentan riesgo de infectarseEl objetivo es mostrar que una reestructuración combinada telemática y presencial de las visitas permite asegurar la asistencia médica, garantizando la protección del personal sanitario y de los pacientes. MATERIAL Y MÉTODO: Estudio descriptivo observacional sobre el seguimiento de pacientes durante la Pandemia COVID combinando la consulta telefónica con la presencial. RESULTADOS: Se realizaron un total de 5031 llamadas telefónicas diferenciando entre Derivaciones, Visitas de atención primaria especializada y Consulta externa hospitalaria.Se registraron como efectivas, tributarias de visita presencial y no se logra contacto telefónico. Y las dividimos en 2 grupos resueltas y no resueltas.Del total de visitas no presenciales telefónicas fueron efectivas un 53%. CONCLUSIONES: La medicina telemática es una opción factible en un servicio de traumatología y de manera adecuada será una opción interesante de mantener tras la pandemia.Futuras líneas de investigación deberían ser abiertas para mejorar la capacidad de resolución de este sistema, analizar su relación coste-efectividad y subsanar los conflictos legales que pudieran existir.

Telephone consultation service in orthopedics during COVID-19 pandemic. / Consultoría telefónica en COT durante la pandemia por COVID-19. ¿Ha venido para quedarse?

BACKGROUND AND AIM: The current COVID-19 pandemic scenario has driven surgical departments to a transformation. The worldwide spread of the disease has led to a public health quarantine where health care professionals are at high risk of infection. In this context, telemedicine has been promoted and scaled up to reduce the risk of transmission. This study aims to demonstrate that a combined framework based on telematics and in-person clinical encounter not only ensures medical care but the safety of healthcare professionals and patients. MATERIAL AND METHOD: Descriptive observational study on the follow-up of patients during the COVID19 Pandemic, combining telephone and traditional. RESULTS: A total of 5031 telephone calls were made, differentiating between medical referrals, specialized primary care visits, and outpatient consultation. They were classified as successful, required an in-person visit, or no successful telephone contact. Furthermore, we divided them into 2 groups: resolved and unresolved. 53% of all telematic visits were successful. CONCLUSIONS: Telematic medical systems are a feasible option in a orthopedics department and an interesting resource to preserve once the pandemic is resolved. Future lines of research should be opened to improve system success, analyze its cost-effectiveness ratio, and correct any legal conflicts that may exist.

Seasonal variations in sperm DNA fragmentation and pregnancy rates obtained after artificial insemination with cooled-stored stallion sperm throughout the breeding season (spring and summer).

The aim of this study was to assess seasonal variations during different periods of the breeding season (spring and summer) on stallion sperm DNA fragmentation and in vivo fertility associated with cooled-stored semen samples. Ejaculates were collected from eleven stallions and assessed for sperm motility (assessed by computer-assisted sperm analysis) and plasma membrane integrity (evaluated under fluorescence microscopy). Sperm DNA fragmentation (evaluated by the Sperm Chromatin Dispersion test) was assessed in cooled-stored semen at 5 °C for up to 24 h. Artificial insemination was performed throughout the breeding season. Mares were inseminated with cooled-stored semen (up to 24 h) every other day until ovulation. Pregnancy rates per cycle were determined detecting the embryonic vesicle by ultrasonography fifteen days after ovulation. Values (mean ± SD) for progressive sperm motility were significantly higher (P < 0.05) in spring (53.57 ± 9.97%) in comparison to summer (41.37 ± 10.81%). No significant differences in plasma membrane integrity were found between seasons (P > 0.05). Sperm DNA fragmentation was significantly lower (P < 0.01) in spring in comparison to summer after 0h (4.81 ± 1.87% vs. 8.77 ± 5.78%), 6h (9.00 ± 3.19% vs. 18.73 ± 8.22%) and 24h (14.6 ± 4.13% vs. 30.14 ± 9.85%) of cooled-storage. Pregnancy rates per cycle were also significantly higher (P < 0.01) in spring (50%) in comparison to summer (37%). There was a moderate negative relationship between positive pregnancies and sperm with fragmented DNA (r = - 0.619; P < 0.001). Semen samples associated with moderate fertility levels (Pregnancy rate < 50%) showed a higher percentage of sperm with fragmented DNA compared to samples obtaining higher fertility levels. In conclusion, seasonal variations were found during the breeding season, obtaining lower sperm DNA fragmentation and higher pregnancy rates in spring. Additionally, samples with the highest proportion of sperm with fragmented DNA showed the lowest fertility levels throughout the breeding season.

