An examination of acute physiological and perceptual responses following blood flow restriction exercise using a traditional research device or novel, automated system.

Objective. To compare the acute physiological and perceptual responses to blood flow restriction (BFR) exercise using a traditional research device or novel, automated system.Methods. Forty-four resistance trained individuals performed four sets of unilateral elbow flexion exercise (30% one-repetition maximum) to volitional failure using two distinct restrictive devices [SmartCuffs PRO BFR Model (SMARTCUFF), Hokanson E20 Rapid Inflation device (HOKANSON)] and with two levels of BFR [40% limb occlusion pressure (LOP), 80% LOP]. Blood pressure (BP), muscle thickness (MT), and isometric strength (ISO) were assessed prior to and following exercise. Perceptual responses [ratings of perceived exertion (RPE), discomfort] were assessed prior to exercise and following each exercise set.Main results. Data are displayed as means (SD). Immediately following exercise with 40% LOP, there were no statistical differences between devices for BP, MT, and ISO. However, only following Set 1 of exercise, RPE was greater with SMARTCUFF compared to HOKANSON (p< 0.05). In addition, only following Set 2 of exercise, discomfort was greater with HOKANSON compared to SMARTCUFF (p< 0.001). Immediately following exercise with 80% LOP, there were no statistical differences between devices for BP, MT, and ISO. However, only following Set 4 of exercise, RPE was greater with HOKANSON compared to SMARTCUFF (p< 0.05). In addition, following all exercise sets, discomfort was greater with HOKANSON compared to SMARTCUFF (p< 0.001). For repetitions completed with 40% LOP there were no statistical differences between SMARTCUFF and HOKANSON across any exercise sets. For repetitions completed with 80% LOP there were no statistical differences between SMARTCUFF and HOKANSON across Set 1 of exercise (p= 0.34), however, for Sets 2-4 of exercise, significantly greater number of repetitions were completed during SMARTCUFF than HOKANSON.Significance. The present study provides valuable insight into the efficacy of a novel, automated BFR system (SMARTCUFF) eliciting comparable acute physiological responses to BFR exercise and in some cases favorable perceptual responses when compared to a traditional research device (HOKANSON).

Change in the swimming pattern of Salmo salar spermatozoa caused by the high temperature of the sperm motility activation medium.

Fish are ectotherms and many have an external reproductive mode. An environmental factor which triggers fish reproductive activity in fish is water temperature. However, climate change is causing increasingly frequent events in which the water temperature varies rapidly; as a result, both in hatchery and in natural conditions, fish sperm are exposed to varying environmental temperatures during their journey toward the egg. This study was based on two experiments: The first experiment was designed to determine how storage at 4 °C for four days affected the sperm functions of Atlantic salmon (Salmo salar) sperm collected by either abdominal massage (stripping/Pure) or testicular dissection (testicular macerate/Macerated). Further, computer-assisted semen analysis (CASA) was used to compare sperm velocity parameters (VCL, VSL, and VAP) and progressivity (STR, LIN, and WOB) after motility activation at different temperatures (8 and 16 °C) of sperm collected by both methods (Pure vs Macerated). The results show that spermatozoa from Macerated samples maintained a higher sperm function when stored at 4 °C for 4 days compared to Pure sperm samples. In the second experiment, CASA determined that all parameters for sperm velocity (VCL, VSL, and VAP) and progressivity (STR (50%/55%), LIN (25%-32%), and WOB (51%-57%) were affected by activation temperature (P < 0.05) and that the motility patterns after activation at 16 °C (P < 0.05), specifically the LIN or STR swimming trajectories of the sperm differed between the two groups. In conclusion, the sperm quality of testicular Macerate was superior to that of Pure sperm abdominal mass, based on the higher quality of various sperm functions during short-term storage. Moreover, there was a significant effect of the temperature of the activation medium on sperm speed and progressivity (motility pattern) in the collected samples of testicular macerate. The sensitivity of Salmo salar spermatozoa to elevated temperature varies markedly between collection methods (Pure and Macerated).

Long-term survive of Aliarcobacter butzleri in two models symbiotic interaction with Acanthamoeba castellanii.

