Boar sperm motility is modulated by CCK at a low concentration of bicarbonate under capacitation conditions.

In a previous study, our group detected the cholecystokinin (CCK) protein in the porcine oviduct. This fact, together with the involvement of CCK in the regulation of sperm protein tyrosine phosphorylation by the modulation of HCO3 - uptake (in mice and humans) suggests a role for CCK during sperm capacitation. Therefore, on the one hand, the expression of CCK receptors (CCK1R and CCK2R) on boar testes has been investigated and probed; on the other hand, boar spermatozoa (from seminal doses of 1-day and 5-day storage) were exposed to different concentrations of CCK (0-control, 25 or 50 µM) in a medium supporting capacitation supplemented with 0, 5 or 25 mmol/L of HCO3 - for 1 h at 38.5°C. Sperm motion (total and progressive motility), kinetic parameters, viability, acrosome status, and mitochondrial activity were determined. No differences between groups (0, 25 or 50 µM of CCK) were observed when HCO3 - was absent in the media (p > .05). However, the results showed that when the media was supplemented with 5 mmol/L HCO3 - in 1-day seminal dose storage, the linearity index (LIN, %), straightness index (STR, %) and oscillation index (WOB, %) (sperm kinetics parameters) increased in the presence of CCK regardless the concentration (p < .05). Nevertheless, CCK in sperm from 5-day storage only increased the WOB parameter in comparison to the control (p < .05). Furthermore, the average amplitude of the lateral displacement of the sperm head (ALH, µm) and curvilinear velocity (VCL, µm/s) decreased when CCK was present, depending on its concentration and sperm aging (1-day vs. 5-days) (p < .05). In the case of the media supporting capacitation supplemented with 25 mmol/L HCO3 - , any differences were observed except for sperm viability in the 5-day seminal doses, which increased in the 50 µM-CCK group compared to the control (p < .05). In conclusion, these data suggest an implication of CCK protein during sperm capacitation under low bicarbonate concentration increasing the sperm linear trajectory.

In vitro supplementation of testosterone or prolactin affects spermatozoa freezability in small ruminants.

In small ruminants, testosterone and prolactin plasma concentrations show circannual fluctuations as an adaptation mechanism to their seasonal breeding behavior. Sperm resistance to the freezing-thawing process shows seasonal fluctuation throughout the year, with lower sperm freezability at the beginning of the breeding season when prolactin and testosterone levels reach their maximum concentration. Nevertheless, whether these hormones directly affect post-thaw sperm quality parameters is still unclear. The objective was to study the effect of testosterone or prolactin added in vitro on sperm freezability in domestic ram (Ovis aries) and buck (Capra hircus). Sperm samples were incubated for 1 h with a range of testosterone (0, 2, 4, or 6 ng/mL; Exp. 1) or prolactin (0, 20, 100, 200, or 400 ng/mL; Exp. 2) concentrations. Samples were cryopreserved by slow freezing in straws at 0 h and after 1 h incubation. Sperm viability, acrosome integrity, motility, and kinetic parameters were assessed at 0 and 1 h in fresh and frozen-thawed samples. Results showed no hormone effect in fresh sperm, whereas these hormones affected post-thaw sperm parameters. In Exp. 1, in vitro incubation with testosterone decreased the post-thaw acrosome integrity of ram sperm (from 68.1 ± 6.3% to 49.6 ± 3.9%; P < 0.05). In Exp. 2, in vitro incubation with prolactin decreased the post-thaw acrosome integrity of ram (from 78.2 ± 3.4% to 66.3 ± 3.5%; P < 0.05) and buck sperm (from 81.7 ± 2.5% to 67.6 ± 3.5%; P < 0.05). Moreover, prolactin increased the post-thaw amplitude of lateral head displacement in ram sperm (from 3.3 ± 0.1 µm to 3.8 ± 0.2 µm; P < 0.05). In conclusion, either testosterone or prolactin added in vitro decreased the post-thaw acrosome integrity of ram and buck sperm. This suggests a destabilization process that could be decreasing sperm freezability when physiological levels of these hormones are high in vivo.

