N-(2-mercaptopropionyl)-glycine enhances in vitro pig embryo production and reduces oxidative stress.

This study evaluated the effects of different concentrations (1, 10, 25, 50, and 100 µM) of the antioxidant N-(2-mercaptopropionyl)-glycine (NMPG), during the culture of in vitro-fertilized porcine oocytes. While the highest concentrations of NMPG (50 and 100 µM) were toxic to the developing embryos during the first two days of culture, 25 µM NMPG achieved cleavage rates that were similar to those achieved by the control but did not sustain blastocyst production by Day 7 of culture. Compared to the control culture medium, the culture medium supplemented with 10 µM NMPG increased (P < 0.05) the rates of blastocyst formation, decreased (P < 0.05) the intracellular levels of reactive oxygen substances, and downregulated (P < 0.05) the expression of the oxidative stress related gene GPX1. In conclusion, these results demonstrated that supplementation of porcine embryo culture medium with 10 µM NMPG can attenuate oxidative stress and increase the yield of in vitro production of blastocysts.

Three-to-5-day weaning-to-estrus intervals do not affect neither efficiency of collection nor in vitro developmental ability of in vivo-derived pig zygotes.

An efficient system to collect large numbers of vital zygotes is a pre-requisite for application of zygote genome-editing technology, including development of efficient models for xenotransplantation using pigs. Owing to the sub-optimal in vitro production of zygotes in pigs, efficient collection of in vivo developed zygotes is required. Timing of ovulation is a key factor to sustain efficiency since the interval between pronuclear formation and the first division is very short in pigs. The weaning-to-estrus interval can, due to its inverse relation with length of estrus and time of ovulation, interfere with ovulation and make it asynchronous, which reduces the probability of obtaining zygotes. This retrospective study compared the effects of three weaning-to-estrus intervals of 3, 4 or 5 days on zygote collection efficiency in a total of 17 trials over a 3-year period including 223 sows. Donor sows in groups of 10-15 animals were super-ovulated with eCG 24 h after weaning and those in estrus at 48-72 h post-eCG were immediately treated with hCG, followed by insemination 6 and 24 h thereafter. Collected structures during laparotomy on Day 2 (Day 0: onset of estrus) were morphologically evaluated and only those with a single cell and two visible polar bodies were considered as zygotes. Zygotes were injected with CRISPR-Cas9 editor mixture and cultured for 6 days to evaluate their developmental ability against non-injected control zygotes. Of all recovered structures (N = 5,468), 67.4%, 30.8% and 1.8% were zygotes, 2-cell embryos and oocytes-degenerated embryos, respectively. The different weaning-to-estrus intervals did not affect either the percentages of collected zygotes (range: 64.1%-70.0%) or the percentages of sows with zygotes at collection time (range: 69.0%-73.3%). The weaning-to-estrus intervals did not affect the in vitro developmental ability of zygotes. After 24 h of culture, 78.1 ±â€¯2.0% and 95.1 ±â€¯0.6 (P < 0.05) of injected (N = 2,345) and non-injected (N = 335) zygotes, respectively, developed to 2-to-4-cell embryo stage. The total efficiency of the system was 64.1 ±â€¯2.2% and 85.8 ±â€¯1.5% (P < 0.05) for injected and non-injected zygotes, respectively. In conclusion, the results indicate that neither the efficiency of collecting in vivo derived porcine zygotes from superovulated sows nor the zygote ability to develop to blastocyst after cytoplasmic genome-editing injection were affected by a weaning-to-estrus interval between 3-to-5 days.

The cytokine platelet factor 4 successfully replaces bovine serum albumin for the in vitro culture of porcine embryos.