Aliarcobacter butzleri (formerly known as Arcobacter butzleri) is an emerging food-borne zoonotic pathogen that establishes in vitro endosymbiotic relationships with Acanthamoeba castellanii, a free-living amoeba. Previously, we described that this bacterium acts as an endocytobiont of A. castellanii, surviving for at least 10 days in absence of bacterial replication. Thus, the aim of this study was to evaluate the ability of A. butzleri to survive as a long-term endosymbiont of A. castellanii for 30 days in two models of symbiotic interaction with A. castellanii: (i) endosymbiotic culture followed by gentamicin protection assay and (ii) transwell co-culture assay. The results allow us to conclude that A. butzleri is capable of surviving as an endosymbiont of A. castellanii for at least 30 days, without multiplying, under controlled laboratory conditions. In addition, in the absence of nutrients and as both microorganisms remain in the same culture, separated by semi-permeable membranes, A. castellanii does not promote the survival of A. butzleri, nor does it multiply. Our findings suggest that the greater survival capacity of A. butzleri is associated with their endosymbiont status inside A. castellanii, pointing out the complexity of this type of symbiotic relationship.

Dietary carbohydrate-to-protein ratio influences growth performance, hepatic health and dynamic of gut microbiota in atlantic salmon (Salmo salar).

Atlantic salmon (Salmo salar) fed a carbohydrate-rich diet exhibit suboptimal growth performance, along with other metabolic disturbances. It is well known that gut microbes play a pivotal role in influencing metabolism of the host, and these microbes can be modified by the diet. The main goal of the present study was to determine the effect of feeding graded levels of digestible carbohydrates to Atlantic salmon on the distal intestine digesta microbiota at 3 sampling times (i.e., weeks 4, 8 and 12), during a 12-week trial. A low carbohydrate-to-high protein diet (LC/HP, 0% wheat starch), a medium carbohydrate-to-medium protein diet (MC/MP, 15% wheat starch) or a high carbohydrate-to-low protein diet (HC/LP, 30% wheat starch) was fed to triplicate fish tanks (27 to 28 fish per tank). We performed an in-depth characterization of the distal intestine digesta microbiota. Further, growth parameters, liver histology and the expression of genes involved in hepatic neolipogenesis in fish were measured. Fish fed a HC/LP diet showed greater hepatosomatic and viscerosomatic indexes (P = 0.026 and P = 0.018, respectively), lower final weight (P = 0.005), weight gain (P = 0.003), feed efficiency (P = 0.033) and growth rate (P = 0.003) compared with fish fed the LC/HP diet. Further, feeding salmon a high digestible carbohydrate diet caused greater lipid vacuolization, steatosis index (P = 0.007) and expression of fatty acid synthase (fas) and delta-6 fatty acyl desaturase (d6fad) (P = 0.001 and P = 0.001, respectively) in the liver compared with fish fed the LC/HP diet. Although, the major impact of feeding a carbohydrate-rich diet to Atlantic salmon in beta diversity of distal intestine digesta microbiota was observed at week 4 (HC/LP vs MC/MP and HC/LP vs LC/HP; P = 0.007 and P = 0.008, respectively) and week 8 (HC/LP vs MC/MP; P = 0.04), no differences between experimental groups were detected after 12 weeks of feeding. Finally, at the end of the trial, there was a negative correlation between lactic acid bacteria (LAB) members, including Leuconostoc and Lactobacillus, with hepatic steatosis level, the hepatosomatic and viscerosomatic indexes as well as the expression of fas and d6fad. Weissella showed negative correlation with hepatic steatosis level and the hepatosomatic index. Finally, further research to explore the potential use of LAB as probiotics to improve liver health in carnivorous fish fed fatty liver-induced diet is warranted.

A bioinformatics analysis of the CatSper channel in the class Actinopterygii.

The regulation of sperm motility is controlled by several variables, including mainly ion concentrations. In fish, Ca2+ concentrations play an important role in the regulation of sperm motility, and several reports highlight the importance of certain Ca2+ channels in the regulation of this cell function. CatSper is a calcium channel scarcely studied in fish. In the species Salmo salar, it has been shown that it is key in the regulation of sperm motility. Taking into account the relevance of this channel in sperm activation in fish, in this study we evaluated the presence and probable functionality of this channel in the class Actinopterygii. For this purpose, a rational bioinformatic analysis was carried out, which had been previously validated using in vitro techniques by our group. The bioinformatic analysis of the present work revealed that the functionality of CatSper of the species of the class Actinopterygii could be exclusive to freshwater and anadromous fish species. The results of this study showed that only some anadromous and freshwater fish species contain 11 subunits of the CatSper channel, which are enough to trigger sperm motility. Consequently, this study provides new data for a better understanding of the sperm activation mechanism in fish.