Effect of boar seminal dose type (cervical compared with post-cervical insemination) on cooling curve, sperm quality and storage time.

Seminal doses used for cervical and post-cervical artificial insemination (CAI and PCAI, respectively) vary in volume, the number of spermatozoa and packaging. The aim was to evaluate the outcomes when there was use of routine processing procedures for CAI- and PCAI-doses. Two different types of seminal doses were processed: 1) CAI: 2.7 × 109 sperm/80 ml; 2) PCAI: 1.5 × 109 sperm/45 ml. In Experiment 1, the cooling curve of seminal doses during processing occurred in two phases: 1st) At room temperature (23.4 ±â€¯0.5 °C) from 0 (just after packaging) to 120 min; 2nd) At refrigeration (15.7 ±â€¯0.8 °C) from 121-240 min. For the PCAI-doses, the time required to reach room temperature was 47 min compared to 107 min for CAI-doses (decreasing velocity of 0.093 °C/min and 0.048 °C/min, respectively). During refrigeration, for the PCAI-doses the time required to reach the desired preservation temperature was 20 min less than for CAI-doses (PCAI: 90 min, 0.074 °C/min; CAI: 110 min, 0.066 °C/min). In Experiment 2, sperm motility, kinetic parameters and acrosome damage for both types of doses were evaluated at 0, 24, 48 and 72 h of refrigeration. Also, morphology, pH, and osmolality were assessed at 0 and 72 h. Values for all these did not differ between CAI- and PCAI-doses. In conclusion, PCAI-doses took less time than CAI-doses to reach the desired temperature, but sperm quality was similar for CAI- and PCAI-doses during storage. Nevertheless, the different cooling curves should be taken into consideration for further investigation.

Effect of temperature and cell concentration on dolphin (Tursiops truncatus) spermatozoa quality evaluated at different days of refrigeration.

The use of cryopreserved dolphin spermatozoa facilitates the exchange of genetic material between aquatic parks and makes spermatozoa accessible to laboratories for studies to further our understanding of marine mammal reproduction. Sperm cryopreservation in the bottlenose dolphin (Tursiops truncatus) has been developed for the exchange of gametes within the ex situ population. The aim of this study was to develop an effective method for refrigeration of bottlenose dolphin spermatozoa diluted in a commercial extender (BTS). In Experiment 1, the effect of temperature (5 compared with 15 °C) on sperm quality was evaluated during 7 days of storage at 100 × 106 spermatozoa/ml. In Experiment 2, the effect of the storage concentration (100 × 106 compared with 20 × 106 spermatozoa/ml) on sperm quality was assessed during 7 days of storage at 5 °C. In Experiment 1, total motility (including % of rapid sperm) was greater at 5 than 15 °C. When the effect of storage concentration was evaluated (Experiment 2), total motility and ALH were greater at the higher storage concentration (100 × 106 spermatozoa/ml). For both experiments, values for viability, acrosome integrity, and normal morphology variables were consistent throughout the 7 days of refrigeration. In Experiment 3, a microbiological study was performed to evaluate the effect of the refrigeration temperature and days of storage on bacterial growth. The results of microbiological analysis indicated there was Staphylococcus aureus in some samples, however, there was no effect of temperature or days of refrigeration. In conclusion, bottlenose dolphin semen can be refrigerated for a short to medium period of storage and there is maintenance of functionality of sperm when stored at 100 × 106 spermatozoa/ml at 5 °C.

Effect of Porcine circovirus 2 (PCV-2) maternally derived antibodies on performance and PCV-2 viremia in vaccinated piglets under field conditions.