The cytokine platelet factor 4 (PF4) enhances differentiation and cell viability of different stem cells lines in vitro. This study investigated whether PF4 addition to customary pig embryo semi-defined culture media can improve their developmental outcome (Experiment 1) and ultimately replace the need for bovine serum albumin (BSA, Experiment 2). Experiment 1 added PF4 (100-1000 ng/mL, 0 = control) to NCSU-23 with 0.4 mg/mL BSA culturing 3430 presumptive zygotes. Experiment 2 added PF4 (100-1000 ng/mL, 0 = Control-PVA) to a BSA-free medium (NCSU-23 with 0.3 mg/mL PVA) culturing 3820 presumptive zygotes. Zygote culture in NCSU-23 with 0.4 mg/mL BSA was used as overall control. All groups of Experiment 1 displayed similar rates of day 2-cleavage (range: 65.0 ±â€¯10.9 to 70.0 ±â€¯5.8%); of day 7-blastocyst rates (range: 46.6 ±â€¯10.0 to 56.4 ±â€¯8.2%) and of total day 7-blastocyst efficiency (range: 32.3 ±â€¯8.3 to 37.2 ±â€¯7.3%). Addition of PF4 did not affect total cell numbers of day 7 blastocysts (range: 44.1 ±â€¯23.2 to 50.5 ±â€¯26.4). In Experiment 2, PF4 accelerated embryo development, increasing (P < 0.01) blastocyst yield compared to 0-PF4, and blastocyst formation by day 5 adding PF4 100-500 ng/mL (range: 29.9 ±â€¯7.8 to 31.8 ±â€¯5.5%; P < 0.05) compared with BSA-control (17.2 ±â€¯8.2%) and PF4 1000 ng/mL (15.5 ±â€¯7.9%); showing similar blastocyst rates (range: 42.0 ±â€¯11.5 to 49.3 ±â€¯10.0%), total efficiency (28.0 ±â€¯8.2 to 32.3 ±â€¯7.1%) total cell numbers (range: 42.6 ±â€¯19.3 to 45.7 ±â€¯23.9) as BSA-controls. In conclusion, although PF4 did not show additive improvement under usual semi-defined, BSA-supplemented embryo media, it successfully replaced BSA sustaining porcine blastocyst production in chemically defined conditions.

Porcine blastocyst viability and developmental potential is maintained for 48 h of liquid storage at 25 °C without CO2 gassing.

Short- and medium-term storage of pig embryos has become relevant for commercial application of non-surgical deep uterine embryo transfer (NsDU-ET) in the light of the strict legal and administrative requirements posed by the International Association for Air Transport (IATA) to allow shipment of liquid nitrogen (LN2) containers and the technical drawbacks when using vitrified embryos. Therefore, this study developed an efficient method for the liquid storage of in vivo-derived porcine blastocysts for a moderate duration (48 h) without controlled CO2 gassing. We evaluated two storage temperatures (25 °C and 37 °C) and three HEPES-supplemented media: the chemically defined media TL-PVA and NCSU-PVA and the semi-defined medium NCSU-BSA. We observed no differences in survival, hatching rate or final developmental stage between the two temperatures, but storage at 25 °C was more efficient to preserve zona pellucida (ZP) integrity. Blastocysts were successfully stored for 24 h in a chemically defined medium. Yet, only 48 h storage in NCSU-BSA medium supported blastocyst development. Although all storage conditions resulted in an embryonic developmental delay, blastocysts stored in NCSU-BSA at either tested temperature could hatch and attain the same final developmental stage as control blastocysts when cultured under standard conditions after storage. Moreover, blastocysts stored at 25 °C for 48 h in NCSU-BSA medium could produce pregnancies after surgical transfer. In conclusion, porcine blastocysts maintain their viability and developmental potential after storage in the semi-defined medium NCSU-BSA for at least 48 h at 25 °C.

High pre-freezing sperm dilution improves monospermy without affecting the penetration rate in porcine IVF.