Chorion Alterations in Eyed-Stage Salmonid Eggs Farmed in La Araucanía, Chile: A Retrospective Study.

The chorion is the primary envelop that protects the fish embryo against mechanical actions, pathogens, and abrupt changes in physical and chemicals conditions of the incubation medium. During embryo development, chorion alterations are not rare, but the occurrence of these is scarcely reported. Increased frequency of chorion alterations can result in increased embryo mortality and thus decreased reproductive performance and losses for fish farms. In this study, we characterize different chorion alterations observed in samples collected over 14 years from 12 salmon and trout farms located in the region of La Araucanía in southern Chile, which sent live eyed-stage embryos ('eyed-eggs') for quality analysis to our laboratory. We found soft chorion as the most common alteration observed, being present in the whole 14-year series analyzed in Atlantic Salmon (Salmo salar) and affecting up to 35.0% of the samples examined in a year. This alteration also affected up to 20.0 and 5.7% of Coho Salmon (Oncorhynchus kisutch) and Rainbow Trout (Oncorhynchus mykiss) samples analyzed in a year, respectively. We also found an increase of other chorion alterations, including perforated and white-spotted chorion in Atlantic and Coho Salmon, in the last 8 years. Among the three species, Rainbow Trout exhibited fewer chorion alterations. As the embryonated eggs analyzed here were obtained from broodstocks maintained under standard industrial conditions, these alterations might be linked to changes in environmental conditions affecting the incubation water that need to be further investigated.

The voltage-gated T-type Ca2+ channel is key to the sperm motility of Atlantic salmon (Salmo salar).

Ca2+ is a key element in the sperm activation process of Salmo salar. However, the molecular mechanisms by which this ion enters the sperm cell have been poorly studied. In this study, we examined, for the first time, the role of the voltage-gated T-type Ca2+ channel in the activation of sperm motility of Salmo salar. Using an in vitro inhibition assay, a significant decrease in total and progressive motility (P < 0.0001) was observed in Salmo salar sperm when they were treated with NNC-55-0396, a highly selective blocker. The in silico analysis showed that this blocker is docked with a strong affinity for the pore of the voltage-gated T-type calcium channel suggesting the blocking of Ca2+ ions. The results show that the T-type voltage-gated Ca2+ channel is key to sperm motility in Salmo salar.

Short-term storage sperm of coho salmon (Oncorhynchus kisutch) at 4 °C: Effect of sperm: Extender dilution ratios and antioxidant butyl-hydroxytoluene (BHT) on sperm function.

Short-term storage of semen is a necessary key procedure in fish; it allows maximizing the use of gametes. Nevertheless, sperm quality decreases during storage has been associated with oxidative stress damage due to an increase in reactive oxygen species (ROS) during storage. This study was designed to optimize a short-term storage protocol for Coho salmon (Oncorhynchus kisutch) spermatozoa, evaluating the effect of extender dilution and the addition of butyl-hydroxytoluene (BHT) antioxidant on sperm function parameters. In the first experiment, fresh semen was diluted in Storfish®: extender dilution (1:2 and 1:3) and a control sample undiluted and stored at 4 °C for 7-days. In both experiments motility (MO), viability and integrity of plasma membrane, mitochondrial membrane potential (MMP) and superoxide anion level (O2-) were evaluated at 0, 3 and 7 days. Result shows that, 1:3 dilution maintained a higher sperm function for a longer period time. In the second experiment, spermatozoa were suspended in Storfish® (1:3) supplemented with two different concentrations of BHT (1.0 mM and 2.0 mM) and a control sample without antioxidant and stored at 4 °C for 7 days. The results demonstrated that, antioxidant-supplemented samples greater MO than control samples (P < 0.05). The viability remained >75% during storage in all groups. MMP was higher in 2.0 mM BHT compared to 1.0 mM and control (P < 0.05), in addition, this concentration reduced O2- level (P < 0.05). In conclusion, sperm: extender dilution 1:3 and adding of 2.0 mM BHT in sperm storage extender may enhance protection sperm function in Oncorhynchus kisutch against effects harmful of the oxidative stress during the in vitro storage.