BACKGROUND: Nowadays, the most common presentation of PCV-2 is the subclinical infection in piglets after weaning. The success of PCV-2 vaccination is associated with the control of the clinical disease as well as the improvement of production parameters. In consequence, the objective of the present study was to analyse the effect of PCV-2 maternally derived antibody (MDA) levels on vaccine efficacy in piglets vaccinated at three weeks of age with a commercial PCV-2 subunit vaccine. The study was performed analysing a database with 6112 wean-to-slaughter piglets from 4 different European regions. RESULTS: Results showed that the use of the vaccine was able to decrease the PCV-2 viremia calculated as area under the curve (AUC = 60.29 ± 3.73), increase average daily weight gain (ADWG = 0.65 ± 0.01 kg/day) and reduce mortality (7%) in vaccinated piglets compared to non-vaccinated ones (AUC of 198.27 ± 6.14, 0.62 ± 0.01 kg/day and 11% respectively). The overall difference of ADWG between both groups was close to 30 g per day (p < 0.05), also when they were split for low and high levels of MDA titres. Moreover, the animals with the highest ADWG were observed in the group of piglets vaccinated with high or extremely high antibody titres (0.66 and 0.65 kg/day respectively). Considering only animals with extremely high antibody titres, both study groups performed similar, however there was a numerical difference of 10 g/day in favour of vaccinated piglets. Likewise, lack of correlation between ADWG and MDA was observed suggesting that no maternal antibody interference was present with the tested vaccine because the vaccinated animals grew faster compared to unvaccinated control animals, regardless of the level of maternal antibodies present at the time of vaccination. CONCLUSIONS: The results of the present study demonstrated that the MDA against PCV-2 transferred through the colostrum intake has a protective effect against this viral infection. The vaccine used in the present study (Ingelvac CircoFLEX®) was effective when applied at three weeks of age and was not affected by the level of MDA at the time of vaccination.

The immune response to Hymenolepis nana in mice decreases tumorigenesis induced by 7,12 dimethylbenz-anthracene.

BACKGROUND: Cancer is a high-impact disease throughout the world. A negative correlation has been established between the development of cancer and the Th2 immune response. Infection by helminth parasites is characterized by the induction of a strong and long-lasting Th2 response. The aim of this work was to evaluate the effect of the immune response induced by the infection with the helminth Hymenolepis nana, on the tumorigenesis induced by dimethylbenz-anthracene (DMBA) in mice. METHODOLOGY: Four different groups of 14 female BALB/c mice were formed; Group A, dimethyl sulfoxide (DMSO) (vehicle) was administered cutaneously, Group B infected with H. nana, group C, cutaneously DMBA and finally Group D infected with H. nana and cutaneous DMBA. The tumor load was determined in those animals that developed cancerous lesions. In all groups were determined: serum concentration of IgE, IFNγ, IL-10, IL-5 and malondialdehyde (MDA). The inflammatory infiltrate was analyzed from skin samples and the expression of the main eosinophilic protein and myeloperoxidase was determined. RESULTS: The group previously infected with H. nana had a reduced amount of tumors with smaller size, in comparison to the group that received only DMBA; this reduction was associated with lower levels of IFNγ and IL-10, while levels of IgE, IL-5 and MDA were higher. Further, the number of eosinophils and neutrophils was statistically higher in the animals that were previously infected with the helminth and developed less tumors. CONCLUSION: The immune response induced by H. nana infection is associated with the reduction of tumors probably due to the activity of eosinophils and neutrophils.

Seminal plasma mitigates the adverse effect of uterine fluid on boar spermatozoa.

