[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.

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

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.ResultadosLa 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%.ConclusionesSe 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. (AU)

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.RESULTSIndia 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%.CONCLUSIONSIt 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. (AU)

[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.

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.

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.

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

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 infectarse. El 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 por COVID-19, combinando la consulta telefónica con la presencial. RESULTADOS: Se realizaron un total de 5.031 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 dos 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

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

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.

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 sperm dosage transportation in stallions: Effect on sperm DNA fragmentation.

Artificial insemination programs for horses usually involve ex vivo handling and transporting of sperm. The present experiment was designed to: (i) assess the effect of transportation on sperm DNA integrity at different time post semen collection, and (ii) evaluate if sperm DNA quality deteriorates rapidly beyond 24 h of cooled storage. After collection, the ejaculates were extended using INRA 96 and semen was prepared for prompt analysis (A0) or 24 h/48 h cooled-shipping (B24 and C48 respectively). Each sample was assessed for sperm DNA fragmentation index (SDFI) at time 0 and after incubation for 2, 6 and 24 h at 37 °C. There was very little difference in SDFI between freshly extended (A0) and 24 h/48 h cooled-transported semen samples (B24/C48) at time 0. After 2 h of incubation at 37 °C, there was an increase in SDFI ranging from 2.7% to 7.5% per hour in freshly extended semen samples (A0: 5.1 ± 1.5), while cooled-transported semen samples had a much greater increase in SDFI, ranging from 5.0% to 20.5% (B24: 14.7 ± 5.6) and from 8.2% to 26.8% (C48: 18.3 ± 7.2) respectively. There were not marked differences in the sperm DNA integrity between 24 and 48 h for transported samples, thus there is the possibility of desirable fertility with use of stallion sperm after 48 h of cooled storage.

Vitrification of stallion sperm using 0.25 ml straws: Effect of volume, concentration and carbohydrates (sucrose/trehalose/raffinose).

Sperm vitrification is a rapid freezing method in which carbohydrates are used as cryoprotectants. The aim of this study was to determine the optimal volume, concentration and type of carbohydrates for stallion sperm vitrification using 0.25 ml straws in comparison to conventional freezing. Ejaculates (n = 54) were collected from six stallions. For vitrification, straws were filled with different volumes (30, 70, 100 µl), sperm concentrations (50, 100, 200 × 106 sperm/ml) and extenders containing sucrose (20, 100, 200 mM), trehalose (50, 100, 200 mM) and raffinose (50, 100, 200 mM) and plunged into LN2. Conventional freezing was performed in 0.5 ml straws frozen in LN2 vapors. Sperm motility, plasma and acrosome membrane integrities and DNA fragmentation were compared among treatments. The use of straws filled with 100 µl at 100 × 106 sperm/ml with the extender containing 100 mM trehalose resulted in greater values for sperm quality than the other concentrations, volumes and carbohydrates. With vitrification, there were greater values (mean ± SEM; P < 0.05) than freezing for progressive motility (48.2 ± 2.3 compared with 37.3 ± 2.2%), plasma membrane integrity (82.8 ± 1.5 compared with 74.1 ± 1.9%), and intact acrosomes (50.2 ± 1.2 compared with 43.1 ± 1.4%); and less DNA fragmentation (6.4 ± 0.7 compared with 8.2 ± 0.3%). In conclusion, stallion sperm can be vitrified in 0.25 ml straws filled with 100 µl of sperm at 100 x 106 sperm/ml using an extender with 100 mM of trehalose, obtaining better sperm quality after warming than conventional freezing.

Massive generalized crystal-storing histiocytosis associated with extracellular crystalline nephropathy: clinical, immunohistochemical, and ultrastructural studies of a unique disorder and review of the literature.

Crystal-storing histiocytosis (CSH) is a rare disorder characterized by the accumulation of nonneoplastic histiocytes containing intracytoplasmic crystallized immunoglobulins. In most cases, there is an associated underlying lymphoplasmacytic neoplasm expressing Ig kappa light chain. About 131 cases of CSH have been identified. There is a localized and a generalized form of CSH and it can involve several sites including bone marrow, lungs, lymph nodes, liver, spleen, gastrointestinal tract, and kidney. Generalized CSH is less frequent and involves multiple organs and tends to have a worst prognosis than localized CSH. Around 20 cases of renal involvement in CSH have been reported so far. Paraprotein-induced crystalline nephropathy can be divided into two categories based on whether the crystals in the kidney are intracellular (including light chain proximal tubulopathy with crystals and CSH) or extracellular (including the crystalline variant of myeloma cast nephropathy and crystalglobulin-induced nephropathy). The former tends to present with slowly worsening kidney dysfunction and generally has a good prognosis, whereas the latter usually presents with rapidly progressive renal failure and is associated with poor renal outcome. We present a case of generalized CSH associated with extracellular crystalline nephropathy with a fulminant and fatal clinical course. Kappa light-chain crystals were found exclusively extracellularly within the tubular lumen, not within the tubular epithelial cells nor the histiocytes, and the massive presence of those precipitates led to the acute renal failure. Consequently, we review the renal involvement in CSH in the literature and discuss the unique mechanism of renal injury in this case.