Effects of selection by the Percoll density gradient method on motility, mitochondrial membrane potential and fertility in a subpopulation of Atlantic salmon (Salmo salar) testicular spermatozoa.

The aim of this study was to evaluate effects of selection using the Percoll density gradient method on motility, mitochondrial membrane potential (ΔΨMit) and fertility in a subpopulation of testicular spermatozoa obtained from Atlantic salmon (Salmo salar). Samples were divided into three groups: Control (C), T1 (45/90 % Percoll®) and T2 (45/60 % Percoll®). Sperm motility was evaluated using CASA (Computer-Assisted Sperm Analysis), ΔΨMit using flow cytometry, and fertility evaluating whether cleavage of fertilised eggs had occurred after 16 h of incubation at 10 °C. Results indicate that motility was greater in T1 (92 ± 2.91 %) and T2 (89 ± 2.88 %) than in the Control (83.2 ± 2.04 %). The percentage of ΔΨMit was 88.3 ± 0.58 % and 85 ± 2% for T1 and T2, respectively, compared to 35 ± 6.24 % for the control. The fertility rates were 76 ± 9.1 % and 70 ± 8.1 % for T1 and T2, respectively, compared with 66 ± 12 % for the control. The kinetic characteristics for T1 were curvilinear velocity (VCL): 92.44 ± 21.12 µm/s, average path velocity (VAP): 85.87 ± 21.83 µm/s; and for T2 VCL was 78.69 ± 17.63 µm/s and VAP was 73.62 ± 17.08 µm/s. The results indicate sperm motility and ΔΨMit were greater in T1 and T2 compared with the control (P < 0.05). Similarly, there was an increase in the fertilisation rate compared to the control. The results from this study are the first where sperm quality variables were evaluated for Salmo salar testicular sperm using the Percoll® density gradient method.

The CatSper channel is present and plays a key role in sperm motility of the Atlantic salmon (Salmo salar).

Among all the Ca2+ channels, CatSper channels have been one of the most studied in sperm of different species due to their demonstrated role in the fertilization process. In fish sperm, the calcium channel plays a key role in sperm activation. However, the functionality of the CatSper channels has not been studied in any of the fish species. For the first time, we studied the relationship of the CatSper channel with sperm motility in a fish, using Atlantic salmon (Salmo salar) as the model. The results of our study showed that the CatSper channel in Salmo salar has chemical-physical characteristics similar to those reported for mammalian CatSper channels. In this work, it was shown that Salmo salar CatSper 3 protein has a molecular weight of approximately 55-kDa similar to Homo sapiens CatSper 3. In silico analyses suggest that this channel forms a heterotetramer sensitive to the specific inhibitor HC-056456, with a binding site in the center of the pore of the CatSper channel, hindering or preventing the influx of Ca2+ ions. The in vitro assay of the sperm motility inhibition of Salmo salar with the inhibitor HC-056456 showed that sperm treated with this inhibitor significantly reduced the total and progressive motility (p < .0001), demonstrating the importance of this ionic channel for this cell. The complementation of the in silico and in vitro analyses of the present work demonstrates that the CatSper channel plays a key role in the regulation of sperm motility in Atlantic salmon.

Effects of cryopreservation on cAMP-dependent protein kinase and AMP-activated protein kinase in Atlantic salmon (Salmo salar) spermatozoa: Relation with post-thaw motility.

Sperm motility in fish with external fertilization is critical for reproductive efficiency in aquaculture, especially in salmonids. Gamete preservation techniques, such as cryopreservation, however, reduce sperm motility and fertilizing capacity. Very few studies have addressed cryodamage from energetic and cell signalling approaches. In this study, cAMP-dependent protein kinase (PKA) and AMP-activated kinase (AMPK) activities were quantified in fresh and cryopreserved spermatozoa of Atlantic salmon (Salmo salar); and the relation with motility was analysed. Results indicate there was a decrease in membrane integrity and motility in post-thawed spermatozoa compared to fresh samples, however, there was about 30% of cells with intact plasma membrane but incapable of motility. The PKA and AMPK activities were less after cryopreservation, indicating that loss of motility may be related to alteration of these key enzymes. Furthermore, PKA and AMPK activities were positively correlated with each other and with motility; and inhibition decreased motility, indicating there is a functional relationship between PKA and AMPK. The PKA inhibition also decreased AMPK activity, but results from protein-protein docking analyses indicated AMPK activation directly by PKA is unlikely, thus an indirect mechanism may exist. There have been no previous reports of these kinase actions in fish spermatozoa, making these findings worthy of assessment when there are future studies being planned, and may serve as base knowledge for optimization of cryopreservation procedures and development of biotechnologies to improve reproduction efficiency in the aquaculture industry.