After natural or artificial insemination, spermatozoa start their journey within the uterus to reach the site of fertilization, but only few of them attain this goal. Part of this spermatozoa loss happens in the uterus, in which uterine fluid (UF) seems to be involved. It is known from other species that UF provokes damage to spermatozoa, which is avoided when seminal plasma (SP) is present. Therefore, the aim of the present study was to evaluate the effect of UF on the quality of ejaculated (previously contacted with SP) and epididymal (without previous contact with SP) boar spermatozoa analyzing motility, kinetic parameters, viability and acrosome integrity in the presence or absence of SP over time. Three experimental groups were evaluated in each source of spermatozoa (ejaculated and epididymal): 1) Control: spermatozoa with 20% of SP; 2) UF: spermatozoa with 20% of UF; and 3) UF-SP: spermatozoa with 20% of SP and 20% of UF. Total and progressive motility, kinetic parameters (VCL, VSL, VAP, LIN, STR, WOB, and BCF), viability and acrosome damage were analyzed at 15, 60, 120 and 180 min after incubation. Total and progressive motility decreased when ejaculated spermatozoa were incubated in UF in contrast to control and UF-SP groups (p < 0.0007), with no differences between control and UF-SP. The VCL decreased in the UF group compared to the control and UF-SP groups in ejaculated spermatozoa (p = 0.0002). The VSL, VAP, LIN and STR kinetic parameters were greater when ejaculated spermatozoa were incubated in the UF-SP group than in the UF group (all: p ≤ 0.02). Acrosome damage increased in ejaculated and epididymal spermatozoa incubated in the UF group compared to the control and UF-SP groups (both: p < 0.0001). Also, the viability of epididymal spermatozoa decreased in the UF group, while it did not change in the control and UF-SP groups (p = 0.0004). The rest of the parameters in either ejaculated or epididymal spermatozoa did not differ between experimental groups, except for WOB when epididymal spermatozoa were used (UF-SP higher than the control group, with UF being similar for both; p = 0.03). In conclusion, both ejaculated and epididymal spermatozoa are affected by UF, suggesting a negative effect on their quality. This negative effect is reduced by the presence of SP, improving the spermatozoa functionality, preserving motility, viability and acrosome integrity.

Post-cervical artificial insemination in porcine: The technique that came to stay.

The porcine industry is of great importance worldwide, and so any technological innovation in one or more of the associated production areas is of interest for meat production. Among such innovations in the reproduction area, post-cervical or intrauterine artificial insemination (PCAI) has emerged as a new approach in artificial insemination (AI). PCAI is gradually replacing traditional cervical insemination (CAI), particularly in countries with intensive pig production industries. This type of insemination, which deposits the semen in the body of the uterus (as opposed to traditional cervical deposition), is increasingly used in the field due to its simplicity and the numerous advantages that it provides at production level (e.g. reduced number of sperm, less time required to perform insemination and faster genetic improvement) and, consequently, from an economic point of view. In addition, since its inception, PCAI has been combined with other reproductive biotechnologies, such as the use of frozen-thawed sperm, fixed-time AI or sperm-mediated gene transfer. However, despite its wide acceptance and application, new approaches for increasing the efficiency of PCAI are constantly being sought, such as the adjustment and standardization in sperm numbers, the conservation of the PCAI semen dose, its association with other biotechnologies (sex-sorted sperm) or its efficacy in young (nulliparous and primiparous) females.

Morphological changes in the porcine cervix: A comparison between nulliparous and multiparous sows with regard to post-cervical artificial insemination.

