Boar sperm protein tyrosine phosphorylation in the presence of egg yolk soluble and low density lipoprotein fractions during cooling.

Increased protein tyrosine phosphorylation and the appearance of a phosphorylated protein of 32 kD (p32) are reported among the capacitation-like changes in cryopreserved boar sperm. Egg yolk freezing extenders are composed by two fractions: insoluble granules and soluble plasma, which contains the low density lipoproteins (LDL) proposed as responsible for the egg yolk cryoprotective action. The aim of this work was to analyze the effects of complete egg yolk and its insoluble, soluble and LDL fractions on boar sperm quality and protein tyrosine phosphorylation after the first stage of a standard cryopreservation protocol. Semen samples in Androstar® Plus diluent were centrifuged and resuspended in the different egg yolk extenders. Temperature was decreased from 17 °C to 5 °C and sperm quality, protein tyrosine phosphorylation and protein pattern were analyzed. Results showed that complete egg yolk as well as soluble and LDL egg yolk fractions maintained sperm quality after temperature decrease. Cooling without any lipid component or in the presence of the insoluble fraction, significantly reduced sperm motility. About sperm protein tyrosine phosphorylation analysis, the p32 band appeared before treatments or after cooling in Androstar® Plus diluent. Complete egg yolk and its insoluble fraction interfered with sperm tyrosine phosphorylation even after cells were extensively washed. Analysis of extenders revealed a high amount of tyrosine phosphorylated proteins in the insoluble fraction, which may have co-precipitate with sperm in experiments. Samples submitted to temperature decrease from 17 °C to 5 °C in the presence of soluble and LDL egg yolk fractions in Androstar® Plus diluent did not show any change in the p32 band associated with sperm capacitation. However, a tyrosine-phosphorylated protein of 33 kD present in clarified egg yolk was also observed in sperm treated with this extender. Protein transference from plasma and LDL egg yolk extenders was also observed in sperm protein profile. Results suggested that soluble and LDL fractions might have a protective action preventing sperm protein tyrosine phosphorylation during cooling from 17 °C to 5 °C. Further studies are needed to expand the knowledge of the LDL protection mechanism as well as to determine the possible benefits of clarified egg yolk in freezing protocols.

Copper-induced cell death and the protective role of glutathione: the implication of impaired protein folding rather than oxidative stress.

Copper (Cu) is a bioelement essential for a myriad of enzymatic reactions, which when present in high concentration leads to cytotoxicity. Whereas Cu toxicity is usually assumed to originate from the metal's ability to enhance lipid peroxidation, the role of oxidative stress has remained uncertain since no antioxidant therapy has ever been effective. Here we show that Cu overload induces cell death independently of the metal's ability to oxidize the intracellular milieu. In fact, cells neither lose control of their thiol homeostasis until briefly before the onset of cell death, nor trigger a consistent antioxidant response. As expected, glutathione (GSH) protects the cell from Cu-mediated cytotoxicity but, surprisingly, fully independent of its reactive thiol. Moreover, the oxidation state of extracellular Cu is irrelevant as cells accumulate the metal as cuprous ions. We provide evidence that cell death is driven by the interaction of cuprous ions with proteins which impairs protein folding and promotes aggregation. Consequently, cells mostly react to Cu by mounting a heat shock response and trying to restore protein homeostasis. The protective role of GSH is based on the binding of cuprous ions, thus preventing the metal interaction with proteins. Due to the high intracellular content of GSH, it is depleted near the Cu entry site, and hence Cu can interact with proteins and cause aggregation and cytotoxicity immediately below the plasma membrane.

Antimicrobial Activity of Starch Hydrogel Incorporated with Copper Nanoparticles.

In order to obtain an antimicrobial gel, a starch-based hydrogel reinforced with silica-coated copper nanoparticles (Cu NPs) was developed. Cu NPs were synthesized by use of a copper salt and hydrazine as a reducing agent. In order to enhance Cu NP stability over time, they were synthesized in a starch medium followed by a silica coating. The starch hydrogel was prepared by use of urea and water as plasticizers and it was treated with different concentrations of silica-coated copper nanoparticles (Si-Cu NPs). The obtained materials were characterized by Fourier transform infrared (FT-IR) spectroscopy, electron paramagnetic resonance (EPR) spectroscopy, scanning electron microscopy (SEM), and rheometry. FT-IR and EPR spectra were used for characterization of Cu NPs and Si-Cu NPs, confirming that a starch cap was formed around the Cu NP and demonstrating the stability of the copper nanoparticle after the silica coating step. SEM images showed Cu NP, Si-Cu NP, and hydrogel morphology. The particle size was polydisperse and the structure of the gels changed along with particle concentration. Increased NP content led to larger pores in starch structure. These results were in accordance with the rheological behavior, where reinforcement by the Si-Cu NP was seen. Antimicrobial activity was evaluated against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacterial species. The hydrogels were demonstrated to maintain antimicrobial activity for at least four cycles of use. A dermal acute toxicity test showed that the material could be scored as slightly irritant, proving its biocompatibility. With these advantages, it is believed that the designed Si-Cu NP loaded hydrogel may show high potential for applications in various clinical fields, such as wound dressings and fillers.

Desbridamiento apropiado y técnica de "pie crusting" en el manejo de herida traumática / Adequate debridement and pie-crusting technique in the management of traumatic injuries

Las heridas crónicas de origen traumático, con exposición de tejidos, requieren de un desbridamiento adecuado, lavado y una pronta cubierta para evitar la infección y la desecación. A veces, incluso deben ser ampliadas para realizar un adecuado desbridamiento quirúrgico; por lo que, al intentar una cobertura completa, el resultado es una herida a tensión, que se complica con inflamación, infección y la dehiscencia que se acentúa aún más si está en una zona de flexión, como la rodilla. Se presenta el caso de una paciente de 28 años, que acudió a emergencias con un antecedente de herida traumática en la rodilla derecha, signos de retraso de la cicatrización, tejido de granulación friable, exposición de la rótula, abundante secreción serosa y dolor al movimiento con rango limitado. Se la trató en un solo tiempo quirúrgico con desbridamiento, irrigación y cobertura completa de la herida mediante la técnica de "pie-crusting". Nivel de Evidencia: IV

Chronic wounds of traumatic origin, with tissue exposure, require adequate debridement, lavage and prompt coverage to prevent infection and desiccation. Wounds may even require to be enlarged in order to perform an adequate surgical debridement. Enlarged wound attempts to perform a complete coverage may result in tension wound closures, which are complicated by inflammation, infection, and dehiscence and aggravated when located on flexure areas, such as on the knee. We report the case of a 28-year female patient, who presented to the Emergency Department with a history of traumatic wound in the right knee and signs of delayed healing, friable granulation tissue, exposed patella, increased serous drainage, and painful limited range of motion. She underwent a single surgical time procedure with debridement, irrigation, and complete coverage of the wound with the help of the pie-crusting technique. Level of Evidence: IV