Effects of pH and sugar supplements on bacteriocin-like inhibitory substance production by Pediococcus pentosaceus.

To improve bacteriocin-like inhibitory substance (BLIS) production by Pediococcus pentosaceus ATCC 43200, the influence of pH as well as the addition of sugars-either prebiotic (inulin) or not (sucrose)-on its metabolism were investigated. This strain was grown at pH 5.0 or 6.0 either in glucose-based MRS medium (control) or after addition of 0.5, 1.0 or 1.5% (w/w) sucrose and inulin (GSI-MRS) in the same percentages. In the control medium at pH 5.0, cell mass concentration after 48 h of fermentation (Xmax = 2.26 g/L), maximum specific growth rate (µmax = 0.180 h-1) and generation time (Tg = 3.84 h) were statistically coincident with those obtained in supplemented media. At pH 6.0 some variations occurred in these parameters between the control medium (Xmax = 2.68 g/L; µmax = 0.32 h-1; Tg = 2.17 h) and the above supplemented media (Xmax = 1.90, 2.52 and 1.86 g/L; µmax = 0.26, 0.33 and 0.32 h-1; Tg = 2.62, 2.06 and 2.11 h, respectively). Lactate production was remarkable at both pH values (13 and 16 g/L) and improved in all supplemented media, being 34 and 54% higher than in their respective control media, regardless of the concentration of these ingredients. Cell-free supernatant of the fermented control medium at pH 5.0 displayed an antimicrobial activity against Enterococcus 101 5.3% higher than that at pH 6.0 and even 20% higher than those of all supplemented media, regardless of the concentration of supplements. BLIS production was favored either at pH 5.0 or in the absence of any additional supplements, which were able, instead, to stimulate growth and lactate production by P. pentosaceus.

Sperm morphology and ultrastructure of Patagonian blenny (Eleginops maclovinus).

In this study, the morphology and ultrastructure of Eleginops maclovinus spermatozoa were characterized through scanning and electron microscopy. Findings revealed that E. maclovinus spermatozoa can be differentiated into three major parts: a spherical head without acrosome (typical for externally fertilizing teleost), a midpiece containing 2-5 spherical mitochondria, and a long flagellum. The mean length of the spermatozoa was 40.08 ± 2.30 µm, with flagella accounting for 38.38 ± 2.08 µm. The head was 1.31 ± 0.17 µm long, and 1.63 ± 0.01 µm wide. The midpiece was 0.39 ± 0.05 µm in length and 0.95 ± 0.12 µm in width. It was located below the nucleus and contained 2 to 5 spherical mitochondria. The mitochondria were separated from the axoneme by a cytoplasmic canal. There was no evidence of the flagellum membrane forming sidefins, and the axoneme was composed of the typical 9 + 2 microtubular doublet structure enclosed by cell membrane. The present study reveals that E. maclovinus sperm can be categorized as a primitive type. This study is the first to provide comprehensive details on the ultrastructure of spermatozoa in E. maclovinus.

Morphology and ultrastructure of pink cusk-eel (Genypterus blacodes, Schneider 1801) spermatozoa by scanning and transmission electron microscopy.

In this study, the morphology and ultrastructure of Genypterus blacodes spermatozoa were characterized through scanning and transmission electron microscopy. Findings revealed that the G. blacodes spermatozoa can be differentiated into three major parts: a spherical head without an acrosome (typical for externally fertilizing fish), a short mid-piece, and a long flagellum. The mean length of the spermatozoa was 57.6 ± 6.08 µm, with flagella accounting for 56.2 ± 7.2 µm. The head was 1.47 ± 0.2 µm long, and 0.89 ± 0.06 µm wide. The mid-piece had a total dimension of 0.72 ± 0.16 µm, and was 0.31 ± 0.02 µm in length and 0.6 ± 0.05 µm in width. It was located lateral to the nucleus and contained 4 or 5 spherical mitochondria. The mitochondria were separated from the axoneme by a cytoplasmic canal. The main piece of the flagellum had short irregular side-fins, and the axoneme was composed of the typical 9 + 2 microtubular doublet structure enclosed by a cell membrane. The present study reveals that G. blacodes sperm can be categorized as a primitive type. This study is the first to provide comprehensive details on the morphology and ultrastructure of spermatozoa in G. blacodes.