In recent decades, new artificial insemination (AI) methods, such as post-cervical AI (PCAI), have been developed in pig. PCAI involves crossing the cervix to deposit the sperm in the uterine body. Although PCAI application in sows is frequent, its application in nulliparous (gilts) females it is still limited due to the difficulty of passing through the cranial part of the cervical lumen. We hypothesized that ageing and parity would modify the cervical canal, facilitating the introduction of AI devices through the cervix. The aim was to compare the morphology of the uterus at different levels between multiparous and nulliparous females. Morphological analysis of the uterus pointed to a longer cervix (25.9 ±â€¯4.6 vs. 21.6 ±â€¯3.3 cm, p < 0.001) and greater length of the part of the reproductive tract involved in PCAI (from rima vulvae to the last cervical cushion) (56.2 ±â€¯6.0 vs. 50.3 ±â€¯5.2 cm, p < 0.001) in multiparous sows compared with nulliparous animals. As regards the structure of the vaginal and uterine parts of the cervix (the part in contact with the vagina and uterine body, respectively), the cross-sectional area, perimeter and total thickness were greater in the uterine part of multiparous than of nulliparous animals (area: 4.07 ±â€¯1.46 vs. 2.46 ±â€¯0.56 cm2, p < 0.01; perimeter: 8.50 ±â€¯1.44 cm vs. 6.28 ±â€¯0.92 cm, p < 0.001; thickness: 10.79 ±â€¯0.96 vs. 8.35 ±â€¯0.62 mm, p < 0.05), but not in the vaginal part. The tissue content analysed in histological cross-sections also showed differences between female groups, a greater content of connective tissue (58.86 ±â€¯10.78 vs. 67.60 ±â€¯13.38%, p < 0.001) and a lower amount of muscle fibres (39.79 ±â€¯10.24 vs. 30.66 ±â€¯13.69%, p < 0.001) being observed in multiparous sows. Finally, silicone casts of the cervical lumen revealed differences between the two groups in the size and shape of the ridges in the lumen trajectory. Parity, which is also influenced by ageing, determines important changes in the size, structure and tissue content of the cervix wall, as well as in the morphology of the cervical canal, which may be responsible for the different levels of performance of PCAI in the female populations. Therefore, the future design of AI strategies and catheters should take into consideration the morphological variations of the cervix lumen, which will depend on age and parity of the females.

Sex difference in counts of α4 and α7 nicotinic acetylcholine receptors in the nasal polyps of adults with or without exposure to tobacco smoke.

OBJECTIVE: To assess counts of α4 and α7 nicotinic acetylcholine receptors in nasal polyps of adults with or without long-term exposure to cigarette tobacco smoke. METHODS: Twenty-two patients with and 22 patients without exposure to cigarette tobacco smoke participated in the study. After endoscopic polypectomy, the fragments of the nasal polyps were analysed by immunohistochemistry. RESULTS: Compared to patients with no exposure, patients with exposure showed higher counts of α4 and α7 nicotinic acetylcholine receptors (t-test, p 0.05). CONCLUSION: Exposure to cigarette tobacco smoke may induce increased counts of α4 and α7 nicotinic acetylcholine receptors in nasal polyps of adults, with lower counts in males than females without exposure to tobacco smoke.

Quality of life of young patients with recurrent respiratory papillomatosis.

OBJECTIVE: To assess quality of life of children and teenagers with recurrent respiratory papillomatosis, according to the evidence of infection by human papillomavirus types 6 and 11, compared with healthy volunteers and patients with chronic otitis media. METHOD: Participants and their parents completed the Pediatric Quality of Life Inventory 4.0. RESULTS: Patients with recurrent respiratory papillomatosis and their parents reported lower quality of life than healthy subjects (p < 0.01), but similar quality of life to patients with chronic otitis media. Those with human papillomavirus type 11 showed the lowest scores among all participants (p < 0.05). CONCLUSION: Young Mexican patients with recurrent respiratory papillomatosis and their parents perceive a poor quality of life, and they may experience limitations in interactions with their peers. Infection by human papillomavirus type 11 may increase the impact of the disease on quality of life.

Mouse model of congenital infection with a non-virulent Toxoplasma gondii strain: Vertical transmission, "sterile" fetal damage, or both?

Congenital transmission of Toxoplasma gondii may occur if the mother gets infected for the first time while pregnant. The risk of mother-to-child transmission depends on the gestation trimester at infection, being lowest in the first and highest in the last. Conversely, fetal damage is frequent and more severe at the beginning of pregnancy. The objective of this study was to evaluate congenital transmission and pathological aspects in the placenta and the fetus using a mouse model of congenital infection of the second gestation third. Forty-five female BALB/c mice were infected intravenously with 2.5-10.0 × 10(6) tachyzoites of the ME49 strain at middle gestation. Samples of maternal spleen and fetal/placental units were taken 72 h later. We determined parasite load and vertical transmission by qPCR, as well as damage macroscopically and by histopathology. With the lowest dose, 18% of the fetuses were infected. Also, 40% of fetuses/litter were altered, while this value was 10% in the control group (P < 0.05). These results are similar to those described in humans in terms of vertical transmission and fetal damage during the second third of gestation. The maternal spleen had 10-1000 times more tachyzoites than the placenta, and the later retained 90-99% of the parasites that could reach the fetus. Nevertheless, we found resorptions, abortions or fetal tissue damage in the presence but also in the absence of parasites. Our data indicate a strong protective effect of maternal organs and the placenta against fetal infection, but extensive damage of the later may led to resorption or abortion without vertical transmission.