The high incidence of polyspermy is still an unresolved problem for the production of in vitro-produced porcine embryos. In this work, we modified the usual sperm processing sequence for in vitro fertilization (IVF), and the spermatozoa from four boars were frozen directly at a low sperm concentration of 20 × 106 sperm/mL (high pre-freezing sperm dilution group; F20), thawed and processed for IVF in three replicates. Spermatozoa from the same boars frozen at a conventional concentration (1000 × 106 sperm/mL) were used as the control group. The post-thaw sperm quality evaluation demonstrated that despite there being no differences in the percentage of motile spermatozoa between groups, the proportion of live spermatozoa with intact acrosomes was significantly higher in the F20 group than in the control. The in vitro penetration rate was also similar between groups; however, the co-incubation of oocytes with F20 sperm increased monospermy, IVF efficiency, cleavage rate and the efficiency of blastocyst formation compared with the results for oocytes co-incubated with control spermatozoa. These results indicate, for the first time, that a high pre-freezing sperm dilution increases monospermy without affecting penetration rates, thereby increasing blastocyst formation.

Sustainable hydrogen production from bio-oil model compounds (meta-xylene) and mixtures (1-butanol, meta-xylene and furfural).

In the present work m-xylene and an equimolecular mixture of m-xylene, 1-butanol and furfural, all of them bio-oil model compounds, were studied in steam reforming (SR) conditions. Three different nickel catalysts, which showed to be active in 1-butanol SR (Ni/Al2O3, Ni/CeO2-Al2O3 and Ni/La2O3-Al2O3), were tested and compared with thermodynamic equilibrium values. Tests were carried out at temperatures from 800 to 600°C at atmospheric pressure with a steam to carbon ratio (S/C) of 5.0. Despite the different bio-oils fed, the amount of moles going through the catalytic bed was kept constant in order to obtain comparable results. After their use, catalysts were characterized by different techniques and those values were correlated with the activity results. All catalysts were deactivated during the SR of the mixture, mainly by coking. The highest hydrogen yields were obtained with Ni/Al2O3 and Ni/CeO2-Al2O3 catalysts in the SR of m-xylene and SR of the mixture, respectively.

Reconstrucción de antebrazo con colgajo DIEP. Caso clínico / DIEP flap for forearm reconstruction. Case report

Las heridas complejas del antebrazo con fracturas asociadas y pérdida circunferencial de piel, suponen un doble reto reconstructivo. Mostramos el tratamiento de una paciente con lesiones combinadas en el miembro superior tras atrapamiento por rodillos fríos industriales mediante el uso de un colgajo libre de perforante del eje epigástrico inferior profundo (DIEP), tras tratamiento de la fractura articular de la extremidad distal de radio guiado por artroscopia. Conseguimos la estabilización de las fracturas y la cobertura completa del defecto. El colgajo DIEP permite la cobertura de áreas extensas con escasa morbilidad en la zona donante y con un correcto resultado estético (AU)

Complex injuries in upper limb like fractures and forearm circumferential skin loss involving are a double reconstructive challenge. We show a case management in a patient with combined injuries in the upper limb because of entrapment by industrial rollers, using deep inferior epigastric perforator free flap (DIEP) after treatment of an articular fracture of distal radius guided by arthroscopy. Was achieved stabilization of fractures and complete coverage of the defect, with good acceptance of the donor site by the patient. DIEP flap provides coverage for large areas with low morbidity and good aesthetic result (AU)

Comparability of river quality assessment using macrophytes: a multi-step procedure to overcome biogeographical differences.

This paper exposes a new methodological approach to solve the problem of intercalibrating river quality national methods when a common metric is lacking and most of the countries share the same Water Framework Directive (WFD) assessment method. We provide recommendations for similar works in future concerning the assessment of ecological accuracy and highlight the importance of a good common ground to make feasible the scientific work beyond the intercalibration. The approach herein presented was applied to highly seasonal rivers of the Mediterranean Geographical Intercalibration Group for the Biological Quality Element Macrophytes. The Mediterranean Group of river macrophytes involved seven countries and two assessment methods with similar acquisition data and assessment concept: the Macrophyte Biological Index for Rivers (IBMR) for Cyprus, France, Greece, Italy, Portugal and Spain, and the River Macrophyte Index (RMI) for Slovenia. Database included 318 sites of which 78 were considered as benchmarks. The boundary harmonization was performed for common WFD-assessment methods (all countries except Slovenia) using the median of the Good/Moderate and High/Good boundaries of all countries. Then, whenever possible, the Slovenian method, RMI was computed for the entire database. The IBMR was also computed for the Slovenian sites and was regressed against RMI in order to check the relatedness of methods (R(2)=0.45; p<0.00001) and to convert RMI boundaries into the IBMR scale. The boundary bias of RMI was computed using direct comparison of classification and the median boundary values following boundary harmonization. The average absolute class differences after harmonization is 26% and the percentage of classifications differing by half of a quality class is also small (16.4%). This multi-step approach to the intercalibration was endorsed by the WFD Regulatory Committee.