Patagonian blenny (Eleginops maclovinus) spermatozoa quality after storage at 4 ºC in Cortland medium.

Patagonian blenny (E. maclovinus) is a marine species recently placed in captivity and which are potentially farmable. Understanding and improving its sperm capacity to withstand short-term storage conditions is a key element of initiating an artificial propagation program for this species. The aim of this study is to evaluate the ultrastructure and quality of E. maclovinus sperm during refrigerated storage. To address this objective, scanning electron microscopy (SEM), cytofluorimetric analysis (membrane integrity; reactive oxygen species generation; mitochondrial membrane potential) and cell respiration/mitochondrial-function analysis (ATP content; oxygen consumption) could be useful for optimizing or improving management for artificial reproduction of this species. Severe damage of plasma membranes was observed by SEM at day 7 and 14 of in vitro storage. Analyses of sperm quality were conducted during the 14-day cold storage period when sperm were in diluted (with Cortland solution) and undiluted conditions. When there were diluted conditions, there was greater preservation of motile capacity (from day-7; P < 0.05), membrane integrity (from day-7; P < 0.05), mitochondrial membrane potential (from day-10; P < 0.05) and ATP stores (from day-3; P < 0.05). Oxygen consumption indicators were 18.6% ±14.7% greater in the undiluted samples from day-3, and 32.1%±2.1% of the total spermatozoa had ample amounts of superoxide anion in both undiluted and diluted semen on day-0. The use of Cortland solution extended the viability of sperm when there were longer storage times. Factors that have a greater effect on the quality of semen during storage are reactive oxygen species generation and ATP depletion. In conclusion, Patagonian blenny spermatozoa can be stored at 4 °C between 7 and 10 days using Cortland solution.

Antioxidant Therapeutic Strategies for Cardiovascular Conditions Associated with Oxidative Stress.

Oxidative stress (OS) refers to the imbalance between the generation of reactive oxygen species (ROS) and the ability to scavenge these ROS by endogenous antioxidant systems, where ROS overwhelms the antioxidant capacity. Excessive presence of ROS results in irreversible damage to cell membranes, DNA, and other cellular structures by oxidizing lipids, proteins, and nucleic acids. Oxidative stress plays a crucial role in the pathogenesis of cardiovascular diseases related to hypoxia, cardiotoxicity and ischemia-reperfusion. Here, we describe the participation of OS in the pathophysiology of cardiovascular conditions such as myocardial infarction, anthracycline cardiotoxicity and congenital heart disease. This review focuses on the different clinical events where redox factors and OS are related to cardiovascular pathophysiology, giving to support for novel pharmacological therapies such as omega 3 fatty acids, non-selective betablockers and microRNAs.

Mitochondria in teleost spermatozoa.

There is an extraordinary diversity of reproductive modes in teleost and this variability is related to the phylogenetic relationships and adaption to very different biotopes. As in all vertebrates, sperm is produced as the end product of the process of spermatogenesis, and regarding teleost the spermatozoa lack an acrosome in almost all species and motility is activated as a response to osmolarity and ion content of the aquatic medium where the sperm is released. In this context, mitochondria possess a fundamental role for fish spermatozoa motility and integrity, hence, fertilizing potential; they are the energy supplier that allows flagellar movement and their dysfunction could play a main role in structural and functional damage to the spermatozoa. The ATP production through oxidative phosphorylation provides not only energy for cell activities, which includes Na+/K+ ATPase pump, endocytosis, protein synthesis and many other cell processes; but also produces reactive oxygen species, that under mitochondrial dysfunction causes oxidative stress. The assessment of mitochondrial function (e.g. through measurement of mitochondrial membrane potential) as well as ATP content (mostly supplied by mitochondrial respiration) can be useful as quality markers of fish spermatozoa. Also quantification of ROS and antioxidant status, strongly influenced by mitochondria, are used as complementary measurements. There is much information about sperm mitochondria and their function but studies of these aspects on fish reproduction are still required for applications in aquaculture. The real role of fish sperm mitochondria under short and long term storage and in vitro manipulation is not fully understood yet. Thus future research should focus on these matters.