Interleukin 10 and dendritic cells are the main suppression mediators of regulatory T cells in human neurocysticercosis.

Neurocysticercosis is caused by the establishment of Taenia solium cysticerci in the central nervous system. It is considered that, during co-evolution, the parasite developed strategies to modulate the host's immune response. The action mechanisms of regulatory T cells in controlling the immune response in neurocysticercosis are studied in this work. Higher blood levels of regulatory T cells with CD4(+) CD45RO(+) forkhead box protein 3 (FoxP3)(high) and CD4(+) CD25(high) FoxP3(+) CD95(high) phenotype and of non-regulatory CD4(+) CD45RO(+) FoxP3(med) T cells were found in neurocysticercosis patients with respect to controls. Interestingly, regulatory T cells express higher levels of cytotoxic T lymphocyte antigen 4 (CTLA-4), lymphocyte-activation gene 3 (LAG-3), programmed death 1 (PD-1) and glucocorticoid-induced tumour necrosis factor receptor (GITR), suggesting a cell-to-cell contact mechanism with dendritic cells. Furthermore, higher IL-10 and regulatory T cell type 1 (Tr1) levels were found in neurocysticercosis patients' peripheral blood, suggesting that the action mechanism of regulatory T cells involves the release of immunomodulatory cytokines. No evidence was found of the regulatory T cell role in inhibiting the proliferative response. Suppressive regulatory T cells from neurocysticercosis patients correlated negatively with late activated lymphocytes (CD4(+) CD38(+) ). Our results suggest that, during neurocysticercosis, regulatory T cells could control the immune response, probably by a cell-to-cell contact with dendritic cells and interleukin (IL)-10 release by Tr1, to create an immunomodulatory environment that may favour the development of T. solium cysticerci and their permanence in the central nervous system.

Four glycoproteins are expressed in the cat zona pellucida.

The mammalian oocyte is surrounded by a matrix called the zona pellucida (ZP). This envelope participates in processes such as acrosome reaction induction, sperm binding and may be involved in speciation. In cat (Felis catus), this matrix is composed of at least three glycoproteins called ZP2, ZP3, and ZP4. However, recent studies have pointed to the presence of a fourth protein in several mammals (rat, human, hamster or rabbit), meaning that a reevaluation of cat ZP is needed. For this reason, the objective of this research was to analyze the protein composition of cat ZP by means of proteomic analysis. Using ZP from ovaries and oocytes, several peptides corresponding to four proteins were detected, yielding a coverage of 33.17%, 71.50%, 50.23%, and 49.64% for ZP1, ZP2, ZP3, and ZP4, respectively. Moreover, the expression of four genes was confirmed by molecular analysis. Using total RNA isolated from cat ovaries, the complementary deoxyribonucleic acids encoding cat ZP were partially amplified by reverse-transcribed polymerase chain reaction. Furthermore, ZP1 was totally amplified for the first time in this species. As far as we are aware, this is the first study that confirms the presence of four proteins in cat ZP.

Boar sperm with defective motility are discriminated in the backflow moments after insemination.