Effect of season on individual stallion semen characteristics.

Horses are long-day seasonal breeding animals, however, with modern stallion reproductive management it is important for collection of semen during periods that are not part of the traditional breeding season. This study was conducted to examine variation in the seminal characteristics of individual stallions in Avila, Spain during 1 year with a particular emphasis on sperm DNA fragmentation. Semen was collected twice per season from a total of 20 stallions. There was a marked seasonal effect on all seminal characteristics, with the greatest on progressive motility, % membrane integrity and least for SDF in the spring months; there was also an interaction effect with respect to individual stallion, indicating that some stallions did not fit this generalised pattern for semen quality. Sperm DNA fragmentation was assessed both immediately after semen collection (T0) and following incubation of extended semen for 24 h (T24) to broadly mimic changes in SDF that might occur in the female reproductive tract. While SDF evaluated at T0 was also generally less in spring, the proportion of stallions with the least SDF values in spring increased from 45% to 60% when assessed at T24, therefore, being consistent with the importance of dynamic SDF assessment in detecting DNA damage that was not detected at T0 or cryptic DNA damage. The results from this study indicate there is individual seasonal variation among stallions in all aspects of seminal characteristics; such variation needs to be considered when prioritising stallions that are to be used for breeding.

Fertilizing capacity of vitrified stallion sperm assessed utilizing heterologous IVF after different semen warming procedures.

The aim of this study was to evaluate the fertilizing capacity of frozen or vitrified stallion sperm after assessing different warming procedures. In Experiment 1, different warming procedures were compared after sperm vitrification: immersion in extender at 43 °C (C), or in a water bath at 37 °C/30 s (W37), 43 °C/10 s (W43) or 60 °C/5 s (W60). With the W60 treatment, there were greater values (P < 0.05) for VCL (83.93 ± 3.6 µm/s) and ALH (3.00 ± 0.2 µm) than freezing and with the C group, and greater values (P < 0.001) for PM (35.33 ± 2.5 %) than with the W43 treatment. In Experiment 2, the fertilizing capacity of vitrified and frozen sperm was assessed utilizing heterologous IVF procedures, using cattle oocytes. Vitrification resulted in greater values (P < 0.05) than freezing for the number of bound sperm (1.36 ± 0.3 and 0.69 ± 0.2, respectively). There were no differences between frozen or vitrified sperm in pronuclear formation (26 hours post-insemination - hpi; 14.08 ± 4.2 % and 22.78 ± 4.8 %, respectively) or cleavage rate (32.77 ± 4.3 % and 39.66 ± 4.6 %, respectively). In conclusion, vitrified stallion sperm warmed in a water bath at 60 ºC had the capacity to penetrate cattle oocytes, leading to pronuclear formation and hybrid embryo cleavage after heterologous IVF.

Avian pollination studies: a simple scanning electron microscopic technique.

Identifying ornithophilous plant species utilized by several different flower-visiting birds is simplified by the scanning electron microscope. The technique involves comparing pollen samples taken from the birds' head feathers with pollen reference standards collected in the same area, which simplifies analysis of pollination patterns in a complex community.

Assessment of sperm DNA fragmentation in stallion (Equus caballus) and donkey (Equus asinus) using the sperm chromatin dispersion test.

Sperm DNA fragmentation (sDF) is an important parameter to assessing sperm quality. Information about sperm quality is not available for donkeys, especially in some breeds at risk of extinction. The objectives of this research were to test the four commercial variants of sperm chromatin dispersion test (SCD; sperm Halomax test), originally developed to assess sDF in boars, bulls, rams and stallions, in order to scrutinize their applicability in the study of sDF in a donkey breed at risk of extinction (Zamorano-Leonesa), for which there is no specific test available to analyze sperm at present. Only the SCD test, originally developed for stallions, produced stable and consistent results, and was deemed suitable to assess DNA fragmentation in sperm samples from donkeys. Image analysis was used to compare differences between the SCD methodology applied to stallion and donkey semen samples processed under the same experimental conditions. The extent of SCD in the SCD test was approximately 20% lower in donkey sperm than in stallion sperm. Yet, the ratio of chromatin sperm dispersion achieved in fragmented and unfragmented nuclei did not differ significantly between species. These data suggest that a similar protein depletion treatment can cause differences in protein removal in equivalent cells from different species and that sperm chromatin may be organized differently in stallions and donkeys.