Molecular aspects of breast cancer resistance to drugs (Review).

Despite continuous advances in the knowledge of breast cancer pathophysiology, this type of neoplasia remains a leading cause of cancer-related death in women worldwide. Carcinogenesis takes a progressive course from somatic mutations, alteration of the DNA repair mechanisms, inhibition of growth suppressors, followed by cell proliferation, tissue invasion and risk of metastasis. Less than 10% of all cancers are hereditary, and in the case of breast cancer only 8%, a phenomenon linked to genetic changes in BRCA1 or BRCA2. All the other cancers can be caused by an infection (15%) or in most cases (75%) the etiology is unknown. Patients with genetic mutations in BRCA1 or BRCA2 have 30-60% likelihood of developing a second primary breast cancer and between 11 and 45% risk of ovarian cancer, HER-2/neu is overexpressed in ~30% of human breast tumors and it has a predictive role in chemotherapy and endocrine therapy.

Protective effects of polyunsatutared fatty acids supplementation against testicular damage induced by intermittent hypobaric hypoxia in rats.

BACKGROUND: Intermittent hypobaric hypoxia (IHH) induces changes in the redox status and structure in rat testis. These effects may be present in people at high altitudes, such as athletes and miners. Polyunsaturated fatty acids (PUFA) can be effective in counteracting these oxidative modifications due to their antioxidants properties. The aim of the work was to test whether PUFA supplementation attenuates oxidative damage in testis by reinforcing the antioxidant defense system. The animals were divided into four groups (7 rats per group): normobaric normoxia (~750 tor; pO2 156 mmHg; Nx); Nx + PUFA, supplemented with PUFA (DHA: EPA = 3:1; 0.3 g kg(-1) of body weight per day); hypoxic hypoxia (~428 tor; pO2 90 mmHg; Hx) and, Hx + PUFA. The hypoxic groups were exposed in 4 cycles to 96 h of HH followed by 96 h of normobaric normoxia for 32 days. Total antioxidant capacity (FRAP) and lipid peroxidation (malondialdehyde, MDA) in plasma and reduced (GSH)/oxidized glutathione (GSSG) ratio, tissue lipid peroxidation (TBARS) and antioxidant enzymes activity were assessed at the end of the study in testis. Also, SIRTUIN 1 and HIF-1 protein expression in testis were determined. RESULTS: IHH increased lipid peroxidation in plasma and HIF-1 protein levels in testis. In addition, IHH reduced FRAP levels in plasma, antioxidant enzymes activities and SIRTUIN 1 protein levels in testis. PUFA supplementation attenuated these effects, inducing the increases in FRAP, in the antioxidant enzymes activity and HIF-1 levels. CONCLUSIONS: These results suggest that the IHH model induces a prooxidant status in plasma and testis. The molecular protective effect of PUFA may involve the induction of an antioxidant mechanism.

Morphometric of blastomeres in Salmo salar.

For Salmo salar, there is a lack of information on the morphology of the first blastomeres formed during embryonic development and which could be used as a diagnostic tool for the first stages of development. The purpose of this investigation, therefore, was to characterize morphometrically the first blastomeres of S. salar. From a pool of eggs incubated at 7.5°C, 100 microphotographs of blastodiscs were extracted and analyzed at different incubation periods: 12, 14, 16, 20 or 24 h. Blastodiscs were characterized morphologically after 16, 20 or 24 h incubation, and classified into symmetric or asymmetric groups according to their morphology. The ratio of length (L) versus width (W) of each blastomere was determined, to establish its symmetry. In addition, 20 microphotographs of blastodiscs of normal appearance were analysed morphologically (control blastodisc: CB) for comparison (20 or 24 h). Results show that the first cleavage ends after 16 h of development. Seven categories were established during blastomere characterization: 47% normal (G1); 27% with dispersed margins (G2); 10% unequal (G3); 9% 'pie-shaped' (G4); 3% amorphous (G5); 2% three equal blastomeres and one different one (G6); and 2% with eccentric cleavage (G7). Although the incidence of abnormal cleavage in S. salar is uncertain, there is a potential for some asymmetries to be corrected during embryogenesis to generate viable individuals. More studies are necessary to correlate these abnormal cleavage patterns with indicators of quality in the later stages of embryogenesis in this species, to establish a quality assessment tool for gametes and/or embryos in salmonid species.