During insemination, a large number of spermatozoa are deposited in the female genital tract, but a very low percentage is able to colonize the site of fertilization. The influx of neutrophils into the uterine lumen and semen reflux (backflow) are known mechanisms that decrease the number of spermatozoa within the uterus. No report has attempted to ascertain whether the backflow is a random or selective process of the spermatozoa. In this work, sows were inseminated using two populations of spermatozoa in the same proportion: (1) unstained spermatozoa with high motility and (2) stained spermatozoa with low, medium, or high motility. Volume, number, and percentage of stained spermatozoa were evaluated in the backflow (collected at 0-15, 16-30, and 31-60 minutes after insemination). This article provides evidence that (1) the motility characteristics of the spermatozoa do not influence the percentage of sows with backflow, the volume and number of spermatozoa in the backflow; (2) the discarding of spermatozoa in the backflow is not specific during the first moments after insemination (0-15 minutes), whereas later (16-60 minutes), spermatozoa with defective motility (low and medium groups) are discarded in a higher proportion than high group in the backflow ([16-30 minutes: low, 85.13 ± 4.32%; medium, 72.99 ± 5.05%; and high, 54.91 ± 2.38%; P < 0.0001; 31-60 minutes: low, 87.16 ± 6.01%; medium, 87.02 ± 4.01%; and high, 59.35 ± 2.86%; P = 0.001]). Spermatozoa with poor motility are discarded in the backflow probably as a selective process, on the part of the female genital tract or as a result of the intrinsic low spermatozoa motility.

Microstructural effects on the wear behavior of a biomedical as-cast Co-27Cr-5Mo-0.25C alloy exposed to pulsed laser melting.

In this work, the effect of pulsed laser melting on the exhibited microstructure and properties of a cast Co-27Cr-5Mo-0.25C alloy was investigated. In particular, properties such as surface hardness and wear behavior of the laser modified microstructure were determined as a function of the implemented laser melting parameters. It was found that laser melting promotes significant grain refinement while preventing the precipitation of coarse carbide phases. Apparently, a refined dendritic grain structure develops which is surrounded by a fine carbide distribution in the interdendritic regions. Moreover, the high-temperature face centered cubic (FCC) phase remains untransformed at room temperature. Hardness measurements and wear testing using a Pin-On-Disk tribological machine indicate that the modified laser surfaces exhibit both, high wear resistance and high microhardness when compared with the untreated as-cast Co-27Cr-5Mo-0.25C alloy. In particular, it was found that the laser modified surfaces exhibit improved wear and friction properties comparable to the ones found in Co-Cr-Mo alloys with a predominantly hexagonal closest packed (HCP) matrix. However, surface defects associated with the laser process can be detrimental for the improved wear performance and they should be considered in identifying the proper laser parameters in alloy melting.

Influence of exposure to tobacco cigarette smoke on the eosinophil count in the nasal mucosa of young patients with perennial allergic rhinitis.

BACKGROUND: To assess the influence of exposure to tobacco cigarette smoke on the eosinophil count and the frequency of apoptosis of eosinophils in the nasal mucosa of teenagers with perennial allergic rhinitis. METHODS: Fifty patients were evaluated (aged 10 to 19 years old): 25 patients with and 25 patients with no recent exposure to tobacco cigarette smoke, by means of The Global Youth Tobacco Survey and cotinine/creatinine ratio. After a clinical evaluation, all the patients replied to a validated questionnaire of the severity of nasal symptoms; then, a nasal sample was processed to identify the eosinophil count and the frequency of apoptosis of eosinophils. RESULTS: Patients with active exposure to tobacco cigarette smoke had higher eosinophil counts than patients with no exposure to the smoke. In the two groups, apoptosis of eosinophils in the nasal mucosa was scarce and no significant correlation was observed between the frequency/severity of the nasal symptoms and the eosinophil count. CONCLUSION: Teenagers with perennial allergic rhinitis and active exposure to tobacco cigarette smoke may show increased eosinophil counts in the nasal mucosa, which might not be related to apoptosis of eosinophils or to the frequency/severity of nasal symptoms.

Effect of 1,25(OH)2D3 on BALB/c mice infected with Leishmania mexicana.