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.

Concentrations of non-permeable cryoprotectants and equilibration temperatures are key factors for stallion sperm vitrification success.

Vitrification is based on rapid freezing by direct exposure of sperm to liquid nitrogen (LN2). This study evaluated the effect of non-permeable CPAs and equilibration temperature on stallion sperm quality after vitrification. In Experiment 1, different concentrations of sucrose (20, 50, 100 mM; mmol/L) and bovine serum albumin (BSA 1%, 5%, 10%) were compared including different temperatures for the equilibration (≈22 °C or 5 °C). Vitrification was performed dropping 30 µl sperm suspension directly into LN2. In Experiment 2, conventional sperm freezing using 2.2% of glycerol in 0.5 ml straws, frozen in LN2 vapours, was compared to the sucrose and BSA extenders (and its combination) producing the most desirable results. Sperm motility, plasma membrane and acrosome integrity were statistically compared between treatments. Vitrification after sperm cooling at 5 °C with sucrose 20 mM (S20) or BSA 1% (BSA1) resulted in the greatest values (mean ±â€¯SEM) for most of the sperm variables assessed. With use of the combination (S20 + BSA1/5 °C), there were greater values (P<0.001) than freezing with glycerol for total (55.67 ±â€¯2.99 vs 35.41 ±â€¯2.96) and progressive sperm motility (38.32 ±â€¯3.05 vs 14.42 ±â€¯1.80), plasma membrane integrity (66.61 ±â€¯2.69 vs 49.16 ±â€¯2.60), intact-acrosomes (49.19 ±â€¯2.60 vs 14.91 ±â€¯1.57) and most of the kinetics assessed, respectively. In conclusion, stallion sperm can be vitrified after cooling at 5 °C using a combination of 20 mM sucrose and 1% BSA based extender and this is a promising alternative compared with conventional sperm freezing using glycerol.

Stallion sperm freezing with sucrose extenders: A strategy to avoid permeable cryoprotectants.

The aim of this study was to assess different concentrations of sucrose-based extenders combined with bovine serum albumin (BSA) as an alternative to stallion sperm cryopreservation with permeable cryoprotectants. Semen samples (n = 16) were collected from six stallions. Sperm was cooled, filled in 0.5 mL straws and frozen in nitrogen vapor. Post-thaw sperm kinetic parameters, plasma and acrosome membrane integrity were statistically compared among treatments. In Experiment 1, extenders containing 1% of BSA and different concentrations of sucrose (mmol/L, M): 0, 50, 100, 250, 350 and 450 mM were compared. Use of sucrose [100 mM (S2)] resulted in greater values for most of the sperm kinetic parameters assessed (P < 0.001). There were no differences for plasma membrane integrity, except for when sucrose was used at 50 and 250 mM concentrations, and plasma membrane integrity was less (P < 0.05) when these concentrations were used than with the other sucrose concentrations. In Experiment 2, the selected sucrose extender (S2) was compared to an extender containing glycerol as permeable cryoprotectant. Use of the S2 extender resulted in a lesser proportion of sperm with denuded-acrosomes (P < 0.05) in comparison to use of glycerol and values for several kinetic parameters were also greater (P < 0.05) with use of S2. There were no significant differences for the other parameters assessed in this study. In conclusion, stallion sperm can be frozen in the absence of permeable cryoprotectants, using a combination of sucrose 100 mM with BSA-1% as alternative agents.

Evaluation of trace element status of organic dairy cattle.

The present study aimed to evaluate trace mineral status of organic dairy herds in northern Spain and the sources of minerals in different types of feed. Blood samples from organic and conventional dairy cattle and feed samples from the respective farms were analysed by inductively coupled plasma mass spectrometry to determine the concentrations of the essential trace elements (cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), iodine (I), manganese (Mn), molybdenum (Mo), nickel (Ni), selenium (Se) and zinc (Zn)) and toxic trace elements (arsenic (As), cadmium (Cd), mercury (Hg) and lead (Pb)). Overall, no differences between organic and conventional farms were detected in serum concentrations of essential and toxic trace elements (except for higher concentrations of Cd on the organic farms), although a high level of inter-farm variation was detected in the organic systems, indicating that organic production greatly depends on the specific local conditions. The dietary concentrations of the essential trace elements I, Cu, Se and Zn were significantly higher in the conventional than in the organic systems, which can be attributed to the high concentration of these minerals in the concentrate feed. No differences in the concentrations of trace minerals were found in the other types of feed. Multivariate chemometric analysis was conducted to determine the contribution of different feed sources to the trace element status of the cattle. Concentrate samples were mainly associated with Co, Cu, I, Se and Zn (i.e. with the elements supplemented in this type of feed). However, pasture and grass silage were associated with soil-derived elements (As, Cr, Fe and Pb) which cattle may thus ingest during grazing.