Least Disturbed Condition for European Mediterranean rivers.

The present report describes a three-step approach that was used to characterize and define thresholds for the Least Disturbed Condition in Mediterranean streams of four different types, regarding organic pollution and nutrients, hydrological and morphological alterations, and land use. For this purpose, a common database composed of national reference sites (929 records) from seven countries, sampled for invertebrates, diatoms and macrophytes was used. The analyses of reference sites showed that small (catchment <100 km(2)) siliceous and non-siliceous streams were mainly affected by channelization, bank alteration and hydropeaking. Medium-sized siliceous rivers were the most affected by stressors: 25-43% of the samples showed at least slight alterations regarding channelization, connectivity, upstream dam influence, hydropeaking and degradation of riparian vegetation. Temporary streams were the least affected by hydromorphological changes, but they were nevertheless affected by alterations in riparian vegetation. There were no major differences between all permanent stream types regarding water quality, but temporary streams showed lower values for oxygenation (DO) and wider ranges for other variables, such as nitrates. A lower threshold value for DO (60%) was determined for this stream type and can be attributed to the streams' natural characteristics. For all other river types, common limits were found for the remaining variables (ammonium, nitrate, phosphate, total P, % of artificial areas, % of intensive and extensive agriculture, % of semi-natural areas in the catchment). These values were then used to select the list of reference sites. The biological communities were characterized, revealing the existence of nine groups of Mediterranean invertebrate communities, six for diatoms and five for macrophytes: each group was characterized by specific indicator taxa that highlighted the differences between groups.

Catalytic reactive distillation process development for 1,1 diethoxy butane production from renewable sources.

Some acetals can be produced from renewable resources (bioalcohols) and seem to be good candidates for different applications such as oxygenated diesel additives. In the present case the production of 1,1 diethoxy butane from bioethanol and butanal is presented. Butanal can be obtained from biobutanol following a partial oxidation or a dehydrogenation process. In this paper innovative process development about the synthesis of the mentioned acetal including catalytic reactive distillation experimental and simulation results will be presented and discussed. Katapak SP modules containing Amberlyst 47 resin were used as structured catalytic packings. This reactive system allowed reaching higher conversions than the equilibrium ones at the same temperatures. All the experimental data gathered allowed to tune a simulation model for the reactive distillation operation which showed a fairly good behavior in order to perform initial 1,1 diethoxy butane production process design studies.

Latissimus dorsi volteado para reconstrucción de carcinoma basocelular gigante en un paciente anciano / Latissimus dorsi turn over flap for giant basal cell carcinoma reconstruction in elderly patient

Los carcinomas basocelulares son tumores cutáneos de crecimiento lento aunque pueden llegar a tener un gran tamaño a lo largo de los años si el entorno socio económico y asistencial del paciente es pobre, como sucede en el caso que presentamos, en el que la lesión ulcerada en la línea media de la espalda llegó a medir 21 x 13 cm de diámetro. El colgajo de Latissimus Dorsi permite gracias a sus dos pedículos y a ser pivotado, una posibilidad reconstructiva en el caso de pacientes ancianos, evitando el uso de colgajos libres (AU)

Basal cell carcinoma are well known as slow local growth cutaneous tumors that can enlarge through years in low socioeconomic and poor health assistance environments, as in the case that we present, with a 21x 13 cm ulcerated lesion located over back midline. The Latissimus Dorsi muscle flap offers a reliable reconstruction in elderly patients avoiding the use of free flaps. This flap, because of its two different pedicles, can be used to reach defects in opposite directions (AU)