The most active metabolite of vitamin D, 1,25(OH)2D3 is a steroid hormone implicated in a wide range of cell functions such as differentiation, proliferation and apoptosis. Leishmania mexicana causes two kinds of cutaneous leishmaniasis: localized or diffuse. In this work we explored the effect of treatment of 1,25(OH)2D3 on a susceptible leishmaniasis mice model. A significant reduction in the lesion size was found in animals treated with 1,25(OH)2D3. Well preserved tissue and presence of large numbers of eosinophils and fibroblasts was found in the group treated with 1,25(OH)2D3. By contrast, destroyed epidermis was observed with large amount of neutrophils and epithelioid macrophages, on infected groups without 1,25(OH)2D3 treatment. The production of pro-inflammatory cytokines in mice infected and treated with 1,25(OH)2D3 was lower than the animals infected without 1,25(OH)2D3 treatment. Interestingly, there were no differences in the number of parasites in both groups. Finally, the amount of collagen was higher in animals with treatment compare with animals without 1,25(OH)2D3 treatment. In summary, mice treated with 1,25 (OH) 2D3 reflect a healing process without elimination of L. mexicana.

Equine spermatozoa stored in the epididymis for up to 96h at 4°C can be successfully cryopreserved and maintain their fertilization capacity.

After injury or death of a valuable male, recovery of epididymal spermatozoa may be the last chance to ensure preservation of its genetic material. The objective of this research was to study the effect of sperm storage, at 4°C up to 96h, in the epididymides obtained from castrated horses and its effect on different functional sperm parameters. Aims were to study the effect of (1) sperm storage on viability and chromatin condensation; (2) pre-incubation of recovered epididymal sperm in the freezing extender, prior cryopreservation, on viability and chromatin condensation; and (3) freezing-thawing on viability, chromatin condensation, ROS generation, protein tyrosine phosphorylation and heterologous fertilization rate (ICSI and IVF using bovine oocytes) of sperm recovered from the epididymis up to 96h post castration. The average volume (720±159µL) and the concentration (6.5±0.4×10(9) spermatozoa/mL) of sperm recovered from the epididymis were not affected by storage. Sperm viability after refrigeration at 4°C for up to72h was similar (P<0.01). The effect of sperm dilution in the freezing media showed similar values up to 48h, while viability was preserved up to 72h (P<0.01). Cryopreserved spermatozoa show similar viability between different storage times. Chromatin condensation was not affected by storage time; however, incubation for 30min in freezing medium and freezing-thawing process induced an increase in the chromatin decondensation. ROS generation was not affected by storage up to 96h. Epididymal storage did not affect sperm protein tyrosine phosphorylation patterns; although the pattern of phosphorylation changed to strong staining of the equatorial segment when the sperm where capacitated in sperm-TALP. Finally, successful and similar pronuclear formation (analyzed by ICSI) and in vitro penetration (evaluated with bovine zone free oocyte) was observed using cryopreserved sperm obtained from prolong epididymal storage at 4°C. In conclusion, cryopreservation of epididymal stallion sperm stored for up to 72h in the epididymis at 4°C, maintain both viability and ability to fertilize in vitro.

Roles of the oviduct in mammalian fertilization.

The oviduct or Fallopian tube is the anatomical region where every new life begins in mammalian species. After a long journey, the spermatozoa meet the oocyte in the specific site of the oviduct named ampulla and fertilization takes place. The successful fertilization depends on several biological processes that occur in the oviduct some hours before this rendezvous and affect both gametes. Estrogen and progesterone, released from the ovary, orchestrate a series of changes by genomic and nongenomic pathways in the oviductal epithelium affecting gene expression, proteome, and secretion of its cells into the fluid bathing the oviductal lumen. In addition, new regulatory molecules are being discovered playing important roles in oviductal physiology and fertilization. The present review tries to describe these processes, building a comprehensive map of the physiology of the oviduct, to better understand the importance of this organ in reproduction. With this purpose, gamete transport, sperm and oocyte changes in the oviductal environment, and other interactions between gametes and oviduct are discussed in light of recent publications in the field.