Reconstrucción de cuero cabelludo mediante colgajo de galea frontal: a propósito de un caso / Scalp reconstruction with galeal frontalis flap: case report

Las grandes pérdidas de sustancia a nivel de cuero cabelludo a menudo presentan dificultad de cobertura debido a la rigidez de los tejidos locales a movilizar, debiéndose recurrir a reconstrucción con colgajos locales y expansión tisular, o bien a la transferencia microvascular de tejido libre. El presente trabajo tiene por objeto presentar el tratamiento para la resolución de una pérdida de sustancia de 6 meses de evolución a nivel temporal izquierdo, de 10 x 12 cm. con exposición de hueso temporal, junto con la restitución de la línea de implantación del pelo mediante utilización de colgajo de galea frontal y expansión tisular posterior. El colgajo de galea frontal tiene una vascularización rica y constante procedente del eje supratroclear, y su anatomía y disección ha sido estudiada y ampliamente descrita. En nuestro caso el colgajo de galea frontal ha resultado de utilidad en la reconstrucción de la zona temporal, permitiendo la cobertura de la bóveda craneal y la reconstrucción del cuero cabelludo mediante expansión tisular (AU)

Large defects of scalp are difficult for reconstruction because of lacking of local tissues. Most of the times, local flaps, tissue expansion and even free microvascular tissue transfer are indicated. This paper aims the treatment of a 6 months 10x12cm left temporal bone exposure, which was solved with a galea frontalis flap followed by scalp expansion for hair line restitution. The galea frontalis flap has a rich constant vascularization, and it´s anatomy and surgical dissection has been described elsewhere. This flap has been useful in our case for temporal area reconstruction, allowing bone coverage and hairline reconstruction with expansion tissue technique (AU)

Treatment of multiple adjacent gingival recessions with the tunnel subepithelial connective tissue graft: a clinical report.

This article describes a surgical periodontal plastic procedure for the coverage of multiple adjacent gingival recessions. This surgical technique is based on the construction of a tunnel under the gingival tissue by means of a sulcular incision beyond the mucogingival line without raising the papillae. A large connective tissue graft obtained from the palatal mucosa is introduced through this tunnel, covering the adjacent gingival recessions. A suturing technique to allow this graft to slip through the tunnel under the gingival tissues and to secure and stabilize the graft covering the recessions is described. Twelve-month postoperative results are presented from 21 teeth that were treated with this technique: 100% root coverage was achieved in 66.7% of the recessions treated, with a mean root surface coverage of 91.6%. This study suggests that the use of this surgical procedure allows the treatment of multiple adjacent recessions in a single procedure with adequate early healing and highly predictable root coverage results.

[Pharmacologic evaluation of penicillin in children with meningococcal meningitis]. / Evaluación farmacológica de la penicilina en niños con meningitis meningocócica.

Levels of penicillin both in blood and CSF are determined in 11 children with meningococcal meningitis. Authors compare these values with MIC. The therapeutic pattern was 500.000 Ul/Kg/day of benzylpenicillin, in six doses, with a single initial doses of 200.000 Ul/Kg. The results recommend not to use a lower doses in order to assure a permanently efficient penicillin concentration.

[Acute epiglottitis. Diagnosis and treatment in eight cases (author's transl)]. / Epiglotitis aguda. Diagnóstico y tratamiento a propósito de ocho casos.

A revision of acute epiglottitis is carried out with reference to eight cases recorded from 1973 to 1981. The ages of the patients varied from 12 to 35 months, with a female predominance (7:1) and a seasonal pattern occurring in winter months. Authors establish the attitude to be taken when faced to a patient clinically suspected of having acute epiglottitis, its' diagnosis and therapeutic protocol. The use of nasotracheal intubation instead of tracheostomy as the best way of obtaining a free airway in these patients at risk of sudden airway obstruction, is specially stressed.